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Jinlei Zhou,1,* Shanggao Xie,1,* Sen Xu,1,* Yuan Zhang,2 Yanlei Li,1 Qice Sun,2 Jun Zhang,1,3 Tingxiao Zhao1 1Center for Plastic & Reconstructive Surgery, Department of Orthopedics, Zhejiang Provincial People’s Hospital (Affiliated People’s Hospital, Hangzhou Medical College), Hangzhou, Zhejiang, People’s Republic of China; 2Department of Rheumatology, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 3Department of Orthopedics, Zhejiang Provincial People’s Hospital Bijie Hospital, Bijie, 551700, People’s Republic of China*These authors contributed equally to this workCorrespondence: Tingxiao Zhao, Center for Plastic & Reconstructive Surgery, Department of Orthopedics, Zhejiang Provincial People’s Hospital (Affiliated People’s Hospital, Hangzhou Medical College), Shangtang Road 158#, Hangzhou, Zhejiang, People’s Republic of China, Email spinezhaotingxiao@163.com Jun Zhang, Department of Orthopedics, Zhejiang Provincial People’s Hospital Bijie hospital, Guanghui Road 112#, Bijie, Guizhou, People’s Republic of China, Email spinecarezhangjun@163.comObjective: Comprehensive analyses of the burden of musculoskeletal disorders (MSDs) are crucial for deepening our understanding of their impact on population health and for evaluating progress toward achieving international health goals.Study Design: A comprehensive analysis was performed using data from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021.Methods: We gathered data on the number of prevalent cases, incidences, deaths, and disability-adjusted life years (DALYs) for MSDs from 1990 to 2021. Subsequently, crude indicators, age-standardized rates (ASRs) and estimated annual percentage change (EAPC) were calculated for MSDs from 1990 to 2021 at the global, regional, national, age and gender levels. Finally, decomposition analysis, frontier analysis, and EAPC cluster analysis were employed to examine changes in the indicators and the factors influencing them.Results: In 2021, there were 367,193,430 incident cases and 1,686,561,517 prevalent cases of MSDs globally. Between 1990 and 2021, the age-standardized incidence rates (ASIR) declined marginally from 4641.5 to 4351.79 cases per 100,000 population [EAPC: − 0.16 (− 0.19 to − 0.13)], while the age-standardized DALY rates (ASDR) increased from 1886.2 to 1908.87 per 100,000 population [EAPC: 0.09 (0.07 to 0.11)]. The ASRs were markedly elevated in the high SDI regions in comparison to the remaining SDI regions. The EAPC and its phase clustering analyses indicated a notable upward trajectory in the burden of MSDs in the low and middle SDI regions, particularly in Central Asia. The prevalence of MSDs is highest among individuals aged 50– 59 years and among women. At the global level, the burden of MSDs is 1.43 to 2.30 times higher for women than for men.Conclusion: From 1990 to 2021, the burden of MSDs has been concentrated in high SDI regions, with a gradual shift towards low and middle SDI regions.Keywords: musculoskeletal disease, global burden of disease, prevalence, incidence, DALYs, epidemiology

Jinjin Xu,1– 3,&ast; Yan Yang,1– 3,&ast; Guixue Guan,1– 3,&ast; Yuan Gao,1– 3 Qian Sun,1– 3 Guangwei Yuan,4 Xiaozuo Zhang,1– 3 Jingyun Yang,5,6 Wen Yang,1– 3 Zuobin Zhu,7 Conghui Han8,9 1Department of Gynecology, The First People’s Hospital of Lianyungang, Lianyungang, Jiangsu, People’s Republic of China; 2Department of Gynecology, The First Affiliated Hospital of Kangda College of Nanjing Medical University, Lianyungang, Jiangsu, 22206, People’s Republic of China; 3Medical University, Lianyungang, Jiangsu, China & Department of Gynecology, Xuzhou Medical University Affiliated Hospital of Lianyungang, Lianyungang, Jiangsu, People’s Republic of China; 4College of Professional Studies, Northeastern University, Boston, MA, USA; 5Rush Alzheimer’s Disease Center, Rush University Medical Center, Chicago, IL, USA; 6Department of Neurological Sciences, Rush University Medical Center, Chicago, IL, USA; 7Department of Genetics, Xuzhou Engineering Research Center of Medical Genetics and Transformation, Key Laboratory of Genetic Foundation and Clinical Application, Xuzhou Medical University, Xuzhou, Jiangsu, People’s Republic of China; 8Department of Urology, Xuzhou Clinical School of Xuzhou Medical University, Xuzhou, Jiangsu, People’s Republic of China; 9Department of Urology, Xuzhou Central Hospital, Xuzhou, Jiangsu, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Wen Yang, Department of Gynecology, The First Affiliated Hospital of Kangda College of Nanjing Medical University, Zhenhua Road, Lianyungang, Jiangsu, 222061, People’s Republic of China, Tel +86-18961325910, Email wen_yang0@163.com Conghui Han, Department of Urology, Xuzhou Clinical School of Xuzhou Medical University, 199 Jiefang South Road, Xuzhou, Jiangsu, 221000, People’s Republic of China, Tel +86-13813461893, Email hanchdoctor@st.btbu.edu.cnObjective: This study investigates the thromboelastography (TEG) changes in patients with unexplained recurrent spontaneous abortion (URSA) to identify effective diagnostic markers for URSA.Methods: We retrospectively analyzed 160 URSA patients from the Gynecology Department of the First People’s Hospital of Lianyungang (June 2017 - June 2020) and compared them with 190 healthy, fertile women without adverse pregnancy histories (control group). TEG parameters were assessed using logistic regression, applying stepwise selection for model optimization. Model performance was evaluated using Receiver Operating Characteristic (ROC) curves, determining sensitivity and specificity. The Youden index identified optimal cut points for predictive probabilities.Results: Significant differences were observed between the URSA and control groups in coagulation reaction time (R), clot formation time (K), clot formation rate (Angle-α), and maximum clot strength (MA) (P< 0.05). Multivariable logistic regression identified R, Angle-α, and MA as independent URSA risk factors. The model demonstrated excellent discrimination (AUC: 0.940; 95% CI: 0.918– 0.962). The optimal cut point of predictive probability (Youden index) was P=0.355, yielding a sensitivity of 0.925 and specificity of 0.795.Conclusion: URSA patients exhibit a hypercoagulable state even when not pregnant. More research is needed to validate our findings and explore the potential clinical implications of anticoagulants in treating URSA.Keywords: unexplained recurrent spontaneous abortion, thromboelastography, prothrombotic state, coagulation function

Hao Huang,1 Zhiping Long,2 Ying Deng,3 Zhicong Huang,3 Zhonghua Lv,4 Qian Sun,4 Hui Liu,4 Hongsheng Liang,5 Fulan Hu6 1Department of Preventive Medicine, Zhuhai Campus of Zunyi Medical University, Zhuhai, Guangdong, People’s Republic of China; 2Department of Epidemiology, Public Health School of Harbin Medical University, Harbin, Heilongjiang, People’s Republic of China; 3Department of Epidemiology and Health Statistics, Fujian Provincial Key Laboratory of Environment Factors and Cancer, School of Public Health, Fujian Medical University, Fuzhou, People’s Republic of China; 4Department of Neurosurgery, Third Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, People’s Republic of China; 5Department of Neurosurgery, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, People’s Republic of China; 6Department of Biostatistics and Epidemiology, School of Public Health, Shenzhen University Medical School, Shenzhen, Guangdong, People’s Republic of ChinaCorrespondence: Fulan Hu, Department of Biostatistics and Epidemiology, School of Public Health, Shenzhen University Medical School, Shenzhen, Guangdong, People’s Republic of China, Email hufu1525@163.com Hao Huang, Department of Preventive Medicine, Zhuhai Campus of Zunyi Medical University, Zhuhai, Guangdong, People’s Republic of China, Email hhxiaowo@163.comPurpose: Mitochondrial metabolism is essential for energy production and the survival of brain cells, particularly in astrocytes. Cuproptosis is a newly identified form of programmed cell death that occurs due to the disruption of mitochondrial metabolism caused by excessive copper toxicity. However, the relationship between cuproptosis-related genes (CRGs) in the tumor microenvironment (TME) and the prognosis of gliomas remains unclear.Patients and Methods: In this study, we utilized 32,293 cells obtained from three in-house single-cell RNA sequencing (scRNA-seq) datasets, along with 6,148 cells acquired from the Chinese Glioma Genome Atlas (CGGA) involving 14 glioma patients, to identify and validate the TME of gliomas.Results: Based on an analysis of 32,293 single cells, we investigated intra-tumor heterogeneity, intercellular communication, and astrocyte differentiation trajectories in gliomas. Our findings revealed that the TGFβ signaling pathway exhibited a higher relative strength in astrocyte subpopulations. Additionally, we identified a novel three-gene signature (CDKN2A, SOX2, and MPC1) was identified for prognostic prediction. Furthermore, glioma patients with a high-risk score demonstrated poorer overall survival (OS) compared to those with a low-risk score in both training and testing datasets (Ptraining set < 0.001; Ptest set = 0.037).Conclusion: Our study revealed the prognostic value of the CRGs in astrocytes exhibiting tumor immunosuppressive characteristics in glioma. We established a novel three-gene prognostic model that offers new insights into the prognosis and treatment strategies for gliomas. Keywords: scRNA-seq, gliomas, astrocyte, cuproptosis, prognosis

Mei Liu,1,&ast; Sili Guo,1,&ast; Xiaohan Li,1,&ast; Yang Tian,1 Yanjie Yu,2 Lili Tang,1 Qimei Sun,1 Ting Zhang,1 Mingwei Fan,3 Lili Zhang,1 Yingjiang Xu,4 Jiajia An,5 Xiangqian Gao,6 Lei Han,7 Lei Zhang1 1Department of Endocrinology and Metabolism, Binzhou Medical University Hospital, Binzhou, Shandong, People’s Republic of China; 2Department of Ultrasound Medicine, Binzhou Medical University Hospital, Binzhou, Shandong, People’s Republic of China; 3Department of Gastroenterology, Binzhou Medical University Hospital, Binzhou, Shandong, People’s Republic of China; 4Department of Interventional Vascular Surgery, Binzhou Medical University Hospital, Binzhou, Shandong, People’s Republic of China; 5Department of Clinical Laboratory, Binzhou Medical University Hospital, Binzhou, Shandong, People’s Republic of China; 6Department of Pathology, Binzhou Medical University Hospital, Binzhou, Shandong, People’s Republic of China; 7Department of Reproductive Medicine, Binzhou Medical University Hospital, Binzhou, Shandong, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Lei Zhang, Department of Endocrinology and Metabolism, Binzhou Medical University Hospital, Binzhou, Shandong, People’s Republic of China, Email binzhouzhanglei@outlook.comBackground: GLP-1 receptor agonists (GLP-1 RA) have been proven to treat several metabolic diseases; however, the effects of GLP-1 RA on polycystic ovary syndrome (PCOS) remain unclear. Here, we aimed to investigate whether semaglutide, a novel GLP-1 RA, could alleviate ovarian inflammation in PCOS mice.Methods: Female C57BL/6J mice were subcutaneously injected with dehydroepiandrosterone for 21 days to establish the PCOS model. Then the mice were randomly divided into three groups: PCOS group (n = 6), S-0.42 group (semaglutide 0.42 mg/kg/w, n = 6), and S-0.84 group (semaglutide 0.84 mg/kg/w, n = 6). The remaining six mice were used as controls (NC). After 28 days of intervention, serum sex hormones and inflammatory cytokine levels were measured. Hematoxylin and eosin staining was used to observe the ovarian morphology. Immunohistochemical staining was used to detect the relative expression of CYP19A1, TNF-α, IL-6, IL-1β, and NF-κB in ovaries. CYP17A1 and StAR were detected using immunofluorescence staining. Finally, the relative expressions of AMPK, pAMPK, SIRT1, NF-κB, IκBα, pIκBα, TNF-α, IL-6, and IL-1β were measured using Western blotting.Results: First, after intervention with semaglutide, the weight of the mice decreased, insulin resistance improved, and the estrous cycle returned to normal. Serum testosterone and IL-1β levels decreased significantly, whereas estradiol and progestin levels increased significantly. Follicular cystic dilation significantly improved. The expression of TNF-α, IL-6, IL-1β, NF-κB, CYP17A1, and StAR in the ovary was significantly downregulated, whereas CYP19A1 expression was upregulated after the intervention. Finally, we confirmed that semaglutide alleviates ovarian tissue inflammation and improves PCOS through the AMPK/SIRT1/NF-κB signaling pathway.Conclusion: Semaglutide alleviates ovarian inflammation via the AMPK/SIRT1/NF‑κB signaling pathway in PCOS mice.Keywords: GLP-1 receptor agonist, polycystic ovary syndrome, semaglutide, inflammation

Yantao Yu,1– 3,&ast; Chenkai Zhang,2,3,&ast; Qiannan Sun,2– 4 Shantanu Baral,2,3 Jianyue Ding,2,3 Fanyu Zhao,2,3 Qing Yao,2,3 Shuyang Gao,2,3 Bin Liu,2– 4 Daorong Wang1– 4 1The Yangzhou School of Clinical Medicine of Dalian Medical University, Yangzhou, 225001, People’s Republic of China; 2General Surgery Institute of Yangzhou, Yangzhou University, Yangzhou, 225001, People’s Republic of China; 3Yangzhou Key Laboratory of Basic and Clinical Transformation of Digestive and Metabolic Diseases, Yangzhou, 225001, People’s Republic of China; 4Northern Jiangsu People’s Hospital, Yangzhou, 225001, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Daorong Wang; Bin Liu, Email 18051062590@yzu.edu.cn; liubin2006cool@126.comBackground: Gastric cancer (GC) is a highly phenotypically heterogeneous disease and is caused by a combination of factors. Retinol binding protein 4 (RBP4) is a member of a family of lipid transport proteins that are involved in the transport of substances between cells and play a crucial role in a variety of cancers. However, the expression and role of RBP4 in GC remain unknown.Methods: In this study, we explored the expression, prognostic significance, immune microenvironment, drug responsiveness and function of associated signaling pathways of RBP4 in GC using web-based bioinformatics tools. Immunohistochemistry and real-time quantitative PCR were utilized to analyze the tissue and cell expression levels of RBP4. CCK-8, colony formation, EDU incorporation, wound healing and transwell assays were applied to demonstrate the effect of RBP4 on GC cell function. Flow cytometric detection of apoptosis after RBP4 knockdown. Nude mice xenograft model elucidates the role of RBP4 for GC in vivo. Related proteins of the RAS signaling pathway were analyzed by employing Western blot assays.Results: RBP4 is highly expressed in GC. RBP4 is closely associated with patient survival and sensitivity to a wide range of antitumor agents. Knockdown of RBP4 promoted apoptosis and inhibited cell proliferation, invasion and migration. RBP4 promotes GC tumorigenesis in vivo. Finally, RBP4 modulates the RAS/RAF/ERK axis.Conclusion: RBP4 may promote gastric carcinogenesis and development through the RAS/RAF/ERK axis and is expected to be a novel target for GC treatment.Keywords: RBP4, gastric cancer, invasion, migration, RAS/RAF/ERK axis

