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Bin Liu,* Bingtian Zhao,* Yan Yin, Yan Jiang, Xue Feng, Lei Wang, Liang Zhai, Guangxin Liu, Dongsheng Shi, Jianwen Qin Department of Respiratory and Critical Medicine, Tianjin Chest Hospital, Tianjin, People’s Republic of China*These authors contributed equally to this workCorrespondence: Dongsheng Shi; Jianwen Qin, Tianjin Chest Hospital, No. 261, Taierzhuang South Road, Jinnan District, Tianjin, 300051, People’s Republic of China, Email shi_ds@126.com; qinjianwen2005@aliyun.com; 5020200603@nankai.edu.cnPurpose: DNA methylation plays a regulatory role in the oncogenesis and tumor progression and is valuable in the diagnosis and prognosis of cancer. While circulating tumor DNA (ctDNA) is widely used in the detection of oncogenic mutations and the guidance of treatment in advanced non-small cell lung cancer (NSCLC), studies of ctDNA methylation remains insufficient. We aim to investigate the methylation profiles of ctDNA in patients with advanced NSCLC undergoing EGFR-tyrosine kinase inhibitor (EGFR-TKI) therapy and to discover novel biomarkers with predictive or prognostic value.Patients and Methods: We recruited 49 patients with EGFR-mutated advanced NSCLC undergoing EGFR-TKI as first-line treatment. Utilizing next-generation sequencing, we examined the somatic mutations and methylation signatures within the tumor-associated genomic regions of ctDNA from pre-treatment blood. Subsequently, we explored the association of these molecular features with the patients’ response to therapy and their progression-free survival (PFS).Results: Genomic mutation profiling revealed no significant association of PFS or best overall response (BOR) and ctDNA status. Evaluation of ctDNA methylation showed a negative correlation between the methylation of small nucleolar RNA (snoRNA) genes and PFS (R=− 0.31, P=0.043). Furthermore, high-level methylation of SNORD3F was associated with poorer PFS (mPFS 346d vs 243d, HR 0.49, 95% CI 0.24– 0.93, P=0.029).Conclusion: Our study explored the prognostic value of ctDNA methylation in patients with advanced NSCLC undergoing targeted therapies and first revealed the predictive role of SNORD3F.Keywords: ctDNA, EGFR, DNA methylation, lung cancer, snoRNA, SNORD3F

Qian Meng,1,* Xiaowei Yang,1,* Zhongcheng Liu,2 Guoxing You,3 Wanyi Chen,3 Bing Zhao,1 Huizi Zhu,4 Liang Xu,1 Yan Zhou,1 Xiang Liu,1 Chunjuan Zhai,5 Rong Wang,1 Lian Zhao,3 Jing Sun1 1Department of Nephrology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, 250021, People’s Republic of China; 2Department of Cardiovascular, The North City Hospital of Jinan, Jinan, Shandong, 250031, People’s Republic of China; 3Academy of Military Medical Sciences, Beijing, 100850, People’s Republic of China; 4Department of Nephrology, Fuyang People’s Hospital, Fuyang, Anhui, 236000, People’s Republic of China; 5Department of Cardiology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, 250021, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jing Sun, Department of Nephrology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 324 Jingwu Road, Jinan, Shandong, 250021, People’s Republic of China, Tel +86 151 6888 6367, Email sunj5708@126.com Lian Zhao, Academy of Military Medical Sciences, 27 Taiping Road, Beijing, 100850, People’s Republic of China, Tel +8613671217095, Email zhaolian@bmi.ac.cnPurpose: Chronic kidney disease (CKD) is associated with persistent systemic inflammation. Reduced red blood cell (RBC) survival in patients with CKD has been identified for several decades. The purpose of this study is to explore whether excessive erythrophagocytosis exists and contributes to systemic inflammation in CKD.Patients and methods: A CKD rat model was induced by 5/6 nephrectomy. Erythrocyte osmotic fragility was determined with hypotonic NaCl solutions. Erythrocyte deformability was evaluated by filterability. RBC cell death was quantified using fluorescence-activated cell sorting analyses of fluorescent annexin V–bound surface phosphatidylserine (PS). Erythrophagocytosis was evaluated in vivo and in vitro. RT-qPCR and immunohistochemistry were used to determine the inflammatory effects after erythrophagocytosis.Results: Erythrocyte osmotic fragility and deformability progressively declined, and the percentage of PS-exposing RBCs progressively increased in CKD rats. Levels of erythrophagocytosis in vivo were evaluated by autologous injection of CFSE-labeled erythrocytes. In comparison with the control group, higher fluorescence intensity of CFSE was detected in the spleen homogenates of rats with CKD. In vitro, more of erythrocytes from 5/6Nx rats were phagocytosed by peritoneal macrophages in comparison to those from control rats. Compared with macrophages phagocytosed control erythrocytes, macrophages phagocytosed CKD erythrocytes exhibited higher mRNA levels of IL-6, CXCL-10, CXCL-11, iNOS, IL-1β, ICAM-1 and MCP-1. Compared with the control group, the red pulp of rats with CKD exhibited higher levels of p-NFκB, IL-6, iNOS and CXCL-10. ELISA results showed significantly increased plasma levels of both IL-6 and CXCL-10 in patients with long-term hemodialysis compared with those in healthy controls (2.30 ± 1.38 pg/mL vs 1.33 ± 0.65 pg/mL, P=0.01; 78.11 ± 27.34 pg/mL vs 37.45 ± 7.08 pg/mL, P=0.001).Conclusion: Our results indicated that excessive erythrophagocytosis may contribute to systemic inflammation in CKD. Keywords: chronic kidney disease, chronic inflammation, erythrocyte, phagocytosis

Xinyi Zhang,1,2,&ast; Zhu Chen,3,&ast; Jun Zheng,4 Chen Feng,5 Bennan Zhao,1 Lijuan Lan,1 Dafeng Liu1 1The First Ward of Internal Medicine, Public Health Clinical Centre of Chengdu, Chengdu, People’s Republic of China; 2Department of Endocrinology & Metabolism, Sichuan University West China Hospital, Chengdu, People’s Republic of China; 3Department of Drug Clinical Trial Center, Public Health Clinical Centre of Chengdu, Chengdu, People’s Republic of China; 4Medical Department, Public Health Clinical Centre of Chengdu, Chengdu, People’s Republic of China; 5Legal Services Division, Public Health Clinical Centre of Chengdu, Chengdu, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Dafeng Liu, The First Ward of Internal Medicine, Public Health Clinical Centre of Chengdu, No. 377 Jingming Road, Jinjiang District, Chengdu, Sichuan, 610066, People’s Republic of China, Email ldf312@126.comAim: Our cohort study aimed to investigate the dynamic changes of lymphocyte subsets and their abilities to predict disease severity and prognosis in primary infection and unvaccinated COVID-19 patients.Methods: A total of 773 cases, including 718 primary infection and unvaccinated COVID-19 patients and 55 controls. COVID-19 patients were assigned to severe and nonsevere groups according to disease severity, as well as survival and death groups according to prognosis. Serum samples were collected to measure the numbers of total lymphocytes and lymphocyte subsets. The differences among different severity groups were analyzed. Spearman correlation was performed to assess associations between lymphocyte subsets and disease severity and prognosis. Meanwhile, receiver operating characteristic (ROC) curves were also analyzed to find optimal cutoff points.Results: At admission, the severe group demonstrated significantly lower total lymphocyte counts and percentages, CD3+ and CD3+CD4+ T cell counts and percentages, CD3+CD8+ T cell counts, CD19+ B cell counts and CD56+ NK cell counts and percentages than the nonsevere group. Meanwhile, compared with the survival group, the death group also had lower total lymphocyte counts and percentages, CD3+, CD3+CD4+ and CD3+CD8+ T cell counts. Additionally, differences in these parameters were also noticed within four weeks after admission. Furthermore, Spearman analysis reported that disease severity was negatively correlated with lymphocyte counts and percentages, CD3+, CD3+CD4+ and CD3+CD8+ T cell counts, CD3+ and CD3+CD4+ T cell percentages (r=− 0.166, − 0.179, − 0.173, − 0.186, − 0.127, − 0.117, − 0.149, respectively)(all P< 0.05). The prognosis of death was also negatively correlated with total lymphocyte counts and percentages, CD3+, CD3+CD4+ and CD3+CD8+ T cell counts (r=− 0.125, − 0.121, − 0.123, − 0.123, − 0.091, respectively)(all P< 0.05).Conclusion: In primary infection and unvaccinated COVID-19 patients total lymphocytes and T cell, B cell and NK cell subsets at COVID-19 onset play valuable roles in predicting disease severity and prognosis.Clinical Trial Registry: Chinese Clinical Trial Register ChiCTR2000034563.Keywords: lymphocyte subsets, coronavirus disease 2019, COVID-19, prediction, severity, prognosis

Bingyang Xu,1 Biao Yu,2 Zining Xu,1 Shuhong Ye,1 Yuxin Qing,1 Hong Sun,3 Bin Zhao,4 Na Wu,5 Jiawen Wu1 1Department of Dermatology, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, People’s Republic of China; 2Department of Dermatology, Taihe Hospital, Hubei University of Medicine, Shi’yan, People’s Republic of China; 3Department of Neurology, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, People’s Republic of China; 4Department of Dermatology, Xi’an International Medical Center Hospital, Xi’an, People’s Republic of China; 5Department of Nursing, Xi’an Jiaotong University Medical School, Xi’an, People’s Republic of ChinaCorrespondence: Jiawen Wu, Department of Dermatology, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, People’s Republic of China, Email jiawen702@126.comPurpose: Psoriasis is not yet completely curable, and its etiology and pathogenesis are unclear. Necroptosis, also known as programmed necrosis, is a regulated mode of necrotic cell death. The interaction between inflammatory diseases and necrotic apoptosis has recently attracted significant attention. We explored the molecular mechanisms of necrotic apoptosis-related genes in psoriasis using bioinformatics methods to identify potential biomarkers for psoriasis.Patients and Methods: In this study, we screened psoriasis differentially expressed genes from the datasets GSE13355 and GSE14905 and took intersections with necrotic apoptosis-related genes for the next analysis. We used multiple machine learning algorithms to screen key genes and perform enrichment analysis. In addition, we performed an immune infiltration analysis. Transcription factors were predicted by the R package “RcisTarget”. We also observed the cellular clustering of key genes in different cell types at the single-cell sequencing level. We used real-time fluorescence-based quantitative-polymerase chain reaction, Western blot, and immunohistochemistry to analyze gene expression in clinical samples. We constructed an imiquimod-induced psoriasis-like dermatitis model in mice for further validation.Results: Seven key genes were screened as follows: AIM2, CARD6, HPSE, MYD88, PYCARD, RAI14, and TNFSF10. Enrichment analysis showed that the key genes were mainly involved in inflammatory pathways. Immune infiltration analysis showed significantly higher levels of CD8 T cells, CD4 initial T cells, and CD4 memory-activated T cells in the disease group’s samples than in the normal patients’ samples. The key gene expression in single cells analyzed showed that PYCARD was significantly expressed in keratinocytes. PYCARD was selected for gene expression analysis; the results showed that its expression was significantly elevated in the skin lesion tissues of patients with psoriasis. We also verified that PYCARD might play a vital role in the development of psoriasis skin lesions using animal experiments.Conclusion: PYCARD plays a vital role in psoriasis development and is a potential biomarker for psoriasis.Keywords: necrotic apoptosis-related gene, bioinformatics, inflammatory disease, immune disease

Juan Li,1,&ast; Shu Wang,2,&ast; Chaoping Ma,1,&ast; Ning Ning,3,&ast; Yingying Huang,1,4,&ast; Min Jiao,1 Jiyuan Zhang,1 Wenwu Sun,3 Jiaoyan Li,3 Bing Zhao,3 Enqiang Mao,3 Zaiqian Che,3 Chengjin Gao1 1Department of Emergency, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China; 2Department of Intensive Care Medicine, Chongqing University Central Hospital, Chongqing Emergency Medical Center, Chongqing, 400016, People’s Republic of China; 3Departments of Emergency, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200025, People’s Republic of China; 4Dementia Research Center, Macquarie University, Sydney, Australia&ast;These authors contributed equally to this workCorrespondence: Chengjin Gao, Department of Emergency, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, 1665 Kongjiang Road, Shanghai, People’s Republic of China, Email gaochengjin@xinhuamed.com.cn Zaiqian Che, Departments of Emergency, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China, Email chezaiqian@163.comPurpose: New-onset atrial fibrillation (NOAF) and sepsis-induced coagulopathy (SIC) are severe complications in septic patients. However, the relationship between NOAF and SIC score has not been clearly defined. This study aims to investigate the association between SIC score and NOAF, as well as their effect on mortality in sepsis.Patients and Methods: This study was a two-center retrospective analysis. Medical data were collected from patients diagnosed with sepsis. The patients were divided into NOAF and non-NOAF groups, and the SIC score was calculated for each group. Univariable and multivariable logistic regression analyses were performed to explore the relationship between the SIC score and NOAF, as well as their effects on mortality. The Kaplan-Meier curve was used to assess the survival rate.Results: A total of 2,280 septic patients were included, with 132 (5.7%) suffering from NOAF. Multivariable logistic regression analyses indicated that age, gender, the Acute Physiology and Chronic Health Evaluation II score (APACHE II), heart rate, renal failure, stroke, chronic obstructive pulmonary disease (COPD), and the SIC score were independent risk factors for NOAF in sepsis. Moreover, NOAF was associated with an increased risk of in-hospital mortality, 28-day mortality, and 90-day mortality. These results were consistent across subgroup analyses.Conclusion: The SIC score was an independent risk factor for NOAF in septic patients, and NOAF was an independent risk factor for predicting mortality.Keywords: sepsis, sepsis-induced coagulopathy, new-onset atrial fibrillation, association, mortality

Xinyu Dai,1,&ast; Rui Zhang,2,&ast; Nan Deng,1 Le Tang,3 Bangshu Zhao1 1Department of Anesthesiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, People’s Republic of China; 2Department of Neurosurgery, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu, People’s Republic of China; 3Department of Psychiatry, the First Affiliated Hospital of Chongqing Medical University, Chongqing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Bangshu Zhao, Department of Anesthesiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, People’s Republic of China, Tel +8615823079815, Email Bangshuzhao@hospital.cqmu.edu.cnAbstract: The prevalence of severe mental disorders has been rising annually. Electroconvulsive therapy (ECT) is considered a valuable treatment option in psychiatry for conditions such as schizophrenia and medication-resistant depression, especially when other treatments have proven insufficient. ECT rapidly improves patients’ mood, alleviates symptoms, and demonstrates significant therapeutic effects. Currently, the form of ECT used in clinical practice is modified electroconvulsive therapy (mECT), which is administered under general anesthesia. Accumulative evidence has confirmed that different anesthetic drugs, anesthetic-ECT time interval, anesthetic depth, and airway management can impact the outcomes of ECT. Therefore, this review aims to summarize the current impact of anesthesia factors on ECT, providing reference for clinical anesthesia during ECT procedures.Keywords: mental disorders, ECT, anesthesia factors

