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Lanying Li,* Xinyuan Zhang,* Qian Li, Wen Zhong, Hua Zou Department of Laboratory Medicine, Chongqing Health Center for Women and Children, Women and Children’s Hospital of Chongqing Medical University, Chongqing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wen Zhong; Hua Zou, Department of Laboratory Medicine, Chongqing Health Center for Women and Children, Women and Children’s Hospital of Chongqing Medical University, 120 Longshan Road, Yubei District, Chongqing, 400016, People’s Republic of China, Tel +8615823827032 ; +8613512371464, Email 23119401@qq.com; 1316933969@qq.comPurpose: Candida vaginitis is widely prevalent worldwide and is one of the common gynecological disorders. The aim of this study is to analyze the sensitivity of recurrent vulvovaginal (RVVC) candidiasis to antifungal drugs and its relationship with vaginal microbiota.Patients and Methods: We Isolated and cultured Candida from RVVC patients, mass spectrometry and broth microdilution method were used to identify and determine MIC values of antifungal drugs. Clinical medical records and vaginal microbiota of RVVC patients were also collected.Results: The main pathogens causing RVVC are predominantly Candida albicans (70.26%), but in recent years, there has been an increasing proportion of Candida glabrata(24.46%). However, only 15.70% of Candida albicans were sensitive to Voriconazole, 35.84% to Fluconazole and 25.60% to Itraconazole. No fluconazole-resistant Candida glabrata was found. Most Candida krusei strains were sensitive to voriconazole (81.80%). More important MIC values of triazoles were increased in Candida species, when exposed to clotrimazole. In addition, we found that the vaginal microecology of candida vaginitis and bacterial vaginitis was significantly different.Conclusion: Triazoles resistant Candida species have emerged, leading to the failure of empirical anti-infective therapy. At the same time, the vaginal microecology of candida vaginitis and bacterial vaginitis was significantly different. In addition, a new breakpoint for Candida from RVVC needs to be established.Keywords: triazole, Candida, vulvovaginal candidiasis, vaginal microbiota

Youqian Li,1,2 Wei Zhong,1,2 Zhidong Liu,1,2 Changjing Huang,1,2 Junyin Peng,1,2 Hanlin Li1,2 1Center for Cardiovascular Diseases, Meizhou People’s Hospital, Meizhou Academy of Medical Sciences, Meizhou, People’s Republic of China; 2Guangdong Provincial Engineering and Technology Research Center for Molecular Diagnostics of Cardiovascular Diseases, Meizhou People’s Hospital, Meizhou Academy of Medical Sciences, Meizhou, People’s Republic of ChinaCorrespondence: Youqian Li, Center for Cardiovascular Diseases, Meizhou People’s Hospital, Meizhou Academy of Medical Sciences, No. 63 Huangtang Road, Meijiang District, Meizhou, People’s Republic of China, Email d_yoyo@163.comBackground: Aldehyde dehydrogenase 2 (ALDH2) is a key catalytic enzyme involved in the aldehyde metabolism that plays an important role in the occurrence and development of acute myocardial infarction (AMI). However, the relationship of ALDH2 polymorphism and susceptibility to AMI may differ among different regions and populations, and it has not yet been reported in Hakka population. The purpose of the present study was to investigate it in this population.Methods: Four hundred and nineteen AMI patients and 636 individuals without AMI were included in the present study. The ALDH2 rs671 polymorphism was genotyped using polymerase chain reaction (PCR)-microarray. Differences in ALDH2 rs671 genotypes and alleles between patients and controls were compared, and the relationship between ALDH2 rs671 genotypes and AMI risk was analyzed.Results: Patients with AMI had a lower frequency of ALDH2 rs671 G/G genotype (43.2% vs 52.7%, p=0.003), and a higher G/A genotype (45.6% vs 38.5%, p=0.025) than controls. And AMI patients had a lower frequency of ALDH2 rs671 G allele (66.0% vs 71.9%), and a higher A allele (34.0% vs 28.1%) (p=0.004) than controls. Logistic regression analysis showed that overweight (body mass index (BMI)≥ 24.0 kg/m2 vs BMI 18.5– 23.9 kg/m2: odds ratio (OR) 2.046, 95% confidence interval (CI): 1.520– 2.754, p< 0.001), history of hypertension (yes vs no: OR 3.464, 95% CI: 2.515– 4.770, p< 0.001), ALDH2 rs671 G/A genotype (G/A vs G/G: OR 1.476, 95% CI: 1.102– 1.976, p=0.009), and A/A genotype (A/A vs G/G: OR 1.656, 95% CI: 1.027– 2.668, p=0.038) maybe the independent risk factors for AMI.Conclusion: Overweight (BMI≥ 24.0 kg/m2), a history of hypertension, and ALDH2 rs671 G/A or A/A genotypes increased the risk of developing AMI in Hakka population.Keywords: aldehyde dehydrogenase 2, acute myocardial infarction, polymorphism, Hakka

Wenjie Xie,1,2,&ast; Yixun Zhang,1,2,&ast; Qianfeng Xu,1,2,&ast; Guowei Zhong,1,2 Jundong Lin,2,3 Huichan He,3 Qiuling Du,2 Huijing Tan,1,2 Muqi Chen,1,2 Zhenjie Wu,1– 3 Yulin Deng,1,2 Zhaodong Han,1– 3 Jianming Lu,1,2,4 Jianheng Ye,1,2,4 Fen Zou,1,2 Yangjia Zhuo,1,2 Weide Zhong1– 4 1Department of Urology, The Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou, 510180, People’s Republic of China; 2Guangdong Key Laboratory of Clinical Molecular Medicine and Diagnostics, Guangzhou First People’s Hospital, School of Medicine, South China University of Technology, Guangzhou, 510180, People’s Republic of China; 3Urology Key Laboratory of Guangdong Province, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510230, People’s Republic of China; 4State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Taipa, Macau, 999078, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yangjia Zhuo; WeideZhong, Email eyyangjiazhuo@scut.edu.cn; zhongwd2009@live.cnPurpose: Current treatment approaches for Prostate cancer (PCa) often come with debilitating side effects and limited therapeutic outcomes. There is urgent need for an alternative effective and safe treatment for PCa.Methods: We developed a nanoplatform to target prostate cancer cells based on graphdiyne (GDY) and a copper-based metal-organic framework (GDY-CuMOF), that carries the chemotherapy drug doxorubicin (DOX) for cancer treatment. Moreover, to provide GDY-CuMOF@DOX with homotypic targeting capability, we coated the PCa cell membrane (DU145 cell membrane, DCM) onto the surface of GDY-CuMOF@DOX, thus obtaining a biomimetic nanoplatform (DCM@GDY-CuMOF@DOX). The nanoplatform was characterized by using transmission electron microscope, atomic force microscope, X-ray diffraction, etc. Drug release behavior, antitumor effects in vivo and in vitro, and biosafety of the nanoplatform were evaluated.Results: We found that GDY-CuMOF exhibited a remarkable capability to load DOX mainly through π-conjugation and pore adsorption, and it responsively released DOX and generated Cu+ in the presence of glutathione (GSH). In vivo experiments demonstrated that this nanoplatform exhibits remarkable cell-killing efficiency by generating lethal reactive oxygen species (ROS) and mediating cuproptosis. In addition, DCM@GDY-CuMOF@DOX effectively suppresses tumor growth in vivo without causing any apparent side effects.Conclusion: The constructed DCM@GDY-CuMOF@DOX nanoplatform integrates tumor targeting, drug-responsive release and combination with cuproptosis and chemodynamic therapy, offering insights for further biomedical research on efficient PCa treatment. Keywords: graphdiyne, biomimetic nanoplatform, cuproptosis, chemotherapy, prostate cancer

Wentao Zhong,1– 3,&ast; Qianyu Wang,2,&ast; Xiaofei Shen,4,&ast; Yuan Lv,2,3 Liang Sun,2 Ran An,5 Hongyan Zhu,5 Huiyun Cai,2 Gang Chen,2,3 Aijun Liu,5 Junfeng Du1– 3 1The Second School of Clinical Medicine, Southern Medical University, Guangdong, 510515, People’s Republic of China; 2Department of General Surgery, The 7th Medical Center, Chinese PLA General Hospital, Beijing, 100700, People’s Republic of China; 3Medical Department of General Surgery, The 1st Medical Center, Chinese PLA General Hospital, Beijing, 100853, People’s Republic of China; 4Division of Gastric Surgery, Department of General Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, 210008, People’s Republic of China; 5Department of Pathology, the 7th Medical Center, Chinese PLA General Hospital, Beijing, 100700, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Aijun Liu, Department of Pathology, The 7th Medical Center, Chinese PLA General Hospital, NO. 5, Nanmen Cang, Beijing, 100700, People’s Republic of China, Email aliu301@126.com Junfeng Du, Department of General Surgery, the 7th Medical Center, Chinese PLA General Hospital, NO. 5, Nanmen Cang, Beijing, 100700, People’s Republic of China, Email dujf66@126.comPurpose: To demonstrate the intrinsic association of Neutrophil extracellular traps (NETs) with outcome and neoadjuvant therapy response of locally advanced rectal cancer (LARC), and the mechanisms.Patients and Methods: We enrolled 240 patients with LARC who underwent surgery without neoadjuvant therapy in two independent sets (training and validation), and 153 patients who received neoadjuvant therapy with biopsy followed by surgery. Immunohistochemistry, immunofluorescence staining and bioinformatics analysis were performed in formalin-fixed paraffin-embedded sections. NETs were identified by costaining for myeloperoxidase and citrullinated histone H3.Results: NETs were associated with recurrence-free survival in the surgical training and validation sets. High-NET density predicted poor postoperative survival of patients with LARC. Multivariate analysis identified NETs, TNM stage, and neutrophil-to-lymphocyte ratio as independent prognostic factors for recurrence-free survival. Low-NETs LARC demonstrated increased CD8+ T cell and lower T regulatory cell infiltration, which indicated a tumor immune microenvironment with strong antitumor capacity. High-NET density was associated with epithelial–mesenchymal transition, which is considered to contribute to tumor progression. In the neoadjuvant therapy cohort, high-NET density on biopsy was significantly associated with reduced likelihood of complete/near complete response.Conclusion: NET was an independent prognostic factor in LARC that were associated with patients’ survival, and NET density in pretreatment biopsies was an independent predictive biomarker of response to neoadjuvant therapy. This biomarker may be helpful in predicting survival in LARC with improved accuracy and selecting patients who will respond to neoadjuvant therapy.Keywords: NETs, tumor microenvironment, locally advanced rectal cancer, outcome