Yixin Li,1– 6,&ast; Qi Sun,2,3,7,&ast; Lingjia Hao,1,8 Han Shan,2,3,7 Zixi Jiang,1– 6 Ying Wang,1– 6 Zeyu Chen,2,3,7 Wu Zhu,1– 6 Shuang Zhao1– 6 1Department of Dermatology, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 2Furong Laboratory (Precision Medicine), Changsha, 410008, People’s Republic of China; 3National Engineering Research Center of Personalized Diagnostic and Therapeutic Technology, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 4Hunan Engineering Research Center of Skin Health and Disease, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 5Hunan Key Laboratory of Skin Cancer and Psoriasis, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 6National Clinical Research Center of Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 7School of Mechanical and Electrical Engineering, Central South University, Changsha, 410083, People’s Republic of China; 8Xiangya School of Medicine, Central South University, Changsha, 410083, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Shuang Zhao; Wu Zhu, Department of Dermatology, Xiangya Hospital of Central South University, 87 Xiangya Road, Kaifu District, Changsha, Hunan Province, People’s Republic of China, Email shuangxy@csu.edu.cn; zhuwuxy@csu.edu.cnIntroduction: Pathological scars, such as hypertrophic scars and keloids, are characterized by the proliferation of fibroblasts and the deposition of collagen that often cause pruritus, pain, and disfigurement. Due to their high incidence and deformity, pathological scars have resulted in severe physical and psychological trauma for patients. Intralesional injection of 5-fluorouracil (5-Fu) is a recommended option for treating pathological scars. However, the efficacy of 5-Fu injection was limited and unstable due to limited drug penetration and short retention time.Methods: Liposomes are promising carriers that have advantages, such as high biocompatibility, controlled release property, and enhanced clinical efficacy. Here, we constructed a transdermal 5-Fu-loaded liposome (5-Fu-Lip) to provide a more effective and safer modality to scar treatment.Results: Compared to 5-Fu, 5-Fu-Lip showed superior ability in inhibiting primary keloid fibroblasts proliferation, migration, and collagen deposition, and also significantly inhibited human umbilical vein endothelial cells (HUVECs) proliferation and microvessel construction. In vivo experiments demonstrated that 5-Fu-Lip can significantly reduce the severity of hypertrophic scars in a rabbit ear wounding model.Discussion: 5-Fu-Lip provides a promising strategy to improve drug efficacy, which has great potential in the treatment of pathological scars.Keywords: liposome, pathological scar, 5-fluorouracil, fibroblast

Yan Jiang,1 Yuanyuan Peng,1 Yingyi Wu,1 Qing Sun,1 Tebo Hua2 1Department of Ultrasound, Ningbo Medical Centre Lihuili Hospital, Ningbo, Zhejiang, People’s Republic of China; 2Department of Thyroid Breast Surgery, Ningbo Medical Centre Lihuili Hospital, Ningbo, Zhejiang, People’s Republic of ChinaCorrespondence: Tebo Hua, Department of Thyroid Breast Surgery, Ningbo Medical Centre Lihuili Hospital, Ningbo, Zhejiang, 315040, People’s Republic of China, Email huatebo@163.comObjective: To develop a clinical-radiomics model using a multimodal machine learning method for distinguishing ductal carcinoma in situ (DCIS) from breast fibromatosis.Methods: The clinical factors, ultrasound features, and related ultrasound images of 306 patients (198 DCIS patients) were retrospectively collected. Patients in the development and validation cohort were 184 and 122, respectively. The independent clinical and ultrasound factors identified by the multivariable logistic regression analysis were used for the clinical-ultrasound model construction. Then, the region of interest of breast lesions was delineated and radiomics features were extracted. Six machine learning algorithms were trained to develop a radiomics model. The algorithm with higher and more stable prediction ability was chosen to convert the output of the results into the Radscore. Further, the independent clinical predictors and Radscore were enrolled into the logistic regression analysis to generate a combined clinical-radiomics model. The receiver operating characteristic curve analysis, DeLong test, and decision curve analysis were adopted to compare the prediction ability and clinical efficacy of three different models.Results: Among the six classifiers, logistic regression model was selected as the final radiomics model. Besides, the combined clinical-radiomics model exhibited a superior ability in distinguishing DCIS from breast fibromatosis to the clinical-ultrasound model and the radiomics model.Conclusion: The combined model by integrating clinical-ultrasound factors and radiomics features performed well in predicting DCIS, which might promote prompt interventions to improve the early diagnosis and prognosis of the patients.Keywords: multimodal machine learning, clinical-ultrasound features, radiomics, ductal carcinoma in situ

Yaxu Feng,&ast; Qing Sun,&ast; Pei Liu, Wei Fan, Bing Fan The State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Wei Fan; Bing Fan, Email weifan@whu.edu.cn; bingfan@whu.edu.cnBackground: Enterococcus faecalis (E. faecalis) is one of the main pathogens responsible for refractory root canal infections in the teeth and shows resistance against various antibacterial managements. Effective control of E. faecalis infection is a prerequisite for successful treatment of refractory apical periodontitis. This study aimed to analyze the antibacterial activity and mechanisms of Au@Ag nanoparticles (NPs) combined with photothermal therapy (PTT) against the original and Ag+-resistant E. faecalis.Methods: Au@AgNPs with optimal shell thicknesses were synthesized and characterized. The antibacterial activity of Au@AgNPs with PTT against the original or Ag+-resistant E. faecalis was evaluated, and the antibiofilm activity was tested on E. faecalis biofilm on the dentin of teeth. The potential antibacterial mechanisms of Au@AgNPs combined with PTT against E. faecalis have also been studied. Moreover, its influence on dentin microhardness and cytotoxicity was assessed.Results: This study revealed that Au@AgNPs combined with PTT showed enhanced antibacterial and antibiofilm effects, no negative effects on dentin microhardness, and low cytotoxicity toward human periodontal ligament cells (hPDLCs). Moreover, Au@AgNPs combined with PTT effectively inhibited the growth of Ag+-resistant E. faecalis. Its antibacterial effects may be exerted through the release of silver ions (Ag+), destruction of the cell membrane, production of reactive oxygen species (ROS) and inhibition of adenosine triphosphate (ATP) production. Hyperthermia generated by Au@AgNPs with PTT reduced membrane fluidity and enhanced Ag+ sensitivity by downregulating fabF expression. The upregulated expression of heat shock genes demonstrated that the Ag+ released from Au@AgNPs compromised the heat adaptation of E. faecalis.Conclusion: PTT significantly enhanced Ag+ sensitivity of the original and Ag+-resistant E. faecalis. Au@AgNPs combined with PTT may have the potential to be developed as a new antibacterial agent to control E. faecalis infections in teeth.Keywords: Au@AgNPs, resistance, photothermal therapy, antibacterial, silver ions, E. faecalis

Lijie Ma, Fang Sun, Kaiyi Zhu, Qiuxia Han, Qianmei Sun Department of Nephrology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, 100020, People’s Republic of ChinaCorrespondence: Qianmei Sun; Qiuxia Han, Email sunqianmei5825@126.com; qiuxiahanamy@163.comPurpose: The serum lipid level is strongly associated with atherosclerosis. However, research on the relationship between lipid-derived indices and acute ischemic stroke (AIS) occurrence in hemodialysis populations is limited. This study aimed to explore the predictive value of lipid-derived indices, including atherogenic index of plasma (AIP), Non- high density lipoprotein cholesterol (Non-HDL-C), Non-HDL-C/HDL-C, and lipoprotein combine index (LCI) in clinical practice for the occurrence and prognosis of AIS in hemodialysis patients.Methods: A total of 451 patients undergoing maintenance hemodialysis were screened and 350 were enrolled in this study. The lipid parameters exhibit a progressive increase across the tertiles, with values rising from Q1 through Q3. Enrolled patients were divided into three groups (Q1, Q2, and Q3) based on tertiles of AIP, Non-HDL-C, Non-HDL-C/HDL-C, and LCI values. Kaplan-Meier curves were performed to investigate the association between the AIP, Non-HDL-C, Non-HDL-C/HDL-C, LCI and AIS-free survival in hemodialysis patients. Chi-square analysis was used to explore the association between the AIP, Non-HDL-C, Non-HDL-C/HDL-C, LCI and AIS outcomes in hemodialysis patients. AIS outcomes were assessed using the modified Rankin Scale (mRS).Results: Kaplan-Meier analysis revealed that the AIS-free survival rates were significantly higher in the Q1 group compared to Q2 and Q3 groups for AIP, Non-HDL-C, Non-HDL-C/HDL-C, and LCI. Log rank tests showed statistically significant differences between the Q1 group and the Q2 and Q3 groups (p < 0.05 for all). The proportion of patients with a good outcome mRS was higher in the Q1 group compared to the Q2-Q3 groups (AIP: 0.818 vs 0.792; Non- HDL-C: 0.866 vs 0.767; Non- HDL-C/HDL-C: 0.867 vs 0.767; LCI: 0.938 vs 0.750).Conclusion: The four lipid-derived parameters are effective predictors of AIS in patients undergoing hemodialysis, and AIP has a strongest correlation with the risk of AIS. Hemodialysis patients with elevated levels of the four lipid-derived indices had a higher incidence of AIS and poorer functional outcomes compared to those with lower levels. Our conclusions may require confirmation by further research in the future.Keywords: hemodialysis, acute ischemic stroke, atherogenic index of plasma, non-high density lipoprotein cholesterol, lipoprotein combine index

Qingran Sun, Yi Wang, Pei Wang, Yuankai Huang, Xiaoyu Xi National Medical Products Administration Key Laboratory for Drug Regulatory Innovation and Evaluation, China Pharmaceutical University, Nanjing, Jiangsu Province, People’s Republic of ChinaCorrespondence: Xiaoyu Xi, Email xixy@cpu.edu.cnPurpose: This study aimed to survey and analyze the preferences for pharmacist-managed clinic among urban residents in China.Materials and Methods: A discrete choice experiment was conducted in Nanjing, China. A D-efficient fractional factorial design was used to generate the questionnaire. Three models were used to investigate each patient’s strength of preference and preference heterogeneity. The relative importance for each treatment attribute was also determined.Results: 156 usable questionnaires (of 228 questionnaires sent out) were received. Respondents preferred pharmacist-managed clinics with the following characteristics: good pharmacists’ knowledge and clinical medication practice competency, lower consultation fees, a dedicated consultation room, physician-pharmacist joint clinic, with pharmacists’ knowledge competency receiving the highest priority. Latent class analysis revealed three classes (Experiential Type, Content Type and Economic Type) were identified based on respondents’ preferences for pharmacist-managed clinics.Conclusion: The respondents were willing to choose a PMC relative to the current situation. When deciding on a pharmacist-managed clinic, residents are driven by pharmacists’ competency, consultation fee, availability of consultation rooms and collaborative care or independent pharmacist service. Differences in patients’ preferences identified in the study provide information on pharmacist-managed clinics that meet residents’ expectations.Keywords: conjoint analysis, discrete choice experiment, pharmacist-managed clinic, preference study, patients

Xinyan Hu,1,&ast; Yanling Sui,1,&ast; Xinhao Yang,1 Zhengyu Yang,2 Qiuting Wang,2 Jiehong Yuan,2 Maoyu Li,1 Xiaotong Ma,1 Chengxuan Qiu,3 Qinjian Sun1 1Key Laboratory of Endocrine Glucose & Lipids Metabolism and Brain Aging, Ministry of Education; Department of Neurology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, People’s Republic of China; 2Department of Neurology, Shandong Provincial Hospital, Shandong University, Jinan, Shandong, People’s Republic of China; 3Aging Research Center, Department of Neurobiology, Care Sciences and Society, Karolinska Institutet-Stockholm University, Stockholm, Sweden&ast;These authors contributed equally to this workCorrespondence: Qinjian Sun, Key Laboratory of Endocrine Glucose & Lipids Metabolism and Brain Aging, Ministry of Education; Department of Neurology, Shandong Provincial Hospital affiliated to Shandong First Medical University, No. 324 Jingwu Weiqi Road, Jinan, Shandong, 250021, People’s Republic of China, Email sqj1210@163.comBackground: The inflammatory response is a pivotal factor in accelerating the progression of atherosclerosis. The high-sensitivity C-reactive protein-to-albumin ratio (CAR) has emerged as a novel marker of systemic inflammation. However, few studies have shown the CAR to be a promising prognostic marker for carotid atherosclerotic disease. This study aimed to analyse the predictive role of the CAR in carotid atherosclerotic disease.Methods: This community-based cohort study recruited 2003 participants from the Rose asymptomatic IntraCranial Artery Stenosis (RICAS) study who were free of stroke or transient ischemic attack. Carotid atherosclerotic plaques and their stability were identified via carotid ultrasound. Logistic regression models were utilized to investigate the association between CAR and the presence of carotid atherosclerotic plaques.Results: The prevalence of carotid atherosclerotic plaques was 38.79% in this study. After adjusting for clinical risk factors, including sex, age, dyslipidemia, hypertension, diabetes mellitus (DM), and smoking and drinking habits, a high CAR-level was independently associated with carotid plaque (odds ratio [OR] of upper: 1.46, 95% confidence interval [CI]: 1.13– 1.90, P = 0.004; P for trend = 0.011). The highest CAR tertile was still significantly associated with carotid plaques among middle-aged (40– 64 years) or female participants. Notably, an elevated CAR may be an independent risk factor for vulnerable carotid plaques (OR of upper: 2.06, 95% CI: 1.42– 2.98, P < 0.001; P for trend < 0.001).Conclusion: A high CAR may be correlated with a high risk of carotid plaques, particularly among mildly aged adults (40– 64 years) or females. Importantly, the CAR may be associated with vulnerable carotid plaques, suggesting that the CAR may be a new indicator for stroke prevention. Keywords: C-reactive protein, albumin, plaque, atherosclerotic, cohort study

Jieyang Yu,1,2 Yanan Zhang,1 Lijia Cai,1 Qingqing Sun,1 Wanru Li,1 Junfang Zhou,1 Jianmin Liang,2 Zan Wang1 1Sleep Centre, Department of Neurology, The First Hospital of Jilin University, Changchun, Jilin Province, People’s Republic of China; 2Department of Pediatric Neurology, The First Hospital of Jilin University, Changchun, Jilin Province, People’s Republic of ChinaCorrespondence: Jianmin Liang, Department of Pediatric Neurology, The First Hospital of Jilin University, Changchun, Jilin Province, 130021, People’s Republic of China, Email liangjm@jlu.edu.cn Zan Wang, Sleep Centre, Department of Neurology, The First Hospital of Jilin University, Changchun, Jilin Province, 130021, People’s Republic of China, Email wangzan@jlu.edu.cn; wangzanprof@163.comPurpose: This study aimed to evaluate nocturnal sleep structure and anxiety, depression, and fatigue in patients with narcolepsy type 1 (NT1).Methods: Thirty NT1 patients and thirty-five healthy controls were enrolled and evaluated using the Epworth sleepiness scale (ESS), Generalized Anxiety Disorder-7, Patient Health Questionnaire-9, Fatigue Severity Scale (FSS), polysomnography, multiple sleep latency test, and brain function state monitoring. Statistical analyses were performed using SPSS Statistics for Windows, version 23.0. Benjamini-Hochberg correction was performed to control the false discovery rate.Results: Apart from typical clinical manifestations, patients with NT1 are prone to comorbidities such as nocturnal sleep disorders, anxiety, depression, and fatigue. Compared with the control group, patients with NT1 exhibited abnormal sleep structure, including increased total sleep time (Padj=0.007), decreased sleep efficiency (Padj=0.002), shortening of sleep onset latency (Padj< 0.001), elevated wake after sleep onset (Padj=0.002), increased N1% (Padj=0.006), and reduced N2%, N3%, and REM% (Padj=0.007, Padj< 0.001, Padj=0.013). Thirty-seven percent of patients had moderate to severe obstructive sleep apnea-hypopnea syndrome. And sixty percent of patients were complicated with REM sleep without atonia. Patients with NT1 displayed increased anxiety propensity (Padj< 0.001), and increased brain fatigue (Padj=0.020) in brain function state monitoring. FSS scores were positively correlated with brain fatigue (Padj< 0.001) and mean sleep latency was inversely correlated with FSS scores and brain fatigue (Padj=0.013, Padj=0.029). Additionally, ESS scores and brain fatigue decreased after 3 months of therapy (P=0.012, P=0.030).Conclusion: NT1 patients had abnormal nocturnal sleep structures, who showed increased anxiety, depression, and fatigue. Excessive daytime sleepiness and fatigue improved after 3 months of treatment with methylphenidate hydrochloride prolonged-release tablets in combination with venlafaxine.Keywords: narcolepsy, sleep structure, excessive daytime sleepiness, anxiety, depression, fatigue