Yukai Hao,1 Baoling Zhao,1 Dongchao Wu,1 Xuejun Ge,1 Jianmin Han2 1Department of Endodontics, Shanxi Province Key Laboratory of Oral Diseases Prevention and New Materials, Shanxi Medical University School and Hospital of Stomatology, Taiyuan, 030001, People’s Republic of China; 2Central Laboratory, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing, 100081, People’s Republic of ChinaCorrespondence: Xuejun Ge, The Department of Endodontics, Shanxi Medical University School and Hospital ofStomatology, 63 Xinjian South Road, Yingze District, Taiyuan, 030001, People’s Republic of China, Tel +8613834568537, Email gxj19722003@163.com Jianmin Han, Central Laboratory, Peking University School and Hospital of Stomatology, 22 Zhongguan Cun South Street, Beijing, 100081, People’s Republic of China, Tel +8613810459668, Email hanjianmin@bjmu.edu.cnIntroduction: Recombinant humanized collagen, as a novel biomaterial, exhibits multiple excellent biological functions, such as inhibition of inflammation, promotion of cell proliferation and vascular proliferation, and promotion of tissue healing. However, there is a lack of conclusive evidence regarding the specific role of recombinant humanized collagen type 17 (rhCol 17) in oral ulcer healing. This study explored whether rhCol 17 could promote the proliferation of human gingival fibroblasts (HGFs) and inhibit its inflammation, and whether it could promote the healing of oral ulcers in rats by inhibiting inflammation and accelerating tissue healing.Methods: At the cellular level, we investigated the effect of rhCol 17 on the proliferation of (HGFs) by CCK8; HGFs were mixed with lipopolysaccharide (LPS) to investigate the effect of rhCol 17 on HGFs in an inflammatory state. Eighteen adult male Sprague-Dawley rats were randomly distributed into three groups: blank control group, carbomer group (carbomer sprayed only), and rhCol 17 group (carbomer containing rhCol 17 sprayed), 1 time/day. The samples were collected at D3 and D5. At completion, histological staining and PCR were carried out to study its effect on the healing of oral ulcers in rats.Results: Through cellular experiments, we found that rhCol 17 possesses good biocompatibility and anti-inflammatory properties, and can effectively promote the proliferation and migration of HGFs, as well as significantly reduce the inflammation level of the cells. The animal experimental results showed that rhCol 17 could significantly reduce the inflammation level, promote collagen deposition and angiogenesis at the ulcer site, thus effectively accelerating the healing of oral ulcers in rats.Conclusion: In summary, the collagen sprays containing rhCol 17 have excellent anti-inflammatory effects and could accelerate tissue healing and are expected to provide a new effective treatment for patients with recurrent oral ulcers.Keywords: oral ulcer, inflammation, wound healing, recombinant humanized collagen

Liping Feng,1,&ast; Wencong He,2,&ast; Zexuan Song,3,&ast; Bing Zhao,3 Chong Teng,3 Eryong Liu,3 Hanfang Zhu,1 Shaojun Pei,4 Lina Liu,5 Yuanyuan Song,3 Yang Zheng,3 Xiangyi Liu,2 Yanlin Zhao,3 Xichao Ou3 1Department of Microbiology, Hinggan League Center for Disease Control and Prevention, Ulanhot, 137499, People’s Republic of China; 2Department of Clinical Laboratory, Beijing Tongren Hospital, Capital Medical University, Beijing, 100176, People’s Republic of China; 3National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Center for Tuberculosis Control and Prevention, Chinese Centre for Disease Control and Prevention, Beijing, 102206, People’s Republic of China; 4School of Public Health, Peking University, Beijing, 100191, People’s Republic of China; 5Blood Transfusion Department, Hinggan League People’s Hospital, Ulanhot, 137400, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xichao Ou; Yanlin Zhao, Email ouxc@chinacdc.cn; zhaoyl@chinacdc.cnBackground: Tuberculosis remains a major public health concern in China, with varying prevalence and drug resistance profiles across regions. This study explores the genetic diversity and drug-resistant profiles of MTB strains in Hinggan League, a high TB burden in Inner Mongolia, China.Methods: This population-based retrospective study, encompassing all culture-positive TB cases from Jun. 2021 to Jun. 2023 in Hinggan League. Drug resistant profiles and genetic diversity of MTB strains were assessed using phenotypic drug susceptibility testing and whole-genome sequencing. Risk factors associated with drug resistance were analyzed using univariate and multivariate logistic regression models.Results: A total of 211 MTB strains were recovered successfully and included into final analysis. Lineage 2.2.1 (88.6%, 187/211) was the dominant sub-lineage, followed by lineage 4.5 (7.1%, 15/211) and lineage 4.4 (4.3%, 9/211). MTB strains exhibited the highest resistance rates to isoniazid (16.1%, 34/211), followed by rifampicin (10.0, 21/211). In addition, the MTB strains also showed relatively high rates of resistance against new and repurposed anti-TB drugs, with resistant rates of 2.4% (5/211) to delamanid and 1.9% (4/211) to bedaquiline. Overall, 25.6% (54/211) of MTB strains were DR-TB, and 14 MTB strains met the definition of MDR-TB, including 7 strains of simple-MDR-TB, 5 of pre-XDR-TB, and 2 of XDR-TB. Genetic analysis revealed that the dominant mutations of isoniazid-, rifampin-, ethambutol-, levofloxacin-/moxifloxacin-, and ethionamide- resistance were katG_Ser315Thr(46.4%), rpoB_Ser450Leu (47.4%), embB_Met306Val (25.0%), gyrA_Asp94Ala (40.0%), and fabG1_c15t (42.9%), respectively. Previously treated patients (AOR = 2.015, 95% CI: 1.052– 4.210) and male patients (AOR = 3.858, 95% CI: 1.416– 10.511) were identified as independent risk factors associated with DR-TB.Conclusion: Our study offers crucial insights into the genetic diversity and drug-resistant profiles of TB strains circulating in Hinggan League. These findings are valuable for DR-TB surveillance and for guiding treatment regimens and public health interventions in the region.Keywords: Mycobacterium tuberculosis, drug resistance, genetic diversity, whole-genome sequencing

Chenkun Wang,1,2,&ast; Jing Gu,2,&ast; Hongwei Li,2,&ast; Bo Zhao,2 Tao Yu,1,2 Chun-Ling Guo,2 Mouxin Huang,2 Weiwei Jiang,1 Qin Ouyang2 1Department of Pharmacy, the Second Affiliated Hospital, Chongqing Medical University, Chongqing, People’s Republic of China; 2Department of Medicinal Chemistry, College of Pharmacy, Third Military Medical University, Chongqing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Weiwei Jiang, Department of Pharmacy, the Second Affiliated Hospital, Chongqing Medical University, No. 1 Yixueyuan Road, Yuzhong District, Chongqing, 400010, People’s Republic of China, Email 304035@cqmu.edu.cn Qin Ouyang, Department of Medicinal Chemistry, College of Pharmacy, Third Military Medical University, No. 30 Gaotanyan St., Shapingba District, Chongqing, 400038, People’s Republic of China, Email ouyangq@tmmu.edu.cnBackground and Objective: GIT1 (G-protein-coupled receptor kinase interacting protein-1) has been found to be highly related with cancer cell invasion and metastasis in many cancer types. β-Pix (p21-activated kinase-interacting exchange factor) is one of the proteins that interact with GIT1. Targeting GIT1/β-Pix complex might be a potential therapeutic strategy for interfering cancer metastasis. However, at present, no well-recognized small-molecule inhibitor targeting GIT1/β-Pix is available. Thus, we aim to discover novel GIT1/β-Pix inhibitors with simple scaffold, high activity and low toxicity to develop new therapeutic strategies to restrain cancer metastasis.Methods: GIT1/β-Pix inhibitors were identified from ChemBridge by virtual screening. Briefly, the modeling of GIT1 was performed and the establishment of GIT1/β-Pix binding pocket enabled the virtual screening to identify the inhibitor. In addition, direct binding of the candidate molecules to GIT1 was detected by biolayer interferometry (BLI) to discover the hit compound. Furthermore, the inhibitory effect on invasion of stomach and colon cancer cells in vitro was carried out by the transwell assay and detection of epithelial-mesenchymal transition (EMT)-related proteins. Finally, the binding mode of hit compound to GIT1 was estimated by molecular dynamics simulation to analyze the key amino residues to guide further optimization.Results: We selected the top 50 compounds from the ChemBridge library by virtual screening. Then, by skeleton similarity analysis nine compounds were selected for further study. Furthermore, the direct interaction of nine compounds to GIT1 was detected by BLI to obtain the best affinitive compound. Finally, 17302836 was successfully identified (KD = 84.1± 2.0 μM). In vitro tests on 17302836 showed significant anti-invasion effect on gastric cancer and colorectal cancer.Conclusion: We discovered a new GIT1/β-Pix inhibitor (17302836) against gastrointestinal cancer invasion and metastasis. This study provides a promising candidate for developing new GIT1/β-Pix inhibitors for tumor treatment.Keywords: hit discovery, virtual screening, GIT1, β-Pix, gastrointestinal cancers, invasion

Wenqing Wang,1,2,&ast; Chuchu Ye,2,3,&ast; Bing Zhao,1 Yingjie Zheng,4 Ge Zhang,5 Jinghua Su,1 Hong Huang,1 Lipeng Hao,1,2 Mingliang Chen6 1Department of Microbiology, Pudong New Area Center for Disease Control and Prevention, Shanghai, People’s Republic of China; 2Fudan University Pudong Institute of Preventive Medicine, Shanghai, People’s Republic of China; 3Department of Infectious Disease Control and Prevention, Pudong New Area Center for Disease Control and Prevention, Shanghai, People’s Republic of China; 4Department of Epidemiology/Key Laboratory for Health Technology Assessment, National Commission of Health and Family Planning/Key Laboratory of Public Health Safety, Ministry of Education, School of Public Health, Fudan University, Shanghai, People’s Republic of China; 5School of Public Health, Dali University, Yunnan, People’s Republic of China; 6Research and Translational Laboratory of Acute Injury and Secondary Infection, and Department of Laboratory Medicine, Minhang Hospital, Fudan University, Shanghai, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Mingliang Chen; Lipeng Hao, Email cmlbright@126.com; hlpmail@126.comBackground: The occurrence and dissemination of hypermucoviscous and hypervirulent Klebsiella pneumoniae (hm-hvKp) isolates in clinical settings are a critical public health problem in the world. However, the data on these isolates in community populations are limited. This study aims to understand the prevalence and molecular characteristics of hm-hvKp isolates in community patients in Shanghai, China.Methods: In 2018, an active surveillance system focused on hm-hvKp in community diarrhoeal cases was implemented in Pudong New Area, Shanghai, China, involving 12 sentinel hospitals. The antimicrobial susceptibility of hm-hvKp isolates from fecal samples was tested, and whole-genome sequencing (WGS) was performed to predict the serotypes and sequence types and to identify antimicrobial resistance determinants, virulence determinants, and phylogenetic clusters.Results: The overall prevalence of hm K. pneumoniae isolates was 2.48% (31/1252), with the proportions of 1.76% (22/1252) for hm-hvKp and 0.72% (9/1252) for hm not hv K. pneumoniae. The prevalence of hm-hvKp isolates among different age groups and different months was statistically significant. All the 22 hm-hvKp isolates were susceptible to 20 antimicrobial agents and only carried blaSHV gene, and KL1 and KL2 accounted for eight (36.36%) cases and seven (31.82%) cases, respectively. The eight ST23/KL1 isolates belonged to the predominant CG23-I clade, which typically possessed the virulence determinants profile of rmpA/rmpA2-iro-iuc-ybt-irp-clb. The five ST86/KL2 isolates were assigned to the global clusters ST86/KL2-1 (n=2), ST86/KL2-2 (n=2), ST86/KL2-3 (n=1), all lack of the clb gene. Shanghai ST23/KL1 and ST86/KL2 isolates were closely related to the global isolates from liver abscesses, blood, and urine.Conclusion: Hm-hvKp is carried by the community population of Shanghai, with ST23/KL1 and ST86/KL2 isolates predominant. Hm-hvKp isolates of different continents, different sources, and different virulence levels were closely related. Ongoing surveillance of hm-hvKp isolates in the community population is warranted.Keywords: hypermucoviscous, hypervirulent, Klebsiella pneumoniae, drug resistance determinants, virulence determinants, whole-genome sequencing

Longjun Sun,1,&ast; Wenjuan Chen,2,&ast; Peixi Zhao,3 Bin Zhao,4 Guangyan Lei,1 Le Han,1,&ast; Yili Zhang2,&ast; 1Department of Thoracic Surgery, Cancer Hospital of Shaanxi Province, Xi’an, 710061, People’s Republic of China; 2Department of Oncology, Cancer Hospital of Shaanxi Province, Xi’an, 710061, People’s Republic of China; 3Department of Department of Pharmacy, Cancer Hospital of Shaanxi Province, Xi’an, 710061, People’s Republic of China; 4Department of Epidemiology, Cancer Hospital of Shaanxi Province, Xi’an, 710061, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Le Han, Department of Thoracic Surgery, Cancer Hospital of Shaanxi Province, Xi’an, 710061, People’s Republic of China, Email millyhan1@sohu.com Yili Zhang, Department of Oncology, Cancer Hospital of Shaanxi Province, Xi’an, 710061, People’s Republic of China, Email doctor_z2023@stu.xjtu.edu.cnIntroduction: Hepatocellular carcinoma (HCC) is a common and deadly malignancy. Traditional Chinese medicine, such as the compound Astragalus (wild Baicalin), has shown promise in improving outcomes for HCC patients. This study aimed to investigate the effects of wild Baicalin on the human hepatoma cell line HepG2 and elucidate the underlying mechanisms, particularly the role of the AKR1B10 and PI3K/AKT signaling pathways.Methods: HepG2 cells were treated with varying concentrations of wild Baicalin. Cell proliferation, apoptosis, migration, invasion, and cell cycle were evaluated using CCK-8, flow cytometry, scratch, Transwell, and clonogenic assays, respectively. Transcriptome sequencing was performed to analyze gene expression changes induced by wild Baicalin. Differentially expressed genes were identified and analyzed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. The expression of AKR1B10 and PI3K was validated by qPCR.Results: Wild Baicalin inhibited HepG2 cell proliferation, induced apoptosis, suppressed migration and invasion, and caused cell cycle arrest in a dose-dependent manner. Transcriptome sequencing revealed 1202 differentially expressed genes, including 486 upregulated and 716 downregulated genes. GO analysis indicated that biological processes were pivotal in the anticancer mechanism of wild Baicalin, while KEGG analysis identified metabolic pathways as the most significantly regulated. AKR1B10 and PI3K, key genes in metabolic pathways, were downregulated by wild Baicalin, which was confirmed by qPCR.Discussion: The findings suggest that wild Baicalin exhibits potent anticancer effects against HepG2 cells by inducing apoptosis, inhibiting proliferation, migration, and invasion, and causing cell cycle arrest. The regulatory effects of wild Baicalin on the AKR1B10 and PI3K/AKT signaling pathways appear to be critical for its inhibitory effects on HCC cell proliferation. These results provide new insights into the mechanism of action of wild Baicalin and support its potential as a therapeutic approach for HCC treatment. Keywords: hepatocellular carcinoma, wild baicalin, transcriptome sequencing, apoptosis