Amandeep Virk,1 Patrick-Julien Treacy,2 Wenjie Zhong,3,4 Norbert Doeuk,1 Celine Doeuk,5 Scott Leslie1– 3 1Department of Uro-Oncology, Chris O’Brien Lifehouse, Camperdown, NSW, Australia; 2The Institute of Academic Surgery at Royal Prince Alfred Hospital, Camperdown, NSW, Australia; 3Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW, Australia; 4Faculty of Medicine and Health, The University of New South Wales, Kensington, NSW, Australia; 5Rozelle Medical Centre, Ultimo, NSW, AustraliaCorrespondence: Amandeep Virk, Chris O’Brien Lifehouse, 119-143 Missenden Road, Camperdown, Sydney, NSW, 2050, Australia, Tel +61421233369, Email amandeep.virk.21@gmail.comBackground: Prostate cancer is often considered a disease of older men and this indeed fits with its peak incidence between 65– 79 years of age. Reports of prostate cancer in men younger than 40 years of age and the outcomes of this age group following treatment are few in the literature. Here, we present the case of an unusual diagnosis of high grade prostate cancer in a very young man and outline early outcomes following treatment with robotic-assisted radical prostatectomy.Case Presentation: A 35-year-old male, intermittently taking finasteride for hair loss, was found to have an elevated prostate-specific antigen (PSA) of 12.5ng/mL leading to an incidental diagnosis of high grade prostate cancer. Targeted trans-perineal prostate biopsy found Gleason 4+5=9 acinar adenocarcinoma, without cribriform architecture but with features suspicious for extracapsular extension. Robotic radical prostatectomy with bilateral pelvic lymph node dissection was performed and found Gleason 4+5=9 adenocarcinoma with focal cribriform architecture, extra prostatic extension and clear margins, stage pT3a N0 M0. PSA was undetectable at 12 months, continence was immediate, and the patient reported strong erections soon after surgery. Family history of prostate cancer and genetic testing were both negative.Conclusion: This case highlights that not all clinically significant cancers will be identified by following PSA screening guidelines starting at 50 years of age (or 40 years of age for men with a family history of prostate cancer). While high grade prostate cancer in a man less than 40 years of age is uncommon, the literature suggests the incidence is increasing. Our case alongside series in the literature indicate that these men have better functional outcomes and equal oncological outcomes with early surgical intervention for localized disease when compared to the older population.Keywords: prostate cancer, prostatectomy, robotic surgery, young men, outcomes, case report

Zhizhuan Guo,1,2 Wenwen Zhong,3 Zhengshou Zou4 1Department of Neurology, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Third Hospital of Shanxi Medical University, Taiyuan, 030032, People’s Republic of China; 2Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People’s Republic of China; 3Department of Rehabilitation Medicine, Huangshi Maternal and Child Health Hospital, Edong Medical Group, Huang Shi, Hubei, 435000, People’s Republic of China; 4Department of Neurology, Huangshi Central Hospital, Edong Medical Group, Huangshi, Hubei, 435000, People’s Republic of ChinaCorrespondence: Zhengshou Zou, Department of Neurology, Huangshi Central Hospital, Edong Medical Group, 141 Tianjin Road, Huangshi, Hubei, 435000, People’s Republic of China, Email zouzhengshouhsch@163.comPurpose: Epilepsy is a serious mental disease, for which oxidative stress and hippocampal neuron death after seizure is crucial. Numerous miRNAs are involved in epilepsy. However, the function of miR-98-5p in oxidative stress and hippocampal neuron death after seizure is unclear, which is the purpose of current study.Methods: Magnesium ion (Mg2+)-free solution was used to establish the in vitro epilepsy model in hippocampal neurons. Oxidative stress was exhibited by measuring malondialdehyde (MDA) level and superoxide Dismutase (SOD) activity using enzyme-linked immune sorbent assay (ELISA) kits. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry were applied for the examination of neuron viability and apoptosis, respectively. Quantitative reverse‐transcription polymerase chain reaction (qRT-PCR) and Western blot were used to evaluate the mRNA and protein levels of miR-98-5p and signal transducer and activator of transcription (STAT3), respectively. The relationship between miR-98-5p and STAT3 was predicted by TargetScan 7.2, and identified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay.Results: miR-98-5p was decreased in the in vitro epileptic model of hippocampal neurons induced by Mg2+-free solution, whose overexpression rescued oxidative stress and neuron apoptosis in epileptic model. Moreover, overexpression of STAT3, one downstream target of miR-98-5p, partially eliminated the effects of miR-98-5p mimic.Conclusion: We shed lights on a pivotal mechanism of miR-98-5p in regulating neuron oxidative stress and apoptosis after seizures, providing potential biomarkers for the diagnosis of epilepsy and therapeutic targets for the treatment of epilepsy.Keywords: epilepsy, oxidative stress, apoptosis, miR-98-5p, STAT3

Qian Fan,1,&ast; Zhongmei Bai,2,&ast; Fabrice Yves Ndjana Lessomo,3,&ast; Bingqing Dong,1 Weiqin Zhong,1 Fenglin Jin,1 Zhiquan Wang3 1Department of Cardiology, The Second Affiliated Hospital of Shandong First Medical University, Taian, Shandong Province, People’s Republic of China; 2Department of Cardiology, The People’s Hospital of Huangmei, Huanggang, Hubei Province, People’s Republic of China; 3Department of Cardiology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei Province, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zhiquan Wang; Fenglin Jin, Email 2839515516@qq.com; cardio7788@163.comBackground: Low volume change and minimal trauma observed during angiography are the reason why physicians often overlook any changes affecting pre-operative electrolytes levels after coronary intervention. However, few studies have addressed the issue of electrolyte changes after the coronary intervention. Therefore, our study investigates coronary angiography’s effect on electrolytes and provides the quick identification of groups more prone to electrolyte changes.Methods: From the department of cardiology of the second affiliated hospital of Shandong’s first medical university, 374 patients undergoing coronary angiography were selected. Pre-intervention and post-intervention serums, sodium (Na+), potassium (K+), chloride (Cl−), magnesium (Mg2+) and renal function were analyzed. The correlation between influential factors was also assessed. The association of hypokalemia with short-major adverse cardiac events (MACE) and arrhythmia was evaluated.Results: Among the 374 subjects including 264 patients who had a simple angiography and 110 patients who received coronary artery interventional therapy. A decrease in potassium levels was found in 81.8% of the patients, and post-interventional hypokalemia was observed in 15.0%. After the intervention, the hypokalemia among males was 2.18 times than that of females, and the pre-operative serum potassium level was 3.5mmol/L≤K+< 4.0mmol/L and was 2.09 times than that of K+≥ 4.0 mmol/L, but was not associated with age and either simple coronary angiography or PCI (percutaneous coronary intervention). Hypernatremia was also prevalent in males under 60 years and with pre-operative hypernatremia. Significant variations were found between hypokalemia and influential factors like hypertension, diabetes, and gastrointestinal disease. We also found that there was no obvious correlation between hypokalemia and recurrent angina, heart failure and death, but significantly increased the risk of some arrhythmias.Conclusion: Male patients are more likely to suffer from electrolyte disturbance after coronary intervention. There is a need to emphasize monitoring and managing electrolyte changes to prevent severe complications in the peri-operative period.Keywords: coronary angiography, electrolytes, hypokalemia, hypernatremia, risk factors

Xiaojia Liu,1,2 Caiyan Lin,1 Wenfei Zhong,1 Zhongwen Yuan,1 Pengke Yan,1 Shixia Guan2 1Department of Pharmacy, Biomedicine Research Center, Guangdong Provincial Key Laboratory of Major Obstetric Diseases, Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, People’s Republic of China; 2School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, People’s Republic of ChinaCorrespondence: Pengke Yan; Shixia Guan, Email gysyypk@126.com; drguan@gzucm.edu.cnBackground: The activation of lymphatic vessel function is the crux to resolving atherosclerosis (AS), a chronic inflammatory disease. Rapamycin (RAPA) recently has attracted considerable attention as a potent drug to induce atherosclerotic plaque attenuation. The objective of this work was to develop a ligand-decorated, RAPA-loaded liposome for lymphatic-targeted delivery of drugs to improve abnormal lymphatic structure and function, resulting in highly effective regression of atherosclerotic plaques.Methods: Hyaluronic acid-decorated, RAPA-loaded liposomes (HA-RL) were fabricated by emulsion-solvent evaporation. The average size, zeta potential, entrapment efficiency were characterized, and the stability and drug release in vitro were investigated. Furthermore, the in vitro and in vivo lymphatic targeting ability were evaluated on lymphatic endothelial cells and LDLR−/− mice, and the efficiency of this nano-system in inducing the attenuation of atherosclerotic plaques was confirmed.Results: HA-RL had a size of 100 nm, over 90% drug encapsulation efficiency, the storage stability was distinguished, demonstrating a slow release from the lipid nano-carriers. The mean retention time (MRT) and elimination half-life (t1/2β) achieved from HA-RL were 100.27± 73.08 h and 70.74± 50.80 h, respectively. HA-RL acquired the most prominent efficacy of lymphatic-targeted delivery and atherosclerotic plaques attenuation, implying the successful implementation of this novel drug delivery system in vivo.Conclusion: HA-RL exhibited the most appreciable lymphatic targeting ability and best atherosclerotic plaques attenuation efficiency, opening a new paradigm and promising perspective for the treatment of arteriosclerosis.Keywords: rapamycin, hyaluronic acid, lymphatic targeting, arteriosclerosis, liposomes