Guangtian Wang,1,2,&ast; Zhihui Li,3,&ast; Gongchen Wang,4,&ast; Qixu Sun,5 Peng Lin,1 Qian Wang,2 Huishu Zhang,6 Yanyan Wang,7 Tongshuai Zhang,8 Feiyun Cui,9 Zhaohua Zhong1,2 1Teaching Center of Pathogenic Biology, School of Basic Medical Sciences, Harbin Medical University, Harbin, Heilongjiang, 150081, People’s Republic of China; 2Department of Microbiology, School of Basic Medical Sciences, Harbin Medical University, Harbin, Heilongjiang, 150081, People’s Republic of China; 3Department of Neurology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, 150086, People’s Republic of China; 4Department of Vascular Surgery, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, 150086, People’s Republic of China; 5Department of Gastroenterology, Penglai People’s Hospital, Yantai, Shandong, 265600, People’s Republic of China; 6Teaching Center of Biotechnology, School of Basic Medical Sciences, Harbin Medical University, Harbin, Heilongjiang, 150081, People’s Republic of China; 7Teaching Center of Morphology, School of Basic Medical Sciences, Harbin Medical University, Harbin, Heilongjiang, 150081, People’s Republic of China; 8Department of Neurobiology, School of Basic Medical Sciences, Harbin Medical University, Harbin, Heilongjiang, 150081, People’s Republic of China; 9Department of Physiology, School of Basic Medical Sciences, Harbin Medical University, Harbin, Heilongjiang, 150081, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zhaohua Zhong; Feiyun Cui, Email zhongzh@hrbmu.edu.cn; feiyun@hrbmu.edu.cnAbstract: Angiogenesis, or the formation of new blood vessels, is a natural defensive mechanism that aids in the restoration of oxygen and nutrition delivery to injured brain tissue after an ischemic stroke. Angiogenesis, by increasing vessel development, may maintain brain perfusion, enabling neuronal survival, brain plasticity, and neurologic recovery. Induction of angiogenesis and the formation of new vessels aid in neurorepair processes such as neurogenesis and synaptogenesis. Advanced nano drug delivery systems hold promise for treatment stroke by facilitating efficient transportation across the the blood-brain barrier and maintaining optimal drug concentrations. Nanoparticle has recently been shown to greatly boost angiogenesis and decrease vascular permeability, as well as improve neuroplasticity and neurological recovery after ischemic stroke. We describe current breakthroughs in the development of nanoparticle-based treatments for better angiogenesis therapy for ischemic stroke employing polymeric nanoparticles, liposomes, inorganic nanoparticles, and biomimetic nanoparticles in this study. We outline new nanoparticles in detail, review the hurdles and strategies for conveying nanoparticle to lesions, and demonstrate the most recent advances in nanoparticle in angiogenesis for stroke treatment. Keywords: angiogenesis, ischemic stroke, nanoparticles, blood brain barrier, neurogenesis, neuroprotection therapy

Qingxiang Liu,1,&ast; Fei Peng,1,&ast; Haitao Liu,2,&ast; Qin Sun,1 Hui Chen,1 Xinyi Xu,1 Zihan Hu,1 Xing Zhou,1 Kai Jin,1 Jianfeng Xie,1 Yingzi Huang,1 Wei Huang,1 Yi Yang1 1Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, 210009, People’s Republic of China; 2School of Life Science, Fudan University, Shanghai, 200000, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Wei Huang; Yi Yang, Email whuang@seu.edu.cn; yiyiyang2004@163.comBackground: Natural killer (NK) cells are key regulators of immune defense in sepsis-induced acute respiratory distress syndrome (ARDS), yet the characteristics of NK cell clusters in ARDS remain poorly understood.Methods: A prospective and observational study enrolled septic patients with ARDS or not was conducted to determine the percentage of NK cells via flow cytometry. The transcriptomes of peripheral blood mononuclear cells (PBMCs) from healthy controls, patients with sepsis only, and patients with sepsis-induced ARDS were profiled. Vitro experiments were performed to confirm the mechanism mediating MX1+NK cell infiltration.Results: A total of 115 septic patients were analyzed, among whom 63 patients developed ARDS and 52 patients did not. Decreased NK percentages were found in sepsis with ARDS patients (%, 7.46± 4.40 vs 11.65± 6.88, P=0.0001) compared with sepsis-only patients. A lower percentage of NK cells showed a significant increase in 28-day mortality. Single-cell sequencing analysis revealed distinct characteristics of NK cells in sepsis-induced ARDS, notably the identification of a unique cluster defined as MX1+NK cells. Flow cytometry analysis showed an elevated percentage of MX1+NK cells specifically in individuals with sepsis-induced ARDS, compared with patients with sepsis only. Pseudo-time analysis showed that MX1+NK cells were characterized by upregulation of type I interferon-induced genes and other pro-inflammatory genes. MX1+NK cells can respond to type I interferons and secrete type I interferons themselves. Ligand–receptor interaction analysis also revealed extensive interaction between MX1+NK cells and T/B cells, leading to an uncontrolled inflammatory response in ARDS.Conclusion: MX1+NK cells can respond to type I interferons and secrete type I interferons themselves, promoting the development of sepsis-induced ARDS. Interfering with the infiltration of MX1+NK cells could be a therapeutic approach for this disease. Due to the limited sample size, a larger sample size was needed for further exploration.Keywords: ARDS, inflammation, natural killer cells, sepsis

Zhe Xiong,1,2 Ying Fang,1,2 Shuangshuang Lu,1 Qiuyue Sun,1,3 Yuhui Sun,1,3 Pengcheng Yang,4,&ast; Jin Huang1,&ast; 1Department of Gastroenterology, The Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University, Changzhou, Jiangsu Province, People’s Republic of China; 2Graduate School of Dalian Medical University, Dalian, Liaoning Province, China; 3Graduate School of Nanjing Medical University, Nanjing, Jiangsu Province, People’s Republic of China; 4Department of Gastroenterology, Hengshanqiao People’s Hospital, Changzhou, Jiangsu Province, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Pengcheng Yang; Jin Huang, Email 496808936@qq.com; hj042153@hotmail.comBackground: Ulcerative colitis (UC) is a nonspecific inflammatory disease confined to the intestinal mucosa and submucosa, and its prevalence significantly increases each year. Disulfidptosis is a recently discovered new form of cell death that has been suggested to be involved in multiple diseases. The aim of this study was to explore the relevance of disulfidptosis in UC.Methods: First, the UC datasets were downloaded from the Gene Expression Omnibus (GEO) database, and UC samples were typed based on upregulated disulfidptosis-related genes (DRGs). Then, weighted gene co-expression network analysis (WGCNA) was performed on the datasets and molecular subtypes of UC, respectively, to obtain candidate signature genes. After validation of the validation set and qRT-PCR, we constructed a nomogram model by signature genes to predict the risk of UC. Finally, single-cell sequencing analysis was used to study the heterogeneity of UC and to demonstrate the expression of DRGs and signature genes at the single-cell level.Results: A total of 7 DRGs were significantly upregulated in the expression profiles of UC, and 180 UC samples were divided into two subtypes based on these DRGs. Five candidate signature genes were obtained by intersecting two key gene modules selected by WGCNA. After evaluation, four signature genes with diagnostic relevance (COL4A1, PRRX1, NNMT, and PECAM1) were eventually identified. The nomogram model showed excellent prediction ability. Finally, in the single-cell analysis, there were eight cell types (including B cells, T cells, monocyte, smooth muscle cells, epithelial cells, neutrophil, endothelial cells and NK cells) were identified. The signature genes were significantly expressed mainly in endothelial cells and smooth muscle cells.Conclusion: In this study, subtypes related to disulfidptosis were identified, and single-cell analysis was performed to understand the pathogenesis of UC from a new perspective. Four signature genes were screened and a prediction model with high accuracy was established. This provides novel insights for early diagnosis and therapeutic targets in UC.Keywords: ulcerative colitis, disulfidptosis, molecular clusters, WGCNA, nomogram

Yanan Zhang,1 Qianqian Chen,1 Qingqing Sun,1 Mingyang Tang,1 Yi Yang,1,2 Zhen-Ni Guo,1,2 Zan Wang1,2 1Sleep Center, Department of Neurology, The First Hospital of Jilin University, Chang Chun, People’s Republic of China; 2Stroke Center, Department of Neurology, The First Hospital of Jilin University, Chang Chun, People’s Republic of ChinaCorrespondence: Zhen-Ni Guo, Stroke Center, Department of Neurology, The First Hospital of Jilin University, Xinmin Street 1&num;, Chang Chun, 130021, People’s Republic of China, Tel +86-431-88782378, Email zhen1ni2@jlu.edu.cn Zan Wang, Sleep Center, Department of Neurology, The First Hospital of Jilin University, Xinmin Street 1&num;, Chang Chun, 130021, People’s Republic of China, Tel +86-431-88782378, Email wangzan@jlu.edu.cnBackground: Restless legs syndrome (RLS) is a prevalent sensorimotor nervous system disorder in patients accompanied with insomnia, blood pressure fluctuation, and sympathetic dysfunction. These symptoms may disrupt cerebral hemodynamics. Dynamic cerebral autoregulation (dCA) describes the temporary response of cerebrovascular system to abrupt fluctuations in blood pressure, which keep cerebral blood flow stable and serve as a marker of cerebrovascular system ability.Objective: This research aimed to assess dCA in RLS patients.Methods: In this study, RLS patients were recruited and subsequently classified into four groups (mild, moderate, severe, and very severe) based on the International RLS Rating Scale (IRLS). Healthy controls matched for age and sex were enrolled. All participants were evaluated dCA by assessing phase difference (PD). A portion of patients with RLS was reassessed for dCA after one month of medication therapy (pramipexole [0.125 mg/day] and gabapentin [300 mg/day]).Results: There were altogether 120 patients with RLS and 30 controls completed the polysomnography and dCA assessment. PD was lower in the moderate, severe, and very severe RLS groups than that in the controls and mild RLS groups. Periodic limb movement index (PLMI), arousal index, and IRLS all showed a linear correlation with PD in RLS patients. Additionally, PD increased in RLS patients after therapy.Conclusion: The dCA was compromised in moderate, severe, and very severe RLS patients and was negatively correlated with the IRLS, arousal index, and PLMI. After 1 month of therapy, dCA improved in RLS patients.Keywords: restless legs syndrome, dynamic cerebral autoregulation, autonomic nerve dysfunction, dopaminergic

Shan Li,1,&ast; Fengdi Zhang,2,&ast; Ritian Lin,1 Qinjuan Sun,1 Lihong Qu,2 Lan Zhong1 1Department of Gastroenterology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, People’s Republic of China; 2Department of Infectious Diseases, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Lan Zhong, Department of Gastroenterology, Shanghai East Hospital, Tongji University School of Medicine, 150, Jimo Road, Pudong New Area, Shanghai, 200120, People’s Republic of China, Email lanzhong@tongji.edu.cn Lihong Qu, Department of Infectious Diseases, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, People’s Republic of China, Email 18916510601@163.comPurpose: The rapid global spread of the SARS-CoV-2 Omicron variant introduces a novel complication: the emergence of IBD (inflammatory bowel disease)-like ulcers in certain patients. This research delves into this new challenge by juxtaposing the clinical manifestations and genetic expression patterns of individuals affected by the Omicron variant of COVID-19 with those diagnosed with IBD. It aims to decode the link between these conditions, potentially shedding light on previously unexplored facets of COVID-19 pathophysiology. This investigation emphasizes gene expression analysis as a key tool to identify wider disease correlations and innovative therapeutic avenues.Patients and Methods: From March to December 2022, patients with SARS-CoV-2 Omicron infection and inflammatory bowel disease and healthy controls were recruited in Shanghai East Hospital, Shanghai, China. The epidemiological and clinical characteristics of the patients were compared. Four RNA sequencing datasets (GSE205244, GSE201530, GSE174159, and GSE186507) were extracted from the Gene Expression Omnibus database to detect mutually differentially expressed genes and common pathways in patients with SARS-CoV-2 infection and inflammatory bowel disease.Results: Compared to patients with active inflammatory bowel disease, patients with SARS-CoV-2 infection were more likely to have elevated interferon-α levels and an increased lymphocyte count and less likely to have high interleukin-6, tumor necrosis factor-α, and C-reactive protein levels and an elevated neutrophil count. A total of 51 common differentially expressed genes were identified in the four RNA-sequencing datasets. Enrichment analysis suggested that these genes were related to inflammation and the immune response, especially the innate immune response and nucleotide oligomerization domain-like receptor signaling pathway.Conclusion: The inflammation and immune-response pathways in COVID-19 and inflammatory bowel disease have several similarities and some differences. The study identifies the NLR signaling pathway’s key role in both COVID-19 and IBD, suggesting its potential as a target for therapeutic intervention and vaccine development.Keywords: viral-induced inflammation, omicron infection, RNA sequencing, gene expression in infection, inflammatory bowel disease

