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Sujing Li,1,* Debiao Kong,1,* Hong Gao2 1The Third Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, People’s Republic of China; 2Department of Acupuncture and Moxibustion, The Third Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, People’s Republic of China*These authors contributed equally to this workCorrespondence: Hong Gao, Department of Acupuncture and Moxibustion, The Third Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, People’s Republic of China, Email qtgh@vip.qq.com

Zhanlin Liao,1,* Liangzhi Huang,1,* Jun Chen,2 Ting Chen,1 Dezhi Kong,1 Qifeng Wei,1 Qiao Chen,1 Bin Deng,1 Yanyan Li,1 Shuai Zhong,1 Zugui Huang1 1Department of Endocrine, Affiliated Nanping First Hospital of Fujian Medical University, Nanping, Fujian, 353006, People’s Republic of China; 2Department of Ophthalmology, Affiliated Nanping First Hospital of Fujian Medical University, Nanping, Fujian, 353006, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zugui Huang, Department of Endocrine, Affiliated Nanping First Hospital of Fujian Medical University, No. 317 Zhongshan Road, Nanping, Fujian, 353006, People’s Republic of China, Email 15159969783@163.comBackground: Type 2 diabetes (T2DM) combined nonalcoholic fatty liver disease (NAFLD) are characterized by metabolic disruptions. Liraglutide has been proved to be effective in T2DM. If LRG could regulate NAFLD combined T2DM has not been reported.Methods: Intraperitoneal injection of 1% streptozotocin (STZ) plus high-sugar and high-fat diet was used to induce NAFLD combined T2DM animal model. Palmitic acid (200 μmol/L) and glucose (25 mmol/L) incubation were used to induce cell model. The cell apoptosis, mRNA and protein expression were measured through flow cytometry, PCR, and Western blotting, respectively.Results: Liraglutide significantly improved the liver injury of NAFLD combined T2DM rats, but Com-C reversed the effect of liraglutide. The decreased AMPK/mTOR signaling pathway in the NAFLD combined T2DM animals was greatly activated by liraglutide. Com-C reversed the protection effects of liraglutide on palmitic acid+glucose induced cell damage.Conclusion: Liraglutide could greatly alleviate the damage caused by NAFLD+T2DM and palmitic acid+glucose. The protection effects of liraglutide were greatly inhibited by suppressing AMPK/mTOR signaling pathway. This research might provide a novel therapeutic strategy for the prevention and treatment of NAFLD combined T2DM disease.Keywords: type 2 diabetes, nonalcoholic fatty liver disease, liraglutide, AMPK/mTOR, autophagy

Dunmin She,1,2,&ast; Wei Xu,3,&ast; Jing Liu,4,&ast; Zikai Zhang,5 Ping Fang,3 Ran Li,3 Dehong Kong,3 Miao Xuan,3 Qi Liu,3 Mo Yuan Pan,6 Yan Wang,1,2 Ying Xue3 1Clinical Medical College, Yangzhou University, Yangzhou, Jiangsu, People’s Republic of China; 2Department of Endocrinology, Northern Jiangsu People’s Hospital Affiliated to Yangzhou University, Yangzhou, Jiangsu, People’s Republic of China; 3Department of Endocrinology and Metabolism, Tongji Hospital, School of Medicine, Tongji University, Shanghai, People’s Republic of China; 4Dalian Medical University, Dalian, Liaoning, People’s Republic of China; 5Department of Oncology, Tongji Hospital, School of Medicine, Tongji University, Shanghai, People’s Republic of China; 6The Second Affiliated Hospital of Qiqihar Medical University, Qiqihar, Heilongjiang, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ying Xue, Department of Endocrinology and Metabolism, Tongji Hospital, School of Medicine, Tongji University, No. 389, Xincun Road, Shanghai, People’s Republic of China, Tel +86-21-66111061, Email doxy0622@163.com Yan Wang, Clinical Medical College, Yangzhou University, No. 98, Nantong West Road, Yangzhou, Jiangsu Province, People’s Republic of China, Email yzyan1972@126.comPurpose: Metabolic syndrome (MetS) is a rising global concern with an increasing prevalence. This study aimed to evaluate the relationship between serum uric acid to creatinine ratio (SUA/Cr) and MetS in adults with overweight/obesity in China.Patients and Methods: We conducted a cross-sectional study comprising 4699 participants with overweight/obesity who underwent physical examinations. Their serum levels of various components, including total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-c), low-density lipoprotein cholesterol (LDL-c), fasting plasma glucose (FPG), creatinine (Cr), and uric acid (UA) were measured. Renal function-normalized SUA was calculated using SUA/Cr. Logistic regression analysis was employed to investigate the association between SUA/Cr and MetS in adults with overweight/obesity.Results: SUA/Cr levels were lower in non-MetS participants (OR: 2.159, 95% CI: 1.82 to 2.56; p < 0.001), and tended to rise with the increasing number of MetS components. Additionally, elevated SUA/Cr levels were associated with a higher risk of hypertension, hyperglycemia, and dyslipidemia.Conclusion: SUA/Cr levels were significantly associated with MetS and its components in Chinese adults with overweight/obesity.Keywords: metabolic syndrome, serum uric acid to creatinine ratio, overweight/obesity

Wei Xu,1,&ast; Zikai Zhang,2,&ast; Kerong Hu,1,&ast; Ping Fang,1 Ran Li,1 Dehong Kong,1 Miao Xuan,1 Yang Yue,3 Dunmin She,4,5 Ying Xue1 1Department of Endocrinology and Metabolism, Tongji Hospital, School of Medicine, Tongji University, Shanghai, People’s Republic of China; 2Department of Oncology, Tongji Hospital, School of Medicine, Tongji University, Shanghai, People’s Republic of China; 3School of Mathematics and Statistics, University of Melbourne, Melbourne, Australia; 4Clinical Medical College, Yangzhou University, Yangzhou, Jiangsu, People’s Republic of China; 5Department of Endocrinology, Northern Jiangsu People’s Hospital Affiliated to Yangzhou University, Yangzhou, Jiangsu, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Ying Xue, Department of Endocrinology and Metabolism, Tongji Hospital, School of Medicine, Tongji University, No. 389, Xincun Road, Shanghai, People’s Republic of China, Tel +86-21-66111061, Email doxy0622@163.com Dunmin She, Clinical Medical College, Yangzhou University, No. 98, Nantong West Road, Yangzhou, Jiangsu, People’s Republic of China, Email sdm1979@126.comPurpose: The objective of this study was to employ machine learning (ML) models utilizing non-invasive factors to achieve early and low-cost identification of MetS in a large physical examination population.Patients and Methods: The study enrolled 9171 participants who underwent physical examinations at Northern Jiangsu People’s Hospital in 2009 and 2019, to determine MetS based on criteria established by the Chinese Diabetes Society. Non-invasive characteristics such as gender, age, body mass index (BMI), systolic blood pressure (SBP), and diastolic blood pressure (DBP) were collected and used as input variables to train and evaluate ML models for MetS identification. Several ML models were used for MetS identification, including logistic regression (LR), k-nearest neighbors algorithm (k-NN), naive bayesian (NB), decision tree (DT), random forest (RF), artificial neural network (ANN), and support vector machine (SVM).Results: Our ML models all showed good performance in the 10-fold cross-validation except for the SVM model. In the external validation, the NB model exhibited the best performance with an AUC of 0.976, accuracy of 0.923, sensitivity of 98.32%, and specificity of 91.32%.Conclusion: This study proposed a new non-invasive method for early and low-cost identification of MetS by using ML models. This approach has the potential to serve as a highly sensitive, convenient, and cost-effective tool for large-scale MetS screening.Keywords: metabolic syndrome, machine learning methods, non-invasive method, naive Bayesian

Di Kong, Danqi Deng Department of Dermatology, Second Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, 650101, People’s Republic of ChinaCorrespondence: Danqi Deng, Fax +86-871-65351214, Email danqid128@sina.comAbstract: We report a case of a patient initially diagnosed with keloid and eventually diagnosed by skin histopathology and immunohistochemistry with primary cutaneous ALK-negative anaplastic large cell lymphoma.Keywords: primary cutaneous ALK-negative anaplastic large cell lymphoma, keloid, misdiagnosis