Wenjia Fan,1,2,&ast; Jie Li,1,2,&ast; Lingxia Chen,2 Wenhao Wu,2 Xi Li,3 Weihong Zhong,1 Hongying Pan2 1College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People’s Republic of China; 2Department of Infectious Disease, Zhejiang Provincial People’s Hospital, Hangzhou, 310014, People’s Republic of China; 3Laboratory Medicine Center, Department of Clinical Laboratory, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital, Hangzhou Medical College, Hangzhou, Zhejiang, 310014, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Hongying Pan; Weihong Zhong, Tel/Fax +86-571-8589-3602 ; +86-571-88813378, Email hypanzjsrmyy@126.com; whzhong@zjut.edu.cnPurpose: Time-consuming culture methods and wet-mount microscopy (WMM) with low sensitivity have difficulties in diagnosing Vulvovaginal candidiasis (VVC). Rapid and highly sensitive polymerase chain reaction coupled with quantum dot fluorescence analysis (PCR-QDFA) for the diagnosis of VVC has not been reported to date. This study was the first to evaluate the performance of PCR-QDFA for diagnosis of Candida strains in the leukorrhea samples from patients with suspected VVC.Patients and Methods: Leukorrhea samples from all visited patients were taken from the vagina using vaginal swabs by clinicians. We evaluated patients admitted with suspected VVC who completed WMM for diagnosis and reported the diagnostic effectiveness of PCR-QDFA and Candida culture (gold standard) when testing leucorrhea samples.Results: A total of 720 leukorrhea samples from 387 VVC-positive patients and 333 VVC-negative patients were included in this study. Of the 387 leukorrhea samples from the VVC-positive patients, 391 Candida strains were identified by culture. 99.23% (388/391) Candida strains were included in the PCR-QDFA list. The 388 Candida strains belonged to four different species of Candida, including C. albicans (n = 273, 70.36%), C. glabrata (n = 85, 21.91%), C. tropicalis (n = 16, 4.12%), and C. krusei (n = 14, 3.61%). PCR-QDFA diagnosed Candida strains in 340/384 (88.54%) of the leucorrhea samples with Candida strains infection. The sensitivity of PCR-QDFA for C. albicans, C. glabrata, C. tropicalis, and C. krusei was 89.01%, 85.88%, 81.25% and 92.86%, respectively. The specificity of PCR-QDFA for C. albicans, C. glabrata, C. tropicalis and C. krusei was 93.69%, 99.37%, 99.71%, and 99.57%, respectively.Conclusion: The highly sensitive and specific PCR-QDFA technique can be exploited as a rapid (approximately 4 h) diagnostic tool for common Candida strains of leucorrhea samples from patients with suspected VVC.Keywords: PCR-QDFA, VVC, NAC, diagnostic performance, Candida culture

Poyu Chen1– 3&ast;, Chuangwei Zhang1– 3&ast;, Ping He,4 Shengyuan Pan,1– 3 Wenjie Zhong,1– 3 Yue Wang,1– 3 Qingyue Xiao,1– 3 Xinyan Wang,1– 3 Wenliang Yu,1– 3 Zhangmin He,1– 3 Xiang Gao,1– 3 Jinlin Song1– 3 1College of Stomatology, Chongqing Medical University, Chongqing, 401147, People’s Republic of China; 2Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing, 401147, People’s Republic of China; 3Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing, 401147, People’s Republic of China; 4Department of Stomatology, Dazhou Central Hospital, Dazhou, SiChuan, 635000, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xiang Gao; Jinlin Song, Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, Chongqing, 401147, People’s Republic of China, Tel/Fax +86 23 88860105 ; Tel/Fax +86 23 88860026, Email xiangg@hospital.cqmu.edu.cn; songjinlin@hospital.cqmu.edu.cnIntroduction: The functional reconstruction of periodontal tissue defects remains a clinical challenge due to excessive and prolonged host response to various endogenous and exogenous pro-inflammatory stimuli. Thus, a biomimetic nanoplatform with the capability of modulating inflammatory response in a microenvironment-responsive manner is attractive for regenerative therapy of periodontal tissue.Methods: Herein, a facile and green design of engineered bone graft materials was developed by integrating a biomimetic apatite nanocomposite with a smart-release coating, which could realize inflammatory modulation by “on-demand” delivery of the anti-inflammatory agent through a pH-sensing mechanism.Results: In vitro and in vivo experiments demonstrated that this biocompatible nanoplatform could facilitate the clearance of reactive oxygen species in human periodontal ligament stem cells under inflammatory conditions via inhibiting the production of endogenous proinflammatory mediators, in turn contributing to the enhanced healing efficacy of periodontal tissue. Moreover, this system exhibited effective antimicrobial activity against common pathogenic bacteria in the oral cavity, which is beneficial for the elimination of exogenous pro-inflammatory factors from bacterial infection during healing of periodontal tissue.Conclusion: The proposed strategy provides a versatile apatite nanocomposite as a promising “inflammation scavenger” and propels the development of intelligent bone graft materials for periodontal and orthopedic applications.Graphical Abstract: Keywords: periodontal bone regeneration, pH-responsive, inflammatory modulation, antibacterial activity, biomimetic nanoplatform

Yale Yang,1 Wuling Zhong,1 Yali Zhang,1 Yalan Cheng,1 Hejin Lai,1 Huimin Yu,1 Ning Feng,1 Yumo Han,2 Rui Huang,1 Qiwei Zhai1,2 1CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, People’s Republic of China; 2School of Life Science and Technology, ShanghaiTech University, Shanghai, People’s Republic of ChinaCorrespondence: Qiwei Zhai, Shanghai, 200031, People’s Republic of China, Tel +86 21 5492 0903, Fax +86 21 5492 0291, Email qwzhai@sibs.ac.cnBackground: Sustained inflammation is implicated in a variety of pathological conditions like infection, obesity and type 2 diabetes. Lipid metabolism is crucial to support immune response during infection of bacteria. However, how sustained inflammation affects lipid metabolism, especially in white adipose tissue remains largely unknown.Methods: Sustained inflammation was induced by daily injection of Lipopolysaccharide (LPS). Tlr4 knockout mice were used to study the mechanism. Inflammation and lipid metabolism were evaluated by quantitative PCR, white blood cell counting, nuclear magnetic resonance, fat cell size quantification, lipolysis and fatty acid uptake assays, respiratory exchange ratio, and energy expenditure.Results: Here, we found that sustained inflammation leads to fat loss in mice with a quick loss and gradual increase manner. Moreover, LPS injection leads to inflammation, anorexia, decreased lipid anabolism, and increased lipid catabolism. Mechanically, we show that LPS induces fat loss, inflammation, anorexia, and alteration of lipid metabolism mainly dependent on Tlr4. Interestingly, sustained inflammation induces less fat loss, especially in epididymal white adipose tissue, than pair-feeding, and pair-feeding has no significant effect on inflammation and leads to less fatty acid uptake, more lipid catabolism and energy expenditure than LPS injection. In addition, we demonstrate that short-term sustained inflammation leads to relative long-term tolerance for LPS-induced anorexia, inflammation and altered lipid metabolism.Conclusion: These findings demonstrate that sustained inflammation induced by LPS leads to tolerable anorexia and fat loss via Tlr4 in mice, and provide new insights into the effect of sustained inflammation on lipid metabolism and subsequent tolerance.Keywords: sustained inflammation, lipopolysaccharide, white adipose tissue, lipid metabolism, tolerance

Jingyu Chen, Yanrong Yin, Yefa Zhang, Xiuquan Lin, Tiehui Chen, Ze Yang, Dengwei Wang, Wenling Zhong Fujian Provincial Center for Disease Control and Prevention, Fuzhou, Fujian Province, People’s Republic of China Correspondence: Wenling Zhong, Fujian Provincial Center for Disease Control and Prevention, No. 386 Chong’an Road, Jin’an District, Fuzhou, Fujian Province, 350012, People’s Republic of China, Tel +86-591-87539007, Email mbzwl@163.comPurpose: Chronic obstructive pulmonary disease (COPD) is one of many major public health problems in China, and its prevalence and associated risk factors in the southeast of China need to be determined to facilitate disease control and prevention.Methods: A multistage stratified cluster sampling method was used to select 5486 participants aged ≥ 40 years from nine COPD monitoring districts in Fujian Province during 2019– 2020. Participants were interviewed using a laptop-based questionnaire and underwent pulmonary function tests. COPD was diagnosed according to the 2019 Global Initiative for Chronic Obstructive Lung Disease (GOLD) criteria.Results: Final analysis was conducted using data from 4999 participants with qualified post-bronchodilator results. The prevalence of COPD was 11.6% (95% confidence interval [CI]: 10.5– 12.7). Risk factors for COPD in the logistic regression model were being male (odds ratio [OR] = 2.83, 95% CI: 2.01– 3.98), > 70 years old (OR = 16.16, 95% CI: 8.14– 32.08), having a low body mass index (BMI) (OR = 1.81, 95% CI: 1.13– 2.89), parental history of respiratory disease (OR = 1.78, 95% CI: 1.50– 2.10), being a current (OR = 2.82, 95% CI: 1.83– 4.36) or former (OR = 2.47, 95% CI: 1.45– 4.19) smoker, and indoor exposure to biomass (OR = 1.28, 95% CI: 1.05– 1.58).Conclusion: The estimated prevalence of COPD in southeast China is high. COPD was strongly associated with sex, aging, a low BMI, parental history of respiratory diseases, smoking, and indoor exposure to biomass in adults aged ≥ 40 years. The government should urgently implement comprehensive measures to reduce the risk factors for COPD.Keywords: COPD, logistic models, biomass, public health, smoking, surveys and questionnaires

Zongbo Zhao,&ast; Jianzhong Xue,&ast; Ziliang Zhuo, Wei Zhong, Hui Liu Department of Neurology and Institute of Neurology, The Affiliated Changshu Hospital of Xuzhou Medical University, Suchow, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Hui Liu, Tel/Fax +860512-52925530, Email laoniu20201111@163.comObjective: To investigate the association between IL7R rs6897932 and multiple sclerosis (MS) in southern Chinese people.Methods: In total, 147 MS patients and 530 healthy controls were recruited according to the revised McDonald criteria. The TaqMan method was used for genotyping.Results: With genetic models, we can observe that the additive model, the dominant model, and the recessive model of IL7R rs6897932 were significantly associated with MS [additive model: p=0.032; dominant model (adjusted): p< 0.001, OR=3.61 (95% CI 2.25– 5.83); recessive model (adjusted): p< 0.001, OR=6.80 (95% CI 3.49– 13.89)].Conclusion: Our results suggest that IL7R rs6897932 is associated with MS in a southern Chinese population. More and larger MS studies to explore the genetic risk factors of MS are warranted.Keywords: single-nucleotide polymorphism, multiple sclerosis, IL7R, interleukin