Ziwen Wang,1,2 Jiaxuan Fang,1,2 Shigao Zu,1,2 Qianru Sun,1,2 Zixin Song,1,2 Jiman Geng,1,2 Dongdong Wang,1,2 Meng Li,1,2 Changtao Wang1,2 1Beijing Key Laboratory of Plant Resource Research and Development, Beijing Technology and Business University, Beijing, 100048, People’s Republic of China; 2School of Light Industry Science and Engineering, Beijing Technology and Business University, Beijing, 100048, People’s Republic of ChinaCorrespondence: Dongdong Wang; Changtao Wang, Beijing Technology and Business University, Beijing, 100048, People’s Republic of China, Tel/Fax +86 158 1135 7756, Email wdd@btbu.edu.cn; wangct@th.btbu.edu.cnPurpose: To produce Panax notoginseng extract as a cosmetic ingredient through Saccharomyces cerevisiae fermentation.Methods: We first compared the total sugar content, polysaccharide content, reducing sugar content, total phenolic content, total saponin content, DPPH free radical, ABTS free radical, hydroxyl free radical scavenging ability and ferric reducing antioxidant power (FRAP) of Panax notoginseng fermented extract (pnFE) and unfermented extract (pnWE). Their potential correlations were analyzed by Pearson’s correlation analysis. Then, the oxidative stress model of H2O2-induced MSFs was used to evaluate the effects of different pnFE on MSF viability, reactive oxygen species (ROS), malondialdehyde (MDA), and the activities of catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) to explore their protective effects on MSFs subjected to H2O2-induced cellular oxidative damage. Finally, their safety and stability were evaluated by using the red blood cell (RBC) test and hen’s egg test-chorioallantoic membrane (HET-CAM) assay, and changes in pH and content of soluble solids, respectively.Results: Compared with pnWE, pnFE has more active substances and stronger antioxidant capacity. In addition, pnFE has a protective effect on H2O2-induced oxidative stress in MSFs with appropriate safety and stability.Conclusion: PnFE has broad application prospects in the field of cosmetics.Keywords: Panax notoginseng, Saccharomyces cerevisiae, fermentation, oxidative stress, fibroblast cells, cosmetic ingredients

Tao Xue,1,&ast; Hongbin Yu,2,&ast; Wenming Feng,2 Yao Wang,2 Shiyong Wu,3 Lili Wang,4 Peiqin Zhu,5 Jianming Guan,6,7 Quan Sun8 1Clinical Research Center, First People’s Hospital Affiliated to Huzhou University, Huzhou, People’s Republic of China; 2Department of Hepatobiliary Surgery, First People’s Hospital Affiliated to Huzhou University, Huzhou, People’s Republic of China; 3Department of Radiology, First People’s Hospital Affiliated to Huzhou University, Huzhou, People’s Republic of China; 4Department of Nursing, Physical Examination Center of Aishan, Huzhou, People’s Republic of China; 5Department of Internal Medicine, Huzhou Traditional Chinese Medicine Hospital Affiliated to Zhejiang Chinese Medical University, Huzhou, People’s Republic of China; 6Department of Ultrasound, First People’s Hospital Affiliated to Huzhou University, Huzhou, People’s Republic of China; 7Department of Ultrasound, Mingzhou Hospital, Huzhou, People’s Republic of China; 8Laboratory of Blood Transfusion Research, Huzhou Center Blood Station, Huzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Quan Sun, Laboratory of Blood Transfusion Research, Huzhou Center Blood Station, Fenghuang Road 577&num;, Huzhou, 313000, People’s Republic of China, Email sunquanhz@163.com Jianming Guan, Department of Ultrasound, First People’s Hospital Affiliated to Huzhou University, Guangchanghou Road 158&num;, Huzhou, 313000, People’s Republic of China, Email guan9300@126.comBackground: The presence of macrovascular invasion (MVI) is associated with poor prognosis in advanced hepatocellular carcinoma (HCC). This study aims to evaluate the efficacy and safety of Cinobufacini therapy via hepatic arterial infusion (HAI) in advanced HCC patients with MVI.Methods: The clinical records of 130 consecutive patients with unresectable advanced HCC and MVI who had received Cinobufacini or cisplatin plus 5-fluorouracil (CF) treatment via HAI were retrospectively analyzed. The therapeutic efficacy, overall survival (OS), progression-free survival (PFS), and adverse events were compared between the two treatment groups.Results: The Cinobufacini group demonstrated significant curative effects on treatment via HAI compared with the CF group, including the objective response rate (44.9% vs 27.9%, P=0.048), the median OS (14.8 months vs 11.1 months, P=0.010), and the median PFS (10.3 months vs 6.0 months, P=0.006). Result in subgroup analysis of portal vein invasion grade supported the efficacy in Cinobufacini treatment, especially in the median OS of Vp1-2 (18.3 months vs 14.3 months, P=0.043) and Vp3 (15.0 months vs 11.4 months, P=0.046), as well as the median PFS of Vp1-2 (14.8 months vs 10.2 months, P=0.028) and Vp3 (10.8 months vs 6.6 months, P=0.033) compared with CF treatment. Cox proportional hazards model and forest plot analysis of factors confirmed the survival benefit from HAI with Cinobufacini over CF (hazard ratio [HR], 0.61; 95% CI: 0.40– 0.91; P=0.010). Multivariable analysis identified portal vein invasion grade (Vp4; HR, 1.78; 95% CI: 1.03– 2.16; P=0.032) and AFP (> 1000; HR, 1.61; 95% CI: 1.08– 1.91; P=0.039) as the independent factors for prognosis. Moreover, the total incidence of adverse events in the Cinobufacini group was significantly lower than in the CF group (60.9% vs 82.0%, P=0.009).Conclusion: Cinobufacini therapy via HAI is a viable strategy for curing advanced HCC with MVI, due to prolonged survival and a superior safety profile.Keywords: hepatocellular carcinoma, macrovascular invasion, cinobufacini, hepatic arterial infusion, efficacy

Lina Wang, Zhenzhen Qu, Qian Sun, Zhuofeng Mao, Peipei Si, Weiping Wang Internal Medicine-Neurology, the Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, People’s Republic of ChinaCorrespondence: Weiping Wang, Internal Medicine-Neurology, the Second Hospital of Hebei Medical University, No. 215 Heping Road, Xinhua District, Shijiazhuang, 050004, People’s Republic of China, Tel +86-13613316623, Email wwphb1223@163.comBackground: The 4-hydroxysesamin (4-HS, a di-tetrahydrofuran lignin) is a modified sesamin that was prepared in the laboratory. This preclinical study was designed to preliminarily investigate the neuroprotective properties of 4-HS.Methods: In vitro, neuronal injury and inflammation were simulated by oxygen-glucose deprivation and lipopolysaccharide (LPS) exposure in mouse hippocampal neuronal HT22 cell line, and treated with 4-HS and/or metformin (MET, MAPK pathway activator for exploring mechanism). CCK-8, flow cytometry, and enzyme-linked immunosorbent assay were performed to evaluate cell viability, apoptosis, and inflammation. Apoptosis- and pathway-related proteins were detected by Western blotting. Middle cerebral artery occlusion (MCAO) was constructed as a stroke model and treated with 4-HS for in vivo confirmation. Histological staining was used for in vivo evaluation of 4-HS properties.Results: The 4-HS showed similar anti-inflammatory activity to sesamin but did not affect the cell viability of HT22 cells. In vitro, 4-HS improved the cell viability, ameliorated neuronal apoptosis, along with the reversion of apoptotic proteins (Bax, cleaved-caspase 3/9, Bcl-2) expression and inflammatory cytokines (IL-6, TNF-α, IL-10) in LPS-treated HT22 cells. The 4-HS suppressed the phosphorylation of ERK, JNK, and p38 but the addition of MET reversed 4-HS-induced changes of phenotype and protein expression in LPS-treated cells. In vivo, 4-HS showed apparent improvement in cerebral infarction, brain tissue morphology, neuronal architecture, apoptosis, and inflammation of MCAO mice, and also showed inhibiting effects on the phosphorylation of ERK, JNK, and p38, confirming in vivo results.Conclusion: In this first pre-clinical study on 4-HS, we preliminarily demonstrated the neuroprotective properties of 4-HS both in cell and animal models, and proposed that the underlying mechanism might be associated with the MAPK pathway.Keywords: sesamin, 4-hydroxysesamin, middle cerebral artery occlusion, ischemic stroke, neuronal apoptosis

Jun Cheng,&ast; Jinjin Pan,&ast; Dongmei Zhao, Xuejiao Ma, Qiulin Sun, Jiabin Li Department of Infectious Disease, The First Affiliated Hospital of Anhui Medical University, Hefei, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jiabin Li, Tel/Fax +86-551-62922281, Email lijiabin@163.comAbstract: The global prevalence of hepatitis C virus (HCV) infection is approximately 3%, with a post-infection chronicity rate of up to 50%– 85%. HCV reactivation can occur when anti-HCV positive individuals receive antineoplastic therapy. In this study, we report a case of an anti-HCV positive patient with negative HCV RNA after 12 weeks of direct antiviral therapy. Two months later, sorafenib was used to treat hepatocellular carcinoma, and HCV reactivation occurred after 8 months of the treatment. HCV RNA was negative after 12 weeks of antiviral treatment with Sofosbuvir-velpatasvir. We also discussed the mechanism of HCV reactivation caused by sorafenib and the antiviral treatment regimen after HCV reactivation with the relevant literature.Keywords: hepatocellular carcinoma, sorafenib, HCV-RNA, anti-HCV therapy, reactivation

Yanbei Duo,1 Shuoning Song,1 Xiaolin Qiao,2 Yuemei Zhang,3 Jiyu Xu,4 Jing Zhang,5 Zhenyao Peng,6 Yan Chen,2 Xiaorui Nie,2 Qiujin Sun,7 Xianchun Yang,7 Ailing Wang,8 Wei Sun,4 Yong Fu,1 Yingyue Dong,1 Zechun Lu,8 Tao Yuan,1,&ast; Weigang Zhao1,&ast; 1Department of Endocrinology, Key Laboratory of Endocrinology of Ministry of Health, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, People’s Republic of China; 2Department of Obstetrics, Beijing Chaoyang District Maternal and Child Health Care Hospital, Beijing, People’s Republic of China; 3Department of Obstetrics, Haidian District Maternal and Child Health Care Hospital, Beijing, People’s Republic of China; 4Core Facility of Instrument, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing, People’s Republic of China; 5Department of Laboratory, Haidian District Maternal and Child Health Care Hospital, Beijing, People’s Republic of China; 6Department of Dean’s Office, Haidian District Maternal and Child Health Care Hospital, Beijing, People’s Republic of China; 7Department of Clinical Laboratory, Beijing Chaoyang District Maternal and Child Health Care Hospital, Beijing, People’s Republic of China; 8National Center for Women and Children’s Health, China CDC, Beijing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Tao Yuan; Weigang Zhao, Email t75y@sina.com; xiehezhaoweigang@163.comObjective: Gestational diabetes mellitus (GDM) is a condition of glucose intolerance, which may be accompanied with inflammation. The levels of hematological parameters during pregnancy can reflect inflammatory conditions in pregnant women. This study aims to describe the dynamic change of blood cell parameters from the first trimester (6– 12 weeks of gestation) to the second trimester (24– 28 weeks of gestation) and to investigate the associations of these biomarkers with the risk of GDM.Methods: This study was a prospective double-center study conducted in Beijing, China (clinical trial number: NCT03246295). Hematological parameters were tested four times during the follow-up. Logistic regression analysis and Receiver Operating Characteristic (ROC) curve analysis were used to explore the association and predictive ability of hematological parameters for GDM.Results: There were 258 of 1027 pregnant women in our study developed GDM. Among the 1027 pregnant women, white blood cells (WBC) gradually increased, and red blood cells (RBC), hemoglobin (HGB), and platelet (PLT) tended to decrease from the first trimester to second trimester. After adjusting for confounding factors, higher levels of RBC, HGB, and PLT in both early and middle pregnancy were positively associated with GDM risk, whereas the level of WBC was associated with GDM risk only in early pregnancy. WBC, RBC, HGB, and PLT in early and middle pregnancy were all correlated with fasting insulin (FINS) in early pregnancy.Conclusion: Higher levels of hematological parameters in early and middle pregnancy were associated with glucose metabolism in early pregnancy and the subsequent risk of GDM.Keywords: hematological parameters, gestational diabetes mellitus, inflammation, insulin resistance, prospective study

Deyong Tan,1– 4 Jianfeng Han,5 Qingzhi Sun,5 Xing Cheng,5 Juan Liu,5 Jia Liu,5 Qing Li,1– 4 Lizhong Dai5 1Department of Clinical Pharmacology, Xiangya Hospital, Central South University, Changsha, 410008 People’s Republic of China; 2Institute of Clinical Pharmacology, Central South University, Hunan Key Laboratory of Pharmacogenetics, Changsha, 410078 People’s Republic of China; 3Engineering Research Center of Applied Technology of Pharmacogenomics, Ministry of Education, Changsha, 410078 People’s Republic of China; 4National Clinical Research Center for Geriatric Disorders, Changsha, Hunan, 410008 People’s Republic of China; 5Sansure Biotech Inc, Changsha, Hunan Province, People’s Republic of ChinaCorrespondence: Qing Li, Department of Clinical Pharmacology, Xiangya Hospital, Central South University, 87 Xiangya Road, Changsha, Hunan, People’s Republic of China, Tel +86 731 84805380, Fax +86 731-82354476, Email liqing9251026@csu.edu.cn Lizhong Dai, Sansure Biotech Inc, No. 680 Lusong Road, Changsha, Hunan, People’s Republic of China, Tel +86 73188883176, Email lizhongd@sansure.com.cnObjective: In this study, we conducted a multi-center research on six common lower respiratory tract pathogens using novel multiplex fluorescence quantitative polymerase chain reaction (PCR), and investigated the additional diagnostic value of this method, to provide a molecular diagnostic basis for clinical practice.Methods: From March 2019 to October 2021, a total of 2047 respiratory sputum samples were collected from Hunan Provincial People’s Hospital (the First Affiliated Hospital of Hunan Normal University), Hunan Provincial Children’s Hospital, Jiangxi Provincial Children’s Hospital, and Wuhan Infectious Disease Hospital. The samples were analyzed using a novel multiplex fluorescence quantitative PCR method for Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, Legionella pneumophila, and Staphylococcus aureus. The results were compared to the results of bacterial culture and sequencing, as well as the results of third-party kits.Results: Compared to the bacterial culture method, 2047 samples were detected with a sensitivity of 100%, a specificity of 72.22%, and an overall compliance rate of 81.91%. Compared to the sequencing method, the positive agreement percentage was 99.88%, the negative agreement percentage was 97.72%, and the overall agreement rate was 98.84%. Compared to similar control reagents, the positive agreement percentage was 100%, negative agreement percentage was 79.79%, and overall compliance rate was 96.19%.Conclusion: The multiplex fluorescence PCR method has the advantages of simultaneously detecting multiple pathogenic bacteria and reducing the duration of pathogen culture identification. Combined detection can increase the detection rate, which has favorable performance and application prospects.Keywords: bacterial infection, lower respiratory tract, melting curve, multiplex PCR, rapid diagnosis

