Search

Jiaying Li,1,2,&ast; Minhui Zhou,2,&ast; Yahan Tong,3 Haibo Chen,1 Ruisi Su,2 Yinghui Tao,2 Guodong Zhang,1 Zhichao Sun1 1Department of Radiology, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou, 310006, People’s Republic of China; 2The First Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou, 310053, People’s Republic of China; 3Department of Radiology, The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou, 310005, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zhichao Sun, Department of Radiology, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), 54 Youdian Road, Hangzhou, People’s Republic of China, Tel/Fax +86-571-87077272, Email sunzhichao@zcmu.edu.cnPurpose: Non-invasive methods are urgently needed to assess the efficacy of transarterial chemoembolization (TACE) and to identify patients with hepatocellular carcinoma (HCC) who may benefit from this procedure. This study, therefore, aimed to investigate the predictive ability of tumor growth patterns and radiomics features from contrast-enhanced magnetic resonance imaging (CE-MRI) in predicting tumor response to TACE among patients with HCC.Patients and Methods: A retrospective study was conducted on 133 patients with HCC who underwent TACE at three centers between January 2015 and April 2023. Enrolled patients were divided into training, testing, and validation cohorts. Rim arterial phase hyperenhancement (Rim APHE), tumor growth patterns, nonperipheral washout, markedly low apparent diffusion coefficient (ADC) value, intratumoral arteries, and clinical baseline features were documented for all patients. Radiomics features were extracted from the intratumoral and peritumoral regions across the three phases of CE-MRI. Seven prediction models were developed, and their performances were evaluated using receiver operating characteristic (ROC) and decision curve analysis (DCA).Results: Tumor growth patterns and albumin-bilirubin (ALBI) score were significantly correlated with tumor response. Tumor growth patterns also showed a positive correlation with tumor burden (r = 0.634, P = 0.000). The Peritumor (AUC = 0.85, 0.71, and 0.77), Clinics_Peritumor (AUC = 0.86, 0.77, and 0.81), and Tumor_Peritumor (AUC = 0.87, 0.77, and 0.80) models significantly outperformed the Clinics and Tumor models (P < 0.05), while the Clinics_Tumor_Peritumor model (AUC = 0.88, 0.81, and 0.81) outperformed the Clinics (AUC = 0.67, 0.77, and 0.75), Tumor (AUC = 0.78, 0.72, and 0.68), and Clinics_Tumor (AUC = 0.82, 0.83, and 0.78) models (P < 0.05 or 0.053, respectively). The DCA curve demonstrated better predictive performance within a specific threshold probability range for Clinics_Tumor_Peritumor.Conclusion: Combining tumor growth patterns, intra- and peri-tumoral radiomics features, and ALBI score could be a robust tool for non-invasive and personalized prediction of treatment response to TACE in patients with HCC.Keywords: hepatocellular carcinoma, radiomics, tumor growth pattern, transarterial chemoembolization

Linbo Tang,1 Xinjing Li,2 Yongbin Wang,1 Yuanhe Tong1 1Department of Radiation Oncology, Longyan First Affiliated Hospital of Fujian Medical University, Longyan, Fujian, 364000, People’s Republic of China; 2Department of Pathology, Longyan First Affiliated Hospital of Fujian Medical University, Longyan, Fujian, 364000, People’s Republic of ChinaCorrespondence: Yuanhe Tong, Department of Radiation Oncology, Longyan First Affiliated Hospital of Fujian Medical University, Longyan, 364000, People’s Republic of China, Email tyhlongyan@126.comPurpose: Inflammatory markers in the blood have been linked to tumor prognosis, but their specific prognostic significance in nasopharyngeal carcinoma (NPC) patients undergoing intensity-modulated radiotherapy (IMRT) is not well established. This study aims to evaluate the prognostic value of the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR) in this patient population.Patients and Methods: A total of 406 non-metastatic NPC patients were included in the study. NLR, PLR, and LMR were stratified according to their average values. The Kaplan-Meier method was used to estimate progression-free survival (PFS) and overall survival (OS). Cox multivariate regression analysis was performed to evaluate the associations of NLR, PLR, and LMR with PFS and OS.Results: Patients with NLR > 2.78 had worse PFS (P = 0.008) and OS (P < 0.001); PLR > 162.48 was related to lower PFS (P = 0.018) but not OS (P = 0.29); LMR > 5.05 showed no significant difference in PFS and OS compared to LMR ≤ 5.05 (P values were 0.13 and 0.94, respectively). Multivariate analysis indicated that NLR was an independent prognostic factor for PFS (HR, 1.674; 95% CI, 1.006– 2.784; P = 0.047) and OS (HR, 4.143; 95% CI, 2.111– 8.129; P = 0.000), while PLR and LMR did not demonstrate significant associations with PFS and OS.Conclusion: This study identifies NLR as a novel and independent prognostic indicator for NPC patients receiving IMRT, offering valuable insights that could inform future clinical decision-making. In contrast, PLR and LMR did not demonstrate significant prognostic value in this context.Keywords: nasopharyngeal carcinoma, inflammatory markers, neutrophil to lymphocyte ratio, platelet to lymphocyte ratio, lymphocyte to monocyte ratio, prognosis

Yan Yang,1,&ast; Yiling Wang,1,&ast; Xinran Jiang,1 Jiahao Mi,1 Dizhang Ge,2 Yuna Tong,3 Yuxuan Zhu1,2 1Department of Pharmacy, Personalized Drug Therapy Key Laboratory of Sichuan Province, Sichuan Academy of Medical Science & Sichuan Provincial People’s Hospital, University of Electronic Science and Technology of China, Chengdu, 610072, People’s Republic of China; 2Department of Pharmacy, People’s Hospital of Aba Tibetan and Qiang Autonomous Prefecture, Aba, 624000, People’s Republic of China; 3Department of Nephrology, The Third People’s Hospital of Chengdu, Chongqing Medical University, Chengdu, 610031, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yuna Tong; Yuxuan Zhu, Email tongyuna1985@163.com; zhuyuxuan6688@163.comIntroduction: Alzheimer’s disease (AD), a neurodegenerative condition, stands as the most prevalent form of dementia. Its complex pathological mechanisms and the formidable blood-brain barrier (BBB) pose significant challenges to current treatment approaches. Oxidative stress is recognized as a central factor in AD, underscoring the importance of antioxidative strategies in its treatment. In this study, we developed a novel brain-targeted nanoparticle, Ce/Zr-MOF@Cur-Lf, for AD therapy.Methods: Layer-by-layer self-assembly technology was used to prepare Ce/Zr-MOF@Cur-Lf. In addition, the effect on the intracellular reactive oxygen species level, the uptake effect by PC12 and bEnd.3 cells and the in vitro BBB permeation effect were investigated. Finally, the mouse AD model was established by intrahippocampal injection of Aβ1-42, and the in vivo biodistribution, AD therapeutic effect and biosafety of the nanoparticles were researched at the animal level.Results: As anticipated, Ce/Zr-MOF@Cur-Lf demonstrated efficient BBB penetration and uptake by PC12 cells, leading to attenuation of H2O2-induced oxidative damage. Moreover, intravenous administration of Ce/Zr-MOF@Cur-Lf resulted in rapid brain access and improvement of various pathological features of AD, including neuronal damage, amyloid-β deposition, dysregulated central cholinergic system, oxidative stress, and neuroinflammation.Conclusion: Overall, Ce/Zr-MOF@Cur-Lf represents a promising approach for precise brain targeting and multi-target mechanisms in AD therapy, potentially serving as a viable option for future clinical treatment.Keywords: metal-organic frameworks, Alzheimer’s disease, drug delivery, brain targeting, oxidative stress

Zemin Ou, Yun You, Hong Yi, Xiaoqian Liu, Yan Tong, Dewen Liu, Jinyu Wang Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, People’s Republic of ChinaCorrespondence: Dewen Liu; Jinyu Wang, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 10700, People’s Republic of China, Email dwliu@icmm.ac.cn; wangjinyu@icmm.ac.cnIntroduction: Parkinson’s disease (PD) is a common neurodegenerative disorder characterized by the degeneration of dopaminergic neurons in the substantia nigra. The precise molecular mechanisms underlying neuronal loss in PD remain unknown, and there are currently no effective treatments for PD-associated neurodegeneration. Echinacoside (ECH) is known for its neuroprotective effects, which include scavenging cellular reactive oxygen species and promoting mitochondrial fusion. However, the blood-brain barrier (BBB) limits the bioavailability of ECH in the brain, posing a significant challenge to its use in PD treatment.Methods: We synthesized and characterized PEGylated ECH liposomes (ECH@Lip) and peptide angiopep-2 (ANG) modified liposomes (ECH@ANG-Lip). The density of ANG in ANG-Lip was optimized using bEnd.3 cells. The brain-targeting ability of the liposomes was assessed in vitro using a transwell BBB model and in vivo using an imaging system and LC-MS. We evaluated the enhanced neuroprotective properties of this formulation in a the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD model.Results: The ECH@ANG-Lip demonstrated significantly higher whole-brain uptake compared to ECH@Lip and free ECH. Furthermore, ECH@ANG-Lip was more effective in mitigating MPTP-induced behavioral impairment, oxidative stress, dopamine depletion, and dopaminergic neuron death than both ECH@Lip and free ECH.Conclusion: The formulation used in our study significantly enhanced the neuroprotective efficacy of ECH in the MPTP-induced PD model. Thus, ECH@ANG-Lip shows considerable potential for improving the bioavailability of ECH and providing neuroprotective effects in the brain.Keywords: nanomedicine, brain delivery, angiopep-2, echinacoside, Parkinson’s disease, liposomes