Delian Kong,1,&ast; Junrong Li,1,&ast; Yingying Lv,1,&ast; Man Wang,2,&ast; Shenghua Li,1 Baoxin Qian,3 Yusheng Yu2 1Department of Neurology, The Affiliated Jiangning Hospital with Nanjing Medical University, Nanjing, Jiangsu, 211100, People’s Republic of China; 2Department of Radiology, The Affiliated Jiangning Hospital with Nanjing Medical University, Nanjing, Jiangsu, 211100, People’s Republic of China; 3Huiying Medical Technology (Beijing); Huiying Medical Technology Co., Ltd, Beijing City, 100192, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Delian Kong; Yusheng Yu, Email xykdl@163.com; yayiba2063@163.comObjective: To develop a radiomics nomogram model based on time-of-flight magnetic resonance angiography (TOF-MRA) images for preoperative prediction of true microaneurysms.Methods: 118 patients with Intracranial Aneurysm Sac (40 positive and 78 negative) were enrolled and allocated to training and validation groups (8:2 ratio). Findings of clinical characteristics and MRA features were analyzed. A radiomics signature was built on the basis of reproducible features by using the least absolute shrinkage and selection operator (LASSO) regression algorithm in the training group. The radiomics nomogram model was constructed by combining clinical risk factors and radiomics signature. In order to compare the classification performance of clinical models, radiomics model and radiomics nomogram model, AUC was used to evaluate them. The performance of the radiomics nomogram model was evaluated by calibration curve and decision curve analysis.Results: Eleven features were selected to develop radiomics model with AUC of 0.875 (95% CI 0.78– 0.97), sensitivity of 0.84, and specificity of 0.68. The radiomics model achieved a better diagnostic performance than the clinic model (AUC = 0.75, 95% CI: 0.53– 0.97) and even radiologists. The radiomics nomogram model, which combines radiomics signature and clinical risk factors, is effective too (AUC = 0.913, 95% CI: 0.87– 0.96). Furthermore, the decision curve analysis demonstrated significantly better net benefit in the radiomics nomogram model.Conclusion: Radiomics features derived from TOF-MRA can reliably be used to build a radiomics nomogram model for effectively differentiating between pseudo microaneurysms and true microaneurysms, and it can provide an objective basis for the selection of clinical treatment plans.Keywords: machine learning, radiomics, microaneurysms, nomogram

Ting Chen,1,2,&ast; Lei Zhang,3,&ast; Wenhua Huang,3,&ast; Huijun Zong,1,2 Qian Li,3 Yuling Zheng,3 Qingyu Lv,3 Decong Kong,3 Yuhao Ren,3 Yongqiang Jiang,3 Yan Li,1,2 Peng Liu3 1The PLA 307 Clinical College of Anhui Medical University, The Fifth Clinical Medical College of Anhui Medical University, Hefei, People’s Republic of China; 2Department of Critical Care Medicine, The Fifth Medical Center, Chinese PLA General Hospital, Beijing, 100071, People’s Republic of China; 3State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yan Li, The PLA 307 Clinical College of Anhui Medical University, The Fifth Clinical Medical College of Anhui Medical University, Hefei, 230032, People’s Republic of China, Email liyanmdhuxi@163.com Peng Liu, Beijing Institute of Microbiology and Epidemiology, Dongdajie Road 20, Beijing, 100071, People’s Republic of China, Tel +86-010-66948487, Email ammsliupeng@163.comBackground: The early identification of pathogens and their antibiotic resistance are essential for the management and treatment of patients affected by ventilator-associated pneumonia (VAP). However, microbiological culture may be time-consuming and has a limited culturability of many potential pathogens. In this study, we developed a rapid nanopore-based metagenomic next-generation sequencing (mNGS) diagnostic assay for detection of VAP pathogens and antimicrobial resistance genes (ARGs).Patients and Methods: Endotracheal aspirate (ETA) samples from 63 patients with suspected VAP were collected between November 2021 and July 2022. Receiver operating characteristic (ROC) curves were established to compare the pathogen identification performance of the target pathogen reads, reads percent of microbes (RPM) and relative abundance (RA). The evaluation of the accuracy of mNGS was performed comparing with the gold standard and the composite standard, respectively. Then, the ARGs were analyzed by mNGS.Results: ROC curves showed that RA has the highest diagnostic value and the corresponding threshold was 9.93%. The sensitivity and specificity of mNGS test were 91.3% and 78.3%, respectively, based on the gold standard, while the sensitivity and specificity of mNGS test were 97.4% and 100%, respectively, based on the composite standard. A total of 13 patients were virus-positive based on mNGS results, while the coinfection rate increased from 27% to 46% compared to the rate obtained based on clinical findings. The mNGS test also performed well at predicting antimicrobial resistance phenotypes. Patients with a late-onset VAP had a significantly greater proportion of ARGs in their respiratory microbiome compared to those with early-onset VAP (P = 0.041). Moreover, the median turnaround time of mNGS was 4.43 h, while routine culture was 72.00 h.Conclusion: In this study, we developed a workflow that can accurately detect VAP pathogens and enable prediction of antimicrobial resistance phenotypes within 5 h of sample receipt by mNGS.Keywords: mechanical ventilation, endotracheal aspirate, pathogen diagnosis, antibiotic resistance, NGS

Ran Li,1 Zhengqin Ye,1 Dunmin She,2,3 Ping Fang,1 Guannan Zong,1 Kerong Hu,1 Dehong Kong,1 Wei Xu,1 Ling Li,1 Yun Zhou,1 Keqin Zhang,1 Ying Xue1 1Department of Endocrinology and Metabolism, Tongji Hospital, School of Medicine, Tongji University, Shanghai, People’s Republic of China; 2Clinical Medical College, Yangzhou University, Yangzhou, People’s Republic of China; 3Department of Endocrinology, Northern Jiangsu People’s Hospital Affiliated to Yangzhou University, Yangzhou, People’s Republic of ChinaCorrespondence: Ying Xue; Keqin Zhang, Department of Endocrinology and Metabolism, Tongji Hospital, School of Medicine, Tongji University, No. 389, Xincun Road, Shanghai, 200065, People’s Republic of China, Tel +86-021-66111061, Email doxy0622@163.com; keqzhang2018@163.comObjective: Although the pathogenesis of non-alcoholic fatty liver disease (NAFLD) has been extensively studied, the role of its underlying pathogenesis remains unclear, and there is currently no approved therapeutic strategy for NAFLD. The purpose of this study was to observe the beneficial effects of Semaglutide on NAFLD in vivo and in vitro, as well as its potential molecular mechanisms.Methods: Semaglutide was used to treat type 2 diabetes mellitus (T2DM) combined with NAFLD mice for 12 weeks. Hepatic function and structure were evaluated by liver function, blood lipids, liver lipids, H&E staining, oil red staining and Sirius staining. The expression of α/β hydrolase domain-6 (ABHD6) was measured by qPCR and Western blotting in vivo and in vitro. Then, dual-luciferase reporter assay was performed to verify the regulation of the upstream miR-5120 on ABHD6.Results: Our data revealed that Semaglutide administration significantly improved liver function and hepatic steatosis in T2DM combined with NAFLD mice. Furthermore, compared with controls, up-regulation of ABHD6 and down-regulation of miR-5120 were found in the liver of T2DM+NAFLD mice and HG+FFA-stimulated Hepa 1– 6 hepatocytes. Interestingly, after Semaglutide intervention, ABHD6 expression was significantly decreased in the liver of T2DM+NAFLD mice and in HG+FFA-stimulated Hepa 1– 6 hepatocytes, while miR-5120 expression was increased. We also found that miR-5120 could regulate the expression of ABHD6 in hepatocytes, while Semaglutide could modulate the expression of ABHD6 through miR-5120. In addition, GLP-1R was widely expressed in mouse liver tissues and Hepa 1– 6 cells. Semaglutide could regulate miR-5120/ABHD6 expression through GLP-1R.Conclusion: Our data revealed the underlying mechanism by which Semaglutide improves hepatic steatosis in T2DM+NAFLD, and might shed new light on the pathological role of miR-5120/ABHD6 in the pathogenesis of T2DM+NAFLD.Keywords: Semaglutide, non-alcoholic fatty liver disease, type 2 diabetes mellitus, α/β hydrolase domain-6, microRNA-5120