Zihan Hu,1,&ast; Danjiang Dong,2,&ast; Fei Peng,1,&ast; Xing Zhou,1 Qin Sun,1 Hui Chen,1,3 Wei Chang,1 Qin Gu,2,&ast; Jianfeng Xie,1,&ast; Yi Yang1,&ast; 1Jiangsu Provincial Key Laboratory of Critical Care Medicine, Department of Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, 210009, People’s Republic of China; 2Department of Critical Care Medicine, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing, People’s Republic of China; 3Department of Critical Care Medicine, The First Affiliated Hospital of Soochow University, Soochow University, Suzhou, 215000, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Jianfeng Xie; Qin Gu, Department of Critical Care Medicine, Zhongda Hospital, School of medicine, Southeast University, 87 Dingjiaqiao Road, Nanjing, 210009, People’s Republic of China, Tel/Fax +00862583262500, Email xie820405@126.com; guqinicu012323@126.comPurpose: Immune dysfunction plays a pivotal role in sepsis pathogenesis. Previous studies have revealed the crucial role of T cells and human leukocyte antigen-DR (HLA-DR) in sepsis. However, the function of natural killer (NK) cells remains unclear. This study aimed to investigate whether NK cells are associated with sepsis prognosis. In addition, we aimed to explore the interrelation and influence between NK and other immunological features in patients with sepsis.Patients and Methods: This retrospective, observational study included patients with sepsis from two hospitals in mainland China. The clinical characteristics and immune results during the early phase were collected. Patients were classified according to the level of immune cells to analyze the relationship between immunological features and 28-day mortality.Results: A total of 984 patients were included in this study. Non-survivors were older and had lower levels of lymphocytes, monocytes, NK cells, HLA-DR, and T cells. Patients were classified into eight groups according to their levels of NK cells, HLA-DR, and T cells. Only patients with decreased NK and T cell counts showed a significant increase in 28-day mortality. An increase in CD8+ T cells was correlated with the alleviation of 28-day mortality only among patients with high NK cell levels.Conclusion: This study provides novel insights into the association between NK cells and 28-day mortality as well as the interrelation between NK cells and other immune cells in sepsis. The relationship between CD8+ T cells and 28-day mortality in sepsis is dependent on NK cell count.Keywords: sepsis, immune dysfunction, natural killer cell, HLA-DR, T cell

Xiaopei Xu, Qingnan Sun, Yang Liu, Dong Wang, Shuo Diao, Hanzhou Wang, Yuling Gao, Tianchao Lu, Junlin Zhou Department of Orthopedic Surgery, Beijing Chaoyang Hospital, Capital Medical University, Beijing, 100020, People’s Republic of ChinaCorrespondence: Junlin Zhou, Department of Orthopedic Surgery, Beijing Chaoyang Hospital, Capital Medical University, 8 Gongren Tiyuchang Nanlu, Chaoyang District, Beijing, 100020, People’s Republic of China, Tel/Fax +86-01085231227, Email junlinzhou_article@outlook.comObjective: The aim was to evaluate the effects of different glenosphere eccentricities on impingement, range of motion (ROM), and muscle length during standard activities in reverse total shoulder arthroplasty (RSA).Methods: In this study, we utilized computational modeling techniques to create native shoulder and shoulder models undergoing RSA and simulate shoulder movements in all abduction-adduction, flexion-extension, and rotation. We tested a total of 36 different glenosphere configurations, which included three different inferior tilts (0°, +10°, +20°) and two different lateral offsets (0 mm and +4 mm), as well as six different glenosphere eccentricities (concentricity, inferior, posterior, anterior, anteroinferior, and posteroinferior). We evaluated the maximum impingement-free ROM, impingement sites, and muscle lengths.Results: All glenosphere configurations exceeded 50% of native shoulder ROM in three planes and total global ROM. In abduction-adduction, there was no significant difference among the different glenosphere eccentricities (p > 0.05). In flexion-extension, the posteroinferior eccentricity had the maximum ROM among the different eccentricities, but no significant difference among the different glenosphere eccentricities (p > 0.05). In rotation, there was a significant difference overall, and anteroinferior eccentricity had a significant advantage over concentricity (p < 0.05). In total global ROM, anteroinferior eccentricity had a significant advantage over concentricity when lateral offset was 0 mm (p < 0.05). In all models of glenosphere eccentricities, only the elongation of the infraspinatus muscle was statistically significant (p < 0.05).Conclusion: Glenosphere eccentricity significantly influenced rotation, total global ROM, and the length of the subscapularis muscle. Among them, anteroinferior offset achieved the maximum ROM in abduction-adduction, rotation, and total global activities. Both anteroinferior and inferior glenoid eccentricity showed significant advantages over the concentricity in rotation and total global ROM.Level of Evidence: Basic Science Study; Computer Modeling.Graphical Abstract: Keywords: reverse shoulder arthroplasty, glenosphere eccentricities, impingement, notching, range of motion, muscle length

Jiawei Qian,1,2 Xiulan Shen,3 Xin Gao,3 Qiuhua Sun2 1Department of Nursing, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, People’s Republic of China; 2School of Nursing, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 3Emergency and Trauma Center, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, People’s Republic of ChinaCorrespondence: Qiuhua Sun, Email sqh807@163.comBackground: Delirium is common in older patients during the postoperative period, increasing the number of adverse events, and grip strength is associated with delirium. Subsyndromal delirium (SSD) is a prostate of clinical delirium; nevertheless, the relationship between grip strength and SSD has not been elucidated. This study aimed to examine the association between grip strength and SSD in the elderly after arthroplasty.Methods: A total of 181 patients were recruited from two Chinese hospitals. SSD and delirium were evaluated before the surgery and the first week after surgery using the confusion assessment method. The Mini-mental State Examination was used to assess patients’ cognitive function, and their grip strength was evaluated with an electronic hand dynamometer before surgery. Logistic regression and ROC curve analysis were conducted to determine the odds ratio and predictive value of grip strength for SSD.Results: The incidence of SSD and postoperative delirium (POD) was 41.44% and 14.36% for the elderly following arthroplasty respectively, and approximately 1/3 of SSD progressed into POD. Older age, declined cognitive function, fall history, and lower grip strength were risk factors for SSD (P< 0.05). The area under the ROC curve of grip strength was 0.863 and 0.900 for males and females respectively, and the cut-off point of it was determined to be 22.050 kg for men and 18.050 kg for women.Conclusion: SSD and POD are common among older people. Decreased grip strength, advanced age, lower cognitive function, and fall history were independent risk factors for SSD, and grip strength was a significant predictor for SSD in aged patients after the arthroplasty.Keywords: subsyndromal delirium, delirium, grip strength, risk factor, joint replacement

Zhe Xiong,1,2 Ying Fang,1,2 Shuangshuang Lu,1 Qiuyue Sun,1,3 Jin Huang1 1Department of Gastroenterology, the Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University, Changzhou, Jiangsu Province, People’s Republic of China; 2Graduate School of Dalian Medical University, Dalian, Liaoning Province, People’s Republic of China; 3Graduate School of Nanjing Medical University, Nanjing, Jiangsu Province, People’s Republic of ChinaCorrespondence: Jin Huang, Email hj042153@hotmail.comBackground: Inflammatory bowel disease (IBD) and periodontitis (PD) are correlated, although the pathogenic mechanism behind their correlation has not been clarified. This study aims to explore the common signature genes and potential therapeutic targets of IBD and PD using transcriptomic analysis.Methods: The GEO database was used to download datasets of IBD and PD, and differential expression analysis was used to identify DEGs. We then conducted GO and KEGG enrichment analyses of the shared genes. Next, we applied 4 machine learning (ML) algorithms (GLM, RF, GBM, and SVM) to select the best prediction model for diagnosing the disease and obtained the hub genes of IBD and PD. The diagnostic value of the signature genes was verified by a validation set and qRT‒PCR experiments. Subsequently, immune cell infiltration in IBD samples and PD samples was analyzed by ssGSEA. Finally, we investigated and validated the response of hub genes to infliximab therapy.Results: We identified 43 upregulated genes as shared genes by intersecting the DEGs of IBD and PD. Functional enrichment analysis suggested that the shared genes were closely associated with immunity and inflammation. The ML algorithm and qRT‒PCR results indicated that IGKC and COL4A1 were the hub genes with the most diagnostic value for IBD and PD. Subsequently, through immune infiltration analysis, CD4 T cells, NK cells and neutrophils were identified to play crucial roles in the pathogenesis of IBD and PD. Finally, through in vivo and in vitro experiments, we found that IGKC and COL4A1 were significantly downregulated during the treatment of patients with IBD using infliximab.Conclusion: We investigated the potential association between IBD and PD using transcriptomic analysis. The IGKC and COL4A1 genes were identified as characteristic genes and novel intervention targets for these two diseases. Infliximab may be used to treat or prevent IBD and PD.Keywords: Inflammatory bowel disease, periodontitis, machine learning, ssGSEA, infliximab, IGKC, COL4A1

Lijie Ma,&ast; Qiuxia Han,&ast; Fang Sun, Kaiyi Zhu, Qianmei Sun Department of Nephrology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, 100020, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Qianmei Sun, Email sunqianmei5825@126.comPurpose: This study aimed to explore whether the mean platelet volume/platelet count ratio (MPR) has predictive value for the occurrence and prognosis of acute ischemic stroke (AIS) in hemodialysis patients.Patients and Methods: A total of 402 patients undergoing maintenance hemodialysis were screened and 259 were enrolled in this study. The receiver operating characteristic curve and area under the curve (AUC) were used to evaluate the predictive power of the models. The patients enrolled in this study were divided into three groups based on the tertiles of the MPR value (Q1, Q2, and Q3). Kaplan-Meier curves were used to investigate the association between the MPR and AIS-free survival in hemodialysis patients. Chi-square analysis was performed to explore the association between the MPR and AIS outcomes in hemodialysis patients. And the AIS outcome was assessed using the modified Rankin Scale (mRS).Results: MPR had a predictive value for the occurrence of AIS (AUC=0.814) in hemodialysis patients with a high sensitivity and specificity. AIS-free survival rates in the MPR Q1, MPR Q2, and MPR Q3 groups were 0.930, 0.701, and 0.360, respectively. The proportion of patients with good outcomes (mRS 0– 2) was significantly greater among patients in the MPR Q1-Q2 group than in the MPR Q3 group (0.844 vs 0.745, p < 0.001).Conclusion: The MPR can be used as a good predictor of AIS in patients undergoing hemodialysis. Patients on hemodialysis with increased MPR levels had a higher incidence of AIS and poorer functional outcomes than those with low MPR levels.Keywords: acute ischemic stroke, hemodialysis, mean platelet volume/ platelet count ratio, predictor, prognosis

Guiling Li,1 Li Wang,2 Heguang Zhang,1 Ying Luan,1 Qi Sun,1 Libo Duo1 1Department of Clinical Laboratory, the Second Hospital Affiliated to Harbin Medical University, Harbin, Heilongjiang, 150086, People’s Republic of China; 2Department of Clinical Laboratory, School of Medicine, Chengdu Women’s and Children’s Central Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, 611731, People’s Republic of ChinaCorrespondence: Libo Duo, Department of Clinical Laboratory, the Second Hospital affiliated to Harbin Medical University, No. 246 XueFu Road, NanGang District, Harbin, Heilongjiang, 150086, People’s Republic of China, Tel +86 045186605382, Email duolibo@163.comObjective: In this study, we constructed ampG knock-out and knock-in strains from a clinically isolated Kp1strain carrying ampR-ampC in its plasmid and compared them with the Kp NTUH-K2044 strain to investigate the relationship between ampG and ampR-ampC-induced expression.Methods: We created the ampG gene deletion mutant strains Kp1-ΔampG and Kp NTUH-K2044-ΔampG with pKO3-km plasmid using homologous recombination technology. We constructed the Kp NTUH-K2044-RC and Kp NTUH-K2044-ΔampG-RC drug resistance model strains with plasmid pACYC184. We constructed the ampG knock-in strains by introducing the ampG genes of Kp1, Enterobacter cloacae 029M, Pseudomonas aeruginosa PAO1, Escherichia coli ATCC25922, and Salmonella typhimurium LT2 into the ampG gene-deleted strains with carrier pet-30a. Real-time polymerase chain reaction (real-time PCR) was used to detect the relative expressions of ampC and ampG mRNAs.Results: Compared with Kp1, the induction phenotype of the ampC of Kp1-ΔampG strain disappeared, the ampC expression was reduced, and the minimal inhibitory concentration (MIC) values of cefoxitin and ceftazidime significant decrease from 128 μg/mL to 1 μg/mL. Based on Kp1, five strain were successfully constructed to complement the ampG genes from five knock-in strain, and all of the above complemented strains showed inducible expression of ampC and restored the expression of ampG to varying degrees, as well as restored resistance to the antimicrobial drugs cefoxitin and ceftazidime (P < 0.05). The ampC and ampG genes were barely expressed in Kp NTUH-K2044-ΔampG-RC when compared with Kp NTUH-K2044-RC. The expressions of ampG and ampC in each knock-in strain were recovered, the induction phenotype of ampC was restored, and the MIC values of cefoxitin and ceftazidime were increased. (P < 0.05).Conclusion: In this study, we found that ampG was an essential regulator for the plasmid-mediated ampC-induced expression in K. pneumoniae.Keywords: ampC inducible expression, AmpG regulator, gene knockout, gene knock-in, Klebsiella pneumoniae

Xinchen Wang,1 Chen Wei,1 Wenjun Fan,1 Lixian Sun,1 Ying Zhang,1 Qiyu Sun,2 Yixiang Liu,1 Jingyi Liu1 1Department of Cardiology, The Affiliated Hospital of Chengde Medical University, The Chengde Institute of Cardiovascular Diseases, Chengde, Hebei, 067000, People’s Republic of China; 2Department of Clinical Laboratory, The Affiliated Hospital of Chengde Medical University, Chengde, Hebei, 067000, People’s Republic of ChinaCorrespondence: Lixian Sun, Department of Cardiology, The Affiliated Hospital of Chengde Medical University, Chengde, Hebei, 067000, People’s Republic of China, Tel +86 314 227 9016, Fax +86 314 227 4895, Email lixiansun01@126.comPurpose: The decreased advanced lung cancer inflammation index (ALI), defined as body mass index (BMI) &ast; albumin (Alb)/neutrophil-to-lymphocyte ratio (NLR), is an independent prognostic risk factor for overall survival in gastric, lung, and colorectal cancers. This study aimed to investigate the value of ALI in predicting the risk of major adverse cardiovascular events (MACEs) in patients with acute coronary syndrome (ACS).Patients and Methods: A total of 1624 patients with ACS undergoing percutaneous coronary intervention (PCI) were consecutively enrolled between January 2016 and December 2018. Follow-up data were collected at 1, 3, 6, and 12 months and annually thereafter. The primary endpoints were MACEs. All endpoints were defined as all-cause mortality, recurrent angina pectoris, restenosis/intra stent thrombosis, stroke, heart failure, and all-cause bleeding.Results: The MACEs group and non-MACEs group showed significant differences in patients with age > 65 years (28 [50.0%] vs 319 [23.7%]), history of heart failure (16 [28.6%] vs 127 [9.4%]), history of ischemic stroke (14 [25.0%] vs 186 [13.8%]), history of cardiogenic shock (6 [10.71%] vs 16 [1.19%]), left ventricular ejection fraction < 40% (8 [14.29%] vs 33 [2.46%]), and ALI < 343.96 (44 [78.65%] vs 680 [50.60%]) (all p< 0.001). The optimal cut-off value for ALI was 334.96. The area under the curve (AUC) of the 1-, 2-, 3-, and 5-year was 0.560, 0.577, 0.665, and 0.749, respectively. The survival rate was significantly lower in the low ALI group than in the high ALI group (log-rank p< 0.001). Low ALI was an independent risk factor for the long-term prognosis of patients with ACS after PCI, univariate HR: 3.671, 95% CI: 1.938– 6.953, p< 0.001; multivariate HR: 3.009, 95% CI: 1.57– 5.769, p=0.001.Conclusion: ALI score less than 334.96 is an independent prognostic risk factor for patients with ACS undergoing PCI and may be a novel marker for clinical practice.Keywords: acute coronary syndrome, advanced lung cancer inflammation index, prognosis, percutaneous coronary intervention