Zhenyu Li,1 Jimin Liu,2 Haishan Cui,3 Wenlong Qi,4 Yangyang Tong,4 Tan Wang4 1College of Traditional Chinese Medicine, Changchun University of Chinese Medicine, Changchun, 130117, People’s Republic of China; 2Department of Respiratory, The Third Affiliated Hospital to Changchun University of Chinese Medicine, Changchun, 130117, People’s Republic of China; 3Department of Traditional Chinese Medicine, The Third Affiliated Hospital to Changchun University of Chinese Medicine, Changchun, 130117, People’s Republic of China; 4Department of Respiratory, The Affiliated Hospital to Changchun University of Chinese Medicine, Changchun, 130021, People’s Republic of ChinaCorrespondence: Tan Wang, Department of Respiratory, The Affiliated Hospital to Changchun University of Chinese Medicine, Changchun, 130021, People’s Republic of China, Email wangtan0215@163.comAbstract: The rising global morbidity and mortality rates of non-small cell lung cancer (NSCLC) underscore the urgent need for more effective treatments. Current therapeutic modalities—including surgery, radiotherapy, chemotherapy, and targeted therapy—face several limitations. Recently, Astragalus membranaceus, a traditional Chinese medicine (TCM), has captured significant attention due to its broad pharmacological properties, such as immune regulation, anti-inflammatory effects, and the modulation of reactive oxygen species (ROS) and enzyme activities. This review delivers a comprehensive summary of the most recent advancements and ongoing applications of Astragalus membranaceus in NSCLC treatment, underlining its potential for integration into existing treatment protocols. It also highlights essential areas for future research, including the elucidation of its molecular mechanisms, optimization of dosage and administration, and evaluation of its efficacy and safety alongside standard therapies, all of which could potentially improve therapeutic outcomes for NSCLC patients.Keywords: non-small cell lung cancer, Astragalus membranaceus, antitumor activity, immunoregulation, cisplatin

Tiantian Liang,1,&ast; Xianbin Liu,1,&ast; Yujun Tong,2 Qian Ding,3 Min Yang,1 Hong Ning1 1Department of Pharmacy, Mianyang Central Hospital, School of Medicine, University of Electronic Science and Technology of China, Mianyang, Sichuan, People’s Republic of China; 2Department of Breast Surgery, Mianyang Central Hospital, School of Medicine, University of Electronic Science and Technology of China, Mianyang, Sichuan, People’s Republic of China; 3Department of Clinical Pharmacy, The Third Hospital of Mianyang, Sichuan Mental Health Center, Mianyang, Sichuan, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Hong Ning; Min Yang, Department of Pharmacy, Mianyang Central Hospital, School of Medicine, University of Electronic Science and Technology of China, No. 12, Changjia Alley, Jingzhong Street, Mianyang, Sichuan, 621000, People’s Republic of China, Email ninghong555@126.com; ym8710@126.comAbstract: Targeted therapy for infantile hemangiomas (IHs) has been extensively studied as they can concentrate drugs, increase therapeutic efficacy and reduce drug dosage. Meanwhile, they can extend drug release times, enhance drug stability, decrease dosing frequency, and improve patient compliance. Moreover, carriers made from biocompatible materials reduced drug immunogenicity, minimizing adverse reactions. However, current targeted formulations still face numerous challenges such as the non-absolute safety of carrier materials; the need to further increase drug loading capacity; the limitation of animal hemangioma models in fully replicating the biological properties of human infantile hemangiomas; the establishment of models for deep-seated hemangiomas with high incidence rates; and the development of more specific targets or markers. In this review, we provided a brief overview of the characteristics of IHs and summarized the past decade’s advances, advantages, and targeting strategies of targeted drug delivery systems for IHs and discussed their applications in the treatment of IHs. Furthermore, the goal is to provide a reference for further research and application in this field.Keywords: targeting, targeted therapies, infantile hemangiomas, drug delivery systems, nanoparticle

Cheng Su,1,&ast; Cai Li,2,&ast; Xinyi Hu,3,&ast; Jing Wang,1,&ast; Linlin Liu,4,&ast; Xianfeng Zhang,4 Yeqing Tong4 1School of Public Health, Huazhong University of Science and Technology, Wuhan, Hubei, People’s Republic of China; 2Infectious Disease Prevention and Control Center, Wuhan Center for Disease Control and Prevention, Wuhan, Hubei, People’s Republic of China; 3Global Study Institute, University of Geneva, Geneva, 1205, Switzerland; 4Infectious Disease Prevention and Control Center, Hubei Center for Disease Control and Prevention, Wuhan, Hubei, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yeqing Tong; Xianfeng Zhang, Email t_yeqing@163.com; 63382251@qq.comAbstract: Angiotensin-converting enzyme 2 (ACE2) is an important regulator of the Renin-Angiotensin System (RAS). Additionally, it has been identified as a functional receptor for the Coronavirus. Research indicates that ACE2 plays a role in the regulation of cardiovascular systems by modulating blood pressure and electrolyte balance. Its role in pulmonary diseases has also garnered significant attention due to the widespread prevalence of Coronavirus. There is solid evidence linking ACE2 to other pulmonary diseases, including chronic obstructive pulmonary disease, acute respiratory distress syndrome, allergic asthma, among others. However, the exact pathological and physiological mechanisms of ACE2 in these diseases remain elusive. Our research aims to review and explore the latest advancements in ACE2-related studies in pulmonary diseases. These findings have the potential to open new avenues for utilizing ACE2 as a potential biomarker for early diagnosis and monitoring of pulmonary diseases.Keywords: ACE2, lung diseases, ARDS, Lung Cancer, COPD

Junwei Zhan,1 Chengfei Peng,2 Yuxin Liu,1 Zhanhua Bi,1 Guoxiu Lu,1 Shanhu Hao,1 Yanan Tong,1 Guoxu Zhang1 1Department of Nuclear Medicine, General Hospital of Northern Theater Command, Shenyang, 110016, People’s Republic of China; 2Cardiovascular Research Institute and Department of Cardiology, General Hospital of Northern Theater Command, Shenyang, 110016, People’s Republic of ChinaCorrespondence: Guoxu Zhang; Yanan Tong, Department of Nuclear Medicine, General Hospital of Northern Theater Command, No. 83, Wenhua Road, Shenyang, 110016, People’s Republic of China, Email zhangguoxu_502@163.com; tongyn1119@foxmail.comObjective: Atrial fibrillation (AF) is a common arrhythmia. This study explored serum miR-29b-3p expression in AF patients and its value in predicting AF recurrence after radiofrequency catheter ablation (RFCA).Methods: Totally 100 AF patients who underwent RFCA were enrolled, with 100 individuals without AF as controls. Serum miR-29b-3p expression in participants was determined using RT-qPCR. The correlation between miR-29b-3p and atrial fibrosis markers (FGF-21/FGF-23) was assessed by Pearson analysis. The diagnostic efficacy of serum miR-29b-3p and FGF-21/FGF-23 in predicting AF recurrence after RFCA was analyzed by the receiver operating characteristic (ROC) curves. The Kaplan-Meier method was adopted to evaluate the effect of miR-29b-3p expression on the incidence of AF recurrence after RFCA. The independent risk factors for AF recurrence after RFCA were analyzed by logistic regression analysis.Results: Serum miR-29b-3p was poorly expressed in AF patients. After RFCA, AF patients showed elevated serum miR-29b-3p expression. Serum miR-29b-3p expression in AF patients negatively correlated with serum FGF-21 and FGF-23 concentrations. The cut-off values of serum miR-29b-3p, FGF-21, and FGF-23 in identifying AF recurrence were 0.860 (sensitivity: 100.00%, specificity: 39.71%), 222.2 pg/mL (sensitivity: 96.88%, specificity: 32.35%) and 216.3 ng/mL (sensitivity: 53.13%, specificity: 70.59%), respectively. Patients with low miR-29b-3p expression had a significantly higher incidence of AF recurrence than patients with high miR-29b-3p expression. Serum miR-29b-3p expression was one of the independent risk factors for AF recurrence after RFCA.Conclusion: Low miR-29b-3p expression in AF patients has certain predictive values and is one of the independent risk factors for AF recurrence after RFCA.Keywords: miR-29b-3p, atrial fibrillation, radiofrequency catheter ablation, recurrence, fibrosis marker, logistic regression analysis

Qiu Huang,&ast; Yue Ge,&ast; Yu He, Jian Wu, Yonghua Tong, Haojie Shang, Xiao Liu, Xiaozhuo Ba, Ding Xia, Ejun Peng, Zhiqiang Chen, Kun Tang Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zhiqiang Chen; Kun Tang, Email zhqchen8366@163.com; tangsk1990@163.comAbstract: Cancer-associated fibroblasts (CAF) are the most abundant stromal cells in the tumor microenvironment (TME), especially in solid tumors. It has been confirmed that it can not only interact with tumor cells to promote cancer progression and metastasis, but also affect the infiltration and function of immune cells to induce chemotherapy and immunotherapy resistance. So, targeting CAF has been considered an important method in cancer treatment. The rapid development of nanotechnology provides a good perspective to improve the efficiency of targeting CAF. At present, more and more researches have focused on the application of nanoparticles (NPs) in targeting CAF. These studies explored the effects of different types of NPs on CAF and the multifunctional nanomedicines that can eliminate CAF are able to enhance the EPR effect which facilitate the anti-tumor effect of themselves. There also exist amounts of studies focusing on using NPs to inhibit the activation and function of CAF to improve the therapeutic efficacy. The application of NPs targeting CAF needs to be based on an understanding of CAF biology. Therefore, in this review, we first summarized the latest progress of CAF biology, then discussed the types of CAF-targeting NPs and the main strategies in the current. The aim is to elucidate the application of NPs in targeting CAF and provide new insights for engineering nanomedicine to enhance immune response in cancer treatment.Keywords: nanoparticles, cancer therapy, cancer-associated fibroblasts, drug delivery, cancer, nanomedicine

Jianing Li,1 Yongsheng Zhang,2 Ying Tong3 1Heilongjiang University of Chinese Medicine, Harbin, People’s Republic of China; 2Harbin Institute of Technology, Harbin, People’s Republic of China; 3First Affiliated Hospital, Heilongjiang University of Chinese Medicine, Harbin, People’s Republic of ChinaCorrespondence: Ying Tong, Email tongying@hljucm.edu.cn