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

Weiwei Zhong,&ast; Chen Chen,&ast; Weixiang Tang Department of Anesthesiology, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui Province, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Weixiang Tang, Department of Anesthesiology, The First Affiliated Hospital of Anhui Medical University, No. 218 Jixi Road, Hefei, 230022, People’s Republic of China, Tel +86-551-62922384, Email vanillatang@163.comPurpose: Nalbuphine is becoming a common analgesic used in hysteroscopic operations. The aim of this study was to identify the median effective dose (ED50) and 95% effective dose (ED95) of nalbuphine combined with propofol in painless hysteroscopy.Patients and Methods: Twenty-five patients aged 18– 60 years with an American Society of Anesthesiologists classification of I–II who were scheduled for painless hysteroscopy were recruited. The initial dose of nalbuphine was set at 0.15 mg/kg and varied by 0.01 mg/kg according to the Dixon sequential method. The ED50/ED95 of nalbuphine combined with propofol for hysteroscopy was calculated by the probit method.Results: The ED50 of nalbuphine was 0.122 (95% confidence interval (CI) 0.092– 0.137) mg/kg, and the ED95 of nalbuphine was 0.153 (95% CI 0.138– 0.361) mg/kg.Conclusion: The ED50/ED95 values of nalbuphine combined with propofol in painless hysteroscopy are 0.122 mg/kg and 0.153 mg/kg, respectively. Nalbuphine at 0.153 mg/kg combined with propofol is effective and safe for painless hysteroscopy.Keywords: nalbuphine, propofol, hysteroscopy, effective dose, ED50, ED95

Junyi Jiang,1– 5,&ast; WeiJun Zhong,1– 4,&ast; WeiHua Huang,1– 4 Yongchao Gao,1– 4 Yijing He,1– 4 Xi Li,1– 4 Zhaoqian Liu,1– 4 Honghao Zhou,1– 4 Yacheng Fu,1,6 Rong Liu,1– 4 Wei Zhang1– 4 1Department of Clinical Pharmacology, Xiangya Hospital, Central South University, Changsha, People’s Republic of China; 2Institute of Clinical Pharmacology, Central South University, Hunan Key Laboratory of Pharmacogenetics, Changsha, People’s Republic of China; 3Engineering Research Center of Applied Technology of Pharmacogenomics, Ministry of Education, Changsha, People’s Republic of China; 4National Clinical Research Center for Geriatric Disorders, Changsha, Hunan, People’s Republic of China; 5Aier Eye Institute, Changsha, Hunan, People’s Republic of China; 6Cofoe Medical Technology Co., Ltd, Changsha, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Wei Zhang; Rong Liu, Department of Clinical Pharmacology, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China, Tel +86 731 84805380, Fax +86 731 82354476, Email csuzhangwei@csu.edu.cn; csuliurong@csu.edu.cnPurpose: To identify more objectively predictive factors of severe outcome among patients hospitalized for coronavirus disease 2019 (COVID-19).Patients and Methods: A retrospective cohort of 479 hospitalized patients diagnosed with COVID-19 in Hunan Province was selected. The prognostic effects of factors such as age and laboratory indicators were analyzed using the Kaplan–Meier method and Cox proportional hazards model. A prognostic nomogram model was established to predict the progression of patients with COVID-19.Results: A total of 524 patients in Hunan province with COVID-19 from December 2019 to October 2020 were retrospectively recruited. Among them, 479 eligible patients were randomly assigned into the training cohort (n = 383) and validation cohort (n = 96), at a ratio of 8:2. Sixty-eight (17.8%) and 15 (15.6%) patients developed severe COVID-19 after admission in the training cohort and validation cohort, respectively. The differences in baseline characteristics were not statistically significant between the two cohorts with regard to age, sex, and comorbidities (P > 0.05). Multivariable analyses included age, C-reactive protein, fibrinogen, lactic dehydrogenase, neutrophil-to-lymphocyte ratio, urea, albumin-to-globulin ratio, and eosinophil count as predictive factors for patients with progression to severe COVID-19. A nomogram was constructed with sufficient discriminatory power (C index = 0.81), and proper consistency between the prediction and observation, with an area under the ROC curve of 0.81 and 0.86 in the training and validation cohort, respectively.Conclusion: We proposed a simple nomogram for early detection of patients with non-severe COVID-19 but at high risk of progression to severe COVID-19, which could help optimize clinical care and personalized decision-making therapies.Keywords: nomogram, COVID-19, SARS-CoV-2, C-reactive protein, predictive model

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

Junmu Peng,1,2,&ast; Jie Qin,1,&ast; Tianji Huang,1 Xiaoji Luo,1 Weiyang Zhong,1 Zhengxue Quan1 1Department of Orthopedic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, People’s Republic of China; 2Department of Orthopedic Surgery, The Ninth People’s Hospital of Chongqing, Chongqing, 400799, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Weiyang Zhong Email 492467112@qq.comBackground: A retrospective study aimed to introduce a new method for improving the diffusion degree of bone cement and to observe its clinical efficacy in percutaneous vertebroplasty treating osteoporotic vertebral compression fractures (OVCFs).Methods: From January 2019 to March 2020, a total of 83 patients were enrolled and reviewed. The patients were divided into two groups according to the operation method. The clinical and radiographic parameters were recorded and compared between these two groups. Those who received percutaneous vertebroplasty with haemorrhage aspiration were recorded as group A (n=42). In group A, the haemorrhage in the vertebral fracture was aspirated compared with conventional percutaneous vertebroplasty. Patients who underwent conventional percutaneous vertebroplasty were classified as group B (n=41).Results: Visual analogue scale (VAS) values and Oswestry Disability Index (ODI) scores showed no significant difference between the two groups preoperatively, postoperatively or at the final follow-up (FU) (P> 0.05). The intraoperative VAS score (bone cement injection) in group A was significantly lower than that in group B (3.83± 0.79 vs 5.44± 1.32, P < 0.01). The local kyphotic angle (LKA) (final follow-up), LKA loss, fractured vertebral anterior height loss (FVAHL) and anterior vertebral height loss ratio (AVHLR) were significantly lower in group A than in group B. The anterior vertebral height ratio (AVHR) at the final FU in group A was higher than that in group B (P=0.013). The distribution of bone cement was significantly different (P=0.034). By analysing the distribution pattern of bone cement, it was found that the values of LKA loss, FVAHL and AVHLR were superior in the type A bone cement distribution to those in types B and C.Conclusion: Compared with traditional surgical methods, bone haemorrhage aspiration could improve the diffusion degree of bone cement and reduce the height loss and deformity of injured vertebrae. This method provides a feasible new scheme for improving the dispersion of bone cement.Keywords: osteoporotic vertebral compression fractures, percutaneous vertebroplasty, bone cement distribution, fracture haemorrhage aspiration

Wenhui Zhong,1,&ast; Chunyu Zhou,2,&ast; Lufei Chen,1 Zhenna Wang,3 Hongxing Lin,1 Kunhai Wu,1 Sujiao Zhang4 1Clinical Laboratory, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, People’s Republic of China; 2Nursing Department, Fujian Medical University Union Hospital, Fuzhou, People’s Republic of China; 3Department of Gynecology, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, People’s Republic of China; 4Department of Pathology, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Wenhui ZhongClinical Laboratory, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, No. 18 Daoshan Road, Fuzhou, 350001, Fujian, People’s Republic of ChinaEmail 289492316@qq.comPurpose: This study assessed the prognostic value of red blood cell distribution width (RDW) and cancer antigen 125 (CA125) in predicting the prognosis of endometrial cancer (EC) patients.Patients and Methods: In this study, we included 525 patients with EC between January 2013 and January 2019. Demographic and clinical indicators were collected, and the receiver operating characteristics curve (ROC) and cutoff values were calculated between the early and advanced stages of EC. Independent risk factors associated with EC prognosis were assessed using Cox regression analyses and the Kaplan–Meier method.Results: Compared to women in the early stage of EC, women with advanced stage had significantly elevated RDW coefficient of variation (RDW-CV) and CA125 levels and lower mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) (both P < 0.05). Consequently, RDW-CV and CA125 were found to be independent risk factors for EC by using ROC curve and multivariate logistic regression analysis. The survival analysis curve was used to assess the diagnostic value of RDW-CV, CA125, and their combination in the prognosis of EC. The results showed that patients with high expression of RDW-CV and CA125 had worse overall survival than those with low expression. Moreover, multivariate Cox regression analysis indicated that RDW-CV+CA125=2 was an independent prognostic factor.Conclusion: These findings suggest that CA125 combined with RDW-CV has a good prognostic value for EC. Thus, the RDW-CV+CA125 score is a promising prognostic marker for the clinical decision-making process regarding EC outcomes.Keywords: CA125, RDW-CV, endometrial cancer, prognosis