Qinghe Zeng,1,2,&ast; Qi Sun,3,&ast; Huihui Xu,1,2,&ast; Jiali Chen,1 Houfu Ling,4 Qinwen Ge,1,2 Kaiao Zou,1,2 Xu Wang,1,2 Hongting Jin,1 Ju Li,4,&ast; Minwei Jin4,&ast; 1Institute of Orthopedics and Traumatology, the First Affiliated Hospital of Zhejiang Chinese Medical University, Zhejiang Provincial Hospital of Chinese Medicine, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 2The First College of Clinical Medicine, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 3Department of Orthopaedic Surgery, Fuyang Orthopaedics and Traumatology Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, People’s Republic of China; 4Department of Orthopaedics, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Minwei Jin; Ju Li, Email jin872@zcmu.edu.cn; juli@zcmu.edu.cnBackground: Intervertebral disc degeneration (IDD) is a major cause of lower back pain (LBP), in which inflammatory is frequently involved. Amygdalin (AMD) is a naturally occurring compound that exerts anti-fibrotic, anti-inflammatory, analgesic, and immunomodulatory effects in various diseases. The purpose of this study was to investigate the therapeutic effects and molecular mechanisms of AMD on Lumbar spine instability (LSI)-induced IDD in mice.Methods: In this study, we first explored the effects of AMD in vivo, and then further explored the mechanism of its effects both in vivo and in vitro. Ten-week-old male C57BL/6J mice were administrated with AMD. At 10 weeks after LSI, spinal were collected for tissue analyses, including histology, micro-CT, and immunohistochemistry for Col2, Mmp-13, TNF-α, and p-P65. Additionally, we also evaluated the mRNA and protein expression level of p-P65 and p-IKBα after being treated with AMD in vitro.Results: Histological staining, micro-CT and immunohistochemical analysis showed that AMD treatment significantly inhibited the expression of TNF-α and Mmp-13, increased the expression of Col2 as well as attenuated the calcification of cartilage endplates, eventually to delayed the progression of IDD. Meanwhile, in vivo and in vitro fluorescence imaging revealed that AMD markedly inhibited the AMD significantly inhibited the LSI-induced increase in TNF-α expression and P65and IKBα phosphorylation.Discussion: Our findings suggest that AMD partly inhibits the activation of NF-κB signaling pathway to reduce the release of inflammatory mediators and delay the degeneration of cartilage endplate in IDD model mice. Therefore, AMD may be a potential candidate for the treatment of IDD.Keywords: intervertebral disc degeneration, amygdalin, cartilage endplate, inflammatory processes, NF-κB pathway

Qing-Wei Gu,&ast; Qi Sun,&ast; Jie Wang, Wen-Sha Gu, Wei Wang, Xiao-Ming Mao Department of Endocrinology, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xiao-Ming Mao, Department of Endocrinology, Nanjing First Hospital, Nanjing Medical University, 68 ChangLe Street, Nanjing, Jiangsu, 210006, People’s Republic of China, Tel +86-18951670295, Email maoxming@163.comPurpose: To investigate the pathogenesis of diabetic kidney disease (DKD) in type 2 diabetes mellitus (T2DM), we evaluated the effects of short-term glycemic variability (GV) on the profile of T cell subpopulations.Methods: A total of 47 T2DM patients with normoalbuminuria, 47 microalbuminuria, and 49 macroalbuminuria were enrolled. The continuous glucose monitoring (CGM) determined the GV of enrolled patients. Flow cytometry was used to determine the proportion of T cell subpopulations.Results: The frequency of T helper (Th) 17 and Th1 cells significantly increased while regulatory T cells (Tregs) significantly decreased in the macroalbuminuria group compared to normoalbuminuria and microalbuminuria groups (P < 0.01). The suppressive function of Tregs was significantly lower in the macroalbuminuria group than the normoalbuminuria group (P < 0.05). Compared with the normoalbuminuria group, the mean amplitude of glucose excursions (MAGE) of the macroalbuminuria group was significantly higher (P< 0.05). Furthermore, there were negative associations between the proportion of Tregs and MAGE.Conclusions: Increased GV could decrease the proportion of Tregs and may impair their function. This may lead to increases in Th1 and Th17 cells, and some inflammatory cytokines, which might contribute to the development and progression of DKD in T2DM.Keywords: type 2 diabetes mellitus, diabetic kidney disease, glycemic variability, regulatory T cells, inflammatory cytokines

Zeng Q, Sun Q, Xu H, et al. J Inflamm Res. 2023;16:3455-3468 The authors have advised there is an error in the author list and affiliations on page 3455. Minwei Jin’s affiliation should read “Department of Orthopaedic Surgery, the First Affiliated Hospital of Zhejiang Chinese Medical University, Zhejiang Provincial Hospital of Chinese Medicine, Hangzhou, People’s Republic of China” The corrected author list and affiliations is: Qinghe Zeng 1,2,*, Qi Sun3,*, Huihui Xu 1,2,*, Jiali Chen1, Houfu Ling4, Qinwen Ge1,2, Kaiao Zou1,2, Xu Wang1,2, Hongting Jin1, Ju Li4,*, Minwei Jin5,* 1Institute of Orthopedics and Traumatology, the First Affiliated Hospital of Zhejiang Chinese Medical University, Zhejiang Provincial Hospital of Chinese Medicine, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 2The First College of Clinical Medicine, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 3Department of Orthopaedic Surgery, Fuyang Orthopaedics and Traumatology Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, People’s Republic of China; 4Department of Orthopaedics, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 5Department of Orthopaedic Surgery, the First Affiliated Hospital of Zhejiang Chinese Medical University, Zhejiang Provincial Hospital of Chinese Medicine, Hangzhou, People’s Republic of China *These authors contributed equally to this work

Hao Qin,1,&ast; Yuxiang Wan,2,&ast; Yuchao Dong,1 Qinying Sun1 1Department of Respiratory and Critical Care Medicine, Shanghai Changhai Hospital, The First Affiliated Hospital of Second Military Medical University, Shanghai, People’s Republic of China; 2Department of Laboratory Diagnosis, Shanghai Changhai Hospital, The First Affiliated Hospital of Second Military Medical University, Shanghai, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yuchao Dong; Qinying Sun, Department of Respiratory and Critical Care Medicine, Shanghai Changhai Hospital, The First Affiliated Hospital of Second Military Medical University, Shanghai, People’s Republic of China, Email dongyc1020@126.com; sqy20031@163.comAbstract: Pulmonary sarcomatoid carcinoma (PSC) is a rare subtype of non-small cell lung cancer (NSCLC), accounting for about 1% of cases. These tumors are characterized by their high malignancy and frequent resistance to chemotherapy, resulting in a worse prognosis compared to other NSCLC subtypes. Currently, there is no established therapeutic strategy for PSC. Recent advancements in targeted therapies have led to the development of ret proto-oncogene (RET) inhibitors, such as selpercatinib and pralsetinib, which have been approved for the treatment of RET fusion-positive NSCLC patients. Despite their effectiveness in RET fusion-positive NSCLC is observed, the efficacy of these inhibitors in PSC remains unclear. In this context, we present a case of metastatic PSC harboring de novo KIF5B-RET fusion. The patient responded to first-line trametinib treatment. These findings suggest that RET inhibitors could be a potential treatment option for metastatic PSC patients with RET fusion-positive tumors.Keywords: pulmonary sarcomatoid carcinoma, RET, pralsetinib, efficacy

Zhe Wang, Ying Zhong, Yidong Zhou, Feng Mao, Xiaohui Zhang, Changjun Wang, Qiang Sun Department of Breast Surgery, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, 100730, People’s Republic of ChinaCorrespondence: Qiang Sun, Email sunqpumch@163.comPurpose: This study aimed to assess the effect of comorbidities on prognosis using the Age-adjusted Charlson Comorbidity Index (ACCI) among the elderly with breast cancer (BC).Methods: This study included 745 patients divided into two groups following the ACCI score (≤ 3 vs > 3). Multivariate logistic regression analysis was conducted for all kinds of outcomes, including BC-specific death (BCSD) and non-breast cancer-specific death (NBCSD). The Kaplan–Meier curves were plotted, and survival analysis was conducted for disease-free survival (DFS), overall survival (OS), BC-specific survival (BCSS), and non-BCSS (NBCSS).Results: A significantly higher NBCSD was found in the high-score (ACCI > 3) group than in the low-score (ACCI < 3) group (p = 0.032). The multivariate logistic regression analysis revealed ACCI score as an independent affecting factor for all-cause death (hazard ratio [HR] = 0.42, 95% confidence interval [CI]: 0.22– 0.83, p = 0.012) and NBCSD (HR = 0.41, 95% CI: 0.20– 0.87, p = 0.020). The Kaplan–Meier curves revealed statistical differences only in NBCSS between the two groups (p = 0.039). Subgroup analysis revealed a worse prognosis in the high-score group for OS and NBCSS among hormone receptor-positive participants and those who without undergoing axillary dissection or receiving chemotherapy (all p < 0.05). Multivariate Cox regression analysis revealed ACCI as an independent prognostic predictor for OS (HR = 2.18, 95% CI: 1.22– 3.92, p = 0.009) and NBCSS (HR = 2.04, 95% CI: 1.02– 4.08, p = 0.044).Conclusion: ACCI was indeed an effective indicator of the effects of comorbidities on survival among elderly patients with BC. However, the co-effect from age and comorbidities was not significant enough on cancer-specific prognosis, although it exerted a significant effect on treatments received.Keywords: comorbidity, elderly, breast cancer, Age-adjusted Charlson Comorbidity Index, prognosis

Min Xu,1,&ast; Ting Liu,2,&ast; Menghua Ye,1 Xiaoxue Tan,1 Qiuhua Sun2 1The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 2The School of Nursing, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Qiuhua Sun, Zhejiang Chinese Medical University, The School of Nursing, 548 Binwen Road, Binjiang District, Hangzhou City, Zhejiang Province, People’s Republic of China, Email sunqiuhua@zcmu.edu.cnBackground: Aplastic anemia is characterized by anemia, hemorrhage and infection, and is accompanied by a variety of complications and psychological burden. Therefore, the quality of life of AA patients is not optimistic. Our team is committed to developing an assessment tool for the quality of life of AA patients, and adopting an iterative strategy of cognitive interview and expert consultation to solve the challenges encountered in item revision.Purpose: We aim to use the strategy of cognitive interview and expert consultation to inform revision of the QLS-AA into a user-friendly tool with unambiguous items and improve the content validity of the scale.Methods: We used an iterative strategy of cognitive interview and expert consultation. Two rounds of cognitive interview were conducted to identify problems with item comprehension, recall and other cognitive processes. As well as, a multi-disciplinary group of expert consultation was consulted to review the rationality of item revisions.Results: In the first round of cognitive interview, 16 participants responded to 107 items. Among them, the most common problems were “clarification” and “item duplication”. Based on the results of the first round of interview, an expert consultation was organized. A total of 16 amendments were put forward by the expert and 14 were adopted. In the second round of cognitive interviews, A total of 5 participants were included and 64 items were evaluated. Two items were suggested to be revised, and the remaining items were accurately understood and recognized by all participants.Couclusion: This study highlights the key issues to consider when incorporating patient perspectives into quality measurement. The revision of QLS-AA through the strategy of cognitive interview and expert consultation may provide valuable insights into the measurement of quality of life in aplastic patients.Trial Registration Number: ChiCTR2100047575.Keywords: cognitive interview, expert consultation, aplastic anemia, quality of life

Panpan Fang,1,&ast; Kaijie Gao,1 Junmei Yang,1 Tiewei Li,1 Weihua Gong,1 Qianqian Sun,1 Yingyuan Wang2,&ast; 1Zhengzhou Key Laboratory of Children’s Infection and Immunity, Children’s Hospital Affiliated to Zhengzhou University, Henan Children’s Hospital, Zhengzhou Children’s Hospital, Zhengzhou, People’s Republic of China; 2Department of Neonatal Intensive Care Unit, Children’s Hospital Affiliated to Zhengzhou University, Henan Children’s Hospital, Zhengzhou Children’s Hospital, Zhengzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Panpan Fang; Yingyuan Wang, Email fang_panpan@163.com; yingyuanxse@163.comIntroduction: Sepsis is the most severe infectious disease with the highest mortality rate, particularly among neonates admitted to the neonatal intensive care unit (NICU). This study retrospectively analyzed the epidemiology, antibiotic resistance profiles, and prevalence of multidrug-resistant (MDR) bacteria isolated from blood or cerebrospinal fluid (CSF) cultures in order to evaluate the appropriateness of initial empirical therapy for neonatal sepsis.Methods: A retrospective study was conducted in the NICU from January 1, 2015, to December 31, 2022. Microbiological data from patients admitted to the NICU were anonymously extracted from the Laboratory of Microbiology database. Neonatal sepsis was classified into two types: early-onset sepsis (EOS), which occurs within the first 72 hours of life, and late-onset sepsis (LOS) for those begins later.Results: A total of 679 bacterial strains, 543 from blood and 136 from CSF, were detected in 631 neonates. Among these, 378 isolates (55.67%) were Gram-positive bacteria, and 301 isolates (44.33%) were Gram-negative bacteria. The most frequently isolated pathogens were Coagulase-negative staphylococci (CoNS) (36.52%), followed by Klebsiella pneumoniae (20.47%) and Escherichia coli (13.84%). In EOS, 121 strains were found, CoNS represented the majority (33.88%), followed by Klebsiella pneumoniae (23.97%) and Escherichia coli (8.26%). Early-onset septicemia exhibited 67 (55.37%) MDR bacteria. In LOS, 558 strains were isolated, CoNS represented the majority of pathogens (37.10%), followed by Klebsiella pneumoniae (19.71%) and Escherichia coli (15.05%). Late-onset septicemia showed 332 (59.50%) MDR bacteria. High rates of MDR were found in CoNS (76.21%), carbapenem-resistant Klebsiella pneumoniae (66.91%), and MRSA (33.33%).Conclusion: The study revealed an alarming prevalence of MDR strains isolated from neonatal sepsis, emphasizing the necessity of finding effective prevention and treatment measures. Colistin can be used for MDR Gram-negative bacteria, while vancomycin and teicoplanin can be considered treatment therapies for staphylococcal infections.Keywords: MDR, EOS, LOS, sepsis, bacterial strains

Jingjun Xie,1 Jinxia Li,2 Qi Sun1 1Department of Acupuncture, The First People’s Hospital of Huzhou, Zhejiang, 313000, People’s Republic of China; 2Department of Acupuncture, Huzhou Hospital of Traditional Chinese Medicine, Affiliated to Zhejiang Chinese Medical University, Zhejiang, 313000, People’s Republic of ChinaCorrespondence: Jinxia Li, Email lijinxia310@163.comObjective: The purpose of this study was to evaluate the clinical efficacy of extracorporeal shock wave combined with warm acupuncture for external humeral epicondylitis.Methods: Eighty-two patients with external humeral epicondylitis were randomly divided into an observation group and a control group. Patients in the control group were treated with extracorporeal shock waves while those in observation group with warm acupuncture on the basis of the control group. Patients in both groups were evaluated by Visual Analogue Scale (VAS), Mayo Elbow Performance Score (MEPS), Disabilities of the Arm, Shoulder and Hand questionnaire (DASH) before and after treatment. The inflammatory factors such as IL-6, IL-10, TNF-ɑ and clinical outcomes were contrasted before and after treatment.Results: There were statistically significant differences in VAS score, MEPS score and DASH score between the two groups before and after treatment (P< 0.05), and the improvement of each score in the observation group was more obvious than that in the control group. After treatment, the inflammatory factors of the two groups were lower than those before treatment, and the difference was statistically significant (P< 0.05). The decrease of inflammatory factors in the observation group was more obvious than that in the control group. The total effective rate of the observation group was higher than that of the control group, and the difference was statistically significant (P< 0.05).Conclusion: Extracorporeal shock wave combined with warm acupuncture could effectively improve the pain symptoms and dysfunction of external humeral epicondylitis and reduce the expression of inflammatory factors, and its effect may be better than that of extracorporeal shock wave treatment alone.Clinical Trial Registration: ChiCTR2200066075.Keywords: extracorporeal shock wave, warm acupuncture, external humeral epicondylitis, a randomized clinical trial