Jinli Xue,1 Mengting Zhu,2 Yanan Guo,3 Demin Kong4 1Research Center for Energy Economics, School of Business Administration, Henan Polytechnic University, Jiaozuo, People’s Republic of China; 2School of Management, Shanghai University, Shanghai, People’s Republic of China; 3School of Financial Management, Zhejiang Financial College, Hangzhou, People’s Republic of China; 4School of Humanities, Shanghai University of Finance and Economics, Shanghai, People’s Republic of ChinaCorrespondence: Mengting Zhu, School of Management, Shanghai University, Shanghai, People’s Republic of China, Tel +86-18682641363, Email jocelemy@163.comPurpose: According to the traditional view, “underdog expectations” induce self-doubt in employees, resulting in negative effects. However, a new study suggests that underdog expectations may encourage employees to work harder, resulting in positive effects. Based on regulatory focus theory, this study constructed a moderated double-mediation model to explain the “double-edged sword effect” of underdog expectations.Subjects and Methods: A three-wave survey method and leader–employee pairing method were used to conduct a questionnaire survey among 346 employees drawn from five enterprises in Shanghai, China. Statistical analysis methods, including hierarchical regression analysis, simple slope analysis, and difference analysis, were used for data analysis. SPSS 24.0, Amoss 24.0, and Mplus 7.4 software were employed to test four proposed hypotheses.Results: Under the positive moderating effect of prevention focus, underdog expectations reduce employee work engagement by adopting an avoidance path of employee feedback-avoiding behaviors(β = 0.090, p < 0.01). Moreover, underdog expectations play a negative role in this situation . Under the positive moderating effect of promotion focus, underdog expectations improve employee work engagement by adopting an approach of proving others wrong (β = 0.189, p < 0.001). Moreover, underdog expectations play a positive role in this situation.Conclusion: The study results refined the double-sided effects of underdog expectations on employee work engagement and provided theoretical and practical implications for managers on how to motivate employees with underdog expectations and how to better convey expectations to subordinates.Keywords: underdog expectations, regulatory focus theory, prove others wrong, feedback-avoiding behaviors

Jie Jia,1,2 Ji-Qun Yang,3 Ying-Rong Du,3 Yu Xu,2 Deshenyue Kong,1,2 Xiu-Ling Zhang,3 Jun-Hong Mao,1,2 Gui-Fang Hu,3 Kun-Hua Wang,1,2,4 Yi-Qun Kuang1,2 1NHC Key Laboratory of Drug Addiction Medicine, First Affiliated Hospital of Kunming Medical University, Kunming Medical University, Kunming, 650032, People’s Republic of China; 2Scientific Research Laboratory Center, First Affiliated Hospital of Kunming Medical University, Kunming, 650032, People’s Republic of China; 3Third People’s Hospital of Kunming City/Drug Rehabilitation Hospital of Kunming City, Kunming, 650041, People’s Republic of China; 4School of Medicine, Yunnan University, Kunming, 650500, People’s Republic of ChinaCorrespondence: Yi-Qun Kuang; Kun-Hua Wang, NHC Key Laboratory of Drug Addiction Medicine, First Affiliated Hospital of Kunming Medical University, Kunming Medical University, Kunming, 650032, People’s Republic of China, Email yq610433@hotmail.com; wangkunhua1964@126.comBackground: Hypertension is a common complication in injection drug users (IDU), especially a high proportion of resistant hypertension occurs among them. However, the involving mechanisms remain largely unknown.Methods: We here investigated the key signaling moieties in resistant hypertension in drug users. Analyses were performed with high-throughput transcriptomic sequencing data of peripheral blood from individuals with drug-sensitive hypertension (Ctrl-DS), IDU with resistant hypertension (IDU-DR), and IDU with sensitive hypertension (IDU-DS).Results: We showed that 17 and 1 genes in IDU-DS, 48 and 4 genes in IDU-DR were upregulated and downregulated compared Ctrl-DS, and 2 and 4 genes were upregulated and downregulated in IDU-DR compared with IDU-DS, respectively (p ≤ 0.01 and |log2(FC)| ≥ 1). Differentially expressed genes (DEGs) between Ctrl-DS and IDU-DS were mainly involved in Gene ontology terms of immunoglobulin complex and blood microparticle. DEGs between IDU-DS and IDU-DR were mainly involved in immune system process and immunoglobulin complex. DEGs between Ctrl-DS and IDU-DR were mainly involved in immunoglobulin complex, blood microparticle and cytoplasmic vesicle lumen. We identified 2 gene clusters (brown modules, MEbrown; turquoise module, MEturquoise) correlated with IDU-DR and a gene cluster (magenta module, MEmagenta) correlated with IDU-DS by weighted gene co-expression network analysis (WGCNA). Functional analysis demonstrated that pathways of focal adhesion and focalin-1-rich granule lumen were involved in the development of IDU-DR, and the cytosolic large ribosomal subunit may relate to IDU-DR. Further, immune cell infiltration analysis demonstrated that the abundance of dendritic cells (DCs), natural Treg cells (nTreg), and exhausted T cells (Tex) in IDU-DR and IDU-DS, naïve CD8+ T cells in IDU-DS was significantly different compared with that in Ctrl-DS. The abundance of cytotoxic T cells (Tc) was significantly different between IDU-DS and IDU-DR.Conclusion: Our findings indicated a potential function of immunoregulation mechanisms for resistant hypertension.Keywords: drug use, hypertension, treatment-resistant, transcriptome, immunoregulation

Leifang Zhang,1 Deyi Kong,1 Junxia Huang,1 Qiongfen Wang,2 Lilin Shao1 1Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, School of Food and Pharmacy, Zhejiang Ocean University, Zhoushan, 316022, People’s Republic of China; 2Zhoushan Institute of Calibration and Testing for Quality and Technical Supervision, Zhoushan, 316012, Zhejiang, People’s Republic of ChinaCorrespondence: Leifang Zhang, Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, School of Food and Pharmacy, Zhejiang Ocean University, 1 Haida South Road, Changzhi Island, Zhoushan, 316022, Zhejiang, People’s Republic of China, Tel +86 18 8570 63534, Fax +86 58 0 254 781, Email zhanglf@zjou.edu.cnBackground: Seawater drowning-induced acute lung injury (ALI) is a severe clinical condition characterized by increased alveolar-capillary permeability, excessive inflammatory response, and refractory hypoxemia. C-phycocyanin (C-PC), a biliprotein found in blue-green algae such as spirulina platensis, is widely used in the food and dietary nutritional supplement fields due to its beneficial pharmacological effects. Previous studies have revealed that C-PC has anti-inflammatory, antioxidant, and anti-apoptotic activities.Purpose: Therefore, this study investigated the protective effect and underlying mechanisms of C-PC on lipopolysaccharide (LPS) and seawater (SW) induced ALI (SW and LPS-induced ALI).Methods: An SW and LPS mouse model of ALI mice was established through intratracheal administration of 5mg/kg LPS and 25% SW. Different doses of C-PC (100, 200 and 400 mg/kg) were administered by intraperitoneal injection for seven days. In addition, gap junction communication in RAW264.7 and MLE-12 cells was determined following stimulation with 25% SW and 10 μg/ml LPS after treatment with C-PC (120 μg/ml). Moreover, the arterial partial pressure of oxygen, lung wet/dry weight ratios, total protein content and MPO levels in the bronchoalveolar lavage fluid (BALF), and the histopathologic and ultrastructure staining of the lung tissues were determined. The oxidative stress index, levels of the pro-inflammatory mediators, epithelial cell viability and apoptosis, and the regulatory effect of C-PC on the NF-κB/NLRP3 axis were investigated.Results: The results showed that C-PC significantly alleviated pathological damages, suppressed oxidative stress, inflammation and apoptosis, and enhanced the viability of epithelial cells in the lung tissues. Furthermore, C-PC was shown to inhibit activation of the NF-κB/NLRP3 pathway and the formation of the NLRP3 inflammasome complex.Conclusions: In conclusion, C-PC shows promising therapeutic value in SW and LPS-induced ALI/ARDS, providing new insight into ALI/ARDS treatment.Keywords: C-phycocyanin, acute lung injury, inflammation, NF-κB/NLRP3, apoptosis