Xiang Li,1 Zigang Ding,1 Yun Tong2 1Department of General Surgery, The Affiliated Hospital of Jiujiang University, Jiujiang, People’s Republic of China; 2Department of Pain, The Affiliated Hospital of Jiujiang University, Jiujiang, People’s Republic of ChinaCorrespondence: Yun Tong, The Affiliated Hospital of Jiujiang University, No. 57 East Xunyang Road, Jiangxi, Jiujiang 332000, People’s Republic of China, Email tongyun1209@163.comPurpose: Papillary thyroid carcinoma (PTC) is the most common subtype of thyroid cancer. Recurrence makes the prognosis for some patients with PTC worse. Increasing evidence have suggested that N6-methyladenosine (m6A) RNA methylation plays an important role in tumorigenesis. However, the significance of m6A-related lncRNAs in the malignant progression of PTC remains unknown. In this study, we explored the significance of M6A-related lncrnas in the malignant progression of PTC.Patients and Methods: Transcriptome and clinical data of PTC were achieved and integrated from The Cancer Genome Atlas (TCGA). Firstly, a Spearman correlation analysis was performed to obtain m6A RNA methylation-associated lncRNAs. Next, We constructed a prognostic signature and assessed the accuracy of the signature by receiver operating characteristic (ROC) curve and Kaplan Meier survival analyses. Furthermore, functional enrichment analysis was performed on the high- and low-risk groups. Finally, we determined prognostic gene expression in clinical samples using quantitative reverse transcription polymerase chain reaction (RT-qPCR).Results: We identified 56 differentially expressed lncRNAs associated with m6A RNA methylation. Univariate Cox and Least Absolute Shrinkage and Selection Operator (LASSO) regression analyses showed that the survival-related lncRNAs associated with m6A RNA methylation were detected, which showed superior calibration and discrimination. Moreover, the biological processes related to energy metabolism were significantly activated in the high-risk group. Finally, the co-expressed genes of lncRNAs in the risk model were significantly enriched in biological processes related to copper ion response. Finally, we validated the expression levels of three prognostic genes in clinical samples using RT-qPCR.Conclusion: Our study revealed m6A RNA methylation-associated lncRNAs were significantly associated with disease-free survival in patients with papillary thyroid cancer, which would improve our understanding of the relationship between m6A RNA methylation-associated lncRNAs and PTC.Keywords: Papillary thyroid carcinoma, m6a RNA methylation, lncrna, PROGNOSIS, disease-free survival

Shaoxing Zhang,1 Yuxin Zhang,1 Ruijie Liu,1 Shuying Yuan,2 Yanwen Chen,1 Wenjie Li,1 Xinrong Lu,3 Yongliang Tong,3 Linlin Hou,4 Li Chen,3 Guiqin Sun1 1School of Medical Technology and Information Engineering, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, People’s Republic of China; 2Clinical Laboratory Department, Jiaxing Maternity and Child Health Care Hospital, Jiaxing, Zhejiang, People’s Republic of China; 3Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Fudan University, Shanghai, People’s Republic of China; 4College of Veterinary Medicine, Qingdao Agricultural University, Qingdao, Shandong, People’s Republic of ChinaCorrespondence: Guiqin Sun, School of Medical Technology and Information Engineering, Zhejiang Chinese Medical University, 548 Binwen Road, Binjiang District, Hangzhou, Zhejiang, People’s Republic of China, Tel +86 13868118601, Fax +86 571-86633307, Email sunguiqin2001@163.com Li Chen, Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, Fudan University, 131 Dongan Road, Xuhui District, Shanghai, People’s Republic of China, Tel +86 15821138980, Fax +86 21-54237381, Email lichen_bk@fudan.edu.cnPurpose: Elizabethkingia meningoseptica (EM) is a multi-drug-resistant bacterium of global concern for its role in nosocomial infection and is generally resistant to aminoglycoside antibiotics. In the whole genome of an EM strain (FMS-007), an aminoglycoside-6-adenyl transferase gene (ant(6)FMS-007) was predicted. This study aimed to characterize the biochemical function of ANT(6)FMS-007 and analyze the relationship between genotype and phenotype of ant(6) in clinical EM isolates, so as to provide evidence for clinical precision drug use. This study could establish a method for the verification of known or unknown functionally resistant genes.Methods: A total of 42 EM clinical isolates were collected from clinical departments during 2015– 2023. The phenotype of aminoglycoside antibiotics was analyzed by broth microdilution (BMD) and Kirby-Bauer (K-B) methods. The whole-length ant(6) from EM clinical isolates was analyzed by polymerase chain reaction (PCR) and sequencing. The biochemical function of predictive ANT(6)FMS-007 from the FMS-007 whole genome was identified by 3D plate experiment and mass spectrometry analysis. Candidate active sites were predicted by multi-species sequence alignment and molecular docking, and other important sites were identified in the comparison of ant(6) genotypes and phenotypes of EM clinical isolates. Drug susceptibility test was used to verify the function of these sites.Results: The predictive ANT(6)FMS-007 protein could inactivate STR by modifying STR with ATP to form STR-AMP. Four active sites (Asp-38, Asp-42, Lys-95, and Lys-213) of ANT(6)FMS-007 were identified. Thirty-one EM clinical isolates (74%) carried the ant(6) gene. Eight EM clinical isolates containing the ant(6) gene had MIC values (

Yan Tong,1 Xin Huang2 1Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong, Hong Kong, People’s Republic of China; 2Department of Ophthalmology, Jiangxi Provincial People’s Hospital, the First Affiliated Hospital of Nanchang Medical College, Nanchang, People’s Republic of ChinaCorrespondence: Xin Huang, Department of Ophthalmology, Jiangxi Provincial People’s Hospital, the First Affiliated Hospital of Nanchang Medical College, No. 152, Ai Guo Road, Dong Hu District, Nanchang, Jiangxi, 330006, People’s Republic of China, Email 334966891@qq.comObjective: To investigate spontaneous neuronal activity changes in patients with central retinal artery occlusion (CRAO) using the resting-state functional magnetic resonance imaging (fMRI) and detect whether these brain functional alterations can represent an objective biomarker of clinical response using a machine learning algorithm.Methods: Eighteen patients with CRAO and eighteen healthy controls (HCs) were recruited. The regional homogeneity (ReHo) method of resting-state fMRI was conducted to evaluate the synchronous brain activity alterations between two groups. Differences of ReHo values between two groups were compared using the independent two-sample t-test. The support vector machine algorithm was to distinguish patients of CRAO from HCs based on the two groups’ whole-brain ReHo patterns. The accuracy, sensitivity, and specificity for the classification were calculated. The classification performance was evaluated using the non-parametric permutation test.Results: Compared to HCs, individuals with CRAO showed significantly lower ReHo in the right cerebellum and precuneus. Meanwhile, significant higher ReHo values were observed in the left superior temporal gyrus, postcentral gyrus, and precentral gyrus in the CRAO group compared to HCs. Furthermore, our results suggested that 77.78% individuals with CRAO could be successfully distinguished from HCs by the machine learning, with a sensitivity of 72.22% and a specificity of 83.33%, respectively. The area of receiver operating characteristic curve was calculated to be 0.85.Conclusion: This study uncovered individuals with CRAO exhibited disturbed synchronous neuronal activities in multiple brain areas using neuroimaging techniques. The ReHo variability could distinguish individuals with CRAO from HCs with high accuracy.Keywords: central retinal artery occlusion, functional magnetic resonance imaging, regional homogeneity, machine learning, support vector machine

Yong Li,1,2,&ast; Senbo Zhu,2,&ast; Junchao Luo,2 Yu Tong,2 Yixuan Zheng,1 Lichen Ji,2 Zeju He,2 Qiangan Jing,1 Jiaqing Huang,2 Yinjun Zhang,1 Qing Bi2 1College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, Zhejiang, 310014, People’s Republic of China; 2Laboratory Medicine Center, Department of Laboratory Medicine, 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: Yinjun Zhang, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, Zhejiang, 310014, People’s Republic of China, Tel +86-13735494556, Email zhangyj@zjut.edu.cn Qing Bi, Laboratory Medicine Center, Department of Laboratory Medicine, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital, Hangzhou Medical College, Hangzhou, Zhejiang, 310014, People’s Republic of China, Tel +86-13588302991, Email bqzjsrmyy@163.comIntroduction: Osteoarthritis (OA) is a common chronic joint disease characterized by articular cartilage degeneration. OA usually manifests as joint pain, limited mobility, and joint effusion. Currently, the primary OA treatment is non-steroidal anti-inflammatory drugs (NSAIDs). Although they can alleviate the disease’s clinical symptoms and signs, the drugs have some side effects. Selenium nanoparticles (SeNPs) may be an alternative to relieve OA symptoms.Materials and Results: We confirmed the anti-inflammatory effect of selenium nanoparticles (SeNPs) in vitro and in vivo experiments for OA disease in this study. In vitro experiments, we found that SeNPs could significantly reduce the expression of nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), the major inflammatory factors, and had significant anti-inflammatory and anti-arthritic effects. SeNPs can inhibit reactive oxygen species (ROS) production and increased glutathione peroxidase (GPx) activity in interleukin-1beta (IL-1β)-stimulated cells. Additionally, SeNPs down-regulated matrix metalloproteinase-13 (MMP-13) and thrombospondin motifs 5 (ADAMTS-5) expressions, while up-regulated type II collagen (COL-2) and aggrecan (ACAN) expressions stimulated by IL-1β. The findings also indicated that SeNPs may exert their effects through suppressing the NF-κB p65 and p38/MAPK pathways. In vivo experiments, the prevention of OA development brought on by SeNPs was demonstrated using a DMM model.Discussion: Our results suggest that SeNPs may be a potential anti-inflammatory agent for treating OA.Keywords: osteoarthritis, NF-κB p65 signaling pathway, p38/MAPK signaling pathway, selenium nanoparticles

Fanghui Li,1,2,&ast; Dongze Li,1,&ast; Jing Yu,1 Yu Jia,3 Jirui Wen,4 Wentao Li,2 Yao Tong,2 Jiang Wu,4 Zhi Wan,1 Yu Cao,1 Qing Zhang,2 Rui Zeng1,2 1Department of Emergency Medicine, Laboratory of Emergency Medicine, West China Hospital, Sichuan University, Chengdu, People’s Republic of China; 2Department of Cardiology, West China Hospital, Sichuan University, Chengdu, People’s Republic of China; 3Department of General Practice and National Clinical Research Center for Geriatrics, International Medical Center, West China Hospital, Sichuan University, Chengdu, People’s Republic of China; 4Deep Underground Space Medical Center, West China Hospital, Sichuan University, Chengdu, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Rui Zeng; Qing Zhang, Department of Cardiology, West China Hospital, Sichuan University, 37 Guoxue Road, Chengdu, Sichuan, 610041, People’s Republic of China, Tel +86-28-85423248, Fax +86-28-85582944, Email zengrui_0524@126.com; qzhang2000cn@163.comBackground: The plasma lipidome profile is likely to improve risk stratification in patients with acute coronary syndrome (ACS) and predict cardiovascular events for secondary disease prevention. Ceramides are involved in the initiation or acceleration of several key pathophysiological processes in atherosclerosis. This study evaluated whether plasma ceramide levels at admission was associated with one-year mortality in patients with ACS.Methods: In total, 826 patients with ACS from a prospective multicenter study for early evaluation of acute chest pain were enrolled. High-performance liquid chromatography with tandem mass spectrometry (LC/MS) was used to measure the plasma levels of eleven ceramides (C16–C26). The primary outcome was all-cause mortality, and the secondary outcome was cardiac mortality during the one-year follow-up. The relationship between the ceramide levels and mortality was evaluated by Cox regression analysis. The receiver operating characteristic (ROC) curve was established to evaluate discrimination of ceramides.Results: Eighty-eight (10.7%) patients died after a 12-month follow-up. Five ceramides (C16:0, C18:0, C20:0, C24:1 and C24:2) and their ratios to Cer(d18:1/24:0) were independently associated with the risk of all-cause death and cardiac death. Combining the Global Registry of Acute Coronary Events (GRACE) score with ceramides and their ratios to Cer(d18:1/24:0) had areas under ROC curves ranging from 0.778– 0.804 (P< 0.001) for all-cause mortality, which was greater than that of the GRACE score alone.Conclusion: Measurements of long-chain ceramides and very-long-chain ceramides may help in identifying a high risk of mortality beyond traditional assessment tools in patients with ACS.Trial Registration: clinicaltrials.gov, identifier: NCT04122573.Keywords: ceramide, acute coronary syndrome, mortality, high-performance liquid chromatography with tandem mass spectrometry