Qianjia Sun,1,2,&ast; Tingjing Zhang,3,&ast; Laeeqa Manji,4 Yashu Liu,1,2 Qing Chang,1,2 Yuhong Zhao,1,2 Yang Ding,5,6 Yang Xia1,2 1Department of Clinical Epidemiology, Shengjing Hospital of China Medical University, China Medical University, Shenyang, People’s Republic of China; 2Key Laboratory of Precision Medical Research on Major Chronic Disease, Shenyang, People’s Republic of China; 3School of Public Health, Wannan Medical College, Wuhu, People’s Republic of China; 4International Educational School, China Medical University, Shenyang, People’s Republic of China; 5Department of Infectious Diseases, Shengjing Hospital of China Medical University, China Medical University, Shenyang, People’s Republic of China; 6Diagnosis and Treatment Centre for Liver Diseases of Liaoning Province, Shenyang, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yang Xia, Department of Clinical Epidemiology, Shengjing Hospital of China Medical University, No. 36, San Hao Street, Shenyang, Liaoning, 110004, People’s Republic of China, Tel +86 24-96615-13650, Email xytmu507@126.com; yxia@cmu.edu.cn Yang Ding, Department of Infectious Diseases, Shengjing Hospital of China Medical University, No. 36, San Hao Street, Shenyang, Liaoning, 110004, People’s Republic of China, Tel +86 24-96615-62211, Email yding0903@sina.comObjective: Recent epidemiological evidence shows that there is an association between serum uric acid (SUA) levels and nonalcoholic fatty liver disease (NAFLD). The purpose of this meta-analysis is to summarize all available evidence and assess the associations between SUA levels and NAFLD.Methods: Using two databases, Web of Science and PubMed, observational studies were applied from the establishment of the databases to June 2022. We used a random effect model to construct the pooled odds ratio (OR) and 95% confidence interval (CI) to appraise the association between SUA levels and NAFLD. The Begg’s test was conducted to appraise publication bias.Results: A total of 50 studies were included, involving 2,079,710 participants (719,013 NAFLD patients). The prevalence and incidence rates (95% CIs) of NAFLD in the patients with hyperuricemia were 65% (57– 73%) and 31% (20– 41%), respectively. Compared to participants with lower levels of SUA, the pooled OR (95% CI) of NAFLD in those with higher levels of SUA was 1.88 (95% CI: 1.76– 2.00). In the subgroup analyses, we found that SUA levels were positively associated with NAFLD in all subgroups, according to study design, study quality, sample size, sex, comparison, age, or country.Conclusion: This meta-analysis shows that increased SUA levels are positively associated with NAFLD. The results suggested that reducing SUA levels can be a potential strategy for the prevention of NAFLD.Registration Number: PROSPERO-CRD42022358431.Keywords: serum uric acid, nonalcoholic fatty liver disease, meta-analysis, updated

Li Wang,1,2 Ruoyan Ying,3 Yidian Liu,1,2 Qin Sun,1– 3 Wei Sha1– 3 1Clinic and Research Center of Tuberculosis, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, People’s Republic of China; 2Department of Tuberculosis, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, People’s Republic of China; 3Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, People’s Republic of ChinaCorrespondence: Wei Sha; Qin Sun, Email shfksw@126.com; sunqinbonjour@163.comBackground: Mycobacterium tuberculosis (MTB) is a global and highly deleterious pathogen that creates an enormous pressure on global public health. Although several effective drugs have been used to treat tuberculosis, the emergence of multidrug-resistant Mycobacterium tuberculosis (MDR-MTB) has further increased the public health burden. The aim of this study was to describe in depth the metabolic changes in clinical isolates of drug-susceptible Mycobacterium tuberculosis (DS-MTB) and MDR-MTB and to provide clues to the mechanisms of drug resistance based on metabolic pathways.Methods: Based on the minimum inhibition concentration (MIC) of multiple anti-tuberculosis drugs, two clinical isolates were selected, one DS-MTB isolate (isoniazid MIC=0.06 mg/L, rifampin MIC=0.25 mg/L) and one MDR-MTB isolate (isoniazid MIC=4 mg/L, rifampin MIC=8 mg/L). Through high-throughput metabolomics, the metabolic profiles of the DS-MTB isolate and the MDR-MTB isolate and their cultured supernatants were revealed.Results: Compared with the DS-MTB isolate, 128 metabolites were significantly altered in the MDR-MTB isolate and 66 metabolites were significantly altered in the cultured supernatant. The differential metabolites were significantly enriched in pyrimidine metabolism, purine metabolism, nicotinate and nicotinamide metabolism, arginine acid metabolism, and phenylalanine metabolism. Furthermore, metabolomics analysis of the bacterial cultured supernatants showed a significant increase in 10 amino acids (L-citrulline, L-glutamic acid, L-aspartic acid, L-norleucine, L-phenylalanine, L-methionine, L-tyrosine, D-tryptophan, valylproline, and D-methionine) and a significant decrease in 2 amino acids (L-lysine and L-arginine) in MDR-MTB isolate.Conclusion: The present study provided a metabolite alteration profile as well as a cultured supernatant metabolite alteration profile of MDR-MTB clinical isolate, providing clues to the potential metabolic pathways and mechanisms of multidrug resistance.Keywords: Mycobacterium tuberculosis, multidrug-resistantMycobacterium tuberculosis, drug-susceptibleMycobacterium tuberculosis, metabolomics, metabolic pathway

JinJian Yao,1,2,&ast; Xiaochun Mao,3,&ast; Qigang Sun,4,&ast; Biao Wu,5 Weiling Yu,6 Yanjing Huang,7 Shuai Luo,1 Jia Zeng,1 Jusheng Lin2 1Department of Emergency, Hainan General Hospital, Haikou, Hainan, People’s Republic of China; 2Institute of Liver and Gastrointestinal Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 3Department of Ophthalmology, Xiangyang Central Hospital Affiliated to Hubei University of Arts and Science, Xiangyang, Hubei, People’s Republic of China; 4Department of Hepatobiliary and Pancreatic Surgery, Hainan General Hospital, Haikou, Hainan, People’s Republic of China; 5Infectious Disease, Hainan General Hospital, Haikou, Hainan, People’s Republic of China; 6Oncology Department, Haikou City People’s Hospital, Haikou, Hainan, People’s Republic of China; 7Oncology Department of Hainan General Hospital, Haikou, Hainan, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: JinJian Yao; Jusheng Lin, Email 114715854@qq.com; jslin2005@163.comPurpose: Liver cirrhosis (LC) and hepatocellular carcinoma (HCC) are progressions affected by genetic predispositions, and persistent hepatitis B virus infection also demonstrates genetic susceptibility. All HBV-related outcomes have been compared in parallel to identify risk polymorphism in HBV progression.Methods: The multiple-stage association study filtered and validated the risk SNPs for HBV progression and explored their association with persistent infection, with a total of 8906 subjects in China from three sites. Cox proportional hazards models and Kaplan–Meier Log rank tests were used to determine the time to the progressive event in relation to the risk SNPs.Results: Rs3825214 in TBX5 replicated a specific association with LC and HCC in 4 progression cohorts and was not related to persistent infection, naivety to HBV infection and natural clearance in 3 persistent cohorts. In combined samples, rs3825214 was associated with an increased risk of LC (P< 0.001; OR = 1.98) and HCC (P< 0.001; OR = 1.68). The results of bioinformatics analysis indicated that rs3825214 genotypes change RNA structure and intron excision ratio. In the follow-up of 571 hospital-based persistent HBV infection patients, ninety-three (16.29%) developed LC, and seventy-four (12.96%) progressed to HCC at a median follow-up of 5.1 years. Rs3825214 was associated with HCC and LC events in Cox proportional hazards models (P< 0.001).Conclusion: We identified and confirmed that genetic variants in TBX5 are significantly associated with susceptibility to and the incidence of LC and HCC.Keywords: TBX5, polymorphism, liver cirrhosis, hepatocellular carcinoma, follow-up, hepatitis B virus

Yuxuan Zhang,1 Qin Sun2 1Department of Social and Behavioural Sciences, City University of Hong Kong, Hong Kong, People’s Republic of China; 2Mental Health Education and Guidance Center, Guangxi International Business Vocational College, Nanning, People’s Republic of ChinaCorrespondence: Qin Sun, Email psysunqin@hotmail.comPurpose: Based on the interpersonal theory of depression and resilience framework theory, this study tested the association between interpersonal sensitivity and depression in college students in the context of the COVID-19 pandemic. Additionally, the mediating role of resilience and the moderating role of the COVID-19 lockdown were investigated.Methods: A total of 5193 South Chinese college students (M=19.27, SD = 1.18) were included in the study. According to which campus they were living on, the subjects were categorized as members of a lockdown group or a nonlockdown group. They completed the interpersonal sensitivity subscales of the Symptom Checklist 90 (SCL-90), Beck Depression Inventory (BDI), and Connor-Davidson Resilience Scale (CD-RISC). SPSS 26.0 statistical software was used to analyse the descriptive statistics, reliability, and correlation. Specifically, a moderated mediation model was analysed by multivariate logistic regression.Results: Interpersonal sensitivity was significantly associated with depression (r = 0.517, p < 0.01), which was mediated by resilience (β = 0.12, 95% CI = [0.10, 0.13]). Lockdown status was shown to have a moderating effect on the relationship between resilience and depression (β = 0.03, t = 2.71, p < 0.01).Conclusion: The high levels of interpersonal sensitivity displayed by South Chinese college students caused low resilience and then facilitated depression. The COVID-19 lockdown strengthened the effect of low resilience on depression. Lower resilience was more strongly associated with higher depression for students under lockdown conditions compared to students who were not in lockdown.Keywords: interpersonal sensitivity, depression, resilience, lockdown, COVID-19

Qingqing Sun,1,&ast; Lixin Shen,1,&ast; Bai-Ling Zhang,2 Jiaoyang Yu,1,3 Fu Wei,1 Yanmei Sun,1 Wei Chen,3,4 Shiwei Wang1 1Key Laboratory of Resources Biology and Biotechnology in Western China, Ministry of Education, Provincial Key Laboratory of Biotechnology of Shaanxi Province, the College of Life Sciences, Northwest University, Xi’an, 710069, People’s Republic of China; 2Department of Laboratory Medicine, The Second Affiliated Hospital of Nanchang University, Nanchang, 330006, People’s Republic of China; 3Clinical Research Center, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, 210003, People’s Republic of China; 4The Clinical Infectious Disease Center of Nanjing, Nanjing, 210003, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Wei Chen; Shiwei Wang, Email njyy039@njucm.edu.cn; wangsw@nwu.edu.cnAbstract: Since bacteriophages (phages) were firstly reported at the beginning of the 20th century, the study on them experiences booming-fading-emerging with discovery and overuse of antibiotics. Although they are the hotspots for therapy of antibiotic-resistant strains nowadays, natural phage applications encounter some challenges such as limited host range and bacterial resistance to phages. Synthetic biology, one of the most dramatic directions in the recent 20-years study of microbiology, has generated numerous methods and tools and has contributed a lot to understanding phage evolution, engineering modification, and controlling phage-bacteria interactions. In order to better modify and apply phages by using synthetic biology techniques in the future, in this review, we comprehensively introduce various strategies on engineering or modification of phage genome and rebooting of recombinant phages, summarize the recent researches and potential directions of phage synthetic biology, and outline the current application of engineered phages in practice.Keywords: phage, synthetic biology, engineering, rebooting, phage therapy, phage application

Ling Chen,1 Qiuwei Sun,1 Junchao Zhang,1 Xizhong Xu,2 Xiaofang Fan1 1Department of Ultrasound Medicine, Affiliated Hospital of Jiangnan University, Wuxi City, Jiangsu Province, People’s Republic of China; 2Department of Gynecology, Affiliated Hospital of Jiangnan University, Wuxi City, Jiangsu Province, People’s Republic of ChinaCorrespondence: Xiaofang Fan, Department of Ultrasound Medicine, Affiliated Hospital of Jiangnan University, No. 200, Huihe Road, Binhu District, Wuxi City, Jiangsu Province, 214062, People’s Republic of China, Tel + 86-13861843829, Email fanxiaolll@126.comBackground: Angiomyofibroblastoma (AMFB) is an uncommon disease with few literature reports, leading to the poor understanding of its diagnosis, treatment, and postoperative follow-up plans among gynecologists.Objective: To study the clinical and pathological features of vulvar AMFB and discuss its treatment and prognosis.Case Summary: The 3 cases were characterized by a gradually increasing painless mass in the vulva. Preoperative diagnosis was difficult and mainly depended on ultrasonic examination. Immunohistochemistry confirmed clear boundaries of AMFB. This condition could be completely cured by surgery, and the prognosis was good.Conclusion: The vulvar AMFB is a rare tumor that is frequently misdiagnosed before surgery. Ultrasound is preferred in auxiliary diagnosis, and surgery remains the best treatment, and long-term follow-up is necessary to avoid recurrence or other complications.Keywords: angiomyofibroblastoma, diagnosis, surgery, follow-up

Chunmei Yan,1 Qiuxia Li,1 Qiang Sun,2 Lu Yang,3 Xing Liu,1 Yuxin Zhao,1 Mingyi Shi,4 Xiaofang Li,1 Kaipei Luo1 1State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, People’s Republic of China; 2Department of Pharmacy, Personalized Drug Therapy Key Laboratory of Sichuan Province, Sichuan Provincial People’s Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu, People’s Republic of China; 3Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, People’s Republic of China; 4School of Intelligent Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, People’s Republic of ChinaCorrespondence: Xiaofang Li; Kaipei Luo, Department of Pharmaceutics of Traditional Chinese Medicine, Chengdu University of Traditional Chinese Medicine, 1166 Liutai Avenue, Chengdu, 611130, People’s Republic of China, Email lixiaofang@cdutcm.edu.cn; luokaipei@126.comAbstract: Malignant tumor, the leading cause of death worldwide, poses a serious threat to human health. For decades, natural product has been proven to be an essential source for novel anticancer drug discovery. Shikonin (SHK), a natural molecule separated from the root of Lithospermum erythrorhizon, shows great potential in anticancer therapy. However, its further clinical application is significantly restricted by poor bioavailability, adverse effects, and non-selective toxicity. With the development of nanotechnology, nano drug delivery systems have emerged as promising strategies to improve bioavailability and enhance the therapeutic efficacy of drugs. To overcome the shortcoming of SHK, various nano drug delivery systems such as liposomes, polymeric micelles, nanoparticles, nanogels, and nanoemulsions, were developed to achieve efficient delivery for enhanced antitumor effects. Herein, this review summarizes the anticancer pharmacological activities and pharmacokinetics of SHK. Additionally, the latest progress of SHK nanomedicines in cancer therapy is outlined, focusing on long circulation, tumor targeting ability, tumor microenvironment responsive drug release, and nanosystem-mediated combination therapy. Finally, the challenges and prospects of SHK nanomedicines in the future clinical application are spotlighted.Graphical Abstract: Keywords: shikonin, cancer therapy, pharmacokinetics, nanomedicine, drug delivery