Dehua Kong,&ast; Wenyu Hong,&ast; Miao Yu, Yanxia Li, YaXin Zheng, Xue Ying Collaborative Innovation Center of Sichuan for Elderly Care and Health & Key Laboratory of Sichuan Province for Specific Structure of Small Molecule Drugs, School of Pharmaceutical Sciences, Chengdu Medical College, Chengdu, 610500, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xue Ying; YaXin Zheng, School of Pharmaceutical Sciences, Chengdu Medical College, Chengdu, 610500, People’s Republic of China, Tel +86 135-7945-5890 ; +86 173-8187-6167, Email yingxueshzu@163.com; zhengyaxinxyz@163.comIntroduction: The inability of many therapeutic molecules to cross the blood–brain barrier (BBB) combined with poor penetration into tumor tissue leads to difficult challenges for treatment of brain tumors. In order to settle these hurdles, we developed a novel multifunctional targeting carrier which enables drugs to transport across the BBB and targets brain tumor tissues.Methods: In the multifunctional targeting liposomes, the natural compound resveratrol (RES) was incorporated into the lipid bilayer membranes of liposomes, while p-aminophenyl-α-D-manno-pyranoside (MAN) and wheat germ agglutinin (WGA) were conjugated to the liposomal surface. Epirubicin (EPI) as an anticancer drug was then loaded into liposomes. Then, liposomes were characterized by evaluation on particle size, zeta potential and apparent morphology. The epirubicin plus resveratrol liposomes modified with WGA and MAN were applied to glioma cells and BBB model in vitro and C6 glioma-bearing rats in vivo.Results: The multifunctional targeting liposomes were round shaped with a smooth surface and uniform particle size. In consideration of the SRB results, the multifunctional targeting liposomes indicated a significant inhibitory effect, suggesting that MAN plus WGA generated robust drug delivery effects into the brain tumor cells. The glioma cells after administering epirubicin plus resveratrol liposomes modified with WGA and MAN displayed the most significant uptake and apoptosis conducted by flow cytometry. In the multifunctional targeting effects assay, the epirubicin plus resveratrol liposomes modified with WGA and MAN exhibited the strongest effects of crossing the BBB and then targeting brain tumor cells. In tumor-bearing rats after applying multifunctional targeting liposomes, the median survival time was evidently observed as being markedly longer than other controls.Conclusion: The epirubicin plus resveratrol liposomes modified with WGA and MAN exhibited strong ability to improve epirubicin and resveratrol transporting across the BBB and therapeutic effect on brain glioma, showing multifunctional targeting capability.Graphical Abstract: Keywords: multifunctional targeting liposomes, blood–brain barrier, apoptosis, resveratrol, brain gliomas

Wei Han,1– 3,* Desong Kong,2,* Qin Lu,4 Wei Zhang,5 Zhimin Fan4 1Nanjing University of Chinese Medicine, Nanjing, People’s Republic of China; 2Chinese Medicine Modernization and Big Data Research Center, Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Nanjing, People’s Republic of China; 3General Surgery Department, Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Nanjing, People’s Republic of China; 4Proctology Department, Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Nanjing, People’s Republic of China; 5Anesthesiology Department, Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Nanjing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhimin FanProctology Department, Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, No. 157 Daming Avenue, Qinhuai District, Nanjing City, Jiangsu Province, 210012, People’s Republic of ChinaTel +86-2552276501Email fanzhimin_zhmf@163.comBackground: Aloperine can regulate miR-296-5p/Signal Transducer and Activator of Transcription 3 (STAT3) pathway to inhibit the malignant development of colorectal cancer (CRC), but the regulatory mechanism is unclear. This study explored the upstream mechanism of Aloperine in reducing CRC damage from the perspective of the circRNA-miRNA-mRNA regulatory network.Methods: After treatment with gradient concentrations of Aloperine (0.1 mmol/L, 0.2 mmol/L, 0.4 mmol/L, 0.8 mmol/L and 1 mmol/L) for 24 hours, changes in CRC cell proliferation and apoptosis were detected by functional experiments. Data of the differential expression of miR-296-5p in CRC patients and healthy people were obtained from Starbase. The effects of Aloperine on 12 differentially expressed circRNAs were detected. The binding of miR-296-5p with NOP2/Sun RNA methyltransferase 2 (circNSUN2) and STAT3 was predicted by TargetScan and confirmed through dual-luciferase experiments. The expressions of circNSUN2, miR-296-5p and STAT3 as well as apoptosis-related genes in CRC cells were detected by qRT-PCR and Western blot as needed. Rescue experiments were conducted to test the regulatory effects of circNSUN2, miR-296-5p and STAT3 on CRC cells.Results: Aloperine at a concentration gradient inhibited proliferation and promoted apoptosis in CRC cells. The abnormally low expression of miR-296-5p in CRC could be upregulated by Aloperine. Among the differentially expressed circRNAs in CRC, only circNSUN2 not only targets miR-296-5p, but also can be regulated by Aloperine. The up-regulation of circNSUN2 offset the inhibitory effect of Aloperine on cancer cells. The rescue experiments finally confirmed the regulation of circNSUN2/miR-296-5p/STAT3 axis in CRC cells.Conclusion: By regulating the circNSUN2/miR-296-5p/STAT3 pathway, Aloperine prevents the malignant development of CRC cells.Keywords: colorectal cancer, aloperine, NOP2/Sun RNA methyltransferase 2, miR-296-5p, signal transducer and activator of transcription 3

Shasha Cui,1,* Yujun Jiang,2,* Qianyu Shi,1,* Lei Zhang,3 Dehua Kong,4 Meijuan Qian,1 Jing Chu3 1Department of Nursing, Nantong Health College of Jiangsu Province, Nantong City, Jiangsu Province, People’s Republic of China; 2Department of Gynecology, The Affiliated Hospital of Qingdao University, Qingdao City, Shandong Province, People’s Republic of China; 3Department of Nursing, Navy Medical University, Shanghai, People’s Republic of China; 4Department of Nursing, The 940th Hospital of Joint Logistics Support Force of Chinese People’s Liberation Army, Lanzhou City, Gansu Province, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jing ChuDepartment of Nursing, Navy Medical University, 800 Xiang Yin Road, Shanghai, 200433, People’s Republic of ChinaTel +86-21-8187 1483Email chujing1999@smmu.edu.cnMeijuan QianDepartment of Nursing, Nantong Health College of Jiangsu Province, 288 Zhen Xing Dong Road, Nantong City, Jiangsu Province, People’s Republic of ChinaEmail 158142474@qq.comBackground: During the Coronavirus Disease 2019 (COVID-19) pandemic, emergency departments and fever clinics nurses acted as gatekeepers to the health care system. To manage the psychological problems that these nurses experience, we should develop appropriate training and intervention programs.Objective: To identify the impact of COVID-19 on the psychology of Chinese nurses in emergency departments and fever clinics and to identify associated factors.Methods: This online cross-sectional study recruited participants through snowball sampling between 13 February and 20 February 2020. Nurses self-administered the online questionnaires, including a general information questionnaire, the Self-Rating Anxiety Scale, the Perceived Stress Scale-14, and the Simplified Coping Style Questionnaire.Results: We obtained 481 responses, of which 453 were valid, an effective response rate of 94.18%. Participants who had the following characteristics had more mental health problems: female gender, fear of infection among family members, regretting being a nurse, less rest time, more night shifts, having children, lack of confidence in fighting transmission, not having emergency protection training, and negative professional attitude.Conclusion: Effective measures are necessary to preserve mental health of nurses in emergency departments and fever clinics. These include strengthening protective training, reducing night shifts, ensuring adequate rest time, and timely updating the latest pandemic situation.Keywords: COVID-19, nurses, psychological health, mental health, stress, anxiety

Daliang Kong, Rui Gu, Chengtao Zhang, Ruofeng Yin Department of Orthopedics, China-Japan Union Hospital, Changchun, Jilin 130031, People’s Republic of ChinaCorrespondence: Ruofeng YinDepartment of Orthopedics, China-Japan Union Hospital, Changchun, Jilin 130031, People’s Republic of ChinaEmail yrf_wind@jlu.edu.cnBackground: Circular RNAs (circRNAs) play an important role in the progression of intervertebral disc (IVD) degeneration (IVDD). Using bioinformatics analysis, we have found that the expression of circRNA hsa_circ_0059955 was significantly downregulated in IVDD tissues. However, the relevant mechanism of hsa_circ_0059955 in the progression of IVDD remains unclear.Methods: CCK-8 and flow cytometry assays were used to evaluate cell proliferation and apoptosis. In addition, Western blot assay was used to detect the expressions of ITCH, p73, CDK2 in nucleus pulposus (NP) cells. Moreover, a puncture-induced IVDD rat model was established to explore the role of hsa_circ_0059955 in IVDD.Results: The level of hsa_circ_0059955 was significantly decreased in IVDD tissues from IVDD patients. Itchy E3 ubiquitin protein ligase (ITCH) is the host gene of hsa_circ_0059955, and downregulation of hsa_circ_0059955 significantly decreased the expression of ITCH in NP cells. In addition, downregulation of hsa_circ_0059955 markedly inhibited proliferation and induced apoptosis and cell cycle arrest in NP cells. Moreover, in vivo study illustrated that overexpression of hsa_circ_0059955 ameliorated IVDD in rats.Conclusion: Downregulation of hsa_circ_0059955 could induce apoptosis and cell cycle arrest in NP cells in vitro, while overexpression of hsa_circ_0059955 attenuated the IVDD in a puncture-induced rat model in vivo. Therefore, hsa_circ_0059955 might serve as a therapeutic target for the treatment of IVDD.Keywords: intervertebral disc degeneration, hsa_circ_0059955, Itchy E3 ubiquitin protein ligase, p73