Junhui He,1,&ast; Dongbo Han,2,&ast; Chunlian Jia,2,&ast; Jiaxiu Xie,1 Fucui Zhu,2 Jie Wei,1 Dongmei Li,1 Dongmei Wei,1 Yi Li,1 Li Tang,3 Guining Wei,1,4 Jing Yan,4 Yuanming Tong,1 Lifang Yang,4,&ast; Xuecai Tan4,&ast; 1Department of Pharmacology, Key Laboratory of Quality Standards, Guangxi Institute of Chinese Medicine & Pharmaceutical Science, Nanning, 530022, People’s Republic of China; 2Department of Pharmacology, Guangxi Medical University, Nanning, 530021, People’s Republic of China; 3Department of Pharmacy, the First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine, Nanning, 530022, People’s Republic of China; 4Guangxi Key Laboratory of Chemistry and Engineering of Forest Products, School of Chemistry and Chemical Engineering, Guangxi Minzu University, Nanning, 530008, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xuecai Tan; Lifang Yang, Guangxi Key Laboratory of Chemistry and Engineering of Forest Products, School of Chemistry and Chemical Engineering, Guangxi Minzu University, Nanning, 530008, People’s Republic of China, Tel +86 0771-5877473, Email gxunxctan@126.com; yanglf1990@163.comPurpose: To investigate the mechanisms of antidepressant action of active fraction of Polyrhachis vicina Rogers (AFPR) through network pharmacology, molecular docking and experimental validation.Methods: GC-MS was used to predict chemical compounds, corresponding databases were used to predict chemical compound targets and depression targets, Cytoscape software was used to construct and analyze the protein interaction network map, DAVID database was used to analyze gene ontology (GO) and KEGG signaling pathway, and AGFR software was used to perform molecular docking. Subsequently, the underlying action mechanisms of AFPR on depression predicted by network pharmacology analyses were experimentally validated in a CORT-induced depression model in vitro and in vivo.Results: A total of 52 potential targets of AFPR on antidepressant were obtained. GO is mainly related to chemical synaptic transmission, signal transduction and others. KEGG signaling pathways are mainly related to cAMP signaling pathway and C-type lectin receptor signaling pathway. The experiment results showed that AFPR significantly increased the expression of PRKACA, CREB and BDNF in mouse brain tissue and PC12 cells. Furthermore, after interfered of cAMP in PC12 cells, the decreased expression of PRKACA, CREB and BDNF was reversed by AFPR.Conclusion: AFPR may exert antidepressant effects through multiple components, targets and pathways. Furthermore, it could improve neuroplasticity via the cAMP signaling pathway to improve depression-like symptoms.Graphical Abstract: Keywords: PC12 cells, BDNF, corticosterone, CAMP signaling pathway, GC-MS

Qi Xin,1 Tonghui Xie,1 Rui Chen,1 Xing Zhang,1 Yingmu Tong,1 Hai Wang,1 Shufeng Wang,2 Chang Liu,1,3 Jingyao Zhang1,3 1Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, 710061, People’s Republic of China; 2Department of General Surgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, 710061, People’s Republic of China; 3Department of SICU, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, 710061, People’s Republic of ChinaCorrespondence: Chang Liu; Jingyao Zhang, Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, 710061, People’s Republic of China, Tel +86-29-85323900, Fax +86-29-85324642, Email liuchangdoctor@163.com; you12ouy@163.comBackground: Sepsis-induced acute kidney injury (S-AKI) is associated with systemic inflammatory responses and coagulation system dysfunction, and it is associated with an increased risk of mortality. However, there was no study to explore the predictive value of inflammatory and coagulation indicators for S-AKI.Methods: In this retrospective study, 1051 sepsis patients were identified and divided into a training cohort (75%, n = 787) and a validation cohort (25%, n = 264) in chronological order according to the date they were admitted. Univariate analyses and multivariate logistic regression analyses were performed to identify the independent predictors of S-AKI. The logistic regression analyses (enter methods) were used to conducted the prediction models. The ROC curves were used to determine the predictive value of the constructed models on S-AKI. To test whether the increase in the AUC is significant, we used a two-sided test for ROC curves available online (http://vassarstats.net/roc_comp.html). The secondary outcome was different AKI stages and major adverse kidney events within 30 days (MAKE30). Stage 3B of S-AKI was defined as both meeting the stage 3 criteria [increase of Cr level by > 300% (≥ 4.0 mg/dL with an acute increase of ≥ 0.5 mg/dL) and/or UO < 0.3 mL/kg/h for > 24 h or anuria for > 12 h and/or acute kidney replacement therapy] and having cystatin C positive. MAKE30 were a composite of death, new renal replacement therapy (RRT), or persistent renal dysfunction (PRD).Results: We discovered that cardiovascular disease, white blood cell (WBC), mean arterial pressure (MAP), platelet (PLT), serum procalcitonin (PCT), prothrombin time activity (PTA), and thrombin time (TT) were independent predictors for S-AKI. The predictive value (AUC = 0.855) of the simplest model 3 (constructed with PLT, PCT, and PTA), with a sensitivity of 77.6% and a specificity of 82.4%, had a similar predictive value comparing with the model 1 (AUC = 0.872) and the model 2 (AUC = 0.864) in the training cohort (P > 0.05). Compared with the model 1 (AUC = 0.888) and the model 2 (AUC = 0.887), the model 3 (AUC = 0.887) had a similar predictive value in the validation cohort. Moreover, model 3 had the best predictive power for predicting S-AKI in the stage 3 (AUC = 0.777), especially in stage 3B (AUC = 0.771). Finally, the model 3 (AUC = 0.843) had perfect predictive power for predicting MAKE30 in sepsis patients.Conclusion: Within 24 hours after admission, the simplest model 3 (constructed with PLT, PCT, and PTA) might be a robust predictor of the S-AKI in sepsis patients, providing information for timely and efficient intervention.Keywords: inflammation, coagulation, prediction model, sepsis, acute kidney injury

Yanping Tong,1,2 Jie Liu,3 Tao Yang,1,2 Jingwen Wang,4 Tianyou Zhao,4 Yuezhi Kang,1,2 Yongping Fan1,2 1Department of Traditional Chinese Medicine, Beijing Tiantan Hospital, Capital Medical University, Beijing, 100070, People’s Republic of China; 2Beijing Integrative Medicine on Encephalopathy Research Institution, Beijing Tiantan Hospital, Capital Medical University, Beijing, 100070, People’s Republic of China; 3Department of Respiratory Medicine, Beijing First Hospital of Integrated Chinese and Western Medicine, Beijing, 100026, People’s Republic of China; 4Department of Neurology, Beijing Miyun District Hospital of Traditional Chinese Medicine, Beijing, 101599, People’s Republic of ChinaCorrespondence: Yongping Fan, Email yongpingf@hotmail.comObjective: To investigate the association of pain with plasma C5a levels and other related inflammatory cytokines in neuromyelitis optica spectrum disorders (NMOSD) patients during remission.Participants and Methods: NMOSD patients (n = 87) and healthy controls (HC; n = 44) were consecutively recruited between January 2017 and April 2018. Plasma complement 5 (C5), C5a, interleukin (IL)-6, tumor necrosis factor (TNF)-α, and IL-1β levels were detected. Visual Analogue Scale (VAS), ID pain scale, 24-item Hamilton Depression Scale (HAMD), Multiple Sclerosis Impact Scale (MSIS-29), and Kurtzke Expanded Disability Status Scale (EDSS) were used to evaluate the degree and types of pain, the existence of depression and anxiety, and the life quality and disability status of patients. Binary logistic regression equation was used to assess the association of pain with plasma C5a levels.Results: Among the 87 NMOSD patients, 40 complained of pain that in 67.5% (27/40) of cases had a neuropathic component (ID pain ≥ 2). Plasma C5a, IL-6, TNF-α, and IL-1β levels were significantly elevated in NMOSD patients than in HC. Plasma C5 levels were negatively correlated with the time from sampling to the last relapse or disease onset. NMOSD patients with pain had higher plasma C5a levels, and they suffered from a higher disability, more anxiety, and worse life quality compared to those patients without pain. In NMOSD patients with pain, there were not significant differences between plasma levels of C5, C5a, IL-6, TNF-α, or IL-1β, regardless of neuropathic pain or not. Binary logistic regression showed that the OR of plasma C5a level was 1.002, with gender and EDSS score were identified as independent factors associated with pain in NMOSD.Conclusion: NMOSD patients during remission had elevated C5a and related inflammatory cytokines levels in peripheral blood. Elevated C5a may have a unique role in the pathogenesis of pain in NMOSD patients.Keywords: neuromyelitis optica spectrum disorders, remission, pain, C5a