Ruijie Wang,1,2,* Wenliang Song,3,* Chengyuan Xie,2,4,* Wenhong Zhong,2 Hui Xu,2,5 Qiuping Zhou,1,2 Yiyu Deng,2 Yimei Hong,2 Xin Li,2 Ming Fang1,2 1Department of Intensive Care Unit, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, School of Medicine, South China University of Technology, Guangzhou, People’s Republic of China; 2Department of Emergency and Critical Care Medicine, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, People’s Republic of China; 3Department of Critical Care Medicine, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510080, People’s Republic of China; 4The Second School of Clinical Medicine, Southern Medical University, Guangzhou, People’s Republic of China; 5Shantou University Medical College, Shantou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Ming FangDepartment of Intensive Care Unit, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, School of Medicine, South China University of Technology, Guangzhou, 510080, People’s Republic of ChinaTel +8613527774075Email drfangming@163.comBackground: Our previous study found that urinary trypsin inhibitor (ulinastatin, UTI) protected tight junctions (TJs) of lung endothelia via TNF-α inhibition, thereby alleviating pulmonary capillary permeability in septic rats. As the activated macrophage is the main source of TNF-α in sepsis, we speculate that UTI may exert the above effects by regulating the functions of macrophages.Methods: Bone-marrow derived macrophages (BMDM) were divided into control, lipopolysaccharide (LPS), UTI+LPS and UTI groups. TNF-α, TGF-β, IL-10, CD86, CD206 and MCP-1 expression were assessed by Western blot. The phagocytosis and migration of BMDM were detected. Pulmonary microvascular endothelial cells (PMVECs) were cultured with the conditioned medium (CM) from each group of BMDM above. Sprague-Dawley rats were divided into sham, cecal ligation and puncture (CLP), and UTI+CLP groups. Western blot and immunofluorescence were used to detected zonula occludens-1 (ZO-1), occludin and claudin-5 expression in PMVECs, as well as TNF-α, TGF-β, iNOS, CD86 and CD206 expression in lungs. Pulmonary capillary permeability was assessed by extravasated Evans blue, lung injury score (LIS), wet-to-dry weight ratio and electron microscope.Results: TNF-α and CD86 expression were increased in LPS-treated BMDM, but were reversed by UTI pretreatment. TGF-β, IL-10 and CD206 expression were the opposite. UTI markedly decreased phagocytosis and migration of LPS-treated BMDM. ZO-1, occludin and claudin-5 expression were markedly decreased in PMVECs of the CM-LPS group, but significantly increased in the CM-UTI+LPS group. TNF-α, iNOS and CD86 expression were increased in the lungs of CLP-rats but decreased with UTI pretreatment, while TGF-β and CD206 expression were the opposite. UTI markedly ameliorated the lung EB leakage, improved LIS, reduced the wet-to-dry ratio and revised the damaged TJs of PMVECs in CLP-rats.Conclusion: UTI effectively inhibits the conversion of M1 macrophage but increases M2, reduces the phagocytosis and migration, which helps to protect endothelia TJs and reduce pulmonary capillary permeability during sepsis.Keywords: ulinastatin, sepsis, macrophage, pulmonary capillary endothelial cells, tight junctions

Wanyi Lin,1,2,* Yaping Yang,1,2,* Wenjing Zhong,1,2 Qun Lin,1,2 Nanyan Rao,1– 3 Gehao Liang,1,2 Yun Ling,1,2 Zihao Liu,1,2 Qing Luo,1,2 Zhenluan Tian,1,2 Chang Gong1,2,4 1Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, People’s Republic of China; 2Department of Breast Surgery, Breast Tumor Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, People’s Republic of China; 3Department of Oncoplastic and Reconstruction Breast Surgery, Breast Tumor Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, People’s Republic of China; 4Bioland Laboratory, Guangzhou Regenerative Medicine and Health Guangdong Laboratory, Guangzhou, 510005, People’s Republic of China*These authors contributed equally to this workCorrespondence: Chang GongDepartment of Breast Surgery, Breast Tumor Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, 107 Yanjiang West Road, Guangzhou 510120, People’s Republic of ChinaEmail gchang@mail.sysu.edu.cnBackground: Vacuum drains have been extensively applied to prevent seroma formation after breast surgery. However, the usage of negative suction drainage is mainly determined by surgeon’s experience and preferences. The aim of this study is to prospectively compare the drain effect after breast surgery between the low and high vacuum drains.Methods: This prospectively randomized trial (from January 2018 to June 2019) involved 188 patients who were subjected to modified radical mastectomy (group A, n=128) or immediate breast reconstruction with implants (group B, n=60). In each group, patients were randomized to receive high vacuum drain (pressure=– 98 kPa) or low vacuum drain (pressure=– 12 kPa) after surgery. Days of drain permanence, which means the duration of drainage, was the primary endpoint.Results: According to the comparison of days of drain permanence, the effect of a low vacuum drain is not inferior to a high vacuum drain in group A (pectoral drain, P< 0.001; axillary drain, P< 0.001) or group B (submuscular drain, P=0.002). The complications frequently occurred on patients with high vacuum drain (11.7%), such as seroma formation. The expense of low vacuum drain was significantly lower than high vacuum drain in both groups (P< 0.01).Conclusion: The drain effect of the low vacuum drain is not inferior to a high vacuum drain in both group A and group B. The low vacuum drain was effective, relatively cheap, and did not increase the incidence of complications; it is therefore more recommended after breast surgery.Keywords: breast surgery, days of drain permanence, postoperative drain

Fan Yang,1 Lu Deng,2 JinPeng Li,3 MuHu Chen,1 Ying Liu,1 YingChun Hu,1 Wu Zhong1 1Department of Emergency Medicine, Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, People’s Republic of China; 2Department of Thyroid Surgery, Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, People’s Republic of China; 3Department of Thyroid and Breast Surgery, Wuhan University Zhongnan Hospital, Wuhan, Hubei 430071, People’s Republic of ChinaCorrespondence: Wu Zhong Department of Emergency MedicineAffiliated Hospital of Southwest Medical University, 25 Taiping Street, Luzhou, Sichuan 646000, People’s Republic of ChinaEmail zhongwu3987@126.comBackground: Renal fibrosis is a frequently occurring type of chronic kidney disease that can cause end-stage renal disease. It has been verified that emodin or HGF can inhibit the development of renal fibrosis. However, the antifibrotic effect of emodin in combination with HGF remains unclear.Methods: Cell viability was detected with CCK8. Gene and protein expression in HK2 cells was detected by qRT-PCR and Western blot, respectively. Moreover, a unilateral ureteral obstruction–induced mouse model of renal fibrosis was established for investigating the antifibrotic effect of emodin in combination with HGF in vivo.Results: HGF notably increased the expression of collagen II in TGFβ-treated HK2 cells. In addition, HGF-induced increase in collagen II expression was further enhanced by emodin. In contrast, fibronectin, αSMA and Smad2 expression in TGFβ-stimulated HK2 cells was significantly inhibited by HGF and further decreased by combination treatment (emodin plus HGF). Moreover, we found that combination treatment exhibited better antifibrotic effects compared with emodin or HGF in vivo.Conclusion: These data demonstrated that emodin plus HGF exhibited better antifibrotic effects compared with emodin or HGF. As such, emodin in combination with HGF may serve as a new possibilty for treatment of renal fibrosis.Keywords: fibrosis, combination, emodin, TGFβ

Dongxing Zhao1,2 ,* Asghar Abbasi2 ,* Harry B Rossiter,2,3 Xiaofen Su,1 Heng Liu,1 Yuhong Pi,1 Li Sang,1 Weiyong Zhong,1 Qifeng Yang,1 Xiongtian Guo,1 Yanyan Zhou,1 Tianyang Li,1 Richard Casaburi,2 Nuofu Zhang1 1State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510120, People’s Republic of China; 2Rehabilitation Clinical Trials Center, The Lundquist Institute for Biomedical Innovation at Harbor-UCLA Medical Center, Torrance, CA, 90502, USA; 3Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, UK*These authors contributed equally to this workCorrespondence: Nuofu ZhangState Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510120, People’s Republic of ChinaEmail nfzhanggird@163.comBackground: We sought to determine whether circulating inflammatory biomarkers were associated with the frequent exacerbator phenotype in stable COPD patients ie, those with two or more exacerbations in the previous year.Methods: Eighty-eight stable, severe, COPD patients (4 females) were assessed for exacerbation frequency, pulmonary function, fraction of expired nitric oxide (FENO); inflammatory variables were measured in venous blood. Logistic regression assessed associations between the frequent exacerbator phenotype and systemic inflammation.Results: Compared with infrequent exacerbators, frequent exacerbators (n=10; 11.4%) had greater serum concentration (median (25th-75th quartile)) of serum amyloid A (SAA; 134 (84– 178) vs 71 (38– 116) ng/mL; P=0.024), surfactant protein D (SP-D; 15.6 (9.0– 19.3) vs 8.5 (3.6– 14.9) ng/mL; P=0.049) and interleukin-4 (IL-4; 0.12 (0.08– 1.44) vs 0.03 (0.01– 0.10) pg/mL; P=0.001). SAA, SP-D and IL-4 were not significantly correlated with FEV1%predicted or FVC %predicted. After adjusting for sex, age, BMI, FEV1/FVC and smoking pack-years, only SAA remained independently associated with the frequent exacerbator phenotype (OR 1.49[1.09– 2.04]; P=0.012). The odds of being a frequent exacerbator was 18-times greater in the highest SAA quartile (≥ 124.1 ng/mL) than the lowest SAA quartile (≤ 44.1 ng/mL) (OR 18.34[1.30– 258.81]; P=0.031), and there was a significant positive trend of increasing OR with increasing SAA quartile (P=0.008). For SAA, the area under the receiver operating characteristic curve was 0.721 for identification of frequent exacerbators; an SAA cut-off of 87.0 ng/mL yielded an 80% sensitivity and 61.5% specificity.Conclusion: In stable COPD patients, SAA was independently associated with the frequent exacerbator phenotype, suggesting that SAA may be a useful serum biomarker to inform progression or management in COPD.Keywords: inflammation, surfactant protein D, interleukin-4