Bingcong Zhao,1,2 Zhigang Li,3 Chuan Shi,4 Yan Liu,5 Yang Sun,3 Bin Li,1,2 Jie Zhang,6 Zhizhong Gong,7 Yuanzheng Wang,8 Xuehong Ma,9 Xinjing Yang,10 Huili Jiang,3 Yuanbo Fu,1,2 Xin Wang,1,2 Yang Li,6 Hengchia Liu,6 Tuya Bao,3 Yutong Fei11– 13 1Department of Acupuncture and Moxibustion, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing, People’s Republic of China; 2Beijing Key Laboratory of Acupuncture Neuromodulation, Beijing, People’s Republic of China; 3School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing, People’s Republic of China; 4Psychological Assessment Center, Peking University Sixth Hospital, Beijing, People’s Republic of China; 5Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, People’s Republic of China; 6Department of Psychosomatic Medicine, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing, People’s Republic of China; 7Division of Medical Affairs, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing, People’s Republic of China; 8Department of Integrative TCM and Western Medicine, Peking University First Hospital, Beijing, People’s Republic of China; 9Department of Acupuncture & Moxibustion, Dongfang Hospital, the Second Clinical Medical College of Beijing University of Chinese Medicine, Beijing, People’s Republic of China; 10Department of Traditional Chinese Medicine, South China Hospital of Shenzhen University, Shenzhen, People’s Republic of China; 11Centre for Evidence-Based Chinese Medicine, Beijing University of Chinese Medicine, Beijing, People’s Republic of China; 12Institute of Excellence in Evidence-Based Chinese Medicine, Beijing University of Chinese Medicine, Beijing, People’s Republic of China; 13Beijing GRADE Centre, Beijing, People’s Republic of ChinaCorrespondence: Tuya Bao, School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, No. 11, Bei San Huan Dong Lu, Chaoyang District, Beijing, 100029, People’s Republic of China, Email tuyab@263.net Yutong Fei, Centre for Evidence-Based Chinese Medicine, Beijing University of Chinese Medicine, No. 11, Bei San Huan Dong Lu, Chaoyang District, Beijing, 100029, People’s Republic of China, Email feiyt@bucm.edu.cnPurpose: Anxious depression (AD) is a common, distinct depression subtype. This exploratory subgroup analysis aimed to explore the effects of acupuncture as an add-on therapy of selective serotonin reuptake inhibitors (SSRIs) for patients with AD or non-anxious depression (NAD).Patients and Methods: Four hundred and sixty-five patients with moderate-to-severe depression from the AcuSDep pragmatic trial were included in analysis. Patients were randomly assigned to receive MA+SSRIs, EA+SSRIs, or SSRIs alone (1:1:1) for six weeks. AD was defined by using dimensional criteria. The measurement instruments included 17-items Hamilton Depression Scale (HAMD-17), Self-Rating Depression Scale (SDS), Clinical Global Impression (CGI), Rating Scale for Side Effects (SERS), and WHO Quality of Life-BREF (WHOQOL-BREF). Comparison between AD and NAD subgroups and comparisons between groups within either AD or NAD subgroups were conducted.Results: Eighty percent of the patients met the criteria for AD. The AD subgroup had poorer clinical manifestations and treatment outcomes compared to those of the NAD subgroup. For AD patients, the HAMD response rate, remission rate, early onset rate, and the score changes on each scale at most measurement points on the two acupuncture groups were significantly better than the SSRIs group. For NAD patients, the HAMD early onset rates of the two acupuncture groups were significantly better than the SSRIs group.Conclusion: For AD subtype patients, either MA or EA add-on SSRIs showed comprehensive improvements, with small-to-medium effect sizes. For NAD subtype patients, both the add-on acupuncture could accelerate the response to SSRIs treatment. The study contributed to the existing literature by providing insights into the potential benefits of acupuncture in combination with SSRIs, especially for patients with AD subtypes. Due to its limited nature as a post hoc subgroup analysis, prospectively designed, high-quality trials are warranted.Clinical Trials Registration: ChiCTR-TRC-08000297.Keywords: anxious depression, non-anxious depression, acupuncture, SSRIs, subgroup analysis

Xiaoqi Chen,1,2,&ast; Zhuang Li,1,&ast; Yanying Feng,3 Zhijun Yang,1,2 Bingbing Zhao1,2 1Department of Gynecologic Oncology, Guangxi Medical University Cancer Hospital, Nanning, Guangxi, 530021, People’s Republic of China; 2Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor (Guangxi Medical University), Ministry of Education, Nanning, Guangxi, 530021, People’s Republic of China; 3Department of Cardiopulmonary Center, Guangxi Medical University Cancer Hospital, Nanning, Guangxi, 530021, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Bingbing Zhao; Zhijun Yang, Department of Gynecologic Oncology, Guangxi Medical University Cancer Hospital, No. 71 Hedi Road, Nanning, Guangxi, 530021, People’s Republic of China, Email zhaobingbing@gxmu.edu.cn; yzj7528@126.comPurpose: Evidence has indicated that PDZD11 is involved in regulating adherens junction. However, the distinct effect of its aberrant expression on epithelial ovarian cancer (EOC) awaits clarification.Methods: In this study, public databases (Gene Expression Omnibus, The Cancer Genome Atlas, and The Genotype-Tissue Expression), online analysis tools (Kaplan-Meier plotter and TIMER), and data analysis methods (Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and the CIBERSORT algorithm) were fully utilized to analyze the differential expression, diagnostic efficiency, prognostic significance, potential function, and correlation with immune infiltration of PDZD11. The differential expression of PDZD11 was tested by immunohistochemistry in EOC tissues (78 cases) and control tissues (37 cases).Results: Our results indicate that PDZD11 was remarkably overexpressed in EOC, which was associated with advanced cancer stages, no lymphatic metastasis status, and poor prognosis. Moreover, PDZD11 played a role in cell adhesion, cell proliferation, and immune responses. Also, PDZD11 was significantly related to the abundances of infiltrating immune cells in EOC, including neutrophils, macrophages, dendritic cells, CD8+ T cells, and CD4+ T cells, and its expression was positively co-expressed with well-known immune checkpoints, including TIGIT, TIM3, LAG3, CTLA4, and PD-1.Conclusion: These results suggest that PDZD11 could be a potential diagnostic and prognostic biomarker associated with immune infiltration in EOC, and our findings might help elucidate the function of PDZD11 in carcinogenesis. Keywords: PDZD11, epithelial ovarian cancer, prognostic biomarker, immune infiltration, immune checkpoint

Meiyang Yang,1,2,&ast; Weijun Chen,1,&ast; Dhanu Gupta,3 Congjin Mei,1 Yang Yang,1 Bingke Zhao,1 Lipeng Qiu,1 Jinghua Chen1 1School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, People’s Republic of China; 2School of Chemical and Material Engineering, Jiangnan University, Wuxi, People’s Republic of China; 3Department of Paediatrics, University of Oxford, Oxford, UK&ast;These authors contributed equally to this workCorrespondence: Lipeng Qiu; Jinghua Chen, School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, 214122, People’s Republic of China, Email qiulp@jiangnan.edu.cn; chenjinghua@jiangnan.edu.cnBackground: New treatment modalities for hepatocellular carcinoma (HCC) are desperately critically needed, given the lack of specificity, severe side effects, and drug resistance with single chemotherapy. Engineered bacteria can target and accumulate in tumor tissues, induce an immune response, and act as drug delivery vehicles. However, conventional bacterial therapy has limitations, such as drug loading capacity and difficult cargo release, resulting in inadequate therapeutic outcomes. Synthetic biotechnology can enhance the precision and efficacy of bacteria-based delivery systems. This enables the selective release of therapeutic payloads in vivo.Methods: In this study, we constructed a non-pathogenic Escherichia coli (E. coli) with a synchronized lysis circuit as both a drug/gene delivery vehicle and an in-situ (hepatitis B surface antigen) Ag (ASEc) producer. Polyethylene glycol (CHO-PEG2000-CHO)-poly(ethyleneimine) (PEI25k)-citraconic anhydride (CA)-doxorubicin (DOX) nanoparticles loaded with plasmid encoded human sulfatase 1 (hsulf-1) enzyme (PNPs) were anchored on the surface of ASEc (ASEc@PNPs). The composites were synthesized and characterized. The in vitro and in vivo anti-tumor effect of ASEc@PNPs was tested in HepG2 cell lines and a mouse subcutaneous tumor model.Results: The results demonstrated that upon intravenous injection into tumor-bearing mice, ASEc can actively target and colonise tumor sites. The lytic genes to achieve blast and concentrated release of Ag significantly increased cytokine secretion and the intratumoral infiltration of CD4/CD8+T cells, initiated a specific immune response. Simultaneously, the PNPs system releases hsulf-1 and DOX into the tumor cell resulting in rapid tumor regression and metastasis prevention.Conclusion: The novel drug delivery system significantly suppressed HCC in vivo with reduced side effects, indicating a potential strategy for clinical HCC therapy. Keywords: hepatocellular carcinoma, engineered bacteria, chemotherapy, immunotherapy, anti-angiogenesis

Bennan Zhao,1,&ast; Qingxiao Liu,2,&ast; Qing Du,3 Jun Kang,1 Rong Tang,4 Yalan Tu,5 Dafeng Liu1 1The First Ward of Internal Medicine, Public Health Clinical Center of Chengdu, Chengdu, People’s Republic of China; 2Department of Neurology, Hospital of Chengdu University of TCM, Chengdu, People’s Republic of China; 3The Second Ward of ICU, Public Health Clinical Center of Chengdu, Chengdu, People’s Republic of China; 4Department of Integrated Chinese and Western Medicine, Public Health Clinical Center of Chengdu, Chengdu, People’s Republic of China; 5Scientific Research and Teaching Department, Public Health Clinical Center of Chengdu, Chengdu, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Dafeng Liu, The First Ward of Internal Medicine, Public Health Clinical Center of Chengdu, Chengdu, People’s Republic of China, Tel +86 1 309 449 4836, Email ldf312@126.com Yalan Tu, Scientific Research and Teaching Department, Public Health Clinical Center of Chengdu, Chengdu, People’s Republic of China, Tel +86 1 501 326 6454, Email tyl950330@163.comPurpose: To date, there are few reports about mpox case series in China, and scarce information is available about the in-vivo kinetics of T-cell responses in the early stage of mpox infection. This study aims to investigate the clinical difference among mpox patients with and without human immunodeficiency virus (HIV) infection.Patients and Methods: A total of 56 patients diagnosed with mpox by Chengdu Center for Disease Control and Prevention (CDC) and hospitalized in Public Health Clinical Center of Chengdu were retrospectively included and divided into an HIV-infected group (n=23) and a non-HIV-infected group (n=33). Clinical characteristics and serum chemistry findings of mpox patients were collected in order to analyze the differences between the HIV-infected group and the non-HIV-infected group.Results: Multiple laboratory abnormalities, including elevated C-reactive protein (69.1%), hypocalcemia (50.9%), elevated CD3+CD8+T counts (47.0%) and inverted ratio of CD3+CD4+T to CD3+CD8+T (64.7%) were common in mpox cases. There were statistically significant differences (all P < 0.05) in age, serum calcium levels, CD3+CD4+T counts, the ratio of CD3+CD4+T to CD3+CD8+T, proportion with > 10 rashes, incidence of proctitis anus and time from rash growth to rash scab shedding between HIV-infected group and non-HIV-infected group. In the early stage of mpox infection, the median of CD3+CD8+T counts in the non-HIV-infected group was significantly higher than that in healthy donors (P< 0.001), and the median of CD3+CD4+T/CD3+CD8+T ratio was significantly lower (P< 0.001). The median of CD3+CD4+T counts in mpox patients co-infected with HIV significantly decreased compared to the pre-infection level (p =0.033).Conclusion: Our study indicates that mpox co-infected with HIV patients have longer lasting rash lesions and a higher incidence of proctitis anus. T-cell responses may be different between HIV-infected and non-HIV-infected individuals in the early stage of mpox infection.Keywords: mpox, HIV, clinical characteristics, complication, T-cell response

Zexuan Song,1 Wencong He,1 Xiaolong Cao,1 Aijing Ma,1 Ping He,1 Bing Zhao,1 Shengfen Wang,1 Chunfa Liu,1,2 Yanlin Zhao1 1National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, Changping, Beijing, 102206, People’s Republic of China; 2Animal Science and Technology College, Beijing University of Agriculture, Huilongguan, Changping, Beijing, 102206, People’s Republic of ChinaCorrespondence: Chunfa Liu; Yanlin Zhao, Email liucf@cau.edu.cn; zhaoyl@chinacdc.cnBackground: Tuberculosis (TB) remains a severe public health problem globally, and it is essential to comprehend the transmission pattern to control tuberculosis. Herein, we evaluated the drug-resistant characteristics, recent transmission, and associated risk factors of TB in Golmud, Qinghai, China.Methods: In this study, we performed a population-based study of patients diagnosed with TB in Golmud from 2013 to 2018. Drug-susceptibility testing and whole-genome sequencing were performed on 133 Mycobacterium tuberculosis strains. The genomic clustering rate was calculated to evaluate the level of recent transmission. Risk factors were identified by logistic regression analysis.Results: Our results showed that 46.97% (62/132) of strains were phylogenetically clustered and formed into 23 transmission clusters, suggesting a high recent transmission of TB in the area. 12.78% (17/133) strains were multidrug-resistant/rifampicin tuberculosis (MDR/RR-TB), with a high drug-resistant burden. Based on drug resistance gene analysis, we found 23 strains belonging to genotype MDR/RR-TB, where some strains may have borderline mutations. Among these strains, 65.2% (15/23) were found within putative transmission clusters. Additionally, risk factor analysis showed that recent transmission of TB happened more in patients with Tibetan nationality or older age.Conclusion: Overall our study indicates that the recent transmissions of MTB strains, especially genotypic MDR/RR strains, drive the tuberculosis epidemic in Golmud, which could contribute to developing effective TB prevention and control strategies.Keywords: tuberculosis, recent transmission, risk factor, whole-genome sequencing

Guanghua Yang,1,&ast; Jiaxi Pu,1,&ast; Sibo Zhu,2 Yong Shi,3 Yi Yang,3 Jiangnan Mao,3 Yongkang Sun,1 Bin Zhao1 1Department of General Surgery, Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai, 200137, People’s Republic of China; 2School of Life Sciences, Fudan University, Shanghai, 200438, People’s Republic of China; 3Cinoasia Institute, Shanghai, 200438, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Bin Zhao, Department of General Surgery, Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine, No. 358, Datong Road, Gaoqiao, Pudong New Area, Shanghai, 200137, People’s Republic of China, Tel +86 13918468421, Email zhaobin0727@sina.com Yongkang Sun, Department of Ultrasound, Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine, No. 358, Datong Road, Gaoqiao, Pudong New Area, Shanghai, 200137, People’s Republic of China, Tel +86 13601659860, Email elangwang@126.comBackground: Thyroidectomy is commonly performed for benign or malignant thyroid tumors, often resulting in hypothyroidism. Levothyroxine (LT4) supplementation is crucial to maintain hormone levels within the normal range and suppress TSH for cancer control. However, determining the optimal dosage remains challenging, leading to uncertain outcomes and potential side effects.Methods: We analyzed clinical examination data from 510 total thyroidectomy patients, including demographic information, blood tests, and thyroid function. Using R, we applied data preprocessing techniques and identified 274 samples with 98 variables. Principal Component Analysis, correlation analysis, and regression analysis were conducted to identify factors associated with optimal LT4 dosage.Results: The analysis revealed that only eight variables significantly influenced the final satisfactory dosage of LT4 in tablets: Benign0/Malignant1 (benign or malignant), BQB (electrophoretic albumin ratio), TP (total protein), FDP (fibrin degradation products), TRAB_1 (thyroid-stimulating hormone receptor antibody), PT (prothrombin time), MONO&num; (monocyte count), and HCV0C (hepatitis C antibody). The resulting predictive model was: .Conclusion: Parameters such as benign/malignant status, TRAB_1, and BQB ratio during medication can serve as observational indicators for postoperative LT4 dosage. The calculated linear model can predict the LT4 dosage for patients after thyroidectomy, leading to improved treatment effectiveness and conserving medical resources.Keywords: thyroid, levothyroxine, dosage, model, prediction