Amina Nazir,1– 3 Chunling Qi,4 Na Shi,4 Xue Gao,4 Qiang Feng,4 Hong Qing,2 Fei Li,3,4 Yigang Tong3 1Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, Shandong Province, People’s Republic of China; 2Key Laboratory of Molecular Medicine and Biotherapy in the Ministry of Industry and Information Technology, Department of Biology, School of Life Sciences, Beijing Institute of Technology, Beijing, People’s Republic of China; 3College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, People’s Republic of China; 4Clinical Laboratory Center, The Affiliated Taian City Central Hospital of Qingdao University, Taian, 271000, People’s Republic of ChinaCorrespondence: Fei Li; Yigang Tong, Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, People’s Republic of China, Email lf-314@163.com; tong.yigang@gmail.comIntroduction: Klebsiella pneumoniae, a multidrug resistant bacterium, that causes nosocomial infections including septicemia, pneumonia etc. Bacteriophages are potential antimicrobial agents for the treatment of antibiotic resistant bacteria.Methods and Results: In this study, a novel bacteriophage IME268 was isolated from hospital sewage against clinical multi-drug resistant Klebsiella pneumoniae. Transmission electron microscopy and genomic characterization of this phage exhibited it belongs to the Webervirus genus, Drexlerviridae family. Phage IME268 possessed a double-stranded DNA genome composed of 49,552bp with a GC content of 50.5%. The phage genome encodes 77 open reading frames, out of 44 are hypothetical proteins while 33 had assigned putative functions. No tRNA, virulence related or antibiotic resistance genes were found in phage genome. Comparative genomic analysis showed that phage IME268 has 95% identity with 87% query cover with other phages in NCBI database. Multiplicity of infection, one step growth curve and host range of phage were also measured.Conclusion: According to findings, Phage IME268 is a promising biological agent that infects Klebsiella pneumoniae and can be used in future phage therapies.Keywords: Klebsiella pneumoniae, bacteriophage, therapy, lytic, drug resistant

Fanxing Meng,1 Yongsheng Xu,1 Yiliang Liu,2 Guozan Zhang,3 Yunze Tong,4 Rong Lin1 1Zhejiang Police College, Hangzhou, Zhejiang, People’s Republic of China; 2College of Economics and Management, Northeast Forestry University, Harbin, Heilongjiang, People’s Republic of China; 3School of Creative Arts and Design, Zhejiang Institute of Mechanical & Electrical Engineering, Hangzhou, Zhejiang, People’s Republic of China; 4College of Computer Science and Technology, Zhejiang University, Hangzhou, Zhejiang, People’s Republic of ChinaCorrespondence: Fanxing Meng, Zhejiang Police College, 555 Binwen Road, Binjiang District, Hangzhou, Zhejiang, People’s Republic of China, Email mengfanxing@zjjcxy.cnPurpose: The issue of employee engagement has increasingly become a focus of concern in public management practice. Based on the theory of purposeful work behavior, integrative theory of employee engagement and Pratt and Ashforth’s typology of work meaningfulness, this study proposes and examines the mediating effects of two types of meaningfulness between transformational leadership and work engagement and the moderating effects of transformational leadership on the relationship between two types of meaningfulness and work engagement.Patients and Methods: By adopting a multilevel cross-sectional design, this study examines assumed mediation and moderation effects. The data collection was conducted anonymously by means of an online survey. A total of 261 local police officers from 32 police stations were recruited in professional training programs as a sample.Results: The analysis reveals that both meaningfulness in work and meaningfulness at work positively mediate the relationship between transformational leadership and work engagement. Transformational leadership moderates the relationship between meaningfulness at work and work engagement rather than the relationship between meaningfulness in work and work engagement. There are no other significant effects of sex, age or length of service.Conclusion: Work meaningfulness transmits and combines the effect of transformational leadership to impact work engagement. These findings not only confirm the critical role of work meaningfulness proposed by the theoretical frameworks of the theory of purposeful work behavior, integrative theory of employee engagement and Pratt and Ashforth’s typology of work meaningfulness, but also further extend and clarify the role of and difference in two substructures of work meaningfulness (ie, work meaningfulness in work and at work) in the context of the linkage between transformational leadership and work engagement.Keywords: transformational leadership, meaningfulness in work, meaningfulness at work, work engagement

Yingmu Tong,1,&ast; Qinglin Li,1,&ast; Sinan Liu,1,2 Tong Liu,1 Kai Qu,1 Chang Liu,1,2 Jingyao Zhang1,2 1Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, Shaanxi, 710061, People’s Republic of China; 2Department of SICU, the First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, Shaanxi, 710061, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Chang Liu; Jingyao Zhang, Department of SICU, The First Affiliated Hospital of Xi’an Jiaotong University, 277 Yanta West Road, Xi`an, 710061, People’s Republic of China, Tel +86-29-85323900, Fax +86-29-85324642, Email liuchangdoctor@163.com; you12ouy@163.comPurpose: We describe the clinical characteristics, isolated microorganisms and antibiotics used in patients with cIAIs during ICU stay.Methods: A retrospective analysis of data from Beth Israel Deaconess Medical Center was performed.Results: A total of 316 patients with cIAIs were included, 57.0% of them were male and the median age was 63 years. A total of 239 patients did have cultures taken, and 74 patients had a positive microbial result. The main pathogens were Escherichia coli, Staphylococci (coagulase negative), Enterococcus sp. and Bacteroides fragilis. The main antibiotics given were vancomycin, metronidazole, piperacillin tazobactam and ciprofloxacin. Univariable and multivariable Cox regression analyses showed that receiving more antibiotics reduced ICU mortality, but the same results were not obtained in the analysis of hospital mortality.Conclusion: The main Gram-positive microorganisms for empirical antimicrobial therapy were Staphylococci (coagulase negative), Enterococcus sp. and Staphylococcus aureus, and Gram-negative microorganisms were Escherichia coli, Bacteroides fragilis and Klebsiella pneumoniae. The use of more antibiotics reduced ICU mortality, but the same results were not obtained in the analysis of hospital mortality.Keywords: intra-abdominal infection, culture, microorganism, antibiotics, intensive care unit, mortality

Qinglin Li,1 Yingmu Tong,1 Hai Wang,1 Jie Ren,1 Sinan Liu,1,2 Tong Liu,1 Kai Qu,1 Chang Liu,1,2 Jingyao Zhang1,2 1Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, Shaanxi, 710061, People’s Republic of China; 2Department of SICU, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, Shaanxi, 710061, People’s Republic of ChinaCorrespondence: Chang Liu; Jingyao ZhangThe First Affiliated Hospital of Xi’an Jiaotong University, 277 Yanta West Road, Xi`an, 710061, People’s Republic of ChinaTel +86-29-85323900Fax +86-29-85324642Email liuchangdoctor@163.com; you12ouy@163.comObjective: The aim of this study was to compare the clinical characteristics and short-term mortality of patients with abdominal and pulmonary sepsis.Design: Retrospective cohort study.Setting: Adult intensive care units (ICUs) at tertiary hospitals.Participants: Adult ICU patients from 2014 to 2015 in the eICU Collaborative Research Database.Interventions: In univariate analysis, we compared the differences in the characteristics of patients in each group. Logistic regression models were used to evaluate the relationships between primary site of sepsis and short-term prognosis.Primary and Secondary Outcome Measures: Hospital and ICU survival.Results: The final dataset included 7023 pulmonary and 2360 abdominal sepsis patients, who accounted for 74.84% and 25.16%, respectively. We compared the results of the baseline characteristics, vital signs and laboratory indicators between the two groups. In the logistic regression models, we found that the hospital and ICU mortality of patients with abdominal sepsis was higher than that with pulmonary sepsis (p < 0.05, OR = 1.15, p < 0.05, OR = 1.19, respectively), although these results were no longer significantly after adjustment for confounders, but in the subgroups with SOFA score ≧ 8, the adjusted hospital mortality rate of patients with abdominal sepsis was 1.30 times higher than that of patients with pulmonary sepsis (p < 0.005, OR = 1.30, 95% CI 1.09– 1.55), while there was no significant difference in the subgroups that SOFA score < 8.Conclusion: In the patients with SOFA score ≧ 8, the adjusted hospital mortality of patients with abdominal sepsis was higher than patients with pulmonary sepsis.Keywords: sepsis, pulmonary, abdominal, eICU-CRD, ICU

Pengfei Li, Fengkai Yang, Yuexin Tong, Ying Chen, Youxin Song Department of Orthopedic, Affiliated Hospital of Chengde Medical University, Chengde, Hebei, 067000, People’s Republic of ChinaCorrespondence: Youxin SongDepartment of Orthopedic, Affiliated Hospital of Chengde Medical University, No. 36 Nanyingzi St, Chengde, Hebei, 067000, People’s Republic of ChinaEmail syxmed@163.comBackground: Modic changes (MC) are generally considered to be related to degenerative disc disease, and there is no uniform standard for surgical methods for lumbar disc herniation (LDH) accompanied by Modic type I changes (MC I). The purpose of this study was to observe the clinical results of percutaneous transforaminal endoscopic decompression (PTED) and transforaminal lumbar interbody fusion (TLIF) for treatment of LDH accompanied by MC I.Methods: Of the 53 consecutive patients included, 29 underwent PTED and 24 underwent TLIF. All patients were followed up for at least 24 months. Preoperative demographic characteristics, perioperative outcomes, and clinical outcomes were recorded. Visual analog scale (VAS) scores, Oswestry disability index (ODI) scores, and modified Macnab criteria were used to assess clinical results.Results: The mean age was 53.7± 9.2 years in the PTED group and 53.6± 9.6 years in the TLIF group. The scores of VAS legs, VAS back and ODI in the two groups after operation were significantly improved compared with those before operation (P< 0.05). Notably, the VAS back pain score and ODI in the PTED group showed an increasing trend with time. And the VAS back pain scores and ODI of the two groups were statistically different at 1 year and 2 years postoperatively (P< 0.05). In addition, compared with the TLIF group, the PTED group showed less operation time, blood loss, and postoperative hospital stay (P< 0.05). At the final follow-up, the excellent rates were 91.7% and 86.2% in the fusion and PTED groups, respectively.Conclusion: Both PTED and TLIF procedures significantly improved the clinical symptoms of single-level LDH patients with MC I. Compared with TLIF, MC I may affect the improvement of low back pain and functional status after PTED.Keywords: PTED, fusion, Modic change, lumbar disc herniation, back pain

Amina Nazir,1,2 Azam Ali,3 Hong Qing,1 Yigang Tong2 1Key Laboratory of Molecular Medicine and Biotherapy in the Ministry of Industry and Information Technology, Department of Biology, School of Life Sciences, Beijing Institute of Technology, Beijing, People’s Republic of China; 2Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, People’s Republic of China; 3Centre for Applied Molecular Biology (CAMB), University of the Punjab, Lahore, PakistanCorrespondence: Hong QingKey Laboratory of Molecular Medicine and Biotherapy in the Ministry of Industry and Information Technology, Department of Biology, School of Life Sciences, Beijing Institute of Technology, Beijing, People’s Republic of ChinaTel +86 13264286266Email hqing@bit.edu.cnYigang TongBeijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, People’s Republic of ChinaEmail tong.yigang@gmail.comAbstract: The bacteriophages have been explored at a huge scale as a model system for their applications in many biological-related fields. Jumbo phages with a large genome size from 200 to 500 kbp were not previously assigned a great value, and characterized by complex structures coupled with large virions with a wide variety of hosts. The origin of most of the jumbo phages was not well understood; however, many other prominent features have been discovered recently. In the current review, we strive to unearth the most advanced characteristics of jumbo phages, particularly their significance and structural organization that holds immense value to the viral life cycle. The unique characteristics of jumbo phages are the basis of variations in different types of phages concerning their organization at the genomic level, virion structure, evolution, and progeny propagation. The presence of tRNA and additional translation-related genes along with chaperonin genes mark the ability of these phages for being independent of host molecular machinery enabling them to have wide host options. A large number of jumbo phages have been isolated from various sources through advanced standard screening methods. The current review has summarized the available data on jumbo phages and discussed the genome orientation of jumbo phages, translational machinery, diversity and evolution of jumbo phages. In the studies conducted, jumbo phages possessed special additional genes that helps to reduce the dependence of jumbo phages on their hosts. Furthermore, their genomes might have evolved from smaller genome phages.Keywords: bacteriophages, tRNA, large genome, wide host range, chaperonin