Pinger Wang,1– 3,&ast; Jianbo Xu,1,&ast; Qi Sun,4,&ast; Qinwen Ge,2,3 Min Qiu,1 Kaiao Zou,2,3 Jun Ying,3,5 Wenhua Yuan,2,3 Jiali Chen,2,3 Qinghe Zeng,2,3 Qi Cui,1 Hongting Jin,2,3 Chunchun Zhang,1 Fanzhu Li1 1College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 2Institute of Orthopaedics and Traumatology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 3The First College of Clinical Medicine, Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China; 4Department of Orthopedic Joint Surgery, Hangzhou Fuyang Hospital of TCM Orthopaedics and Traumatology, Hangzhou, People’s Republic of China; 5Department of Orthopedic Surgery, the First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Chunchun Zhang, College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310053, People’s Republic of China, Tel/Fax +86 571-86613684, Email 20081026@zcmu.edu.cn Fanzhu Li, College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310053, People’s Republic of China, Tel/Fax +86 571-86613684, Email lifanzhu@zcmu.edu.cnBackground: Knee osteoarthritis (KOA) is the primary prevalent disabling joint disorder among osteoarthritis (OA), and there is no particularly effective treatment at the clinic. Traditional Chinese medicine (TCM) herbs, such as Eucommia ulmoides Oliv. and Glycyrrhiza uralensis Fisch. (E.G.) couplet medicines, have been reported to exhibit beneficial health effects on KOA, exact mechanism of E.G. nevertheless is not fully elucidated.Purpose: We assess the therapeutic effects of E.G. on KOA and explore its underlying molecular mechanism.Methods: UPLC-Q-TOF/MS technique was used to analyze the active chemical constituents of E.G. The destabilization of the medial meniscus model (DMM) was employed to evaluate the chondroprotective action of E.G. in KOA mice using histomorphometry, μCT, behavioral testing and immunohistochemical staining. Additionally, network pharmacology and molecular docking were used to predict potential targets for anti-KOA activities of E.G., which was further verified through in vitro experiments.Results: In vivo studies have shown that E.G. could significantly ameliorate DMM-induced KOA phenotypes including subchondral bone sclerosis, cartilage degradation, gait abnormality and thermal pain reaction sensibility. E.G. treatment could also promote extracellular matrix synthesis to protect articular chondrocytes, which was indicated by Col2 and Aggrecan expressions, as well as reducing matrix degradation by inhibiting MMP13 expression. Interestingly, network pharmacologic analysis showed that PPARG might be a therapeutic center. Further study proved that E.G.-containing serum (EGS) could up-regulate PPARG mRNA level in IL-1β-induced chondrocytes. Notably, significant effects of EGS on the increment of anabolic gene expressions (Col2, Aggrecan) and the decrement of catabolic gene expressions (MMP13, Adamts5) in KOA chondrocytes were abolished due to the silence of PPARG.Conclusion: E.G. played a chondroprotective role in anti-KOA by inhibiting extracellular matrix degradation, which might be related to PPARG.Graphical Abstract: Keywords: couplet medicines, UPLC-Q-TOF/MS, network pharmacology, cartilage degeneration, knee osteoarthritis, PPARG

SuWen Yang,1– 3,&ast; Bing Wang,2,4,5,&ast; Jing Li,6 Xue Zhao,7 Yan Zhu,8 Qian Sun,9 HongMei Liu,2,4,5 XiaoJun Wen1 1School of Basic Medical Sciences, Basic Medical School, Guizhou Medical University, Guiyang, 550025, People’s Republic of China; 2Engineering Research Centre of Medical Biotechnology, Guizhou Medical University, Guiyang, Guizhou, 550025, People’s Republic of China; 3People’s Hospital of Kaiyang, Guiyang, 550300, People’s Republic of China; 4Key Laboratory of Biology and Medical Engineering, Guizhou Medical University, Guiyang, Guizhou, 550025, People’s Republic of China; 5School of Biology and Engineering, Guizhou Medical University, Guiyang, Guizhou, 550025, People’s Republic of China; 6Department of Microbial Immunology, The First Affiliated Hospital of Guizhou Medical University, Guiyang, 550004, People’s Republic of China; 7Department of Clinical Laboratory, The First People’s Hospital of Guiyang, Guiyang, 550002, People’s Republic of China; 8Department of Clinical Laboratory, The Fourth People’s Hospital of Guiyang, Guiyang, 550002, People’s Republic of China; 9Department of Emergency Medicine, The First People’s Hospital of Guiyang, Guiyang, 550002, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: XiaoJun Wen; HongMei Liu, Guizhou Medical University, Guiyang, 550025, People’s Republic of China, Email 2547285543@qq.com; hmliu@gmc.edu.cnPurpose: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common pathogens of community- and hospital-acquired infections, and its prevalence is increasing globally. Guiyang is the capital city of Guizhou Province, Southwest China; as the transport and tourism centre of Southwest China, Guizhou Province is bordered by Yunnan, Sichuan, Chongqing, and Guangxi Provinces. Although MRSA prevalence is increasing, little is known about its aspects in the area. The purpose of this study was to analyse MRSA molecular characteristics, antimicrobial resistance, and virulence genes in Guiyang.Methods: In total, 209 MRSA isolates from four hospitals (2019– 2020) were collected and analysed by antimicrobial susceptibility testing and molecular classification by the MLST, spa, and SCCmec typing methods. Isolate antibiotic resistance rates were detected by a drug susceptibility assays. PCR amplification was used to detect the virulence gene-carrying status.Results: Twenty-four STs, including 4 new STs (ST7346, ST7347, ST7348, and ST7247) and 3 new allelic mutations, were identified based on MLST. The major prevalent ST type and clone complex were ST59 (49.8%) and CC59 (62.7%), respectively. Spa type t437 (42.1%) and SCCmec IV (55.5%) were identified by spa and SCCmec typing methods as the most important types. Drug sensitivity data showed that the multidrug resistance rate was 79.0%. There were significant differences in multidrug resistance rates and virulence gene-carrying rates for seb, hla, hlb, cna and bap between ST59 and non-ST59 types.Conclusion: ST59-SCCmecIV-t437 is a major epidemic clone in Guiyang that should be monitored by local medical and health institutions. The situation differs from other adjacent or middle provinces of China, which may be due to the special geographical location of the region and the trend in antibiotic use or lifestyle. This study provides empirical evidence for local medical and health departments to prevent and control the spread of MRSA.Keywords: methicillin-resistant Staphylococcus aureus, MRSA, multilocus sequence type, MLST, Staphylococcal protein A type, SPA, Staphylococcal chromosomal cassette mec type, SCCmec, multidrug-resistant organism, virulence gene

Qiuxue Sun,1,&ast; Hailian Xu,2,&ast; Wenyue Zhang,3 Yuqiu Zhou,1 Yumei Lv1 1School of Nursing, Harbin Medical University (Daqing), Daqing, People’s Republic of China; 2School of Medical, Quzhou College of Technology, Quzhou, People’s Republic of China; 3Department of Rehabilitation, People’s Hospital of Daqing, Daqing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yumei Lv, School of Nursing, Harbin Medical University (Daqing), No. 39 Xinyang Road, Longfeng District, Daqing, People’s Republic of China, Email lymktz@163.comBackground: Subthreshold depression (SD) is known to be a major risk factor for the development of post-stroke depression (PSD). Appropriate intervention to prevent the transition from SD to PSD is thus imperative. As a form of short-term psychotherapy adapted for individuals with cognitive and communication impairments, behavioral activation therapy (BAT) may be a suitable choice. However, the effects of BAT on psychological outcomes in stroke patients with SD has not been established. This study investigated the feasibility and effectiveness of BAT in reducing the development of depressive symptoms in this patient population.Methods: A double-blind, randomized controlled trial was performed. Seventy participants were randomized to either a BAT group (n=35) or a control group (n=35). Participants in the BAT group received a six-week BAT intervention. The feasibility of BAT was assessed by the number of sessions attended by participants, and the acceptability of BAT to participants and the incidence of adverse events were recorded. The primary clinical outcome measure was the Center for Epidemiological Studies Depression Scale (CES-D) and the 17-item Hamilton Depression Scale (HAMD-17) at baseline, six weeks, and three months after the group allocation. Secondary outcomes included behavioral activation and the incidence of depression.Results: The intervention was feasible and acceptable, with 94.3% of participants in the BAT group (33 of 35) attending at least five sessions. No adverse events were reported in either group. Compared with the control group, the BAT group showed significant improvements in the CES-D (F=67.689, P< 0.05), HAMD-17 (F=4.170, P< 0.05), and behavioral activation (F=25.355, P< 0.05) scores after intervention, and these differences were maintained at the three-month assessment.Conclusion: BAT appears to be feasible and efficacious for reducing depressive symptoms and increasing behavioral activation among stroke patients with SD. The findings of this study may contribute to the primary prevention of PSD.Keywords: stroke, subthreshold depression, post-stroke depression, behavioral activation therapy

Qixian Sun,1 Xinrong Tao,1– 4 Bing Li,1 Hangbing Cao,1 Haoming Chen,1 Yuanjie Zou,1 Huihui Tao,1– 4 Min Mu,1– 4 Wenyang Wang,1 Keyi Xu1– 4 1Center for Medical Research, Medical School, Anhui University of Science and Technology, Huainan, People’s Republic of China; 2Key Laboratory of Industrial Dust Control and Occupational Health, Ministry of Education, Anhui University of Science and Technology, Huainan, People’s Republic of China; 3Key Laboratory of Industrial Dust Deep Reduction and Occupational Health and Safety, Anhui Higher Education Institutes, Anhui University of Science and Technology, Huainan, People’s Republic of China; 4Engineering Laboratory of Occupational Safety and Health, Anhui Province, Anhui University of Science and Technology, Huainan, People’s Republic of ChinaCorrespondence: Xinrong Tao, Medical School, Anhui University of Science and Technology, Huainan, People’s Republic of China, Email xrtao1116@hotmail.comBackground: Silicosis is a severe pulmonary disease caused by inhaling dust containing crystalline silica. The progression of silicosis to pulmonary fibrosis is usually unavoidable. Recent studies have revealed positivity for the overexpression of C-X-C chemokine receptor type 4 (CXCR4) in pulmonary fibrosis and shown that the CXCR4 inhibitor AMD3100 attenuated pulmonary fibrosis after bleomycin challenge and paraquat exposure. However, it is unclear whether AMD3100 reduces crystalline silica-induced pulmonary fibrosis.Methods: C57BL/6 male mice were instilled intranasally with a single dose of crystalline silica (12 mg/60 μL) to establish an acute silicosis mouse model. Twelve hours later, the mice were injected intraperitoneally with 5 mg/kg AMD3100 or control solution. Then, the mice were weighed daily and sacrificed on day 7, 14, or 28 to collect lung tissue and peripheral blood. Western blotting was also applied to determine the level of CXCR4, while different histological techniques were used to assess pulmonary inflammation and fibrosis. In addition, the level of B cells in peripheral blood was measured by flow cytometry.Results: CXCR4 and its ligand CXCL12 were upregulated in the lung tissues of crystalline silica-exposed mice. Blocking CXCR4 with AMD3100 suppressed the upregulation of CXCR4/CXCL12, reduced the severity of lung injury, and prevented weight loss. It also inhibited neutrophil infiltration at inflammatory sites and neutrophil extracellular trap formation, as well as reduced B-lymphocyte aggregates in the lung. Additionally, it decreased the recruitment of circulating fibrocytes (CD45+collagen I+CXCR4+) to the lung and the deposition of collagen I and α-smooth muscle actin in lung tissue. AMD3100 also increased the level of B cells in peripheral blood, preventing circulating B cells from migrating to the injured lungs.Conclusion: Blocking CXCR4 with AMD3100 delays pulmonary inflammation and fibrosis in a silicosis mouse model, suggesting the potential of AMD3100 as a drug for treating silicosis.Keywords: CXCR4, silicosis, pulmonary fibrosis, inflammation, AMD3100, mice

Zhichun Zhang,1 Yixuan Han,2 Qiuyue Sun,1 Yipeng Wang,3 Lichao Sun4 1Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, People’s Republic of China; 2Department of Rheumatology and Immunology, Affiliated Kailuan General Hospital of North China University of Science and Technology, Tangshan, People’s Republic of China; 3Department of Breast Surgery, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, People’s Republic of China; 4State Key Laboratory of Molecular Oncology, National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, People’s Republic of ChinaCorrespondence: Lichao Sun; Yipeng Wang, National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100021, People’s Republic of China, Tel/Fax +86 10-67781331, Email sunlichao@cicams.ac.cn; yidoctor99@126.comBackground: DPY30 is a common subunit of the human SET1/MLL complex and is an essential protein required for the activity of SET1/MLL methyltransferase. DPY30 regulates the histone H3K4 modification, and dysfunction of DPY30 might contribute to the regulation of cancer immune evasion. However, the functions and regulation of DPY30 in the expression of programmed cell death ligand 1 (PD-L1) is still not completely explored.Methods: Various online databases were used for data processing and visualization, including UALCAN, Oncomine, cBioPortal, SangerBox, TISIDB, TIMER, and GEPIA databases. The expression of DPY30 and PD-L1 in melanoma tissues were evaluated by IHC. Chromatin Immunoprecipitation (ChIP), RT-PCR and flow cytometry were used to elucidate the underlying molecular mechanism of PD-L1 expression regulation and its function.Results: The mRNA level of DPY30 in melanoma was higher than in normal tissues. The expression of DPY30 was positively associated with TMB, neoantigens and PD-L1 expression. Furthermore, DPY30 expression showed significant positive correlations with immune suppressor cells and ICP genes involved in T-cell exhaustion. IHC showed that the positive rates of DPY30 and PD-L1 in melanoma tissues were 62% and 58%, respectively. Correlation analysis revealed that DPY30 over-expression was positively associated with PD-L1 expression. Silencing of DPY30 by specific siRNA significantly inhibited PD-L1 expression. ChIP analysis revealed that H3K4me3 levels were enriched in the proximal PD-L1 promoter region in tumor cells. Inhibition of DPY30 still suppressed the PD-L1 level in IFN-γ treated MMAC-SF cells. Furthermore, the apoptosis of PD1+ T-cells in co-culture with MMAC-SF cells by knockdown of DPY30 were markedly reduced.Conclusion: This study shows the roles of DPY30 in regulating the cancer immune evasion in melanoma. Targeting the DPY30-H3K4me3 axis might be an alternative approach to enhance the efficacy of checkpoint immunotherapy.Keywords: DPY-30, H3K4me3, PD-L1, melanoma, immunotherapy