Yi Yao,1,&ast; Juan Li,2,&ast; Wenwu Sun,1,&ast; Chaoping Ma,2,&ast; Ning Ning,1 Jiaoyan Li,1 Tongtian Ni,1 Menglu Gui,1 Ying Chen,1 Bing Zhao,1 Jiyuan Zhang,2 Jiyue Zhu,3 Chengjin Gao,2 Huiqiu Sheng,1 Enqiang Mao1 1Departments of Emergency, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200025, People’s Republic of China; 2Department of Emergency, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China; 3Department of General Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Huiqiu Sheng, Departments of Emergency, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200025, People’s Republic of China, Tel +86-15801971839, Email shenghq803@163.com Chengjin Gao, Department of Emergency, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China, Tel +86-18221088783, Email gaochengjin@xinhuamed.com.cnPurpose: Both nonthyroidal illness syndrome (NTIS) and disseminated intravascular coagulation (DIC) are commonly occurred in sepsis. The objective of this study is to evaluate the association between NTIS and DIC, as well as their impacts on the mortality in adults with sepsis.Patients and methods: A total of 1219 septic patients in two Chinese academic centers from October 2012 and October 2022 were enrolled in analysis. We conduct logistic regression models to analyze the independent risk factors for DIC. Modified Poisson regression models are used to estimate the relative risk (RR) of NTIS on the 28 days mortality in septic patients with DIC. Correlation analysis between thyroid function parameters and coagulation parameters is performed with Pearson coefficient be reported.Results: DIC is diagnosed on 388 (31.8%) of all the 1219 enrolled septic patients within 72 hours after admission. In multivariate logistic regression models, NTIS (OR 3.19; CI 2.31– 4.46; p< 0.001) is a statistically significant independent risk factor for DIC after adjustment for potential confounders. Twenty-eight days mortality is significantly higher in DIC patients complicated with NTIS compared with the other DIC patients (23.2% vs 14.0%, p=0.024). This result is also robust in different modified Poisson regression models (Model 1: RR 1.46; CI 1.25– 1.70; p< 0.001; Model 2: RR 1.35; CI 1.14– 1.60; p< 0.001; Model 3: RR 1.18; CI 1.02– 1.37; p=0.026). Correlation analysis reveals that the thyroid function parameters of FT3, FT4 and TSH only have weak correlations with coagulation parameters of platelet count, fibrinogen, FDP, D-dimers, PT, APTT and INR in sepsis.Conclusion: NTIS is an independent risk factor for DIC in adults with sepsis. DIC patients complicated with NTIS have significantly higher severity and higher rate of mortality.Keywords: sepsis, low T3 syndrome, nonthyroidal illness syndrome, thyroid function, disseminated intravascular coagulation

Loy Eid,&ast; Xiyuan Mao,&ast; Binfan Zhao, Jiayi Mao, Shutong Qian, Yuyu Zhang, Ying Zhang, Lu Zhang, Liying Cheng, Yuguang Zhang, Xiaoming Sun Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yuguang Zhang; Xiaoming Sun, Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai, People’s Republic of China, Email zhangyg18@126.com; drsunxm@126.comBackground: Facial fat compartments and their role in facial aging have gained increased recognition and are playing a significant role in facial rejuvenation. The superficial fat compartments glide inferiorly during the aging process, leading to the flattening and elongation of the face and the appearance of facial bulges, folds, and grooves.Patients and Methods: Ultrasound imaging of the facial soft tissues was performed on nine female volunteers to demonstrate the change in superficial facial fat compartments from an upright to supine position. The net suture jowl and medial cheek fat compartment repositioning technique was operated on 165 Asian patients between September 2020 and July 2021. Volume and projection change of malar and jowl regions, as well as change in elevation of malar protrusion were measured 1, 3, and 6 months postoperatively using a three-dimensional imaging system.Results: Ultrasound measurements confirmed the medial and middle cheek, nasolabial, and jowl fat compartments changed in thickness during positional changes with age-related differences. Postoperative three-dimensional imaging showed volume and projection increase in the malar region (2.23mL and 1.11mm) and decrease in the jowl region (− 0.18mL and − 0.52mm) by the 6-month follow-up date, and malar projection saw a superior displacement of 3.08mm.Conclusion: The superficial fat glide inferiorly within their compartments under the force of gravity and naturally reposition themselves when the effect of gravity is reversed. The net suture technique offers a minimally invasive method for lifting the jowl fat, volumizing the mid-cheek and achieving facial rejuvenation by repositioning the superficial fat compartments.Keywords: fat compartments, facial rejuvenation, face lift, facial aging

Shi Chen,1,&ast; Jiaqi Qiang,1,2,&ast; Bin Zhao,3 Ran Tian,4 Tao Yuan,1 Ming Li,1 Mei Li,1 Zhaoqi Gu,5 Liping Yang,6 Huijuan Zhu,1 Hui Pan,1,7 Yan Tang,3 Yuxiu Li1 1Key Laboratory of Endocrinology of National Health Commission, Department of Endocrinology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, People’s Republic of China; 2Eight-Year Program of Clinical Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, People’s Republic of China; 3Department of Pharmacy, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, People’s Republic of China; 4Department of Cardiology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, People’s Republic of China; 5Department of Radiotherapy, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, People’s Republic of China; 6Department of Pharmacy, Handan First Hospital, Handan, Hebei Province, People’s Republic of China; 7State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yuxiu Li, Key Laboratory of Endocrinology of National Health Commission, Department of Endocrinology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, No. 1 Shuaifuyuan, Wangfujing, Dongcheng District, Beijing, 100730, People’s Republic of China, Email liyuxiu@medmail.com.cn Yan Tang, Department of Pharmacy, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, No. 1 Shuaifuyuan, Wangfujing, Dongcheng District, Beijing, 100730, People’s Republic of China, Email tangyan@pumch.cnAbstract: The sulfhydryl group of clopidogrel metabolite could induce insulin autoimmune syndrome (IAS) with hypoglycemia as the major symptom. For patients with cardiovascular disease taking clopidogrel for vascular protection, this adverse event hypoglycemia increases the risk of cardiovascular events. However, discontinuing clopidogrel leaves patients without appropriate antiplatelet therapy. Treating IAS with glucocorticoids is also risky for these patients’ primary cardiovascular diseases. Early recognition and appropriate treatment of clopidogrel-induced IAS (CIAS) would be beneficial for patients. This research aimed to discover the clinical features and investigate optimal therapeutic management of CIAS. We systematically searched for cases of CIAS in PubMed and Embase and performed data mining in Food and Drug Administration Adverse Event Reporting System (FAERS). In the CIAS series, clinical features were summarized and compared to 287 IAS cases, including demographic information, HLA alleles, onset, and symptoms. The therapeutic effect of glucocorticoids was compared between the receiving group and the not-receiving group. The possibilities of common antiplatelet drugs to induce hypoglycemia/IAS were investigated with chemical structure and FAERS reports. A CIAS series of 51 patients was established. CIAS had an onset age of 74.8± 8.6 years old, 92.2% male, and a balanced proportion of East Asians and non-East Asians. Confusion occurred more frequently in CIAS than in IAS from various causes, while the other symptoms and hypoglycemia types were similar. The recovery time was approximately the same whether using glucocorticoids/immunotherapy in CIAS or not. Among common antiplatelet drugs, ticagrelor and rivaroxaban were unlikely to induce hypoglycemia/IAS. Clopidogrel is a distinctive cause of IAS featuring an elderly male presenting confusion as the symptom of hypoglycemia. Glucocorticoids/immunotherapy might not be necessary for the long-term recovery of CIAS. To balance the risks of hypoglycemia and cardiovascular events, substituting clopidogrel with ticagrelor and rivaroxaban might be considered.Keywords: autoimmune hypoglycemia, antiplatelet therapy, glucocorticoids, cardiovascular diseases, ticagrelor

Bi Zhao,1 Yuee Liu,2 Yi Yang,3 Jihui He1 1Department of Gastroenterology, Jinshan Branch of Shanghai Sixth People’s Hospital, Shanghai, 201500, People’s Republic of China; 2Department of Emergency, Changhai Hospital, Shanghai, 200433, People’s Republic of China; 3Department of Emergency, Ruijin Hospital and Luwan Branch, School of Medicine, Shanghai Jiaotong University, Shanghai, 200020, People’s Republic of ChinaCorrespondence: Bi Zhao, Department of Gastroenterology, Jinshan Branch of Shanghai Sixth People’s Hospital, Shanghai, 201500, People’s Republic of China, Email biabia0120@126.comBackground: The aim of this study was to explore the relationship between systemic immune-inflammation (SII) index with non-alcoholic fatty liver disease (NAFLD) in the general population of the United States (U.S.).Methods: We conducted a cross-sectional study of subjects in the National Health and Nutrition Examination Survey 2017– 2018. For the analysis of the association between SII index and risk of NAFLD, the restricted cubic spline (RCS) plot, we performed multivariable logistic regression models and subgroup analysis. In addition, generalized additive models with smooth functions were conducted for the relationship between the SII index and the ZJU index, the BARD score, and the NAFLD fibrosis score.Results: There were a total of 1197 individuals in our study. Taking into account known confounding variables, compared with the lowest quartiles, the odds ratios with 95% confidence intervals for NAFLD across the quartiles were 0.923 (0.585, 1.455), 0.563 (0.351, 0.901), and 1.061 (0.669, 1.682), respectively. As shown by the RCS plot, the SII index was linked with NAFLD risk in a U-shaped pattern. Based on the results of subgroup analysis, SII index and NAFLD risk were U-curve correlated among participants in all age groups, male or female, with or without hypertension, with diabetes mellitus, and with a BMI of < 30 or > 30 kg/m2. The SII index was linearly positive with the ZJU index but negative with the NAFLD fibrosis score. However, the SII index and BARD score showed a trend of first decreasing, then increasing, and then decreasing.Conclusion: The U-shaped relationships exist between SII index and risk of NAFLD, which highlighted that we should focus on the dynamic change of SII index.Keywords: cross-sectional study, non-alcoholic fatty liver disease, systemic immune-inflammation index, United States

Menglu Gui,1,&ast; Jun Huang,2,&ast; Huiqiu Sheng,1,&ast; Ying Chen,1 Zhitao Yang,1 Li Ma,1 Daosheng Wang,3 Lili Xu,1 Wenwu Sun,1 Junling Liu,4 Yanyan Xu,4 Erzhen Chen,1 Bing Zhao,1 Enqiang Mao1 1Department of Emergency in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China; 2Department of Cardiovascular Medicine, State Key Laboratory of Medical Genomics, Shanghai Key Laboratory of Hypertension, Shanghai Institute of Hypertension, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China; 3Department of Laboratory Medicine in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China; 4Department of Biochemistry and Molecular Cell Biology, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Enqiang Mao; Bing Zhao, Department of Emergency in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China, Tel +8613501747906 ; +8618017117871, Email maoeq@yeah.net; zhaobing124@163.comBackground: Platelet activation in the early stage of pancreatitis is the key step developing into pancreatic necrosis. Studies suggested that vitamin C (Vit C) can inhibit platelet activity by targeting CXCL12/CXCR4 pathway. High-dose Vit C were showed to reduce pancreatic necrosis in severe acute pancreatitis (SAP) but the mechanism remains unclear. Here we speculate high-dose Vit C reduce pancreatic necrosis by inhibiting platelet activation through downregulating CXCL12/CXCR4 pathway.Methods: The pancreatic microcirculation of rats was observed by intravital microscopy. The platelet activity of SAP rats treated with or without high-dose Vit C was analyzed by platelet function test. Besides, the activity of platelets preincubated with high-dose Vit C or vehicle from SAP patients was also evaluated. Then, the TFA (CXCR4 agonist) and rCXCL12 were used to neutralize the effect of high-dose Vit C in SAP rats treated with high-dose Vit C. Meanwhile, the levels of enzymes and inflammatory cytokines in rat plasma, and rats’ pancreatic histopathology and mortality were assessed.Results: Platelets from animals and patients with SAP are more sensitive to agonists and are more easily activated. Administration of high-dose Vit C significantly ameliorated excessive activation of platelets in SAP rats, ultimately increasing the microvessel density and inducing microthrombus and blood stasis; these results were consistent with clinical sample analysis. Moreover, high-dose Vit C significantly inhibited the release of amylase, lipase, TNF-α, and IL-6 in SAP rat plasma, reducing pancreatic damage and the mortality of SAP rats. However, using TFA and rCXCL12 significantly reversed the effect of high-dose Vit C on excessive activation of platelets, aggravating microcirculation impairment and pancreatic damage.Conclusion: The present study suggests that high-dose Vit C can ameliorate pancreatic necrosis by improving microcirculation disorders of SAP. For the first time, the underlying mechanism is related with inhibiting platelet activation through the CXCL12/CXCR4 pathway.Keywords: severe acute pancreatitis, microcirculation impairment, platelets activity, vitamin C, CXCL12/CXCR4 pathway

Lili Sun, Cong Wang, Mingqing Zhang, Xiang Li, Bin Zhao Department of Emergency, Beijing Jishuitan Hospital, Beijing, People’s Republic of ChinaCorrespondence: Bin Zhao, Department of Emergency, Beijing Jishuitan Hospital, No. 31 of Xinjiekou East Street, Xicheng District, Beijing, 100035, People’s Republic of China, Tel +86-1058516417, Fax +86-1058516931, Email binzhao1492@163.comBackground: Guidelines exist for the surgical treatment of hip fractures, but the association between the surgical timing and the incidence of postoperative complications and other important outcomes in elderly patients with hip fracture remains controversial.Objective: This study aims to explore the association between the surgical timing and the prognoses in elderly patients with hip fracture.Methods: A total of 701 elderly patients (age ≥ 65 years) with hip fractures who were treated in our hospital from June 2020 to June 2021 were selected. Patients who underwent surgery within 2 d of admission were assigned to the early surgery group, and those who underwent surgery after 2 d of admission were assigned to the delayed surgery group. The prognosis indices of the patients in the two groups were recorded and compared.Results: The length of postoperative hospitalisation in the early surgery group was significantly lower than that in the delayed surgery group (P < 0.001). The European quality of life questionnaire (EQ-5D) utility in the delayed surgery group was significantly lower than that in the early surgery group at 30 days and 6 months after operation (P< 0.05). Compared with the delayed surgery group, the incidence of pulmonary infection, urinary tract infection (UTI) and deep vein thrombosis (DVT) in the early surgery group were significantly lower. There were no significant differences between the two groups in terms of mortality and excellent rates of the HHS at six months after the operation. In addition, the early surgery group had a lower readmission rate than the delayed surgery group [34 (9.5%) vs 56 (16.3%), P = 0.008].Conclusion: Earlier surgery can reduce the incidence of pulmonary infections, UTI, DVT and readmission rate among elderly patients with hip fractures, shorten postoperative hospitalisation.Keywords: Hip fracture, elderly patients, timing of surgery, prognosis

Mina Wang,1,2,&ast; Xiaobai Xu,1,&ast; Bingcong Zhao,1 Lu Liu,1 Luopeng Zhao,1 Fan Zhang,1 Xu Ji,1 Fang Yuan,1 Qiuyu Xia,1 Shaosong Wang,1 Wei Tian,1 Linpeng Wang,1 Bin Li1 1Department of Acupuncture and Moxibustion, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing, People’s Republic of China; 2Graduate School, Beijing University of Chinese Medicine, Beijing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Bin Li; Xiaobai Xu, Email libin@bjzhongyi.com; xu_xiaobai1990@sina.comPurpose: Knee osteoarthritis (KOA) is regarded as one of the leading musculoskeletal diseases. Although the efficacy is under exploration, fire needling therapy is considered an effective alternative for KOA. This trial aims to investigate the effectiveness of different frequencies of fire needling therapy in attenuating pain and promoting function in KOA patients.Methods: This is a study protocol for a pilot, three-arm, single-center, randomized controlled trial. A total of 90 participants with KOA will be recruited and randomly assigned to the high-frequency fire needling group (3 sessions per week, for 6 weeks), the low-frequency fire needling group (1 session per week, for 6 weeks) or the positive control group (Diclofenac Diethylamine Emulgel, 3 times per day, for 6 weeks) in a 1:1:1 ratio. Participants will accomplish the trial at Week 14 after a follow-up evaluation. The response rate will be set as the primary outcome that the proportion of participants obtaining a minimal clinically important difference, which is identified as ≥ 2 units on the numerical rating scale (NRS) and ≥ 6 units on the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) function score at Week 6 compared with Week 0. Secondary outcomes are NRS, WOMAC, Brief Pain Inventory, Short-Form Health Survey-12, Timed Up and Go Test, and pain threshold.Discussion: This is the first standardized protocol comparing fire needling therapy and positive control drugs. This trial may provide reliable evidence for the effectiveness of fire needling therapy and dose–effect property of it in KOA. Trial registration: The trial has been registered on Chinese Clinical Trial Registry (Registered number: ChiCTR2100043041), registered on 4 February 2021.Keywords: knee osteoarthritis, fire needling therapy, different frequencies, dose–effect property