Yao Tong,&ast; Chengrong Bao,&ast; Yi-Qiong Xu,&ast; Lei Tao,&ast; Yao Zhou, Lei Zhuang, Ying Meng, Hui Zhang, Jingjing Xue, Weijun Wang, Lele Zhang, Qingbo Pan, Zhenzhen Shao, Tianran Hu, Qian Guo, Qingsheng Xue, Han Lu, Yan Luo Department of Anesthesiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yan Luo; Han Lu Email ly11087@rjh.com.cn; luhan0301@163.comBackground: Acute lung injury (ALI) is a severe respiratory disease with high rates of morbidity and mortality. Many mediators regarding endogenous or exogenous are involved in the pathophysiology of ALI. Here, we have uncovered the involvement of integrins and matrix metalloproteinases, as critical determinants of excessive inflammation and endothelial permeability, in the regulation of ALI.Methods: Inflammatory cytokines were measured by quantitative real-time PCR for mRNA levels and ELISA for secretion levels. Endothelial permeability assay was detected by the passage of rhodamine B isothiocyanate-dextran. Mice lung permeability was assayed by Evans blue albumin (EBA). Western blot was used for protein level measurements. The intracellular reactive oxygen species (ROS) were evaluated using a cell-permeable probe, DCFH-DA. Intratracheal injection of lipopolysaccharide (LPS) into mice was conducted to establish the lung injury model.Results: Exogenous MMP-9 significantly aggravated the inflammatory response and permeability in mouse pulmonary microvascular endothelial cells (PMVECs) treated by LPS, whereas knockdown of MMP-9 exhibited the opposite phenotypes. Knockdown of integrin β 3 or β 5 in LPS-treated PMVECs significantly downregulated MMP-9 expression and decreased inflammatory response and permeability in the presence or absence of exogenous MMP-9. Additionally, the interaction of MMP-9 and integrin β 5 was impaired by a ROS scavenger, which further decreased the pro-inflammatory cytokines production and endothelial leakage in PMVECs subjected to co-treatment (LPS with exogenous MMP-9). In vivo studies, exogenous MMP-9 treatment or knockdown β 3 integrin significantly decreased survival in ALI mice. Notably, knockdown of β 5 integrin alone had no remarkable effect on survival, but which combined with anti-MMP-9 treatment significantly improved the survival by ameliorating excessive lung inflammation and permeability in ALI mice.Conclusion: These findings support the β 3/5 integrin-MMP-9 axis as an endogenous signal that could play a pivotal role in regulating inflammatory response and alveolar-capillary permeability in ALI.Keywords: acute lung injury, endothelial cells, integrin, MMP-9

Yujie Lu,1,&ast; Yiwei Tong,1,&ast; Jiahui Huang,1 Lin Lin,2 Jiayi Wu,1 Xiaochun Fei,3 Xiaosong Chen,1 Kunwei Shen1 1Department of General Surgery, Comprehensive Breast Health Center, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China; 2Department of Clinical Laboratory, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, 200025, People’s Republic of China; 3Department of Pathology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Xiaosong Chen; Kunwei ShenDepartment of General Surgery, Comprehensive Breast Health Center, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of ChinaEmail chenxiaosong0156@hotmail.com; kwshen@medmail.com.cnBackground: It remains uncertain whether the 21-gene recurrence score (RS) of a primary tumor has selective prognostic value for locoregional recurrence (LRR) or distant metastasis (DM). The current study aimed to compare the distribution and single-gene expression on the RS panel in breast cancer patients with LRR versus DM.Methods: Consecutive early breast cancer patients who had been operated on at the Comprehensive Breast Health Center, Ruijin Hospital from January 2009 to December 2016 were retrospectively reviewed. Patients were divided into LRR, DM, and no-recurrence groups according to the first reported recurrent event. Comparison and subgroup analysis of 21-gene RS, RS category, and single-gene expression on the RS panel were conducted among patients with different recurrence status.Results: A total of 1,287 patients were included, with median follow-up of 61.5 months, and 27, 47, and 1,213 patients were classified as LRR, DM, and no recurrence groups, respectively. RS was significantly diversely distributed among the three groups (P< 0.001). No-recurrence patients (median 22) presented much lower RS than LRR (median 39, P< 0.001) and DM (median 30, P< 0.001) patients. LRR patients had lower PR (P< 0.001), BCL2 (P=0.010), and CEGP1 (P< 0.001) expression, and DM patients had higher STMY3 (P=0.019) expression than no-recurrence patients. Moreover, CEGP1 expression was significantly lower in the LRR group than the DM one (P=0.028).Conclusion: RS was differently distributed between recurrent and nonrecurrent patients. PR, BCL2, CEGP1, and STMY 3 expression was associated with LRR and DM, while CEGP1 was lower in the LRR group than DM patients, warranting further clinical evaluation.Keywords: breast cancer, 21-gene recurrence assay, gene expression, locoregional recurrence, distant metastasis

Li Cai,1,2 Lixian Wang,3 Xuhua Guan,4 Lei Wang,4 Xinyi Hu,5 Yang Wu,4 Yeqing Tong,4 Peigang Wang1 1School of Health Sciences, Wuhan University, Wuhan, 430071, People’s Republic of China; 2Wuhan Center for Disease Control and Prevention, Wuhan, 430015, People’s Republic of China; 3Xiaonan Center for Disease Control and Prevention, Xiaogan, 432100, People’s Republic of China; 4Hubei Center for Disease Control and Prevention, Wuhan, Hubei, 430079, People’s Republic of China; 5Global Study Institute, University of Geneva, Geneva, SwitzerlandCorrespondence: Peigang Wang Email wpg926@163.comYeqing Tong Tel +8627 87652010Email t_yeqing@163.comObjective: The study aimed to timely grasp the epidemiologic status of rabies in Central China from 2013 to 2018 and provide scientific evidence for the implementation of follow-up prevention and control measures.Methods: We initiated a retrospective observational and descriptive study of bite-related injuries data and rabies disease data in Hubei province from 2013 to 2018, managed by the Center for Disease Control and Prevention.Results: A total of 2,028,691 individuals were exposed to bites from 2013 to 2018, of which 221 were diagnosed with rabies and deceased. Among those cases, the incubation periods of rabies varied from 3 days to 18,406 days, which has been shown to be statistically associated with where the infected person was exposed and whether the wound care has been conducted.Conclusion: Epidemiological studies have shown that from 2013 to 2018, the current situation of rabies in Central China is still severe. The case fatality rate keeps virtually 100%. The rural population is still the most vulnerable group to rabies, characterized by a high exposure ratio and low treatment rate as well as poor vaccination compliance. Hoewever, larger populations are warranted to validate our findings.Keywords: rabies, epidemiology, China

Yingying Xu,1 Huanyu Zhao,2 Yusuo Tong,1 Wanwei Wang,1 Jing Huang,1 Weiguo Zhu1 1Department of Radiation Oncology, The Affiliated Huaian No.1 People’s Hospital of Nanjing Medical University, Huai’an, Jiangsu, 223300, People’s Republic of China; 2Department of Pathology, The Affiliated Huaian No.1 People’s Hospital of Nanjing Medical University, Huai’an, Jiangsu, 223300, People’s Republic of ChinaCorrespondence: Weiguo Zhu Email zhuweiguo_1270@126.comBackground: Basaloid squamous cell carcinoma (BSCC) is a rare variant of squamous cell carcinoma (SCC) of the esophagus. This study aimed to assess the discrepancy in clinicopathological characteristics and protein expression between esophageal BSCC and typical esophageal SCC.Study Design: We reviewed 40 cases of esophageal BSCC. As controls, 63 well-differentiated SCC (WSCC) patients, 70 moderately differentiated SCC (MSCC) patients, and 51 poorly differentiated SCC (PSCC) patients were selected. The clinicopathologic characteristics and immunoreactivity of Ki-67, p53, p63, and epidermal growth factor receptor (EGFR) were then evaluated in the BSCC and typical SCC patients.Results: The 5-year survival rates for the BSCC patients were 27.5%. The prognostic outcomes of the BSCC group were similar to those of the PSCC and MSCC groups but worse than that of the WSCC group, with a significant difference (P=0.045). Ki-67 expression was significantly higher in the BSCC group than that in the WSCC group (P < 0.05). Meanwhile, there were no significant differences in the expression of the other molecular markers (p53, p63, and EGFR) between the typical SCC and BSCC groups (P > 0.05). The median survival time of esophageal the BSCC patients with low p53 expression was significantly longer than that of the patients with high p53 expression (P=0.026). Further, the median survival time of the esophageal BSCC patients with high p63 expression was significantly longer than that of the patients with low p63 expression (P=0.041). Meanwhile, Ki-67 and EGFR expressions were not correlated with OS in the BSCC group.Conclusion: Esophageal BSCC has a more clinically virulent course. Notably, p53 and p63 expression are associated with prognosis in BSCC. These findings conject that evaluation of multiple cancer biomarkers might be a promising auxiliary diagnostic indicator in BSCC.Keywords: basaloid squamous cell carcinoma, Ki-67, p53, p63, EGFR