Weimin Dong,1 Yong Luo,2 Guian Zhang,1 Hui Zhang,1 Yuxiang Liang,1 Yangjia Zhuo,1 Yingke Liang,1 Fen Zou,1 Weide Zhong1,3,4 1Department of Urology, Guangdong Key Laboratory of Clinical Molecular Medicine and Diagnostics, Guangzhou First People’s Hospital, School of Medicine, South China University of Technology, Guangzhou, Guangdong 510180, People’s Republic of China; 2Department of Urology, The Second People’s Hospital of Foshan, Affiliated Foshan Hospital of Southern Medical University, Foshan 528000, People’s Republic of China; 3Department of Urology, Huadu District People’s Hospital, Southern Medical University, Guangzhou 510800, People’s Republic of China; 4Urology Key Laboratory of Guangdong Province, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou Medical University, Guangzhou 510230, People’s Republic of ChinaCorrespondence: Weide Zhong; Fen ZouDepartment of Urology, Guangdong Key Laboratory of Clinical Molecular Medicine and Diagnostics, Guangzhou First People’s Hospital, School of Medicine, South China University of Technology, Guangzhou 510180, People’s Republic of ChinaTel +86 20-81048312Fax +86 20-8337332Email zhongwd2009@live.cn; zoufen2008@126.comIntroduction: Although carbon nanospheres (CNPs) are promising nanomaterials in cancer treatment, how they affect prostate cancer (PCa) remains unclear.Methods: In this study, scanning electron microscopy (SEM), X-ray diffraction (XRD), and Raman spectroscopy were used to confirm the successful synthesis of CNPs. CCK-8, flow cytometry, Transwell, wound healing, Western blot and immunohistochemistry (IHC) assays were performed to evaluate the antitumor effect of CNPs toward the two kinds of prostate cancer cell lines PC3 and DU145.Results: Our results showed that CNPs inhibited cell growth, invasion, and migration and induced apoptosis and autophagy in PCa cells. Multifactor detection of a single Akt phosphorylation pathway and Western blot results suggested the suppression of 4E-BP1 in PCa cells after incubation with CNPs. The results from animal experiments also suggested the antitumor effect of CNPs and reduced 4E-BP1 expression in PCa tissue samples from BALB/c nude mice administered a local subcutaneous injection of CNPs.Keywords: carbon nanosphere, prostate cancer, antitumor, 4E-BP1

Yun Zhu,* Ao Lin,* Yi Zheng, Xiaoli Xie, Qiuming He, Wei Zhong Department of Pediatric Surgery, Guangzhou Institute of Pediatrics, Guangdong Provincial Key Laboratory of Research in Structural Birth Defect Disease, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou, Guangdong 510623, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wei ZhongDepartment of Pediatric Surgery, Guangzhou Institute of Pediatrics, Guangdong Provincial Key Laboratory of Research in Structural Birth Defect Disease, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou, Guangdong 510623, People’s Republic of ChinaTel/ Fax +86-18902268667Email zhongwei@gwcmc.orgBackground: Hirschsprung disease (HSCR) is a rare congenital gastrointestinal disease characterized by the absence of intestinal submucosal and myometrial ganglion cells. Recently, researches indicated that miR-100 regulated the growth, differentiation and apoptosis of neurons, and affected the functions of HSCR-associated pathways. While miR-100 rs1834306 A>G polymorphism was shown to modify the susceptibility to tumors, the association between this polymorphism and HSCR susceptibility is still unknown.Methods: This was a case–control study consisting of 1470 HSCR cases and 1473 controls from southern China. DNA was genotyped by TaqMan real-time PCR. Odds ratios (ORs) and 95% confidence intervals (CIs) were used as statistical indicators.Results: We found that miR-100 rs1834306 G allele and GG genotype significantly increased HSCR susceptibility (GG vs AA: adjusted OR=1.31, 95% CI=1.04– 1.64, P=0.020; G vs A: adjusted OR=1.12, 95% CI=1.01– 1.25, P=0.041; GG vs AA/AG: adjusted OR=1.30, 95% CI=1.07– 1.59, P=0.010). In the stratified analysis, miR-100 rs1834306 GG genotype carriers had higher risk to develop HSCR in all clinical subtypes when compared with those with AA/AG genotypes, and OR was rising with HSCR aggravation (SHSCR: adjusted OR=1.28, 95% CI=1.03– 1.59, P=0.029; LHSCR: adjusted OR=1.48, 95% CI=1.06– 2.07, P=0.020; TCA: adjusted OR=2.12, 95% CI=1.22– 3.69, P=0.008).Conclusion: Our findings suggested that miR-100 rs1834306 A>G polymorphism was associated with increased HSCR susceptibility in southern Chinese children. Furthermore, miR-100 rs1834306 GG genotype had a greater genetic pathopoiesis in severe HSCR.Keywords: Hirschsprung disease, miR-100, polymorphism, susceptibility

Zhiduan Cai,1,* Yulin Deng,1,* Jianheng Ye,2 Yangjia Zhuo,3 Zezhen Liu,2 Yingke Liang,3 Hui Zhang,3 Xuejin Zhu,2 Yong Luo,3 Yuanfa Feng,4 Ren Liu,2 Guo Chen,5 Yongding Wu,3 Zhaodong Han,3 Yuxiang Liang,3 Funeng Jiang,3 Weide Zhong1– 4,6 1Guangdong Provincial Institute of Nephrology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, People’s Republic of China; 2Department of Urology, Guangdong Key Laboratory of Clinical Molecular Medicine and Diagnostics, Guangzhou First People’s Hospital, Guangzhou Medical University, Guangzhou 510180, People’s Republic of China; 3Department of Urology, Guangdong Key Laboratory of Clinical Molecular Medicine and Diagnostics, Guangzhou First People’s Hospital, School of Medicine, South China University of Technology, Guangzhou 510180, People’s Republic of China; 4Urology Key Laboratory of Guangdong Province, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou Medical University, Guangzhou 510230, People’s Republic of China; 5Department of Urology, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong 510632, People’s Republic of China; 6School of Medicine, Jinan University, Guangzhou, Guangdong 510632, People’s Republic of China*These authors contributed equally to this workCorrespondence: Weide ZhongGuangdong Provincial Institute of Nephrology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, People’s Republic of ChinaEmail zhongwd2009@live.cnPurpose: Citrate synthase (CS) is a rate-limiting enzyme in the citrate cycle and is capable of catalyzing oxaloacetate and acetyl-CoA to citrate. CS has been uncovered to be upregulated in a variety of cancers, and its expression and clinical significance in prostate cancer (PCa) remain unknown.Methods: In this study, we examined the association between CS expression level and clinicopathological features of prostate cancer patients in a TMA cohort and the public cancer database (The Cancer Genome Atlas-Prostate Adenocarcinoma, TCGA-PRAD). The CS knockdown cell lines were constructed to study the effects of CS downregulation on proliferation, colony formation, migration, invasion, and cell cycle of prostate cancer cells in vitro. And the effect of CS downregulation on tumor growth in mice was studied in vivo. In addition, the metabolomics and mitochondrial function were detected in the CS knockdown cell lines.Results: CS expression level in PCa tissues was higher than that in normal tissues (P < 0.05). CS upregulation was significantly associated with high Gleason score (P < 0.05), advanced pathological stage (P < 0.001), and biochemical recurrence (P < 0.001). Functionally, decreased expression of CS inhibited PCa cell proliferation, colony formation, migration, invasion and cell cycle in vitro, and inhibited tumor growth in vivo. In addition, CS downregulation exerted potential inhibitory effects on the lipid metabolism and mitochondrial function of PCa cells.Conclusion: In conclusion, these findings suggested that CS upregulation may contribute to the aggressive progression and poor prognosis of PCa patients, which might be partially associated with its influences on the cell lipid metabolism and mitochondrial function.Keywords: prostate cancer, citrate synthase, prognosis, lipid metabolism

Xu Zhou,1 Jun Chen,2 Hua Tao,1,2 Yujie Cai,1 Lidan Huang,1 Haihong Zhou,2 Yanyan Chen,1 Lili Cui,1 Wangtao Zhong,2 Keshen Li3 1Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524001, People’s Republic of China; 2Department of Neurology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong 524001, People’s Republic of China; 3Institute of Neurology, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, 524001 China; Stroke Center, Neurology & Neurosurgery Division, Clinical Medicine Research Institute & The First Affiliated Hospital, Jinan University, Guangzhou, Guangdong 510630, People’s Republic of ChinaCorrespondence: Hua Tao; Keshen LiAffiliated Hospital of Guangdong Medical University No. 57, Renmindadaonan Road, Xiashan District, Zhanjiang, Guangdong 524001, People’s Republic of ChinaTel/ Fax +86 759 2387 427; +86 759 2386 772 Email taohua1943@126.com; likeshen1971@126.comIntroduction: To confront the resistance to existing antiepileptic drugs, studies have gradually begun to investigate alternative pathologies distinct from the traditional treatments that overwhelmingly target ion channels. Microglia activation is the first inflammatory response in the brain, in which miR-155-5p plays a key proinflammatory role and thus represents a promising target for inflammatory modulation in epilepsy pathologies.Methods: In this study, a pentetrazol-induced acute seizure model was established, and the seizure degree was evaluated within 60 min after pentetrazol administration. Animals were then sacrificed for hippocampal tissue collection for biological experiments.Results: Intranasal delivery of miR-155-5p antagomir (30 min before pentetrazol administration) increased the percentage of animals with no induced seizures by 20%, extended the latency to generalized convulsions, and decreased seizure severity. In addition, miR-155-5p antagomir treatment alleviated hippocampal damage and decreased the expression of typical inflammatory modulators (TNF-α, IL-1β and IL-6). Further research revealed that intranasal delivery of miR-155-5p antagomir significantly decreased the relative level of miR-155-5p and increased the expression of its targets LXRα and SOCS1 in IBA1-labeled microglial cells in the hippocampus.Conclusion: These findings demonstrate that intranasal delivery of miR-155-5p antagomir alleviated acute seizures, likely by blocking hippocampal inflammation. However, other potential mechanisms of the effects of miR-155-5p antagomir and its long-term safety for epilepsy treatment remain to be investigated.Keywords: intranasal delivery, epilepsy, pentetrazol, inflammatory response, miR-155-5p