Xiaonan Meng,1,2,&ast; Jie Sun,3,&ast; Qi Liu,4,&ast; Yueping Huang,5 Xianwen Qiu,6 David Jung Seto,7,8 Ying Li,3 Liping Wang,2 Chunying Li,2 Sen Gao,9 Haikuo Yu,10 Jiping Zhao,1 Baixiao Zhao1 1Department of Acupuncture and Moxibustion, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, People’s Republic of China; 2Department of Acupuncture and Moxibustion, Beijing Huguosi TCM Hospital, Affiliated with Beijing University of Chinese Medicine, Beijing, People’s Republic of China; 3Department of Integrated Chinese and Western Medicine Rehabilitation, Beijing Xiaotangshan Hospital, Beijing, People’s Republic of China; 4Department of General Internal Medicine, Guang’anmen Hospital, China Academy of Chinese Medical Sciences, Beijing, People’s Republic of China; 5School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing, People’s Republic of China; 6Department of Acupuncture and Moxibustion, Beijing Shichahai Community Healthcare Center, Beijing, People’s Republic of China; 7Division of Integrative Medicine, Department of Medicine, Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles, CA, USA; 8Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, CA, USA; 9Department of Rehabilitation, Beijing Huguosi TCM Hospital, affiliated with Beijing University of Chinese Medicine, Beijing, People’s Republic of China; 10Rehabilitation Department, Xuanwu Hospital Capital Medical University, Beijing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jiping Zhao; Baixiao Zhao, Email zjp7883@sina.com; baixiao100@vip.sina.comBackground: Post-stroke shoulder-hand syndrome (PS-SHS), a common neurological comorbidity after stroke episodes, poses a grave threat on patients’ functional recovery. Preliminary trials have demonstrated that the acupuncture and moxibustion treatment, including a dermal acupuncture tapping method known as plum blossom needling (PBN) can improve pain and motor dysfunctions in patients with PS-SHS. However, there are few reports describing simultaneous moxibustion treatment in combination with PBN. Hence, a novel plum blossom needle device with mild moxibustion (PBNMM) was developed to evaluate its potential efficacy and safety in patients with stage 1 PS-SHS.Materials and Methods: This multicenter, sham-controlled, randomized controlled trial (RCT) will recruit 102 eligible patients with stage 1 PS-SHS from three clinical centers, randomly allocated in a ratio of 1:1:1 to the PBNMM group, PBNMM with no moxa smoke (PBNMM-NMS) group and sham control group. Patients in each group will receive a 30-minute treatment once per day for 4 weeks, with 5 consecutive sessions per week, for a total of 20 sessions. The primary outcome measure will be defined as the decreased scores from baseline in the visual analog scale (VAS) assessment at week 4. Secondary outcome measures will include scores on the Fugl-Meyer Assessment of the Upper Extremity Scale (FMA-UE), the Modified Barthel Index (MBI), and the somatosensory evoked potential (SEP) records. All outcomes will be evaluated at baseline and weeks 4, 5, 6 and 10, and the intention-to-treat analysis will be applied.Conclusion: This study aims to provide robust evidence for the efficacy and safety of the PBNMM for PS-SHS treatment, as well as the specific impact of moxibustion smoke itself in dealing with PS-SHS.Clinical Trial Registration: Chinese Clinical Trial Registry No. ChiCTR2200062441. Registered on 7 August 2022.Keywords: plum blossom needling, mild moxibustion, novel device, post-stroke shoulder-hand syndrome, protocol, randomized controlled trial

Jianyi Li,1,&ast; Keke Li,2,&ast; Yukun Du,1,&ast; Xiaojie Tang,3,&ast; Chenjing Liu,1 Shannan Cao,3 Baomeng Zhao,2 Hai Huang,3 Hongri Zhao,3 Weiqing Kong,1 Tongshuai Xu,3 Cheng Shao,1 Jiale Shao,1 Guodong Zhang,4 Hongbo Lan,5 Yongming Xi1 1Department of Orthopaedic Surgery, the Affiliated Hospital of Qingdao University, Qingdao, People’s Republic of China; 2Yantai Campus of Binzhou Medical University, Yantai, People’s Republic of China; 3Yantai Affiliated Hospital of Binzhou Medical University, Yantai, People’s Republic of China; 4Tengzhou Central People’s Hospital, Tengzhou, People’s Republic of China; 5Shandong Engineering Research Center for Additive Manufacturing Qingdao University of Technology, Qingdao, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yongming Xi, Department of Orthopaedic Surgery, the Affiliated Hospital of Qingdao University, Qingdao, 266071, People’s Republic of China, Email xym700118@163.com Hongbo Lan, Shandong Engineering Research Center for Additive Manufacturing Qingdao University of Technology, Qingdao, 266520, People’s Republic of China, Email hblan99@126.comBackground: Successful treatment of infectious bone defect remains a major challenge in the orthopaedic field. At present, the conventional treatment for infectious bone defects is surgical debridement and long-term systemic antibiotic use. It is necessary to develop a new strategy to achieve effective bone regeneration and local anti-infection for infectious bone defects.Methods: Firstly, vancomycin / poly (lactic acid-glycolic acid) sustained release microspheres (VAN/PLGA-MS) were prepared. Then, through the dual-nozzle 3D printing technology, VAN/PLGA-MS was uniformly loaded into the pores of nano-hydroxyapatite (n-HA) and polylactic acid (PLA) scaffolds printed in a certain proportion, and a composite scaffold (VAN/MS-PLA/n-HA) was designed, which can not only promote bone repair but also resist local infection. Finally, the performance of the composite scaffold was evaluated by in vivo and in vitro biological evaluation.Results: The in vitro release test of microspheres showed that the release of VAN/PLGA-MS was relatively stable from the second day, and the average daily release concentration was about 15.75 μg/mL, which was higher than the minimum concentration specified in the guidelines. The bacteriostatic test in vitro showed that VAN/PLGA-MS had obvious inhibitory effect on Staphylococcus aureus ATCC-29213. Biological evaluation of VAN/MS-PLA/n-HA scaffolds in vitro showed that it can promote the proliferation of adipose stem cells. In vivo biological evaluation showed that VAN/MS-PLA/n-HA scaffold could significantly promote bone regeneration.Conclusion: Our research shows that VAN/MS-PLA/n-HA scaffolds have satisfying biomechanical properties, effectively inhibit the growth of Staphylococcus aureus, with good biocompatibility, and effectiveness on repairing bone defects. The VAN/MS-PLA/n-HA scaffold provide the clinic with an application prospect in bone tissue engineering.Graphical Abstract: Keywords: osteogenesis, dual-nozzle 3D printing technology, drug-loaded microspheres, vancomycin, nano-hydroxyapatite

Chen Jiang,1,&ast; Stefanos Mastrotheodoros,2,3 Yihong Zhu,4,&ast; You Yang,5 Souheil Hallit,6– 8 Bing Zhao,4 Yuping Fan,1 Mengyi Huang,1 Cen Chen,1 Haiyan Ma,1,9 Runtang Meng1,9,&ast; 1School of Public Health, Hangzhou Normal University, Hangzhou, People’s Republic of China; 2Department of Psychology, University of Crete, Rethymno, Greece; 3Department of Youth and Family, Utrecht University, Utrecht, the Netherlands; 4The Affiliated Hospital of Hangzhou Normal University, Hangzhou Normal University, Hangzhou, People’s Republic of China; 5School of Nursing, Hangzhou Normal University, Hangzhou, People’s Republic of China; 6School of Medicine and Medical Sciences, Holy Spirit University of Kaslik, Jounieh, Lebanon; 7Applied Science Research Center, Applied Science Private University, Amman, Jordan; 8Research Department, Psychiatric Hospital of the Cross, Jal Eddib, Lebanon; 9Engineering Research Center of Mobile Health Management System, Ministry of Education, Hangzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Runtang Meng; Haiyan Ma, School of Public Health, Hangzhou Normal University, No. 2318, Yuhangtang Road, Yuhang District, Hangzhou, Zhejiang, 311121, People’s Republic of China, Email mengruntang@hznu.edu.cn; mahaiyan@hznu.edu.cnPurpose: Stress may relate to an increased risk of psychological and physical disorders. Thus, a brief and efficient measurement instrument for researchers to measure stress is essentially needed.Participants and Methods: To assess measurement properties of the validated Chinese version of the Perceived Stress Questionnaire-13 (PSQ-C-13), we conducted a two-wave longitudinal study from September to December, 2021 with a convenient sample of medical students.Results: A two-factor (constraint and imbalance) structure showed good fit indices (Comparative Fit Index [CFI] = 0.972, Tucker-Lewis Index [TLI] = 0.966, Root Mean Square Error of Approximation [RMSEA] = 0.062). Spearman correlations with the Chinese Perceived Stress Scale-10 illustrated that convergent validity of the PSQ-C-13 was relatively satisfactory (r = 0.678 [baseline], 0.753 [follow-up]). Measurement invariance was supported across subgroups (gender, age, home location, single-child status, monthly households’ income, and part-time status) and time points. Internal consistency was sound (Cronbach’s α = 0.908 [baseline], 0.922 [follow-up]; McDonald’s ω = 0.909 [baseline], 0.923 [follow-up]). Stability between time points was good (Intraclass Correlation Coefficient = 0.834).Conclusion: The two factors of the PSQ-C-13 including constraint and imbalance may adequately measure the level of stress on participants. The PSQ-C-13 is a convenient and efficient instrument that contains valid and reliable psychometric properties.Keywords: Perceived Stress Questionnaire, longitudinal survey, medical students, psychometrics

Menglu Gui,1,&ast; Bing Zhao,1,&ast; Jun Huang,2,&ast; Erzhen Chen,1 Hongping Qu,3 Enqiang Mao1 1Department of Emergency, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 2Department of Cardiovascular Medicine, State Key Laboratory of Medical Genomics, Shanghai Key Laboratory of Hypertension, Shanghai Institute of Hypertension, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 3Department of Critical Care Medicine, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Enqiang Mao, Department of Emergency, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, 197 Ruijin 2nd Road, Huangpu District, Shanghai, People’s Republic of China, Tel +86 13501747906, Email maoeq@yeah.netAbstract: Ischemia superimposed upon pancreatic edema leads to acute necrotizing pancreatitis. One possible mechanism contributing to ischemia is intravascular thrombogenesis since fibrin deposits have been detected in pancreatic capillaries by electron microscope. Current experimental and clinical data provided compelling evidence that the disorders in the blood coagulation system play a critical role in the pathogenesis of severe acute pancreatitis (SAP). This leads to microcirculatory failure of intra- and extrapancreatic organs and multiple organ failure and increases the case fatality rate. However, the mechanism of coagulopathy underlying SAP is not yet clear, although some anticoagulant drugs have entered clinical practice showing improvement in prognosis. Thus, enhanced understanding of the process might improve the treatment strategies with safety and high efficacy. Herein, the pathogenesis of the coagulation system of SAP was reviewed with a focus on the coagulation pathway, intercellular interactions, and complement system, thereby illustrating some anticoagulant therapies and potential therapeutic targets.Keywords: coagulation, acute pancreatitis, endothelial cells, complement, anticoagulant therapy

Yan Li, Xiaoping Ma, Zhenhui Zhao, Li Li, Chunyan Gao, Dan Liu, Bingyu Li, Bing Zhao Department of Breast Cancer, Affiliated Tumor Hospital of Xinjiang Medical University, The Clinical Research Center of Breast Tumor and Thyroid Tumor in Xinjiang Uygur Autonomous Region, Urumqi, Xinjiang, 830011, People’s Republic of ChinaCorrespondence: Bing Zhao, Affiliated Tumor Hospital of Xinjiang Medical University, No. 789 Suzhou East Street, Urumqi, Xinjiang, 830011, People’s Republic of China, Tel +86 991 781 9142, Email doctorcrystal@163.comIntroduction: According to the latest global cancer data released by WHO in 2020, the incidence of breast cancer (BC) has been the most prevalent, and the mortality rate of female malignant tumor ranks the first.Methods: To evaluate toxicity and efficacy regarding oral Pyrotinib for elderly patients with advanced HER2-positive breast cancer (BC) in Xinjiang, 45 elderly patients having advanced HER2-positive BC with age ≥ 65 years and receiving Pyrotinib-based combined therapy from January 2019 to May 2021 in Xinjiang were enrolled in this study. PFS, CBR, ORR and drug-related adverse events (AE) of oral Pyrotinib in the patients were retrospectively analyzed. All 45 patients completed the efficacy evaluation.Results: Total ORR and CBR of the whole group was 37.8% and 77.8%, respectively. There were 14 patients with brain metastases (31.1%), with a median PFS of 6.8 months (95% CI: 5.4~9.8). In terms of the number of treatment lines, mPFS for line 1– 2 was 8.3 months (95% CI: 6.3~11.4), and mPFS for line ≥ 3 was 3.3 months (95% CI: 2.7~5.1). At the final maintenance dose, mPFS at standard doses of 400mg, 320mg and 240mg were 9.1 months (95% CI: 4.1~9.5), 8.3 months (95% CI: 4.3~12.2) and 4.8 months (95% CI: 2.1~7.5), respectively.Discussion: Applying Pyrotinib in elderly patients, the main adverse reaction was diarrhea, accounting for 88.9% (40/45). Pyrotinib is safe and effective for elderly patients with advanced HER2 positive BC.Keywords: HER2, breast cancer, Pyrotinib, toxicity, adverse events, elderly

Yang Zheng,1 Hui Xia,1 Xundi Bao,2 Bing Zhao,1 Ping He,1 Yanlin Zhao1 1National Tuberculosis Reference Laboratory, Chinese Center for Disease Control and Prevention, Beijing, People’s Republic of China; 2Laboratory Department, Anhui Chest Hospital, Anhui, People’s Republic of ChinaCorrespondence: Yanlin Zhao, National Center for TB Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155 Chang Bai Road, Changping District, Beijing, 102206, People’s Republic of China, Tel +86-10-58900517, Fax +86-10-58900556, Email zhaoyl@chinacdc.cnPurpose: The drug resistance of Mycobacterium tuberculosis constitutes a major public health threat. Existing approaches make it challenging to detect low levels of drug-resistant TB, also known as heteroresistance (HR), in a population. The recently found droplet digital PCR (ddPCR) is a sensitive method for determining the precise amount of nucleic acid in a sample. We used ddPCR to test the Mycobacterium tuberculosis heteroresistance because it delivers more exact quantitative data without the need for a reference curve.Patients and Methods: A TaqMan-MGB probe mutation detection assay was developed in order to determine the mutant and wild-type sequences of the isoniazid resistance katG (315) gene. We produced heteroresistant MTB combinations, which were subsequently identified by ddPCR, qPCR, and MeltPro/INH. In addition, 21 clinical sputum samples with positive smears were used to validate each method’s capacity to determine HR in sputum.Results: We discovered that ddPCR can detect mutant sequences in as few as 0.01% of a combination. DeepMelt TB/INH, which is less sensitive in comparison, cannot detect HR with high resolution and requires a mutation rate of 50% to identify. qPCR likewise has a high resolution of 0.02%, but unlike ddPCR, it cannot determine the exact number of mutations. Our assay is applicable to sputum as well. ddPCR found a katG 315 substitution in two sputums with extremely low values of HR (0.26% and 0.14%). In 21 samples of clinical sputum, the HR prevalence of INH was 9.5%.Conclusion: This work demonstrates that a well-designed ddPCR HR detection test can detect low levels of HR with high accuracy and consistency and gives new information for the clinical diagnosis of drug resistance.Keywords: Mycobacterium tuberculosis, drug-resistant, isoniazid, heteroresistance