Chen-xing Qi,1,&ast; Xin Huang,2,&ast; Yan Tong,1 Yin Shen1,3 1Eye Center, Renmin Hospital of Wuhan University, Wuhan, 430060, Hubei, People’s Republic of China; 2Department of Ophthalmology, Jiangxi Provincial People’s Hospital, Nanchang 330006, People’s Republic of China; 3Medical Research Institute, Wuhan University, Wuhan, 430071, Hubei, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yin ShenEye Center, Renmin Hospital of Wuhan University, No. 238, Jie Fang Road, Wu Chang District, Wuhan, 430060, Hubei, People’s Republic of ChinaTel +86 13871550513Email yinshen@whu.edu.cnObjective: The purpose of the study was to find the differences in intrinsic functional connectivity (FC) patterns of the primary visual area (V1) among diabetic retinopathy (DR), diabetes mellitus (DM), and healthy controls (HCs) applying resting-state functional magnetic resonance imaging (rs-fMRI).Patients and Methods: Thirty-five subjects with DR (18 males and 17 females), 22 DM (10 males and 12 females) and 38 HCs (16 males and 22 females) matched for sex, age, and education underwent rs-fMRI scanning. Seed-based FC analysis was performed to find the alterations in the intrinsic FC patterns of V1 in DR compared with DM and HCs.Results: The study found that DR patients had a significant lower FC between the bilateral calcarine (CAL)/left lingual gyrus (LING) (BA 17/18) and the left V1, and between the bilateral CAL/left LING (BA 17/18) and the right V1 compared with the HCs. Meanwhile, patients with DR exhibited higher FC strength between the left V1 and the bilateral Caudate/Olfactory/Orbital superior frontal gyrus (OSFG), and between the bilateral Caudate/Olfactory/OSFG (BA 3/4/6) and the right V1. Compared with DM group, patients with DR showed increased FC strength between the right CAL (BA 17/18) and the right V1. DM group exhibited lower FC strength between the left fusiform and the left V1, and between the bilateral CAL and the right V1 when compared with HCs. Moreover, DM group was observed to have higher FC strength between the left superior frontal gyrus and the left V1.Conclusion: Our findings indicated that DR patients exhibited FC disruptions between V1 and higher visual regions at rest, which may reflect the aberrant information communication in the V1 area of DR individuals. The findings offer important insights into the neuromechanism of vision disorder in DR patients.Keywords: intrinsic functional connectivity, diabetic retinopathy, primary visual area

Xin Zheng,1,2 Yanyan Qi,1,2 Hongjun Zhou,2,3 Haiqiong Kang,2,3 Yanming Tong,1,2 Lina Bi1,2 1Department of Endocrinology, China Rehabilitation Research Center, Beijing, People’s Republic of China; 2School of Rehabilitation Medicine, Capital Medical University, Beijing, People’s Republic of China; 3Department of Spinal Cord Injury Rehabilitation, China Rehabilitation Research Center, Beijing, People’s Republic of ChinaCorrespondence: Xin ZhengDepartment of Endocrinology, China Rehabilitation Research Center, No. 10 Jiaomen North Road, Fengtai District, Beijing, 100068, People’s Republic of ChinaTel +86-13811568977Email zhengxincrrc@126.comHongjun ZhouDepartment of Spinal Cord Injury Rehabilitation, China Rehabilitation Research Center, No. 10 Jiaomen North Road, Fengtai District, Beijing, 100068, People’s Republic of ChinaTel +86-010-87563115Email zhjcrrc@163.comBackground: Spinal cord injury (SCI) can lead to disuse osteoporosis. The most vulnerable sites for fragility-induced fractures are the distal femur (DF) and proximal tibia (PT). The aim of this study was to evaluate changes in bone mineral density (BMD) at the DF and PT, as well as related factors, during the first year of SCI.Patients and Methods: Thirty-six SCI patients within 12 months of their injury were selected, as were 36 healthy controls. The dual-energy X-ray absorptiometry was used to measure BMDs at the DF, PT, and hip of all subjects. According to the duration of SCI when receiving DXA scan, 36 SCI patients were divided into three subgroups. The BMDs of overall patients and subgroups were compared to those of controls. Biochemical markers of bone metabolism were detected in SCI patients.Results: The BMDs at the DF, PT, and hips of overall SCI patients were significantly lower than those of controls. The percentage difference of BMD between SCI patients and controls at the DF and PT was higher than at the hip. The BMD at the PT of SCI within 6 weeks post-injury was lower than that of controls. The BMDs at the DF and PT of SCI during 6 weeks-3 months post-injury were lower than those of controls. Whereas there was no difference in the BMD at the hip during the first 3 months of SCI. Age and 25OHD were the influencing factors of DF BMD. Age and gender were found to influence PT BMD.Conclusion: The rapid loss of BMD at the PT and DF during the first year of SCI occurred significantly earlier than that of the hip. It is recommended to monitor the BMD of DF and PT in early-stage SCI patients, combined with detection of biochemical markers of bone metabolism.Keywords: spinal cord injury, bone mineral density, distal femur, proximal tibia, disuse osteoporosis

Fengjuan Tian,* Jing Li,* Amina Nazir, Yigang Tong Beijing Advanced Innovation Center for Soft Matter Science and Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yigang Tong Email tong.yigang@gmail.comAbstract: Bacterial biofilms can enhance bacteria’s viability by providing resistance against antibiotics and conventional disinfectants. The existence of biofilm is a serious threat to human health, causing incalculable loss. Therefore, new strategies to deal with bacterial biofilms are needed. Bacteriophages are unique due to their activity on bacteria and do not pose a threat to humans. Consequently, they are considered safe alternatives to drugs for the treatment of bacterial diseases. They can effectively obliterate bacterial biofilms and have great potential in medical treatment, the food industry, and pollution control. There are intricate mechanisms of interaction between phages and biofilms. Biofilms may prevent the invasion of phages, and phages can kill bacteria for biofilm control purposes or influence the formation of biofilms. At present, there are various measures for the prevention and control of biofilms through phages, including the combined use of drugs and the application of phage cocktails. This article mainly reviews the function and formation process of bacterial biofilms, summarizes the different mechanisms between phages and biofilms, briefly explains the phage usage for the control of bacterial biofilms, and promotes phage application maintenance human health and the protection of the natural environment.Keywords: biofilm, phage, drug-resistant bacteria, antibiotic substitute, bacterial biofilm control

Yao Tong,1,* Xiaoyan Xie,2,* Xiaodan Mao,1,3 Huifang Lei,1 Yaojia Chen,1 Pengming Sun1– 3 1Laboratory of Gynecologic Oncology, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou 350001, People’s Republic of China; 2Department of Gynecology, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou 350001, People’s Republic of China; 3Fujian Key Laboratory of Women and Children’s Critical Diseases Research, Fujian Maternity and Child Health Hospital, Fuzhou 350001, People’s Republic of China*These authors contributed equally to this workCorrespondence: Pengming SunLaboratory of Gynecologic Oncology, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University, No. 18 Daoshan Road, Fuzhou 350001, Fujian, People’s Republic of ChinaTel +86 591 8755 8732Fax +86 591 8755 1247Email sunfemy@hotmail.comPurpose: We investigated the relationship between metabolic syndrome (MetS) and its components and erythrocyte parameters such as red blood cell count (RBC) and hemoglobin (Hb) levels, and their association with the risk of deep myometrial invasion in endometrioid endometrial carcinoma (EEC).Patients and Methods: A total of 473 women diagnosed with EEC between January 2012 and December 2018 were included in the study. MetS was diagnosed using a modified version of the Chinese Diabetes Society 2004 criteria. Demographic and clinical characteristics were stratified by MetS and compared. Logistic regression analyses were performed to calculate the odds ratios (ORs) of myometrial invasion.Results: The prevalence of MetS was 18.8% (89/473). RBC and Hb levels were significantly higher in EEC patients with MetS. RBC and Hb values increased linearly with the number of MetS components. The OR of the MetS risk increased across the RBC quartiles (OR=2.212, 95% confidence interval [Cl]=1.125-4.350, P˂0.05). Among EEC patients with MetS, the OR of deep myometrial invasion increased with the decrease of the RBC quartiles (P=0.037). Similarly, in EEC patients with obesity and hyperglycemia or with ≥ 2 MetS criteria, the lowest RBC level (Q1) was a risk factor for deep myometrial invasion compared with the reference RBC group (Q4, P˂0.05). However, this trend was not evident in EEC patients without MetS and its components.Conclusion: EEC with MetS and its components had higher RBC and Hb levels. The association between the erythrocyte parameters and myometrial invasion differed with MetS. RBC was identified as a risk factor of myometrial invasion in EEC patients with MetS and its components.Keywords: metabolic syndrome, endometrioid endometrial carcinoma, red blood cell count, hemoglobin, myometrial invasion

Yin Zhang,1,2 Xinji Chen,1 Yu Tong,3 Junchao Luo,3 Qing Bi1,2 1Department of Orthopedic Surgery, Zhejiang Provincial People’s Hospital and People’s Hospital of Hangzhou Medical College, Hangzhou, Zhejiang 310014, People’s Republic of China; 2The First Affiliated Hospital of Bengbu Medical University, Bengbu, Anhui 233004, People’s Republic of China; 3The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, People’s Republic of ChinaCorrespondence: Qing BiDepartment of Orthopedic Surgery, Zhejiang Provincial People’s Hospital, No. 158 Shangtang Road, Hangzhou, Zhejiang, People’s Republic of ChinaEmail bqzjsrmyy@163.comAbstract: Osteoarthritis (OA) is a common degenerative disease that affects the vast majority of the elderly and may eventually embark on the road of the total knee arthroplasty (TKA), although controversy still exists in the medical community about the best therapies for osteoarthritis. Compared with physical therapy, oral analgesics and other non-operative treatments, intra-articular injection is more safe and effective. Moreover, intra-articular injection is much less invasive and has fewer adverse reactions than surgical treatment. This article reviews mechanism, benefits and adverse reactions of corticosteroids (CS), hyaluronic acid (HA), platelet-rich plasma (PRP), mesenchymal stem cell (MSCs), stromal vascular fraction (SVF) and other new therapies (for example: gene therapy). The application prospect of intra-articular injection was analyzed according to the recent progress in drug research.Keywords: osteoarthritis, intra-articular injection, treatment, prospect, review