Yi Zheng,* Zhenjian Zhuo,* Xiaoli Xie, Lifeng Lu, Qiuming He, Wei Zhong Department of Pediatric Surgery, Guangzhou Institute of Pediatrics, Guangdong Provincial Key Laboratory of Research in Structural Birth Defect Disease, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou 510623, Guangdong, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wei ZhongDepartment of Pediatric Surgery, Guangzhou Institute of Pediatrics, Guangdong Provincial Key Laboratory of Research in Structural Birth Defect Disease, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, 9 Jinsui Road, Guangzhou 510623, Guangdong, People’s Republic of ChinaTel/Fax +86-18902268667Email zhongwei_gwcmc@163.comBackground: Hirschsprung disease (HSCR) is a congenital disease that arises from defective intestinal neural system. LncRNA HOTTIP is a critical gene in various diseases, including HSCR. No epidemiological studies have explored the correlation between lncRNA HOTTIP single nucleotide polymorphisms (SNPs) and HSCR risk. We here lead as a pioneer to explore whether SNPs in lncRNA HOTTIP impact the risk of HSCR and HSCR subtypes in an unrelated Chinese population.Methods: We used the TaqMan method to genotype rs3807598 C>G of the lncRNA HOTTIP gene using 1470 HSCR cases and 1473 healthy controls. Of them, 1441 cases and 1434 controls were successfully genotyped. We adopted odds ratios (ORs) and 95% confidence intervals (CIs) to quantify the relationship.Results: We got an unexpected outcome that lncRNA HOTTIP SNP rs3807598 C>G could not modify the risk of HSCR (CG vs. CC: adjusted OR=0.89, 95% CI=0.74– 1.07; GG vs. CC: adjusted OR=1.10, 95% CI=0.89– 1.37; GG/CG vs CC: adjusted OR=0.95, 95% CI=0.80– 1.13; and GG vs. CC/CG: adjusted OR=1.19, 95% CI=0.99– 1.43). What’s more, risk effect of lncRNA HOTTIP rs3807598 C>G is still not obvious in stratification analysis by HSCR subtype.Conclusion: Our studies did not provide statistical evidence of a correlation between lncRNA HOTTIP SNP rs3807598 C>G and susceptibility of HSCR in the Chinese population that is being studied. Further validation study with a larger sample size covering multi-ethnic groups is warranted.Keywords: Hirschsprung disease, HOTTIP, polymorphism, susceptibility, Chinese

Hongge Liang1, Di Ma2, Yan Xu1, Jing Zhao1, Minjiang Chen1, Xiaoyan Liu1, Wei Zhong1, Junling Li2, Mengzhao Wang1 1Department of Respiratory and Critical Care Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing, People’s Republic of China; 2Department of Oncology, Chinese Academy of Medical Sciences Cancer Institute and Hospital, Beijing, People’s Republic of ChinaCorrespondence: Wei ZhongDepartment of Respiratory and Critical Care Medicine, Peking Union Medical College Hospital, No. 1 Shuanfuyuan Wangfujing Dongcheng District, Beijing 100730, People’s Republic of ChinaTel +86 0 106 915 5039Email zw_pumch@126.comMengzhao WangDepartment of Respiratory and Critical Care Medicine, Peking Union Medical College Hospital, No. 1 Shuanfuyuan Wangfujing Dongcheng District, Beijing 100730, People’s Republic of ChinaTel +86 0 106 915 5154Email mengzhaowang@sina.comBackground: Targeted therapy is an important treatment for advanced non-small cell lung cancer (NSCLC) patients with specific genetic mutations, crizotinib can prolong survival in advanced NSCLC patients with echinoderm microtubule-associated protein-like 4–anaplastic lymphoma kinase (EML4-ALK) rearrangement. We performed a retrospective analysis to investigate the association between the lactate dehydrogenase (LDH) levels and progression-free survival (PFS) in patients with EML4-ALK rearrangement NSCLC receiving treatment with crizotinib.Methods: Advanced (stage IIIb–IV) NSCLC patients with EML4-ALK rearrangement receiving treatment with crizotinib were enrolled between January 2007 and January 2016 at Peking Union Medical College and Cancer Hospital Chinese Academy of Medical Sciences.Results: Overall, 212 patients were enrolled. Kaplan–Meier univariate analysis showed that elevated pre-treatment LDH level (7.9 vs 14.1 months, HR =1.251, CI: 1.008–1.553, P=0.004) was significantly associated with shorter PFS, while the post-treatment mean-LDH level (13.3 vs 14.3 months, HR=1.439, 95% CI: 0.994–2.082, P=0.970) was not significantly associated with PFS. Cox proportional hazards model also identified that pre-treatment LDH level (HR=2.085, 95% CI: 1.150–3.781, P=0.016) was associated with the PFS. Logistic regression analysis showed that post-treatment LDH level was associated with creatine kinase (OR=6.712, 95% CI 3.395–13.273, P250 U/L) was significantly associated with shorter PFS in patients with EML4-ALK rearrangement NSCLC. Post-treatment elevated serum LDH level was not significantly associated with PFS, which related to adverse events including muscle damage and anemia.Keywords: non-small cell lung cancer, lactate dehydrogenase, crizotinib, echinoderm microtubule-associated protein-like 4–anaplastic lymphoma kinase, progression-free survival

Shi-yin Xiao,1 Ji Zhang,2,* Zheng-quan Zhu,3,* You-ping Li,4,* Wei-ying Zhong,5 Jian-bin Chen,6 Zhen-yu Pan,7 Hai-chen Xia3 1Department of Neurosurgery, Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, China; 2Department of Neurosurgery, State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center, Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, China; 3Department of Neurosurgery, Tumor Hospital Affiliated of Xinjiang Medical University, Xinshi District, Urumqi, Xinjiang 830011, China; 4Department of Neurosurgery, First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330046, China; 5Department of Neurosurgery, Qilu Hospital, Shandong University, Jinan 250012, China; 6Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China; 7Department of Radiation-Oncology, First Hospital of Jilin University, Changchun 130021, China *These authors contributed equally to this work Objective: Surgical resection serves an important role in the multidisciplinary treatment of cerebral metastases (CMs). Conventional white-light, microsurgical, and circumferential stripping of CMs is standard neurosurgical procedure, but is associated with a high recurrence rate. Based on this outcome, there is an urgent need for a new surgical strategy, such as fluorescence-guided resection, for CMs, in order to achieve total removal. Methods: A retrospective study was carried out in 38 patients clinically and pathologically diagnosed with breast cancer brain metastasis at three medical centers from May 2012 to June 2016. The study comprised group 1 (fluorescein-guided surgery) and group 2 (standard microsurgery). In group 1, 5 mg/kg of fluorescein sodium was injected intravenously after an allergy test and before general anesthesia for 17 patients. A yellow 560 filter was employed for microsurgical tumor resection. Group 2 consisted of 21 patients for whom fluorescein was not administered. Results: Surgical outcomes were assessed concerning the extent of resection and Karnofsky performance status. Gross total resection was achieved in these patients, with high fluorescence markedly enhancing tumor visibility. The extent of resection had a powerful influence on performance status. Overall survival after CM was 24.1 months in patients given fluorescein and was 22.8 months in the nonfluorescein group. Conclusion: Fluorescein-guided surgery is a simple, safe, and practical method to resect breast cancer brain metastasis, and leads to a higher proportion of resection compared to common microsurgery. This offers a tremendous advantage when navigating a tiny tumor, and improves the quality of life of patients with CM. Keywords: fluorescein sodium, breast cancer brain metastasis, fluorescence-guided surgery, extent of resection, KPS

Yi Kong,1,2,* Rui Xu,1,* Mohammad Ali Darabi,2 Wen Zhong,3 Gaoxing Luo,1 Malcolm MQ Xing,1,2 Jun Wu1 1Institute of Burn Research, Southwest Hospital, State Key Laboratory of Trauma, Burns, and Combined Injury, Chongqing Key Laboratory for Diseases Proteomics, Third Military Medical University, Chongqing, People’s Republic of China; 2Department of Mechanical Engineering, Biochemistry and Medical Genetics, Children’s Hospital Research Institute of Manitoba, 3Department of Biosystem Engineering, University of Manitoba, Winnipeg, MB, Canada *These authors contributed equally to this work Abstract: Polymeric ultrathin membranes that are compatible with cells offer tremendous advantages for tissue engineering. In this article, we report a free-standing nanomembrane that was developed using a layer-by-layer self-assembly technique with a safe and sacrificial substrate method. After ionization, two oppositely charged polyelectrolytes, alginate and chitosan, were alternately deposited on a substrate of a solidified gelatin block to form an ultrathin nanomembrane. The space between the two adjacent layers was ~200 nm. The thickness of the nanomembrane was proportional to the number of layers. The temperature-sensitive gelatin gel served as a sacrificial template at 37°C. The free-standing nanomembrane promoted bone marrow stem cell adhesion and proliferation. Fluorescence-activated cell sorting was used to analyze green-fluorescent-protein-positive mesenchymal stem cells from the wounds, which showed a significantly high survival and proliferation from the nanomembrane when cells were transplanted to mouse dorsal skin that had a full-thickness burn. The bone-marrow-stem-cell-loaded nanomembrane also accelerated wound contraction and epidermalization. Therefore, this methodology provides a fast and facile approach to construct free-standing ultrathin scaffolds for tissue engineering. The biocompatibility and free-standing nature of the fabricated nanomembrane may be particularly useful for stem cell delivery and wound healing. Keywords: nanomembrane, layer-by-layer, cell delivery, wound healing

Xiaoyan Zhang, Xiaofei Wang, Weitong Zhong, Xiaoqing Ren, Xianyi Sha, Xiaoling FangKey Laboratory of Smart Drug Delivery, Ministry of Education and People’s Liberation Army of China, School of Pharmacy, Fudan University, Shanghai, People’s Republic of China Abstract: Since elevated expression of matrix metalloproteinase (MMP)-2 and MMP-9 is commonly observed in several malignant tumors, MMPs have been widely reported as key factors in the design of drug delivery systems. Several strategies have been proposed to develop MMPs-responsive nanoparticles to deliver chemotherapeutics to malignant solid tumors. A stimuli-responsive drug delivery system, which could be cleaved by MMPs, was proposed in this study. By inserting an MMP-2/9 cleavable oligopeptide GPVGLIGK-NH2 (GK8) as spacer between α-tocopherol succinate (α-TOS) and methoxy-polyethylene glycol molecular weight (MW 2000 Da) activated by N-hydroxysuccinimide (mPEG2K-NHS), mPEG2K-GK8-α-TOS (TGK) was synthesized as the primary ingredient for MMP-2/9-sensitive micelles composed of D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) and TGK (n:n =40:60, TGK micelles). mPEG2K-α-TOS (T2K) was similarly synthesized as nonsensitive control. The TGK micelles showed better stability than nonsensitive micelles composed of TPGS and T2K (n:n =40:60, T2K micelles) owing to the inserted peptide. Fluorescence resonance energy transfer results indicated that TGK micelles could be successfully cleaved by MMP-2/9. Effective drug release was demonstrated in the presence of collagenase type IV, a mixture of MMP-2 and MMP-9. Compared with nonsensitive micelles, docetaxel (DTX)-loaded TGK micelles showed a fold higher cellular uptake in HT1080 cells. While the half-maximal inhibitory concentration (IC50) of TGK and T2K micelles were similar (P>0.05) in MCF-7 cells (MMP-2/9 underexpression), the IC50 values of the aforementioned micelles were 0.064±0.006 and 0.122±0.009 µg/mL, respectively, in HT1080 cells (MMP-2/9 overexpression). The MMP-2/9-sensitive micelles also demonstrated desired tumor targeting and accumulation ability in vivo. The results of in vivo antitumor effect evaluation indicate that TGK micelles are potent against solid tumors while maintaining minimum systemic toxicity compared with T2K micelles and DTX. Keywords: matrix metalloproteinases, stimuli responsive, nanocarriers, TPGS, FRET