Linfa Chen,1,2,&ast; Xinglan Chen,1,3,&ast; Yajun Wang,4,&ast; Shengnan Li,1,5 Shaoting Huang,1,5 Zhaochun Wu,1,5 Jiawen He,1,5 Shaofeng Chen,1,5 Fu Deng,1,5 Peiyi Zhu,1,5 Wangtao Zhong,1,3 Bin Zhao,1,5 Guoda Ma,1,4 You Li1,5 1Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical University, Zhanjiang, People’s Republic of China; 2Department of Neurology, Huizhou Third People’s Hospital, Guangzhou Medical University, Huizhou, People’s Republic of China; 3Department of Neurology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, People’s Republic of China; 4Maternal and Children’s Health Research Institute, Shunde Maternal and Children’s Hospital, Guangdong Medical University, Shunde, People’s Republic of China; 5Institute of Neurology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: You Li, Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524001, People’s Republic of China, Email youli805@163.com Guoda Ma, Maternal and Children’s Health Research Institute, Shunde Maternal and Children’s Hospital, Guangdong Medical University, Shunde, 528300, People’s Republic of China, Email sihan1107@126.comBackground: The S100/calgranulin gene appears to modulate neuroinflammation following cerebral ischemia and could be a valuable biomarker for stroke prognosis, according to growing research. This study aimed at evaluating the correlation between calgranulin gene variants and susceptibility to ischemic stroke (IS) in the Southern Chinese population.Methods: Using an enhanced multi-temperature ligase detection reaction genotyping, 310 IS patients and 324 age-matched healthy controls were genotyped to identify five calgranulin gene variants.Results: According to the obtained results, the S100A8 rs3795391, rs3806232, and S100A12 rs2916191 variants were linked to a higher risk of IS, while the S100A9 rs3014866 variant was associated with a lower risk of IS. Moreover, the T-T-C-A-T, T-T-C-G-T, or C-C-C-G-C haplotypes have been linked to a greater risk of developing IS, according to haplotype analysis. The occurrence of the variant C allele there in S100A8 rs3795391, rs3806232, and S100A12 rs2916191 variants may impart a greater risk of stroke in the LAA subtype, according to further stratification by IS subtypes, while the T allele of the S100A9 rs3014866 variant may be linked to a reduced risk of stroke of all subtypes. Furthermore, patients with the variant C allele of the S100A8 rs3795391, rs3806232, and S100A12 rs2916191 variants presented with increased circulating S100A8 and S100A12 levels and larger infarct volumes relative to those with the major TT genotype.Conclusion: Our findings suggest that calgranulin gene variants are linked to IS susceptibility, implying that the calgranulin gene may be a potential biomarker for IS prevention and personalized treatment.Keywords: ischemic stroke, calgranulin, single-nucleotide polymorphism, risk

Linfa Chen,1,2,&ast; Ying Wang,1,3,&ast; Yajun Wang,4,&ast; Shaoting Huang,1,3 Zhaochun Wu,1,3 Jiawen He,1,3 Wangtao Zhong,3 Bin Zhao,1,3 Guoda Ma,1,4 You Li1,5 1Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, 524001, People’s Republic of China; 2Department of Neurology, Huizhou Third People’s Hospital, Guangzhou Medical University, Huizhou, Guangdong, 516002, People’s Republic of China; 3Department of Neurology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, 524001, People’s Republic of China; 4Maternal and Children’s Health Research Institute, Shunde Maternal and Children’s Hospital, Guangdong Medical University, Shunde, Guangdong, 528300, People’s Republic of China; 5Institute of Neurology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, 524001, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: You Li, Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, 524001, People’s Republic of China, Email youli805@163.com Guoda Ma, Maternal and Children’s Health Research Institute, Shunde Maternal and Children’s Hospital, Guangdong Medical University, Shunde, Guangdong, 528300, People’s Republic of China, Email sihan1107@126.comBackground: Emerging evidences suggest that the angiotensin receptor type 1 (AT1R) contributes heavily to the pathogenesis of atherosclerotic cerebral infarction (ACI). Herein, we examined a potential link between AT1R gene polymorphisms and ACI risk among a Southern Han Chinese population.Methods: The rs3772616, rs275645, and rs377262 AT1R polymorphisms were genotyped in 689 ACI patients and 712 healthy controls, using the iMLDR-TM assay.Results: The genotypic and allelic frequencies of AT1R rs3772616 differed tremendously between ACI patients and healthy controls, and the rs3772616 T allele is a risk allele for ACI. However, the rs275645 and rs377262 allelic and genotypic frequency distributions were comparable between ACI patients and controls. In addition, the G-T-T haplotype was linked to an enhanced risk of ACI. We, next, classified our study subjects based on environmental factors and revealed that the rs3772616 T allele was strongly associated with an elevated ACI risk in males, hypertensive individuals, and those over 65 years old. In addition, we observed a marked link between the rs3772616 T allele and enhanced AT1R levels.Conclusion: Based on our research, there is a strong correlation between the AT1R rs3772616 polymorphism and enhanced ACI risk. Hence, the AT1R rs3772616 polymorphism can serve as a potential therapeutic target and bioindicator for ACI development.Keywords: angiotensin receptor type 1, polymorphism, atherosclerotic cerebral infarction, case-control

Lin Zhu,&ast; Wanyi Lian,&ast; Zhiwen Yao, Xiao Yang, Ziyi Wang, Yupei Lai, Shiting Xu, Bingcheng Zhao, Kexuan Liu Department of Anesthesiology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Kexuan Liu; Bingcheng Zhao, Department of Anesthesiology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People’s Republic of China, Tel/Fax +86 020 61641881, Email liukexuan705@163.com; zhaobch@mail2.sysu.edu.cnPurpose: Intestinal ischemia/reperfusion (I/R) injury is an unresolved clinical challenge due to its high prevalence, difficulty in diagnosis, and lack of clinically effective therapeutic agents. Ferroptosis is a novel form of cell-regulated death that has been shown to play a role in various I/R models and has been shown to be immune-related. Further unraveling the molecular mechanisms associated with ferroptosis and immunity in intestinal I/R injury may lead to the discovery of potentially effective drugs.Methods: We obtained differentially expressed mRNAs (DEGs) in mouse intestinal tissues following intestinal I/R injury or sham surgery. Then, we extracted ferroptosis-related DEGs (FRGs) and immune-related DEGs (IRGs) from the DEGs. In addition, we performed functional analysis of FRGs and IRGs. Next, we used transcriptome sequencing from patients with intestinal I/R injury to validate the results. Then, we constructed transcription factors (TFs)-gene networks and gene-drug networks using mouse and human co-expressed FRGs (coFRG) and mouse and human co-expressed IRGs (coIRG). We also analyzed the composition of immune cells to reveal correlations between FRGs signatures and immune cells in the mouse and human gut. Finally, we validated these results through animal experiments.Results: We extracted 61 FRGs and 294 IRGs from mouse samples and performed PPI and functional analyses. We extracted 45 FRGs and 200 IRGs from human samples for validation, and identified 24 coFRGs,100 coIRGs and 6 hub genes (HSPA5, GDF15, TNFAIP3, HMOX1, CXCL2 and IL6) in both. We also predicted potential TF-gene networks for coFRGs and coIRGs, as well as predicted gene-drug pairs for hub genes. In addition, we found that the immune cells were altered in the early stages of intestinal I/R injury and that FRGs were closely associated with immune cells in mice and humans. Finally, we validated the hub genes in mouse samples.Conclusion: In conclusion, we identified ferroptosis and immunity-related genes to predict their correlations in intestinal I/R injury. We also predicated potential TF-genes network and potential therapeutic targets (HSPA5, GDF15, TNFAIP3, HMOX1, CXCL2 and IL6) to provide clues for further investigation of intestinal I/R injury.Keywords: intestinal ischemia-reperfusion injury, ferroptosis, immunity, RNA-seq, bioinformatics

Tingfeng Xu,1,&ast; Liying Ren,1,&ast; Minjun Liao,1,2,&ast; Bigeng Zhao,1 Rongyu Wei,1 Zhipeng Zhou,3 Yong He,4 Hao Zhang,3 Dongbo Chen,5 Hongsong Chen,5 Weijia Liao1 1Laboratory of Hepatobiliary and Pancreatic Surgery, Affiliated Hospital of Guilin Medical University, Guilin, 541001, Guangxi, People’s Republic of China; 2Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology and Hepatology Unit, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, People’s Republic of China; 3Department of Radiology, Affiliated Hospital of Guilin Medical University, Guilin, 541001, Guangxi, People’s Republic of China; 4Department of Radiology, the Second Affiliated Hospital of Guilin Medical University, Guilin, 541001, Guangxi, People’s Republic of China; 5Peking University People’s Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Disease, Beijing, 100044, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Weijia Liao, Laboratory of Hepatobiliary and Pancreatic Surgery, Affiliated Hospital of Guilin Medical University, Guilin, 541001, Guangxi, People’s Republic of China, Tel +86 773 2833021, Fax +86 773 2822703, Email liaoweijia288@163.com Hongsong Chen, Peking University People’s Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Disease, No. 11 Xizhimen South Street, Beijing, 100044, People’s Republic of China, Tel +86 10 88325724, Email chenhongsong2999@163.comPurpose: Microvascular invasion (MVI) impairs long-term prognosis of patients with hepatocellular carcinoma (HCC). We aimed to develop a novel nomogram to predict MVI and patients’ prognosis based on radiomic features of contrast-enhanced CT (CECT).Patients and Methods: HCC patients who underwent curative resection were enrolled. The radiomic features were extracted from the region of tumor, and the optimal MVI-related radiomic features were selected and applied to construct radiomic signature (Rad-score). The prediction models were created according to the logistic regression and evaluated. Biomarkers were analyzed via q-PCR from randomly selected HCC patients. Correlations between biomarkers and radiomic signature were analyzed.Results: A total of 421 HCC patients were enrolled. A total of 1962 radiomic features were extracted from the region of tumor, and the 11 optimal MVI-related radiomic features showed a favor predictive ability with area under the curves (AUCs) of 0.796 and 0.810 in training and validation cohorts, respectively. Aspartate aminotransferase (AST), tumor number, alpha-fetoprotein (AFP) level, and radiomics signature were independent risk factors of MVI. The four factors were integrated into the novel nomogram, named as CRM, with AUCs of 0.767 in training cohort and 0.793 in validation cohort for predicting MVI, best among radiomics signature alone and clinical model. The nomogram was well-calibrated with favorable clinical value demonstrated by decision curve analysis and can divide patients into high- or low-risk subgroups of recurrence and mortality. In addition, gene BCAT1, DTGCU2, DOCK3 were analyzed via q-PCR and serum AFP were identified as having significant association with radiomics signature.Conclusion: The novel nomogram demonstrated good performance in preoperatively predicting the probability of MVI, which might guide clinical decision.Keywords: hepatocellular carcinoma, microvascular invasion, preoperative noninvasive diagnosis, contrast-enhanced CT, biologic correlation

Hui Xia,1 Yuanyuan Song,1 Yang Zheng,1 Shengfen Wang,1 Bing Zhao,1 Wencong He,1 Dongxin Liu,2 Xichao Ou,1 Yang Zhou,1 Yanlin Zhao1 1National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, People’s Republic of China; 2Institute of Hepatology, National Clinical Research Center for Infectious Disease, Shenzhen Third People’s Hospital, Shenzhen, People’s Republic of ChinaCorrespondence: Yanlin Zhao, National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, No. 155, Changbai Road, Changping District, Beijing, People’s Republic of China, Tel +86 10-58900517, Email zhaoyl@chinacdc.cnObjective: To compare the ability of detection of borderline rifampicin resistance in Mycobacterium tuberculosis between molecular assay and phenotypic drug susceptibility tests.Methods: Fifty-seven isolates with His445Leu, Asp435Tyr, Leu452Pro, Leu430Pro, His445Asn, Ile491Phe, and His445Ser mutations in rpoB gene identified by whole-genome sequencing conferring borderline rifampicin resistance were included. Molecular-based Xpert MTB/RIF, phenotypic Löwenstein–Jensen (L-J) medium-based drug susceptibility test (DST) with a critical concentration of 40.0μg/mL and minimal inhibitory concentration (MIC) assay were performed to detect borderline rifampicin resistance.Results: When using Xpert MTB/RIF, 48/57 (84.2%) isolates exhibited resistance to rifampicin. 25/57 (43.9%) and 33/57 (57.9%) isolates showed rifampicin resistance by L-J medium-based DST with 4 and 6 weeks of incubation, respectively. 30/57 (52.6%) and 40/57 (70.2%) strains were resistant to rifampicin by MIC method at cutoff values of 1.0 and 0.5μg/mL, respectively. The detection rate of rifampicin resistance of Xpert MTB/RIF was significantly higher than that of phenotypic methods (p < 0.001). Of the 57 isolates with borderline rpoB mutations, 5 (8.8%) had MICs of 0.25 or 0.12μg/mL, 22 (38.6%) had MICs of 0.5μg/mL or 1.0μg/mL, and 30 (52.6%) other isolates showed MICs ≥ 2.0μg/mL.Conclusion: Molecular-based Xpert MTB/RIF showed superior ability to detect borderline rifampicin resistance over phenotypic DST methods. Extending the incubation time of L-J DST or lowering the cutoff value of the MIC method can improve borderline rifampicin resistance detection.Keywords: Mycobacterium tuberculosis, rifampicin, borderline resistance, phenotypic drug susceptibility test

Bingjiao Zhao,1,2 Qingqing Chen,2 Liru Zhao,3 Jiaqi Mao,3 Wei Huang,2 Xinxin Han,2 Yuehua Liu1,2 1Department of Orthodontics, Shanghai Stomatological Hospital, Fudan University, Shanghai, 200001, People’s Republic of China; 2Shanghai Key Laboratory of Craniomaxillofacial Development and Diseases, Fudan University, Shanghai, 200001, People’s Republic of China; 3Department of Orthodontics, School of Stomatology, Hebei Medical University, Shijiazhuang, 050017, People’s Republic of ChinaCorrespondence: Yuehua LiuDepartment of Orthodontics, Shanghai Stomatological Hospital, Fudan University, 356 East Beijing Road, Shanghai, 200001, People’s Republic of China, Tel +86-63298475, Fax +86-63614515, Email liuyuehua@fudan.edu.cnPurpose: Small extracellular vesicles (sEVs) are natural biocarriers for biomolecule transfer between cells and promising therapeutic strategies for bone defect repair. In this study, human periodontal ligament stem cell (PDLSC)-derived sEVs (P-EVs) were immobilized in Matrigel to establish a topical cell-free transplantation strategy for bone repair.Methods: PDLSCs were cultured and P-EVs were isolated from the culture supernatant. In a rat bilateral calvarial defect model, P-EV/Matrigel was plugged into one defect and PBS/Matrigel was applied to the other. Bone repair in vivo was assessed by micro-computed tomography, histomorphometry, and immunohistochemical staining. In vitro, we investigated the effects of P-EVs on the proliferation and migration capabilities of bone marrow mesenchymal stem cells (BMMSCs) and explored the potential mechanism of action.Results: The in vivo study showed that P-EV/Matrigel accelerated bone tissue repair by increasing cell infiltration when compared with the control. In vitro, P-EVs enhanced proliferation and migration of BMMSCs via increased phosphorylation of AKT and extracellular signal-regulated kinase 1/2 (ERK1/2). The role of P-EV-induced adenosine receptor signaling in AKT and ERK1/2 phosphorylation was a key mediator during enhanced BMMSC migration.Conclusion: These results are the first to demonstrate that P-EVs accelerated the repair of bone defects, partially through promoting cell proliferation and migration. P-EV/Matrigel, which combines topical EV-implantation and extracellular matrix scaffolds, provides a new cell-free strategy for bone tissue repair.Keywords: small extracellular vesicles, periodontal ligament stem cells, osteogenesis, cell-free therapy, bone defects