Hongfeng Wang,1,* Yuna Shang,2,* Xiaoniao Chen,3 Zhongyan Wang,2 Dashuai Zhu,4 Yue Liu,4 Chuyue Zhang,1 Pu Chen,1 Jie Wu,1 Lingling Wu,1 Deling Kong,2 Zhimou Yang,2 Zongjin Li,4 Xiangmei Chen1 1Department of Nephrology, Chinese PLA General Hospital, Chinese PLA Institute of Nephrology, State Key Laboratory of Kidney Diseases, National Clinical Research Center for Kidney Diseases, Beijing Key Laboratory of Kidney Diseases, Beijing 100039, People’s Republic of China; 2State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Key Laboratory of Bioactive Materials, Ministry of Education, Nankai University, Tianjin 300071, People’s Republic of China; 3Department of Ophthalmology, Chinese PLA General Hospital, Beijing 100039, People’s Republic of China; 4Department of Pathophysiology, Nankai University School of Medicine, Tianjin, 300071, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xiangmei ChenChinese PLA General Hospital, Beijing 100039, People’s Republic of ChinaEmail xmchen301@126.comZhimou YangNankai University, Tianjin 300071, People’s Republic of ChinaEmail yangzm@nankai.edu.cnPurpose: By providing a stem cell microenvironment with particular bioactive constituents in vivo, synthetic biomaterials have been progressively successful in stem cell-based tissue regeneration by enhancing the engraftment and survival of transplanted cells. Designs with bioactive motifs to influence cell behavior and with D-form amino acids to modulate scaffold stability may be critical for the development and optimization of self-assembling biomimetic hydrogel scaffolds for stem cell therapy.Materials and Methods: In this study, we linked naphthalene (Nap) covalently to a short D-form peptide (Nap-DFDFG) and the C domain of insulin-like growth factor-1 (IGF-1C) as a functional hydrogel-based scaffolds, and we hypothesized that this hydrogel could enhance the therapeutic efficiency of human placenta-derived mesenchymal stem cells (hP-MSCs) in a murine acute kidney injury (AKI) model.Results: The self-assembling peptide was constrained into a classical β-sheet structure and showed hydrogel properties. Our results revealed that this hydrogel exhibited increased affinity for IGF-1 receptor. Furthermore, cotransplantation of the β-IGF-1C hydrogel and hP-MSCs contributed to endogenous regeneration post-injury and boosted angiogenesis in a murine AKI model, leading to recovery of renal function.Conclusion: This hydrogel could provide a favorable niche for hP-MSCs and thereby rescue renal function in an AKI model by promoting cell survival and angiogenesis. In conclusion, by covalently linking the desired functional groups to D-form peptides to create functional hydrogels, self-assembling β-sheet peptide hydrogels may serve as a promising platform for tissue-engineering and stem cell therapy.Keywords: self-assembly, β-sheet, hydrogel, D-form peptide, C domain of insulin-like growth factor, IGF-1C, mesenchymal stem cells, MSCs, acute kidney injure, AKI

Zhang Y, Liu X, Kong D, et al. Int J Chron Obstruct Pulmon Dis. 2020;15:213–223. The authors have advised due to an error at the time of figure assembly, Figure 4 on page 218 is incorrect. The correct Figure 4 is shown in Download Article. The authors apologize for this error and advise it does not affect the results of the paper. Read the original article

Yongqiao Zhang,1 Xiaole Liu,1 Dehui Kong,1 Jia Fu,1 Yanbo Liu,1 Yakun Zhao,1 Hui Lian,1 Xiaoyi Zhao,2 Jun Yang,3 Zhongjie Fan1 1Department of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, People’s Republic of China; 2Department of Physical Medicine and Rehabilitation, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, People’s Republic of China; 3Institute for Environmental and Climate Research, Jinan University, Guangzhou 511443, People’s Republic of ChinaCorrespondence: Zhongjie FanDepartment of Cardiology, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, People’s Republic of ChinaTel +86 139 0119 9579Email Fan@pumch.cnPurpose: To evaluate the associations between acute exacerbations of chronic obstructive pulmonary disease (AECOPD) hospitalizations and daily mean temperature (Tmean) as well as daily apparent temperature (AT), and to explore the practical values of these two indices in policymaking and patient education.Methods: Daily AECOPD hospitalizations and Meteorological data in Beijing were obtained between 2013 and 2016. Distributed lag non-linear model was adopted to investigate the association between daily ambient temperature and AECOPD hospitalizations. The cumulative effects of cold/hot temperature were abstracted. For the extreme and moderate low-temperature effect estimates, we, respectively, computed the RR of AECOPD hospitalizations at the 1st and 10th percentiles of temperature in comparison with that at the 25th percentile of temperature. For the extreme and moderate high temperature effect estimates, we, respectively, computed the RR of AECOPD hospitalizations at the 99th and 90th percentiles of temperature in comparison with that at the 75th percentile of temperature.Results: During the study period, 143, 318 AECOPD hospitalizations were collected. A reverse J-shape relationship was found between temperature and AECOPD hospitalizations. When comparing the effect of Tmean, higher RRs were associated with increases in AT on AECOPD hospitalizations but a lower value of Akaike’s Information Criterion for quasi-Poisson (Q-AIC). The RR of extremely low temperature of Tmean and AT were 1.55 (95% CI: 1.21,2.00) and 2.08 (95% CI: 1.44,3.01), respectively. Moderate low temperature also had an adverse impact on AECOPD hospitalizations. No associations were found between high temperature and AECOPD risk. We found the females and those aged < 65 years to be more susceptible to temperature change.Conclusion: Lower temperature is associated with a higher risk for AECOPD hospitalizations. Ambient temperature is probably a better predictor in terms of quantifying risk than mean temperature when studying temperature impact on health.Keywords: ambient temperature, distributed lag non-linear model, hospitalization

Demiao Kong,1,2,* Yi Zhan,3,* Canzhao Liu,4 Yerong Hu,1 Yangzhao Zhou,1,4 Jiawen Luo,1 Lu Gu,1 Xinmin Zhou,1 Zhiwei Zhang1,4 1Department of Cardiovascular Surgery, The Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, China; 2Department of Thoracic Surgery, Guizhou Provincial People’s Hospital, Guiyang, Guizhou 550002, China; 3Department of Dermatology, The Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, China; 4Department of Medicine, University of California San Diego, La Jolla, CA 92093, USA*These authors contributed equally to this workCorrespondence: Zhiwei ZhangDepartment of Cardiovascular Surgery, The Second Xiangya Hospital, Central South University, No. 139 Middle Renmin Road, Changsha, Hunan 410011, People’s Republic of ChinaTel +86 13787083210 Email somebody_zhang@csu.edu.cnBackground: Emery-Dreifuss muscular dystrophy, caused by mutations in genes such as emerin (EMD) or lamin A/C (LMNA), is a disorder affecting the joints, muscles, and heart, with a wide spectrum of patient phenotypes including muscle wasting and cardiac conduction defects.Methods and results: Here we report a multi-generation family from the Hunan Province of China. Affected family members displayed an uncommon clinical presentation of serious cardiac conduction abnormalities at an early age and a high incidence of sudden cardiac death along with mild skeletal muscular atrophy and joint contracture. Clinical analysis of affected members provided evidence of X-linked recessive inheritance. Consequently, using Sanger sequencing of X chromosome exomes, we identified a novel duplication mutation (c.405dup/p.Asp136X) in the EMD gene as the cause for the disease in this family. This variant is a novel mutation that has not been previously reported in Pubmed, Clinvar or other cases reported in the Human Gene Mutation Database.Conclusion: Our finding expands the mutation spectrum of Emery-Dreifuss muscular dystrophy and provides a rationale for EMD mutation testing in cases of X-linked inherited cardiac conduction disease and sudden cardiac death, even in those lacking pathognomonic neuromuscular features.Keywords: Emery-Dreifuss muscular dystrophy, emerin, sudden cardiac death, cardiac conduction abnormalities, mutation