Liangpeng Ge,1–5,* Qingtao Li,2,3,* Junzi Jiang,2,3 Xiaoyan You,1 Zuohua Liu,1 Wen Zhong,6 Yong Huang,1 Malcolm MQ Xing2,31Chongqing Academy of Animal Sciences, Chongqing, People’s Republic of China; 2Department of Mechanical and Manufacturing Engineering, Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, MB, Canada; 3Manitoba Institute of Child Health, Winnipeg, MB, Canada; 4Key Laboratory of Pig Industry Sciences, Ministry of Agriculture,Chongqing, People’s Republic of China; 5Chongqing Key Laboratory of Pig Industry Sciences, Chongqing, People’s Republic of China; 6Department of Textile Sciences, Faculty of Human Ecology, University of Manitoba, Winnipeg, MB, Canada*These authors contributed equally to this workAbstract: Pelvic organ prolapse (POP) is a serious health issue affecting many adult women. Complications of POP include pelvic pressure, pelvic pain, and problems in emptying their bowels or bladder. Sometimes, POP may even cause urinary outflow obstruction and lead to bladder or kidney infections. Currently, synthetic and naturally derived materials have been chosen for treatment of POP to reduce the high recurrence rates after surgical interventions. However, existing materials for POP treatment cannot meet the clinical requirements in terms of biocompatibility, mechanics, and minimal risk of rejection. Especially, erosion in synthetic polymers and rapid degradation in natural polymers limit their further applications in clinics. To address these concerns, we report a novel POP replacement using core–sheath polystyrene/gelatin electrospun nanofiber mesh. The outside gelatin sheath provides a hydrophilic surface and implantable integrity between host and guest, while the inner PS core offers the necessary mechanical support. The composite mesh shows graft accommodation in pelvic submucosa after implantation in vivo, as shown in hematoxylin–eosin staining and T helper cell phenotype and macrophage phenotype stainings. Qualitative analysis of inducible nitric oxide synthase, arginase, interferon-γ, and interleukin-10 gene expressions also indicates that the implanted composite mesh switches to accommodation mode 2 weeks postimplantation. Thus, these novel core–sheath polystyrene/gelatin nanofibrous membranes are promising in pelvic reconstruction.Keywords: core-sheath nanofibrous patch, polystyrene, gelatin, pelvic reconstruction

Wenbing Wan,1–3,* Shiwen Zhang,2–4,* Liangpeng Ge,2,3,5 Qingtao Li,1 Xingxing Fang,1 Quan Yuan,4 Wen Zhong,6 Jun Ouyang,1 Malcolm Xing1,2,7 1Department of Anatomy, Guangdong Provincial Medical Biomechanical Key Laboratory, Southern Medical University, Guangzhou, People’s Republic of China; 2Department of Mechanical Engineering, University of Manitoba, Winnipeg, MB, Canada; 3Manitoba Institute of Child Health, Winnipeg, MB, Canada; 4Sichuan University, Chengdu, People’s Republic of China; 5Chongqing Academy of Animal Sciences, Chongqing, People’s Republic of China; 6Department of Textile Sciences, University of Manitoba, Winnipeg, MB, Canada; 7Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, MB, Canada *These authors contributed equally to this work Abstract: Bone tissue engineering through seeding of stem cells in three-dimensional scaffolds has greatly improved bone regeneration technology, which historically has been a constant challenge. In this study, we researched the use of adipose-derived stem cell (ADSC)-laden layer-by-layer paper-stacking polycaprolactone/gelatin electrospinning nanofibrous membranes for bone regeneration. Using this novel paper-stacking method makes oxygen distribution, nutrition, and waste transportation work more efficiently. ADSCs can also secrete multiple growth factors required for osteogenesis. After the characterization of ADSC surface markers CD29, CD90, and CD49d using flow cytometry, we seeded ADSCs on the membranes and found cells differentiated, with significant expression of the osteogenic-related proteins osteopontin, osteocalcin, and osteoprotegerin. During 4 weeks in vitro, the ADSCs cultured on the paper-stacking membranes in the osteogenic medium exhibited the highest osteogenic-related gene expressions. In vivo, the paper-stacking scaffolds were implanted into the rat calvarial defects (5 mm diameter, one defect per parietal bone) for 12 weeks. Investigating with microcomputer tomography, the ADSC-laden paper-stacking membranes showed the most significant bone reconstruction, and from a morphological perspective, this group occupied 90% of the surface area of the defect, produced the highest bone regeneration volume, and showed the highest bone mineral density of 823.06 mg/cm3. From hematoxylin and eosin and Masson staining, the new bone tissue was most evident in the ADSC-laden scaffold group. Using quantitative polymerase chain reaction analysis from collected tissues, we found that the ADSC-laden paper-stacking membrane group presented the highest osteogenic-related gene expressions of osteocalcin, osteopontin, osteoprotegerin, bone sialoprotein, runt-related transcription factor 2, and osterix (two to three times higher than the control group, and 1.5 times higher than the paper-stacking membrane group in all the genes). It is proposed that ADSC-laden layer-by-layer paper-stacking scaffolds could be used as a way of promoting bone defect treatment. Keywords: paper-stacking, layer-by-layer membrane, bone regeneration, adipose-derived stem cells, calvarial defect

Yanzuo Chen,1 Xianyi Sha,1 Wei Zhang,1,2 Weitong Zhong,1 Zhuoyang Fan,1 Qiuyue Ren,1 Liangcen Chen,1 Xiaoling Fang1 1Key Laboratory of Smart Drug Delivery, Ministry of Education and PLA, Department of Pharmaceutics, School of Pharmacy, Fudan University, Shanghai, People’s Republic of China 2Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, USA Abstract: A Pluronic polymeric mixed micelle delivery system was developed in this study by using Pluronic P105 and F127 block copolymers to encapsulate the antitumor compound, methotrexate (MTX). The MTX-loaded Pluronic P105/F127 mixed micelle exhibited the spherical shape with about 22 nm in diameter, high encapsulation efficiency (about 85%) and pH-dependent in vitro drug release. In this study, A-549 and KBv cell lines were selected as multidrug resistance tumor cell models, while H-460 and KB cell lines were chosen as sensitive tumor cells. The MTX-loaded Pluronic P105/F127 mixed micelle exhibited significant higher in vitro cytotoxicity in multidrug resistant tumor cells than that of control (MTX injection) mainly because of higher cellular uptake of MTX. The pharmacokinetic studies indicated that the Pluronic micelles significantly prolonged systemic circulation time of MTX compared to MTX injection. Moreover, a much stronger antitumor efficacy in KBv tumor xenografts nude mice was observed in the MTX-loaded Pluronic P105/F127 mixed micelle group, than MTX. Collectively, Pluronic P105/F127 mixed micelles could significantly enhance the antitumor activity of MTX and might be a promising drug delivery platform for multidrug resistance modulation. Keywords: multidrug resistance, drug delivery system, micelles, Pluronic, methotrexate

Jun Chen1,2, Beth Zhou1–3, Qi Li1,2, Jun Ouyang4, Jiming Kong2,4,5, Wen Zhong3,6, Malcolm MQ Xing1,2,4,71Department of Mechanical Engineering, Faculty of Engineering, University of Manitoba, Winnipeg, MB, Canada; 2Manitoba Institute of Child Health, Winnipeg, MB, Canada; 3Department of Textile Sciences, Faculty of Human Ecology, University of Manitoba, Winnipeg, MB, Canada; 4School of Basic Medical Science, Southern Medical University, Guangzhoug, China; 5Department of Human Anatomy and Cell Sciences, 6Department of Medical Microbiology, Faculty of Medicine, 7Department of Biochemistry and Medical Genetics, Faculty of Medicine, University of Manitoba, Winnipeg, MB, CanadaAbstract: By mimicking the native extracellular matrix, electrospun nanofibrous scaffolds (ENSs) can provide both chemical and physical cues to modulate cell adherence and differentiation and to promote tissue regeneration while retaining bioresorbable and biocompatible properties. In this study, ENSs were developed to deliver multiple biomolecules by loading them into the core-sheath structure and/or by conjugating them to the nanofiber surfaces. In this work, poly(L-lactide)-poly(ethylene glycol)-NH2 and poly(L-lactide) were emulsion electrospun into nanofibers with a core-sheath structure. A model drug, tetracycline hydrochloride, was loaded within the nanofibers. Amino and carboxyl reactive groups were then activated on the fiber surfaces using saturated water vapor exposure and base hydrolysis, respectively. These reactive groups allowed the surface of the ENS to be functionalized with two other bioactive molecules, fluorescein isothiocyanate- and rhodamine-labeled bovine serum albumins, which were used as model proteins. The ENSs were shown to retain their antimicrobial capacity after two functionalization reactions, indicating that multifunctional nanofibers can potentially be developed into functional wound dressings or periodontal membranes or used in more complicated tissue systems where multiple growth factors and anti-infection precautions are critical for the successful implantation and regeneration of tissues.Keywords: multifunctional nanofibrous scaffold, drug delivery, antibiotics