Yan Fu, Xue Liu, Beichen Cui, Cong Wang, Zhiwei Liu, Bin Zhao Emergency Department, Beijing Jishuitan Hospital, Beijing, People’s Republic of ChinaCorrespondence: Bin ZhaoEmergency Department, Beijing Jishuitan Hospital, No. 31, Xinjiekou East Street, Xicheng District, Beijing, 100035, People’s Republic of ChinaTel +86-10-5851-6417Email zhaobin60@aliyun.comObjective: The objective of this retrospective study was to evaluate the clinical characteristics of diabetic ketoacidosis (DKA) in type 2 diabetes mellitus (T2DM) patients with acute pancreatitis (AP) in a cohort from China and to identify simple laboratory parameters to discriminate high-risk patients.Methods: Patients diagnosed with AP and T2DM from January 2015 to December 2020 were retrospectively enrolled. They were divided into DKA group and non-DKA group. We calculated etiologies of AP, compared demographic and clinical features, laboratory findings on admission and clinical course. The receiver operating characteristic (ROC) curve was used to explore the ability of clinical parameters to identify high-risk patients.Results: A total of 136 patients were enrolled, of which 19.9% (27 patients) concomitant with DKA. Compared with patients without DKA, patients with DKA were younger and more obese with higher levels of serum triglycerides (TGs), blood glucose and hemoglobin A1c (HbA1C). There was a significant difference in etiology between the two groups. Compared with the non-DKA group, the DKA group showed higher rate of HTG, lower rates of gallstones and alcohol abuse. And patients in the DKA group were more likely to be admitted to intensive care unit (ICU) and have longer hospital stays. ROC analyses showed that blood glucose concentration at a cutoff value of 21.75 mmol/L resulted in the highest Youden index with sensitivity and specificity at 81.5% and 94.5%, respectively, for identifying concomitant DKA (AUC = 0.949).Conclusion: The prevalence of concomitant DKA in T2DM patients with AP was 19.9%, concomitant DKA patients tend to be younger, more obese and have higher blood glucose and TGs levels on admission. We found that blood glucose > 21.75mmol/L could potentially be used as a simple laboratory parameter to identify high-risk patients.Keywords: type 2 diabetes mellitus, acute pancreatitis, diabetic ketoacidosis

Honglei Hu,1,2,&ast; Shuting Zheng,1,2,&ast; Meirong Hou,1,2 Kai Zhu,1,2 Chuyao Chen,1 Zede Wu,1,2 Li Qi,3 Yunyan Ren,1 Bin Wu,4 Yikai Xu,1 Chenggong Yan,1,5 Bingxia Zhao2,6 1Department of Medical Imaging Center, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, People’s Republic of China; 2Guangzhou Key Laboratory of Tumor Immunology Research, Cancer Research Institute, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, People’s Republic of China; 3Guangdong Provincial Key Laboratory of Medical Image Processing, Guangdong Province Engineering Laboratory for Medical Imaging and Diagnostic Technology, School of Biomedical Engineering, Southern Medical University, Guangzhou, 510515, People’s Republic of China; 4Institute of Respiratory Diseases, Respiratory Department, The Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524001, People’s Republic of China; 5Guangdong Provincial Key Laboratory of Shock and Microcirculation, School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, People’s Republic of China; 6Experimental Education/Administration Center, School of Basic Medical Science, Southern Medical University, Guangzhou, 510515, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Bingxia Zhao; Yikai Xu, Tel +86 20 61647272; +86 20 62787333, Email bingxiaz@gmail.com; yikaixu917@gmail.comBackground: Radiotherapy (RT) is clinically well-established cancer treatment. However, radioresistance remains a significant issue associated with failure of RT. Phototherapy-induced radiosensitization has recently attracted attention in translational cancer research.Methods: Cu-Sb-S nanoparticles (NPs) coated with ultra-small Au nanocrystals (Au@Cu-Sb-S) were synthesized and characterized. The biosafety profiles, absorption of near-infrared (NIR) laser and radiation-enhancing effect of the NPs were evaluated. In vitro and in vivo spectral computed tomography (CT) imaging and photoacoustic (PA) imaging were performed in 4T1 breast cancer-bearing mice. The synergetic radio-phototherapy was assessed by in vivo tumor inhibition studies.Results: Au@Cu-Sb-S NPs were prepared by in situ growth of Au NCs on the surface of Cu-Sb-S NPs. The cell viability experiments showed that the combination of Au@Cu-Sb-S+NIR+RT was significantly more cytotoxic to tumor cells than the other treatments at concentrations above 25 ppm Sb. In vitro and in vivo spectral CT imaging demonstrated that the X-ray attenuation ability of Au@Cu-Sb-S NPs was superior to that of the clinically used Iodine, particularly at lower KeV levels. Au@Cu-Sb-S NPs showed a concentration-dependent and remarkable PA signal brightening effect. In vivo tumor inhibition studies showed that the prepared Au@Cu-Sb-S NPs significantly suppressed tumor growth in 4T1 breast cancer-bearing mice treated with NIR laser irradiation and an intermediate X-ray dose (4 Gy).Conclusion: These results indicate that Au@Cu-Sb-S integrated with spectral CT, PA imaging, and phototherapy-enhanced radiosensitization is a promising multifunctional theranostic nanoplatform for clinical applications.Keywords: Au@Cu-Sb-S nanoparticle, phototherapy, radiotherapy, spectral CT, photoacoustic imaging

Junting Huang,1,* Yating Zhan,1,* Lili Jiang,1,* Yuxiang Gao,1,* Binyu Zhao,1,* Yuxiao Zhang,1 Wenjie Zhang,1 Jianjian Zheng,1 Jinglu Yu2,3 1Key Laboratory of Diagnosis and Treatment of Severe Hepato-Pancreatic Diseases of Zhejiang Province, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, People’s Republic of China; 2Department of Laboratory Medicine, Lishui Municipal Central Hospital, Lishui, People’s Republic of China; 3The Fifth Affiliated Hospital of Wenzhou Medical University, Lishui, Zhejiang, 323000, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jinglu YuDepartment of Laboratory Medicine, Lishui Municipal Central Hospital, The Fifth Affiliated Hospital of Wenzhou Medical University, No. 289 Kuocang Road, Lishui, Zhejiang, 323000, People’s Republic of ChinaEmail yujinglu2482@163.comJianjian ZhengKey Laboratory of Diagnosis and Treatment of Severe Hepato-Pancreatic Diseases of Zhejiang Province, The First Affiliated Hospital of Wenzhou Medical University, No. 2 Fuxue Lane, Wenzhou, Zhejiang, People’s Republic of ChinaEmail 120378196@qq.comAim: This study was done to determine biomarkers for the prognostic prediction of hepatocellular carcinoma (HCC).Materials and Methods: In the Gene Expression Omnibus, the gene expression profiles of HCC were downloaded. Biomarkers were identified by weighted gene co-expression network analysis and protein–protein interaction network analysis.Results: There were 24 modules, which were characterized by the high correlation with HCC. Meanwhile, through enrichment analysis, differentially expressed genes were largely participated in the ubiquitination and autophagy processes. Moreover, PRC1, TOP2A and CKAP2L may be the hub genes involved in HCC tumorigenesis, and their biomarker roles were further demonstrated via Gene Expression Profiling Interactive Analysis (GEPIA) and Oncomine databases. In addition, the levels of PRC1, TOP2A and CKAP2L were obviously up-regulated in the sera of HCC patients.Conclusion: PRC1, TOP2A and CKAP2L may serve as biomarkers for the prognostic prediction of HCC patients.Keywords: hepatocellular carcinoma, biomarker, WGCNA, PPI, prognosis

Qianxue Mou,1 Ke Yao,1 Meng Ye,1 Bowen Zhao,1 Yuanyuan Hu,1 Xiaotong Lou,1 Huixia Li,2 Hong Zhang,1 Yin Zhao1 1Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People’s Republic of China; 2Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Xi’an Jiaotong University Health Science Center, Xi’an, Shaanxi, People’s Republic of ChinaCorrespondence: Hong ZhangDepartment of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People’s Republic of ChinaEmail tjyksys@163.comCorrespondence: Yin ZhaoDepartment of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People’s Republic of ChinaEmail zhaoyin85@hust.edu.cnPurpose: Optic nerve injury (ONI) causes neuroinflammation and neurodegeneration leading to visual deficits. The response of microglia has emerged as an impactful component of etiology in neurodegeneration. This study aimed to investigate the effect of SIRT1-mTORC1 signaling pathway in microglia regulation after ONI.Methods: Cx3Cr1-CreERT2/RaptorF/F and Cx3Cr1-CreERT2/Sirt1F/F mice were used to delete Raptor and Sirt1 in microglia, respectively. Optic nerve crush (ONC) model was established to mimic ONI. PLX5622, a highly specific inhibitor of the colony-stimulating factor 1 receptor (CSF1R), is used to eliminate microglia in optic nerve. Ionized calcium binding adaptor molecule 1 (Iba1) immunostaining was used to detect microglial activation. Retinal ganglion cells (RGCs) were quantified by Nissl staining and retinal whole-mount immunostaining with RNA-binding protein with multiple splicing (RBPMS). Axonal damage was valued by transmission electron microscopy (TEM).Results: Microglial activation emerged on day 3 post ONC and was earlier than RGCs loss which occurred at day 5 after injury. Depleting microglia with PLX5622 could attenuate the loss of RGCs and axon damage after ONC. Gain- and loss-of-function studies revealed that SIRT1 determined the activation of microglia in optic nerve. In addition, microglia-specific deletion of Raptor resulted in decreased microglial activation. Interestingly, activating mTORC1 with CCT007093 could reverse the function of SIRT1 in regulating the process of microglial activation mediated RGCs loss.Conclusion: Our study reveals a potential novel mechanism of SIRT1-mTORC1 pathway in microglia regulation, and indicates a therapeutic potential for the protection of RGCs in ONI.Keywords: microglia, retinal ganglion cells, optic nerve injury, SIRT1, mTORC1

Qingqing Chen,1,2 Xinxin Han,2 Meihua Chen,2,3 Bingjiao Zhao,1,2 Bingjing Sun,1,2 Liangyan Sun,1,2 Weihua Zhang,1,2 Liming Yu,1,2 Yuehua Liu1,2 1Department of Orthodontics, Shanghai Stomatological Hospital, Fudan University, Shanghai, People’s Republic of China; 2Shanghai Key Laboratory of Craniomaxillofacial Development and Diseases, Fudan University, Shanghai, People’s Republic of China; 3Department of Periodontology, Shanghai Stomatological Hospital, Fudan University, Shanghai, People’s Republic of ChinaCorrespondence: Yuehua Liu; Liming YuShanghai Stomatological Hospital, Fudan University, No. 356 East Beijing Road, Shanghai, 200001, People’s Republic of ChinaTel +86 2163298475Fax +86 2163614515Email liuyuehua@fudan.edu.cn; ylmphoebe@126.comPurpose: Obesity is a worldwide metabolic disease and a critical risk factor for several chronic conditions. Obstructive sleep apnea (OSA) is an important complication of obesity. With the soaring morbidity of obesity, the prevalence of OSA has markedly increased. However, the underlying mechanism of the high relevance between obesity and OSA has not been elucidated. This study investigated the effects of obesity on the structure and function of the genioglossus to explore the possible mechanisms involved in OSA combined with obesity.Methods: Six-week-old male C57BL/6J mice were fed high-fat diet (HFD, 60% energy) or normal diet (Control, 10% energy) for 16 weeks. The muscle fibre structure and electromyography (EMG) activity of genioglossus were measured. The ultrastructure and function of mitochondrial, oxidative damage and apoptosis in genioglossus were detected by transmission electron microscopy (TEM), qPCR, Western blotting, immunohistochemistry and TUNEL staining. We further studied the influence of palmitic acid (PA) on the proliferation and myogenic differentiation of C2C12 myoblasts, as well as mitochondrial function, oxidative stress, and apoptosis in C2C12 myotubes.Results: Compared with the control, the number of muscle fibres was decreased, the fibre type was remarkably changed, and the EMG activity had declined in genioglossus. In addition, a HFD also reduced mitochondria quantity and function, induced excessive oxidative stress and increased apoptosis in genioglossus. In vitro, PA treatment significantly inhibited the proliferation and myogenic differentiation of C2C12 myoblasts. Moreover, PA decreased the mitochondrial membrane potential, upregulated mitochondrial reactive oxygen species (ROS) levels, and activated the mitochondrial-related apoptotic pathway in myotubes.Conclusion: Our findings suggest that a HFD caused genioglossus injury in obese mice. The mitochondrial dysfunction and the accompanying oxidative stress were involved in the genioglossus injury, which may provide potential therapeutic targets for OSA with obesity.Keywords: obesity, obstructive sleep apnea, genioglossus, muscle, mitochondria

Huanyu Guo,1,&ast; Wenwei Jiang,2,&ast; Bo Zhao,1 Yanhua Xiong,3 Zhenya Lu1 1Department of FSTC Clinic of The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, 310009, People’s Republic of China; 2Department of Internal Medicine of Traditional Chinese Medicine, Huzhou Central Hospital, Huzhou, 310003, People’s Republic of China; 3Department of Internal Medicine of Traditional Chinese Medicine, Zhejiang Hospital, Hangzhou, 310007, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zhenya LuDepartment of FSTC Clinic of The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, 310009, People’s Republic of ChinaTel +86 13575458480Email 1199026@zju.edu.cnPurpose: To develop a predictive model for the risk of metabolic syndrome (MetS).Patients and Methods: Totally, 1556 residents without MetS were finally included in 2006 and they were observed for 8 years to check who developed MetS. Univariate and multivariate logistic regression analyses was adopted to explore the risk factors of MetS and develop the predictive model that used the medical examination information of MetS risk after 8 years. The receiver operating characteristic (ROC) curve was drawn to assess the predictive capacity of the model.Results: The risk of MetS in overweight, prehypertension, hypertension subjects were 4.610 [95% confidence interval (CI): 2.415 to 8.800], 2.759 (95% CI: 1.519 to 5.011) and 3.589 (95% CI: 1.672 to 7.706) times higher than that in controls, respectively. The risk of MetS in people with high-density lipoprotein (HDL) < 1.10 mmol/L was 3.716-fold in comparison with HDL ≥ 1.55 mmol/L [odds risk (OR) = 3.716, 95% CI: 1.483 to 9.313]. Individuals with fatty liver had a higher risk of MetS (OR = 2.577, 95% CI: 1.472 to 4.512). The AUC of the predictive model was 0.831 (95% CI: 0.798 to 0.865), with the sensitivity of 0.898 (95% CI: 0.831 to 0.941) and the specificity of 0.676 (95% CI: 0.651 to 0.700).Conclusion: The model performed well predictive power for the risk of MetS, which may provide a reference for clinicians to identify high-risk groups early.Keywords: metabolic syndrome, predictive model, 8-year, Chinese cohort