Jie Qin, Ping Wang, Taile Jing, Debo Kong, Sunyi Ye, Dan Xia, Shuo WangDepartment of Urology, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, People’s Republic of ChinaBackground: To report our experience with retroperitoneal robot-assisted laparoscopic upper pole heminephrectomy in adult patients with duplex kidneys.Methods: We retrospectively reviewed the medical records of 7 patients who underwent retroperitoneal robot-assisted laparoscopic upper pole heminephrectomy at our institution between September 2014 and July 2017. Of the robot-assisted laparoscopic procedures, 5 were on the left and 2 on the right side.Results: All patients underwent robot-assisted laparoscopic surgery successfully in a totally retroperitoneal manner without conversion to open surgery. The mean operative time was 175 mins (range 140–270). The mean estimated blood loss was 84 mL (range 20–200). The mean postoperative hospital stay was 7 days (range 5–9). No major intraoperative and postoperative complications occurred. All patients had a resolution of their presenting symptoms after surgery at a mean follow-up of 24 months (range 14–38).Conclusion: Our initial clinical experience suggests that robot-assisted laparoscopic upper pole heminephrectomy using a retroperitoneal approach for a duplex kidney appears to be safe with acceptable perioperative outcomes.Keywords: robotic, laparoscopy, duplex kidney, heminephrectomy

Zhang Y, Zhou J, Yang C, et al. Int J Nanomedicine. 2016;11:1119–1130. The authors have advised due to an error at the time of figure assembly, Figure 4 on page 1124 is incorrect. The correct Figure 4 is shown in Download Article. Page 1126, Table 1, the authors have advised a mistake was made in calculating the Drug loading content (DLC) in the manuscript. The correct Table 1 is shown in Download Article. The authors apologize for these errors and advise it does not affect the results of the paper. Read the original article

Run Liu,1,* Bi-Xia Liu,2,* Mingyue Ma,1,* Dan Kong,3,* Guanglin Li,4 Junle Yang,1 Xiaoping Wu,1 Jiyong Zheng,3 Yan Dong1 1Department of Radiology, The Affiliated Xi’an Central Hospital of Xi’an Jiaotong University, Xi’an, 710004 Shaanxi, People’s Republic of China; 2Department of ICU, Jiangxi Provincial Cancer Hospital, Nanchang, 330029 Jiangxi, People’s Republic of China; 3Department of Medical Imaging, The Affiliated Huaian No. 1 People’s Hospital of Nanjing Medical University, Huaian, 223300 Jiangsu, People’s Republic of China; 4Honghui Hospital, Xi’an Jiaotong University, Xi’an, 710054 Shaanxi, People’s Republic of China *These authors contributed equally to this work Objective: To identify whether the amplitude of low-frequency fluctuations (ALFF) analysis has the potential to serve as a biological marker to detect alcohol-induced spontaneous brain activities and distinguish the patients with alcohol dependence from the healthy subjects. Methods: We utilized the ALFF analysis to report on the alcohol-induced spontaneous brain activities in 29 patients with alcohol dependence (9 female, 20 male) relative to 29 status-matched healthy subjects (11 female, 18 male). Receiver operating characteristic curve was used to test the ability of the ALFF analysis in discriminating the patients with alcohol dependence from the healthy subjects. Pearson correlation was used to evaluate the relationships between the signal value of those ALFF differences in brain areas and behavioral characteristics. Results: Alcohol-induced brain differences located in the right inferior parietal lobule and right supplementary motor area with significant higher ALFF values, and in the left precuneus and bilateral cerebellum posterior lobe with lower ALFF values. The movement-related areas were significantly correlated with each other (P

Jianwei Duan,1,* Chao Liu,1,* Xiaoyu Liang,1 Xuanling Li,1 Youlu Chen,1 Zuoguan Chen,2 Xiaoli Wang,1 Deling Kong,1,3 Yongjun Li,2 Jing Yang1 1Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Chinese Academy of Medical Science and Peking Union Medical College, Tianjin, China; 2Department of Vascular Surgery, Beijing Hospital, National Center of Gerontology, Chinese Academy of Medical Science and Peking Union Medical College, Beijing, China; 3Key Laboratory of Bioactive Materials, Ministry of Education, Nankai University, Tianjin, China *These authors contributed equally to this work Background: To address the issue of delivery of proteins, a six-arm copolymer, six-arm poly(ε-caprolactone)–poly(ethylene glycol) (6S-PCL-PEG), was synthesized by a simple two-step reaction. Thereafter, the application of 6S-PCL-PEG as a protein carrier was evaluated. Materials and methods: A six-arm copolymer, six-arm poly(ε-caprolactone) (6S-PCL), was synthesized by ring-opening polymerization, with stannous octoate as a catalyst and inositol as an initiator. Then, poly(ethylene glycol) (PEG) was linked with 6S-PCL by oxalyl chloride to obtain 6S-PCL-PEG. Hydrogen-1 nuclear magnetic resonance spectrum, Fourier-transform infrared spectroscopy, and gel-permeation chromatography were conducted to identify the structure of 6S-PCL-PEG. The biocompatibility of the 6S-PCL-PEG was evaluated by a cell counting kit-8 assay. Polymeric nanoparticles (NPs) were prepared by a water-in-oil-in-water double emulsion (W1/O/W2) solvent evaporation method. The size distribution and zeta potential of NPs were determined by dynamic light scattering. Transmission electron microscopy was used to observe the morphology of NPs. Drug-loading capacity, encapsulation efficiency, and the release behavior of ovalbumin (OVA)-loading NPs were tested by the bicinchoninic acid assay kit. The stability and activity of OVA released from NPs were detected and the uptake of NPs was evaluated by NIH-3T3 cells. Results: All results indicated the successful synthesis of amphiphilic copolymer 6S-PCL-PEG, which possessed excellent biocompatibility and could formulate NPs easily. High drug-loading capacity and encapsulation efficiency of protein NPs were observed. In vitro, OVA was released slowly and the bioactivity of OVA was maintained for over 28 days. Conclusion: 6S-PCL-PEG NPs prepared in this study show promising potential for use as a protein carrier. Keywords: six-arm PCL-PEG, copolymer synthesis, protein carrier, sustained release

Rui Zhou,* Tao Xu,* XiaoHong Liu, YuanShou Chen, DeYing Kong, Hong Tian, Mingxia Yue, Dujuan Huang, Junwei Zeng Department of Physiology, Zunyi Medical College, Zunyi, People’s Republic of China *These authors contributed equally to this work Objective: The dorsal horn P2Y13 receptor is involved in the development of pain behavior induced by peripheral nerve injury. It is unclear whether the expression of proinflammatory cytokines interleukin (IL)-1β and IL-6 at the spinal dorsal horn are influenced after the activation of P2Y13 receptor in rats with diabetic neuropathic pain (DNP). Methods: A rat model of type 1 DNP was induced by intraperitoneal injection of streptozotocin (STZ). We examined the expression of P2Y13 receptor, Iba-1, IL-1β, IL-6, JAK2, STAT3, pTyr1336, and pTyr1472 NR2B in rat spinal dorsal horn. Results: Compared with normal rats, STZ-diabetic rats displayed obvious mechanical allodynia and the increased expression of P2Y13 receptor, Iba-1, IL-1β, and IL-6 in the dorsal spinal cord that was continued for 6 weeks in DNP rats. The data obtained indicated that, in DNP rats, administration of MRS2211 significantly attenuated mechanical allodynia. Compared with DNP rats, after MRS2211 treatment, expression of the P2Y13 receptor, Iba-1, IL-1β, and IL-6 were reduced 4 weeks after the STZ injection. However, MRS2211 treatment did not attenuate the expression of the P2Y13 receptor, Iba-1, IL-1β, and IL-6 at 6 weeks after the STZ injection. MRS2211 suppressed JAK2 and STAT3 expression in the early stage, but not in the later stage. Moreover, pTyr1336 NR2B was significantly decreased, whereas pTyr1472 NR2B was unaffected in the dorsal spinal cord of MRS2211-treated DNP rats. Conclusion: Intrathecal MRS2211 produces an anti-nociceptive effect in early-stage DNP. A possible mechanism involved in MRS2211-induced analgesia is that blocking the P2Y13 receptor downregulates levels of IL-1β and IL-6, which subsequently inhibit the activation of the JAK2/STAT3 signaling pathway. Furthermore, blocking the activation of the P2Y13 receptor can decrease NR2B-containing NMDAR phosphorylation in dorsal spinal cord neurons, thereby attenuating central sensitization in STZ-induced DNP rats.Keywords: diabetic neuropathic pain, dorsal horn, P2Y13 receptor

Qiang Yang,1,* Bin-Hong Teng,2,* Li-Na Wang,3 Kun Li,2 Chen Xu,2 Xin-Long Ma,1 Yang Zhang,1 De-Ling Kong,3 Lian-Yong Wang,3 Yan-Hong Zhao2 1Department of Spine Surgery, Tianjin Hospital, Tianjin , People’s Republic of China; 2School and Hospital of Stomatology, Tianjin Medical University,Tianjin, People’s Republic of China; 3The Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Sciences, Nankai University, Tianjin, People’s Republic of China *These authors contributed equally to this work Abstract: A 3-D scaffold that simulates the microenvironment in vivo for regenerating cartilage is ideal. In this study, we combined silk fibroin and decellularized cartilage extracellular matrix by temperature gradient-guided thermal-induced phase separation to produce composite scaffolds (S/D). Resulting scaffolds had remarkable mechanical properties and biomimeticstructure, for a suitable substrate for attachment and proliferation of adipose-derived stem cells (ADSCs). Moreover, transforming growth factor β3 (TGF-β3) loaded on scaffolds showed a controlled release profile and enhanced the chondrogenic differentiation of ADSCs during the 28-day culture. The S/D scaffold itself can provide a sustained release system without the introduction of other controlled release media, which has potential for commercial and clinical applications. The results of toluidine blue, Safranin O, and immunohistochemical staining and analysis of collagen II expression showed maintenance of a chondrogenic phenotype in all scaffolds after 28-day culture. The most obvious phenomenon was with the addition of TGF-β3. S/D composite scaffolds with sequential delivery of TGF-β3 may mimic the regenerative microenvironment to enhance the chondrogenic differentiation of ADSCs in vitro. Keywords: cartilage tissue engineering, composite scaffold, silk fibroin, decellularized cartilage extracellular matrix, adipose-derived stem cells, transforming growth factor β3 (TGF-β3)

Shaofeng Lou,1 Xiuyuan Zhang,2 Mingming Zhang,2 Shenglu Ji,1 Weiwei Wang,2 Ju Zhang,1 Chen Li,2 Deling Kong1,2 1Key Laboratory of Bioactive Materials, Ministry of Education, College of Life Sciences, Nankai University, 2Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Chinese Academy of Medical Science, Tianjin, People’s Republic of China Abstract: A reductase-cleavable and thermo-responsive star-shaped polymer nanogel was prepared via an “arm-first” atom transfer radical polymerization approach. The nanogel consists of a thermo- and redox-sensitive core and a zwitterionic copolymer block. The dual sensitive core is composed of poly(N-isopropylacrylamide) that is formed by disulfide crosslinking of N-isopropylacrylamide. The zwitterionic copolymer block contains a poly(sulfobetaine methacrylate) component, a known anti-adsorptive moiety that extends blood circulation time, and a lactose motif of poly(2-lactobionamidoethyl methacrylamide) that specifically targets the asialoglycoprotein receptors (ASGP-Rs) of hepatoma. Doxorubicin (DOX) was encapsulated into the cross-linked nanogels via solvent extraction/evaporation method and dialysis; average diameter of both blank and DOX-loaded nanogels was ~120 nm. The multi-responsiveness of nanogel drug release in different temperatures and redox conditions was assessed. After 24 h, DOX release was only ~20% at 30°C with 0 mM glutathione (GSH), whereas over 90% DOX release was observed at 40°C and 10 mM GSH, evidence of dual responsiveness to temperature and reductase GSH. The IC50 value of DOX-loaded nanogels was much lower in human hepatoma (HepG2) cells compared to non-hepatic HeLa cells. Remarkably, DOX uptake of HepG2 cells differed substantially in the presence and absence of galactose (0.31 vs 1.42 µg/mL after 48 h of incubation). The difference was non-detectable in HeLa cells (1.21 vs 1.57 µg/mL after 48 h of incubation), indicating that the overexpression of ASGP-Rs leads to the DOX-loaded lactosylated nanogels actively targeting hepatoma. Our data indicate that the lactose-decorated star-shaped nanogels are dual responsive and hepatoma targeted, and could be employed as hepatoma-specific anti-cancer drug delivery vehicle for cancer chemotherapy. Keywords: glycopolymer, multi-responsive nanogel, hepatoma targeting, drug delivery, arm-first ATRP

Xiaowei Zhu,* Chao Liu,* Jianwei Duan, Xiaoyu Liang, Xuanling Li, Hongfan Sun, Deling Kong, Jing Yang Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Chinese Academy of Medical Science and Peking Union Medical College, Tianjin, People’s Republic of China *These authors contributed equally to this work Purpose: Synthesis of star-shaped block copolymer with oxalyl chloride and preparation of micelles to assess the prospect for drug-carrier applications.Materials and methods: Three-arm star block copolymers of poly(lactic-co-glycolic acid) (3S-PLGA)–polyethylene glycol (PEG) were synthesized by ring-opening polymerization, then PEG as the hydrophilic block was linked to the terminal hydroxyl of 3S-PLGA with oxalyl chloride. Fourier-transform infrared (FT-IR) spectroscopy, gel-permeation chromatography (GPC), hydrogen nuclear magnetic resonance (1H-NMR) spectra, and differential scanning calorimetry were employed to identify the structure and properties of 3S-PLGA-PEG. Rapamycin (RPM)-loaded micelles were prepared by solvent evaporation, and pyrene was used as the fluorescence probe to detect the critical micelle concentration of the copolymer. The particle size, distribution, and ζ-potential of the micelles were determined by dynamic light scattering, and the morphology of the RPM-loaded micelles was analyzed by transmission electron microscopy. High-performance liquid chromatography was conducted to analyze encapsulation efficiency and drug-loading capacity, as well as the release behavior of RPM-loaded micelles. The biocompatibility of material and the cytostatic effect of RPM-loaded micelles were investigated by Cell Counting Kit 8 assay.Results: FT-IR, GPC, and 1H-NMR suggested that 3S-PLGA-PEG was successfully synthesized. The RPM-loaded micelles prepared with the 3S-PLGA-PEG possessed good properties. The micelles had good average diameter and encapsulation efficiency. For in vitro release, RPM was released slowly from 3S-PLGA-PEG micelles, showing that 3S-PLGA-PEG-RPM exhibited a better and longer antiproliferative effect than free RPM.Conclusion: In this study, we first used oxalyl chloride as the linker to synthesize 3S-PLGA-PEG successfully, and compared with reported literature, this method shortened the reaction procedure and improved the reaction yield. The micelles prepared with this material proved suitable for drug-carrier application. Keywords: block copolymer, RPM, micelles, cytostatic effect

Yumin Zhang,1,* Junhui Zhou,2,* Cuihong Yang,1 Weiwei Wang,3 Liping Chu,1 Fan Huang,1 Qiang Liu,1 Liandong Deng,2 Deling Kong,3 Jianfeng Liu,1 Jinjian Liu1 1Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Institute of Radiation Medicine, Chinese Academy of Medical Science and Peking Union Medical College, 2Department of Polymer Science and Technology, School of Chemical Engineering and Technology, Tianjin University, 3Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Chinese Academy of Medical Science and Peking Union Medical College, Tianjin, People’s Republic of China *These authors contributed equally in this work Abstract: Although the shortcomings of small molecular antitumor drugs were efficiently improved by being entrapped into nanosized vehicles, premature drug release and insufficient tumor targeting demand innovative approaches that boost the stability and tumor responsiveness of drug-loaded nanocarriers. Here, we show the use of the core cross-linking method to generate a micelle with enhanced drug encapsulation ability and sensitivity of drug release in tumor. This kind of micelle could increase curcumin (Cur) delivery to HeLa cells in vitro and improve tumor accumulation in vivo. We designed and synthesized the core cross-linked micelle (CCM) with polyethylene glycol and folic acid-polyethylene glycol as the hydrophilic units, pyridyldisulfide as the cross-linkable and hydrophobic unit, and disulfide bond as the cross-linker. CCM showed spherical shape with a diameter of 91.2 nm by the characterization of dynamic light scattering and transmission electron microscope. Attributed to the core cross-linking, drug-loaded CCM displayed higher Nile Red or Cur-encapsulated stability and better sensitivity to glutathione than noncross-linked micelle (NCM). Cellular uptake and in vitro antitumor studies proved the enhanced endocytosis and better cytotoxicity of CCM-Cur against HeLa cells, which had a high level of glutathione. Meanwhile, the folate receptor-mediated drug delivery (FA-CCM-Cur) further enhanced the endocytosis and cytotoxicity. Ex vivo imaging studies showed that CCM-Cur and FA-CCM-Cur possessed higher tumor accumulation until 24 hours after injection. Concretely, FA-CCM-Cur exhibited the highest tumor accumulation with 1.7-fold of noncross-linked micelle Cur and 2.8-fold of free Cur. By combining cross-linking of the core with active tumor targeting of FA, we demonstrated a new and effective way to design nanocarriers for enhanced drug encapsulation, smart tumor responsiveness, and elevated tumor accumulation. Keywords: core cross-linking, folic acid targeting, self-assembling, curcumin, drug delivery, micelles