Shuiping Dai,1,&ast; Bennan Zhao,2,&ast; Dafeng Liu,2 Yongzhao Zhou,3 Yaling Liu,2 Lijuan Lan,2 Yalun Li,3 Wenxin Luo,3 Yilan Zeng,2 Weimin Li3 1Center of Gerontology and Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan, People’s Republic of China; 2Department of Comprehensive Internal Medicine, The Public and Health Clinical Center of Chengdu, Chengdu, Sichuan, People’s Republic of China; 3Department of Respiratory and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Weimin Li; Yilan Zeng Email weimin003@163.com; 2499081791@qq.comPurpose: Some studies have shown that patients with coronavirus disease 2019 (COVID-19) still have sequelae after discharge. However, little is known about the long-term physical and psychological sequelae of patients, especially factors that influenced the prognosis.Patients and Methods: Patients with COVID-19 were followed up for 6 months. The psychological status of patients was evaluated by DASS-21 questionnaire, while physical functions were determined using medical history, laboratory examination, thoracic computed tomography (CT), and echocardiography.Results: Fifty patients infected with COVID-19 were enrolled, and 11 (22%) patients still showed symptoms related to COVID-19. The mean contents (cells/ul) of CD3+ cells, CD4+ and CD8+ T, B lymphocytes and NK cells of the survivors elevated significantly after 6-month discharge (P < 0.001). The frequency of ground-glass opacities and consolidations decreased from 90% to 42% (P < 0.001), and 54% to 20%, (P = 0.001), respectively, while the changes of reticulation and bronchiectasis were insignificant (P > 0.05). The frequency of left ventricular diastolic dysfunction decreased from 40% to 15% (P = 0.002). Depression was observed in 5 (12.5%) participants, stress in 3 (7.5%), anxiety in 6 (15%), and among them 1 (2.5%) showed extremely severe anxiety. Covariation analysis elucidated age might be a risk factor (OR: 1.09, 95% CI: 1.01– 1.18, P = 0.038), while NK cell was a good prognostic factor for pulmonary recovery. The comorbidities were significantly positive correlated with persist pulmonary damage (r = 0.33, P = 0.020). Compared with patients with antiviral therapy, patients without antiviral therapy had higher anxiety score (3 vs 0, P = 0.033).Conclusion: After 6-month discharge, the persisting cardiopulmonary damage was observed in recovery patients, and psychological implications should not be ignored. Age, comorbidities, NK cell and antiviral therapy might be associated with the prognosis of COVID-19.Keywords: COVID-19, follow-up, cardiopulmonary, psychology, influence factors

Benhui Zhao,1,&ast; Xingtong Chen,1,&ast; Qian Chen,1,2 Gaoming Li,3 Zhe Chen,4 Ziheng Yang,1 Li Gu,1 Xudong Xiao,5 Zhen Wang,6 Jiaolin Ning,1 Bin Yi,1 Kaizhi Lu,1 Hongyan Zhang,7 Jianteng Gu1 1Department of Anesthesiology, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, People’s Republic of China; 2Anaesthetics, Pain Medicine and Intensive Care, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, Chelsea & Westminster Hospital, London, UK; 3Department of Health Statistics, Third Military Medical University (Army Medical University), Chongqing, People’s Republic of China; 4Quality Management and Control Department, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, People’s Republic of China; 5Department of Anesthesiology, Xinqiao Hospital, Third Military Medical University (Army Medical University), Chongqing, People’s Republic of China; 6Department of Anesthesiology; 7Hospital Office, Daping Hospital, Third Military Medical University (Army Medical University), Chongqing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jianteng Gu; Hongyan Zhang Email jiantenggu@hotmail.com; etengyan@163.comBackground: Blood pressure fluctuation is very common during non-cardiac surgery in elderly. This retrospective study was to analyse whether intraoperative hypotension in elderly and other risk factors relate to the postoperative mortality.Methods: A total of 118 cases (Observational group), who underwent noncardiac surgery in three medical centers between September 2014 and March 2017, and died in the hospital after the noncardiac surgery. With 1:2 ratio of propensity matching, 236 survival cases (Control group) were selected for comparison analyses with the death cases. Intraoperative blood pressure and perioperative parameters from both groups were collected from electronic anaesthesia charts. Data were analysed with univariate logistic regression analysis where variables with p values less than 0.05 were analysed with multivariate logistic regression analysis. The receiver operating characteristic (ROC) curve was constructed.Results: There are five risk factors related to postoperative death in elderly patients: ASA grade, COPD, emergency surgery, general anesthesia, 60 < MAP ≤ 65mmHg (OR > 1), and one factor may reduce the risk of postoperative mortality, which is PACU therapy (OR < 1). Compared with the Control group, the Observational group had a higher proportion of cerebral hernia, kidney injury and trauma (p < 0.001). The intraoperative blood transfusion volume and intraoperative blood loss volume were higher in the Observational group than the Control group (p < 0.001). The proportion of using vasoactive drugs was higher in the Observational group (p < 0.001), and there was more urine output during the operation in the Observational group (p = 0.005).Conclusion: The intraoperative MAP of geriatric patients lower than 65mmHg is highly related to the postoperative mortality. Elderly patients with emergency surgery, high ASA grade and a history of COPD have an increased risk of postoperative mortality. General anesthesia is a risk factor for postoperative death in elderly patients, and the PACU therapy is a protective factor to avoid postoperative death.Trial Registration: This study has been retrospectively registered in the Chinese Clinical Trials Registry (ChiCTR2000038912, 10/10/2020).Keywords: intraoperative hypotension, geriatric patients, COPD, postoperative mortality, non-cardiac surgery

Binbin Zhao,1,&ast; Yanhua Wu,2,&ast; Xiaoying Jin,1 Lihong Yang,2 Jian Yang,1,2 Xiancang Ma,1 Bin Yan1,2 1Department of Psychiatry, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, People’s Republic of China; 2Department of Clinical Research Center, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Bin YanDepartment of Clinical Research Center, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, People’s Republic of ChinaTel +86 2985323614Email yanbin.1987@hotmail.comObjective: Sleep disorders are associated with the prevalence of stroke. However, there is limited evidence regarding the association between objectively measured sleep characteristics and ischemic stroke.Methods: Ischemic stroke was assessed during the mean follow-up period of 11 years in the Sleep Heart Health Study. Sleep parameters such as wake after sleep onset (WASO) and sleep efficiency (SE) were objectively measured based on in-home polysomnography records. Multivariable Cox regression analysis was utilized to examine the relationship between objective sleep characteristics and ischemic stroke incidence.Results: This study involved 4204 participants (1978 males and 2226 females, 63.8± 11.1 years). The incidence of ischemic stroke increased in individuals with long WASO, poor SE, and short sleep duration. Multivariable Cox regression analysis showed that WASO within the fourth quartile (hazard ratio [HR] 3.771, 95% confidence interval [CI] 1.805– 7.877, P< 0.001), third quartile (HR 3.009, 95% CI 1.433– 6.317, P=0.004), and second quartile (HR 3.108, 95% CI 1.470– 6.568, P=0.003) had a higher incidence of ischemic stroke than WASO within the first quartile. Poor SE (< 80.0%) was also found to be a predictor for ischemic stroke (HR 2.220, 95% CI 1.244– 3.960, P=0.007). Additionally, a short sleep duration (< 6 h) was associated with an increased risk of ischemic stroke (HR 1.725, 95% CI 1.026– 2.899, P=0.040).Conclusion: Our results revealed a relationship between WASO, SE, and sleep duration and ischemic stroke. Therefore, these sleep characteristics may be adequate predictors for the incidence of ischemic stroke.Keywords: sleep characteristics, ischemic stroke, polysomnography, Sleep Heart Health Study

TengFei Yang,1 Bo Zhao,2 Dongmei Pei1 1Department of Health Management, Shengjing Hospital of China Medical University, Shenyang, People’s Republic of China; 2Department of Pulmonary and Critical Care Medicine, Shengjing Hospital of China Medical University, Shenyang, People’s Republic of ChinaCorrespondence: Dongmei PeiDepartment of Health Management, Shengjing Hospital of China Medical University, No. 36, Sanhao Street, Heping District, Shenyang, 110004, People’s Republic of ChinaEmail peidm1111@hotmail.comBackground: Nonalcoholic fatty liver disease (NAFLD) is the commonest form of chronic liver disease worldwide and its prevalence is rapidly increasing. Screening and early diagnosis of high-risk groups are important for the prevention and treatment of NAFLD; however, traditional imaging examinations are expensive and difficult to perform on a large scale. This study aimed to develop a simple and reliable predictive model based on the risk factors for NAFLD using a decision tree algorithm for the diagnosis of NAFLD and reduction of healthcare costs.Methods: This retrospective cross-sectional study included 22,819 participants who underwent annual health examinations between January 2019 and December 2019 at Physical Examination Center in Shengjing Hospital of China Medical University. After rigorous data screening, data of 9190 participants were retained in the final dataset for use in the J48 decision tree algorithm for the construction of predictive models. Approximately 66% of these patients (n=6065) were randomly assigned to the training dataset for the construction of the decision tree, while 34% of the patients (n=3125) were assigned to the test dataset to evaluate the performance of the decision tree.Results: The results showed that the J48 decision tree classifier exhibited good performance (accuracy=0.830, precision=0.837, recall=0.830, F-measure=0.830, and area under the curve=0.905). The decision tree structure revealed waist circumference as the most significant attribute, followed by triglyceride levels, systolic blood pressure, sex, age, and total cholesterol level.Conclusion: Our study suggests that a decision tree analysis can be used to screen high-risk individuals for NAFLD. The key attributes in the tree structure can further contribute to the prevention of NAFLD by suggesting implementable targeted community interventions, which can help improve the outcome of NAFLD and reduce the burden on the healthcare system.Keywords: nonalcoholic fatty liver disease, J48 algorithm, decision tree, risk factors

Peng Chen,1 Mei-Mei Hao,2 Yong Chen,3 Hong Zhang,4 Zhe Wang,5 Bin Zhao,1 Yani Xue,1 Yumei Chai,1 Yong-Feng Huang,1 Jiang Zhu1 1Department of Neurology, The First Hospital of Yulin, Yulin, People’s Republic of China; 2Department of Neurology, Yan’an People’s Hospital, Yan’an, People’s Republic of China; 3Department of Endoscopy Center, Shaanxi Cancer Hospital, Xi’an, Shannxi Province, People’s Republic of China; 4Department of Vascular Surgery, Shaanxi Provincial People’s Hospital, Xi’an, Shannxi Province, People’s Republic of China; 5Department of General Practice, Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, Shannxi Province, People’s Republic of ChinaCorrespondence: Jiang ZhuDepartment of Neurology, The First Hospital of Yulin, No. 93, Yuxi Street, Gaoxin District, Yulin, Shaanxi, 719000, People’s Republic of ChinaEmail 13709204858@163.comAbstract: The Percheron artery (artery of Percheron, AOP) is a rare variant vessel. Its acute occlusion can cause a bilateral symmetrical thalamic stroke; typical symptoms of bilateral paramedian thalamic infarcts due to occlusion of AOP are vertical gaze palsy, memory impairment, confusion, drowsiness, hypersomnolence, or coma. We present the MR imaging findings in two cases with cerebral infarction caused by Percheron artery occlusion. Due to the difficulty in the diagnosis of acute Percheron arterial infarction, early conservative treatment is used. The prognosis of the disease is poor, with few patients completely rehabilitating. Therefore, clinicians must understand the characteristics of the disease, provide early diagnosis and administer timely and effective treatment to reduce the patient’s disability rate and fatality rate and therefore improve the quality of life of patients. The patient’s prognosis has extraordinary significance.Keywords: thalamic infarcts, artery of Percheron, MR imaging

Zongying Liang, Baoshan Zhao, Jishen Hou, Jingxiong Zheng, Guohua Xin Department of Thoracic Surgery, The Affiliated Hospital of Chengde Medical University, Chengde, 067000, People’s Republic of ChinaCorrespondence: Guohua XinDepartment of Thoracic Surgery, The Affiliated Hospital of Chengde Medical University, No. 36, Nanyingzi Street, Shuangqiao District, Chengde, 067000, People’s Republic of ChinaEmail iqbqth@163.comBackground: Circular RNA hsa_circ_0003340 (circ-OGDH) has been uncovered to be involved in esophageal squamous cell carcinoma (ESCC) progression. However, the mechanism by which circ-OGDH regulates ESCC progression is unclear.Methods: Expression levels of circ-OGDH, microRNA (miR)-615-5p, and PDX1 (pancreatic and duodenal homeobox 1) mRNA were evaluated with quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation, apoptosis, migration, invasion, and cell cycle progression of ESCC cells were analyzed by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide), colony formation, flow cytometry, and transwell assays. Measurement of glutamine consumption, α-KG (α-ketoglutarate) production, and ATP (Adenosine Triphosphate) content using corresponding kits. Protein levels were analyzed by Western blotting. The targeting relationship between circ-OGDH or PDX1 and miR-615-5p was verified by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. The function of circ-OGDH in ESCC was confirmed by animal experiments.Results: Circ-OGDH was upregulated in ESCC. Circ-OGDH inhibition reduced ESCC growth in vivo and accelerated cell apoptosis, cell cycle arrest, repressed cell proliferation, migration, invasion, and reduced cell glutamine metabolism in ESCC cells in vitro. MiR-615-5p was downregulated in ESCC, while PDX1 had an opposite result. Circ-OGDH sponged miR-615-5p to regulate PDX1 expression. MiR-615-5p inhibitor neutralized the repressive effect of circ-OGDH knockdown on malignancy and glutamine metabolism of ESCC cells. PDX1 overexpression counteracted the inhibitory impact of miR-615-5p mimic on malignancy and glutamine metabolism of ESCC cells.Conclusion: Circ-OGDH sponged miR-615-5p to elevate PDX1 expression, thus elevating glutamine metabolism and promoting tumor growth in ESCC. The study offered evidence to support circ-OGDH as a promising target for ESCC therapy.Keywords: ESCC, circ-OGDH, miR-615-5p, PDX1, glutamine

Yue Zhong,1– 3 Yong Yang,2,4 Lei He,2,4 Yang Zhou,2 Niangmei Cheng,2 Geng Chen,2,4 Bixing Zhao,2,4 Yingchao Wang,2,4 Gaoxiong Wang,5,6 Xiaolong Liu2– 4 1College of Life Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, People’s Republic of China; 2The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, Fujian, 350025, People’s Republic of China; 3Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian, 350002, People’s Republic of China; 4College of Biological Science and Engineering and Mengchao Med-X Center, Fuzhou University, Fuzhou, Fujian, 350116, People’s Republic of China; 5Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, Fujian, 362001, People’s Republic of China; 6Quanzhou Maternal and Child Health Hospital, Children’s Hospital, Quanzhou, Fujian, 362017, People’s Republic of ChinaCorrespondence: Yingchao WangThe United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao, Hepatobiliary Hospital of Fujian Medical University, Fuzhou, 350025, People’s Republic of ChinaEmail yingchaowang@live.comGaoxiong WangDepartment of Hepatobiliary Surgery, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, 362001, People’s Republic of ChinaEmail wanggaoxiong2013@163.comBackground: The aberrant expressions of lncRNAs have been frequently demonstrated to be closely associated with the prognosis of patients in many cancer types including hepatocellular carcinoma (HCC). Integration of these lncRNAs might provide accurate evaluation of HCC. Therefore, this study aims to develop a novel prognostic evaluation model based on the expression of lncRNAs to predict the survival of HCC patients, postoperatively.Patients and Methods: RNA sequencing (RNA-seq) analysis was performed for 61 HCC patients (training cohort) to screen prognosis-associated lncRNAs with univariate Cox regression and Log rank test analyses. Multivariate Cox regression analysis was then applied to establish the final model, which was further verified in a validation cohort (n=191). Moreover, performance of the mode was assessed with time-dependent receiver operating characteristic curve (tdROC), Harrell’s c-index, and Gönen & Heller’s K.Results: After a serial statistical computation, a novel risk scoring model consisting of four lncRNAs and TNM staging was established, which could successfully divide the HCC patients into low-risk and high-risk groups with significantly different OS and RFS in both training and validation cohorts. tdROC analysis showed that this model achieved a high performance in predicting OS and 2-year RFS in both cohorts. Gene Set Enrichment Analysis revealed that HCC tumor tissues with high-risk score have stronger capacities in immune escape and resistance to treatment.Conclusion: We successfully established a novel prognostic evaluation model, which exhibited reliable capacity in predicting the OS and early recurrence of HCC patients with relatively higher accuracy.Keywords: hepatocellular carcinoma, long noncoding RNA, risk score, prognosis, survival