The effect of edible films based on carrot puree, chitosan, corn starch, gelatin, glycerol and cinnamaldehyde on fresh-cut carrots was studied during storage. Several parameters, such as firmness, colour, weight loss, total carotenoids, total phenols, polyphenol oxidase (PPO) activity and peroxidase (POD) activity in coated carrots were determined at regular intervals and then compared with the uncoated carrots throughout the storage period. Significant and expected changes were observed in all carrot samples that were compared. The coating treatment significantly (P < 0.05) delayed the senescence, reduced the deterioration of exterior quality and retained total carotenoids well compared with control (P < 0.05). In addition, significant inhibition of PPO activity (P < 0.05) and POD activity (P < 0.05) as well as reduced accumulation of polyphenols (P < 0.05) were observed for all coated samples. All of these favourable responses induced by coating treatment on minimally processed fresh-cut carrots showed beneficial physiological effects, which would give some useful references to the fresh-cut fruit and vegetable processing industry and satisfy people’s requirements allowing for extending product shelf life without negatively affecting the sensory quality or acceptability.

Linhua Zhang,1 Dunwan Zhu,1 Xia Dong,1 Hongfan Sun,1 Cunxian Song,1 Chun Wang,2 Deling Kong1 1Tianjin Key Laboratory of Biomaterials, Institute of Biomedical Engineering, Peking Union Medical College and Chinese Academy of Medical Sciences, Tianjin, People’s Republic of China; 2Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN, USA Abstract: The purpose of this study was to develop a novel lipid–polymer hybrid drug carrier comprised of folate (FA) modified lipid-shell and polymer-core nanoparticles (FLPNPs) for sustained, controlled, and targeted delivery of paclitaxel (PTX). The core-shell NPs consist of 1) a poly(ε-caprolactone) hydrophobic core based on self-assembly of poly(ε-caprolactone)–poly(ethylene glycol)–poly(ε-caprolactone) (PCL-PEG-PCL) amphiphilic copolymers, 2) a lipid monolayer formed with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy (polyethylene glycol)-2000] (DSPE-PEG2000), 3) a targeting ligand (FA) on the surface, and were prepared using a thin-film hydration and ultrasonic dispersion method. Transmission electron microscopy and dynamic light scattering analysis confirmed the coating of the lipid monolayer on the hydrophobic polymer core. Physicochemical characterizations of PTX-loaded FLPNPs, such as particle size and size distribution, zeta potential, morphology, drug loading content, encapsulation efficiency, and in vitro drug release, were also evaluated. Fluorescent microscopy proved the internalization efficiency and targeting ability of the folate conjugated on the lipid monolayer for the EMT6 cancer cells which overexpress folate receptor. In vitro cytotoxicity assay demonstrated that the cytotoxic effect of PTX-loaded FLPNPs was lower than that of Taxol®, but higher than that of PTX-loaded LPNPs (without folate conjugation). In EMT6 breast tumor model, intratumoral administration of PTX-loaded FLPNPs showed similar antitumor efficacy but low toxicity compared to Taxol®. More importantly, PTX-loaded FLPNPs showed greater tumor growth inhibition (65.78%) than the nontargeted PTX-loaded LPNPs (48.38%) (P

Yongxia Chen,1,3,* Ziying Yang,1,* Chao Liu,1 Cuiwei Wang,1 Shunxin Zhao,2 Jing Yang,1 Hongfan Sun,1 Zhengpu Zhang,2 Deling Kong,1 Cunxian Song1 1Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Chinese Academy of Medical Science and Peking Union Medical College, 2Institute of Polymer Chemistry, Nankai University, 3Center for Medical Device Evaluation of Tianjin, Tianjin, People's Republic of China *These authors contributed equally to this work Background: Star-shaped polymers provide more terminal groups, and are promising for application in drug-delivery systems. Methods: A new series of six-arm star-shaped poly(lactic-co-glycolic acid) (6-s-PLGA) was synthesized by ring-opening polymerization. The structure and properties of the 6-s-PLGA were characterized by carbon-13 nuclear magnetic resonance spectroscopy, infrared spectroscopy, gel permeation chromatography, and differential scanning calorimetry. Then, paclitaxel-loaded six-arm star-shaped poly(lactic-co-glycolic acid) nanoparticles (6-s-PLGA-PTX-NPs) were prepared under the conditions optimized by the orthogonal testing. High-performance liquid chromatography was used to analyze the nanoparticles' encapsulation efficiency and drug-loading capacity, dynamic light scattering was used to determine their size and size distribution, and transmission electron microscopy was used to evaluate their morphology. The release performance of the 6-s-PLGA-PTX-NPs in vitro and the cytostatic effect of 6-s-PLGA-PTX-NPs were investigated in comparison with paclitaxel-loaded linear poly(lactic-co-glycolic acid) nanoparticles (L-PLGA-PTX-NPs). Results: The results of carbon-13 nuclear magnetic resonance spectroscopy and infrared spectroscopy suggest that the polymerization was successfully initiated by inositol and confirm the structure of 6-s-PLGA. The molecular weights of a series of 6-s-PLGAs had a ratio corresponding to the molar ratio of raw materials to initiator. Differential scanning calorimetry revealed that the 6-s-PLGA had a low glass transition temperature of 40°C–50°C. The 6-s-PLGA-PTX-NPs were monodispersed with an average diameter of 240.4±6.9 nm in water, which was further confirmed by transmission electron microscopy. The encapsulation efficiency of the 6-s-PLGA-PTX-NPs was higher than that of the L-PLGA-PTX-NPs. In terms of the in vitro release of nanoparticles, paclitaxel (PTX) was released more slowly and more steadily from 6-s-PLGA than from linear poly(lactic-co-glycolic acid). In the cytostatic study, the 6-s-PLGA-PTX-NPs and L-PLGA-PTX-NPs were found to have a similar antiproliferative effect, which indicates durable efficacy due to the slower release of the PTX when loaded in 6-s-PLGA. Conclusion: The results suggest that 6-s-PLGA may be promising for application in PTX delivery to enhance sustained antiproliferative therapy. Keywords: PTX, polymer, drug delivery, nanoparticles, PLGA, antiproliferative therapy

Incompletely known fish assemblages and species diversity are substantial obstacles in fish conservation, particularly when their aquatic habitats are under threat due to rapid human-induced changes. Fish assemblages and diversity in three tributaries of the upper Irrawaddy River in China (the Dulong, Daying and Ruili rivers) were examined based on field collections and literature resources. The newly compiled fish assemblage recorded 85 species (in 8 orders, 20 families and 51 genera) distributed in the upper Irrawaddy. The fish compositions in the Daying (67 species, 44 genera, 19 families, 7 orders) and Ruili rivers (65 species, 44 genera, 19 families, 8 orders) were more similar to each other and more speciose than that in the Dulong River (14 species, 10 genera, 4 families, 3 orders). Two indices of taxonomic diversity (the average taxonomic distinctness (Δ+), and the variation in taxonomic distinctness (Λ+)) were used to discriminate four collections spanning a ten-year period. A decrease in taxonomic diversity and an increase in unevenness of the fish assemblages were found in both the Daying River and Ruili rivers, which indicated that the impacts were accumulated gradually during this decade, when dams and the spread of non-native species were major threats. Comparatively speaking, the Dulong River is still in a near-natural state, and thus the fish community has experienced less disturbance. In situ conservation (nature reserves and tributary protection) and ex situ conservation (artificial propagation and release) should be combined and managed to promote fish conservation in the future.