Yao Tong, Peipei Wei, Shuang Wang, Qiuying Sun, Yanzheng Cui, Ning Ning, Sitong Chen, Xin He Department of Thoracic Surgery, Cancer Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, People’s Republic of ChinaCorrespondence: Xin He Tel/ Fax +86 10-87787160Email hexinzlyy@163.comPurpose: No final conclusion has yet been reached on characteristics of postoperative pain and pain-related factors after video-assisted thoracoscopic surgery (VATS). This study was designed to explore features of acute severe pain and chronic post-surgical pain (CPSP), and the pain-related factors of VATS.Patients and Methods: Data of patients who underwent VATS for lung cancer in Cancer Hospital, Chinese Academy of Medical Sciences between March 2017 and January 2019 were reviewed in this retrospective study. A numerical rating scale (NRS) was used for evaluating the intensity of postoperative pain including no pain (NRS=0), mild pain (NRS=1– 3), moderate pain (NRS=4– 6), and severe pain (NRS=7– 10). Pain intensity was assessed daily within a week after operation, and also evaluated at 3 months postoperatively.Results: One hundred and five (3.4%) of the 3072 patients enrolled experienced severe pain (NRS=7– 10) on the 1st day after operation, and 17 (0.6%) on the 2nd day. Smoking history, three-port VATS, prolonged operation time, and without patient-controlled analgesia (PCA) were correlated to increased incidence of severe pain. Among all patients, 237 (7.7%) cases generated CPSP, and VATS type, operation time, duration of drainage, and severe pain on the 1st day were four independent risk factors related to CPSP.Conclusion: Patients seemed to experience a lower incidence of acute severe pain and CPSP after VATS than traditional open surgery. Acute severe pain was correlated with smoking history, VATS type, operation time, and PCA; VATS type, operation time, duration of drainage, and severe pain on the 1st day postoperatively were four independent risk factors of CPSP.Keywords: VATS, acute severe pain, chronic post-surgical pain, NRS, risk factor

Amina Nazir,1,2,* Yachao Zhao,2,* Manli Li,2 Rakia Manzoor,3,4 Rana Adnan Tahir,1 Xianglilan Zhang,2 Hong Qing,1 Yigang Tong2 1Key Laboratory of Molecular Medicine and Biotherapy in the Ministry of Industry and Information Technology, Department of Biology, School of Life Sciences, Beijing Institute of Technology, Beijing, People’s Republic of China; 2State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, People’s Republic of China; 3State Key Laboratory of Molecular Development Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, People’s Republic of China; 4School of Life Science, University of Chinese Academy of Sciences, Beijing 100101, People’s Republic of China*These authors contributed equally to this workCorrespondence: Hong QingKey Laboratory of Molecular Medicine and Biotherapy in the Ministry of Industry and Information Technology, Department of Biology, School of Life Sciences, Beijing Institute of Technology, Beijing, People’s Republic of ChinaTel +8613264286266Email hqing@bit.edu.cnYigang TongState Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, People’s Republic of ChinaEmail tong.yigang@gmail.comBackground: Multidrug-resistant plasmids carrying replication genes have been widely present in various strains of Klebsiella pneumoniae. RepA and repB1 were found in plasmids belong to the IncFIB, but their detailed structural and genomic characterization was not reported yet. This is the first study that delivers structural and functional insights of repA- and repB1-carrying IncFIB plasmids.Methods: Klebsiella pneumoniae strains A1705, 911021, and 1642 were isolated from the human urine samples and bronchoalveolar fluids collected from different hospitals of China. Antibacterial susceptibility and plasmid transfer ability were tested to characterize the resistant phenotypes mediated by the pA1705-qnrS, p911021-tetA, and p1642-tetA. The complete nucleotide sequences of these plasmids were determined through high-throughput sequencing technology and comparative genomic analyses of plasmids belong to the same incompatibility group were executed to extract the genomic variations and features.Results: The pA1705-qnrS, p911021-tetA, and p1642-tetA are defined as non-conjugative plasmids, having two replication genes, repA and repB1 associated with IncFIB family, and unknown incompatible group, respectively. Comparative genomic analysis revealed that relatively small backbones of IncFIB plasmids integrated massive accessory module at one “hotspot” that was located between orf312 and repB1. These IncFIB plasmids exhibited the distinct profiles of accessory modules including one or two multidrug-resistant regions, many complete and remnant mobile elements comprising integrons, transposons and insertion sequences. The clusters of resistant genes were recognized in this study against different classes of antibiotics including β-lactam, phenicol, aminoglycoside, tetracycline, quinolone, trimethoprim, sulfonamide, tunicamycin, and macrolide. It has been observed that all resistant genes were located in multidrug resistance regions.Conclusion: It is concluded that multidrug-resistant repA and repB1-carrying IncFIB plasmids are a key source to mediate the resistance through mobile elements among Klebsiella pneumoniae. Current findings provide a deep understanding of horizontal gene transfer among plasmids of the IncFIB family via mobile elements that will be utilized in further in vitro studies.Keywords: plasmids, repA, repB1, multidrug resistance, structural genomics, bioinformatics

Qiaoyu Zhang,1,* Binhua Dong,1,* Lihua Chen,1 Tingting Lin,2 Yao Tong,1 Wenyu Lin,1 Haifeng Lin,1 Yuqin Gao,1 Fen Lin,1 Pengming Sun1,3 1Laboratory of Gynecologic Oncology, Fujian Provincial Maternity and Children’s Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, People’s Republic of China; 2Department of Gynecology, The First Affiliated Hospital of Xiamen University, Xiamen, People’s Republic of China; 3Department of Gynecology, Fujian Provincial Maternity and Children’s Health Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, People’s Republic of China*These authors contributed equally to this workCorrespondence: Pengming SunLaboratory of Gynecologic Oncology, Fujian Provincial Maternity and Children’s Hospital, Affiliated Hospital of Fujian Medical University, Fuzhou, People’s Republic of ChinaTel +86 591 8755 8732Fax +86 591 8755 1247Email sunfemy@hotmail.comObjective: To assess the clinical value of the PCR-reverse dot blot human papillomavirus genotyping test during follow-up of patients with CIN grade 2 or worse (CIN 2+).Methods: Four hundred patients with CIN 2+ receiving treatment from January 2008 to January 2017 were included in our study. Postoperative follow-up procedures comprised HPV examination and cervical cytology every 3– 6 months for the first 2 years and then followed up every 6– 12 months. A pathology examination was performed when there was a positive funding for HPV 16/18 or an abnormal ThinPrep cytology test (TCT) with or without positive for HR-HPV according to the American Society for Coloscopy and Cervical Pathology (ASCCP) guidelines.Results: The median follow-up period was 27.10± 12.47 months (ranging from 3 to 50 months). During follow-up, 12.00% (48/400) of the women developed residual/recurrent disease. The highest risk in CIN 2+ and CIN 3+ residual/recurrence was HPV-16/-18 (hazard ratio (HR)=12.898, 95% CI= 6.849– 24.289; HR= 20.726, 95% CI= 9.64– 44.562, respectively). Among the different follow-up methods, type-specific (TP) HR-HPV persistent infection showed the highest cumulative incidence risk (CIR) (84.62%, 95% CI=73.29– 95.94) and HR (5.38, 95% CI= 2.596– 11.149) during the 4-year follow-up period. At the CIN 2+ and CIN 3+ endpoints, TP-HPV testing had relatively high sensitivity (84.62%, 95% CI=73.29– 95.94 and 89.28%, 95% CI= 77.83– 100.00, respectively) and specificity (78.07%, 95% CI= 72.70– 83.44 and 75.73%, 95% CI= 70.30– 81.17, respectively). However, at the CIN 2+/CIN 3+ endpoint, TCT follow-up had a sensitivity of 60.42%/62.16% (95% CI=46.58– 72.25/46.54– 77.79) and specificity of 90.18%/88.72% (95% CI=86.95– 93.41/85.35– 92.10).Conclusion: TP HR-HPV follow-up can provide a reliable and sensitive clinical reference for CIN 2+ postoperative patients.Keywords: papillomavirus, genotype, cell biology, histology, postoperative

Xin Huang, Yan Tong, Chen-Xing Qi, Yang-Tao Xu, Han-Dong Dan, Yin ShenEye Center, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei, People’s Republic of ChinaCorrespondence: Yin ShenEye Center, Renmin Hospital of Wuhan University, No 238, Jie Fang Road, Wu Chang District, Wuhan 430060, Hubei, People’s Republic of ChinaTel +86 1 387 155 0513Email yinshen@whu.edu.cnObjective: There is increasing neuroimaging evidence that type 2 diabetes patients with retinal microvascular complications show abnormal brain functional and structural architecture and are at an increased risk of cognitive decline and dementia. However, changes in the topological properties of the functional brain connectome in diabetic retinopathy (DR) patients remain unknown. The aim of this study was to explore the topological organization of the brain connectome in DR patients using graph theory approaches.Methods: Thirty-five DR patients (18 males and 17 females) and 38 healthy controls (HCs) (18 males and 20 females), matched for age, sex, and education, underwent resting-state magnetic resonance imaging scans. Graph theory analysis was performed to investigate the topological properties of brain functional connectome at both global and nodal levels.Results: Both DR and HC groups showed high-efficiency small-world network in their brain functional networks. Notably, the DR group showed reduction in the clustering coefficient (P=0.0572) and local efficiency (P=0.0151). Furthermore, the DR group showed reduced nodal centralities in the default-mode network (DMN) and increased nodal centralities in the visual network (VN) (P

WeiHong Ren,1,2 XuRan Zhang,2 WenBo Li,2 Qian Feng,2 HuiJie Feng,2 Yan Tong,2 Hao Rong,2 Wei Wang,2 Dai Zhang,2 ZhenQiang Zhang,3 ShiChun Tu,4 XiaoYan Zhu,1 QinXian Zhang11Department of Histology and Embryology, College of Basic Medicine, Zhengzhou University, Zhengzhou, Henan Province, People’s Republic of China; 2Department of Laboratory Medicine, The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, Henan Province, People’s Republic of China; 3Immunology Laboratory of Chinese Medicine, Henan University of Chinese Medicine, Zhengzhou, Henan Province, People’s Republic of China; 4Neurodegenerative Disease Center, Scintillon Institute, San Diego, CA, USABackground: Myeloid-derived suppressor cells (MDSCs) promote immunosuppression in the tumor microenvironment, support tumor growth and survival, and may contribute to immunotherapy resistance. Recent studies showed that tumor-derived exosomes (TDEs) can induce MDSCs accumulation and expansion, the mechanisms of which are largely unknown.Methods: The morphologies and sizes of the exosomes was observed by using a JEM-1400 transmission electron microscope. MicroRNA(miR)-107 and ARG1, DICER1, PTEN, PI3K, AKT, mTOR, and NF-kB mRNAs were quantified by quantitative reverse tanscription PCR. Dual-Luciferase Reports Assay were used to examine the expression of genes which was targeted by miR-107. The expression of proteins were analyzed by using western blot.Results: MiR-107 was not only overexpressed in gastric cancer cells but also enriched in their secreted TDEs. Also, these miR-107 enriched TDEs could be taken up by HLA-DR-,CD33+,MDSCs, where miR-107 was able to target and suppress expression of DICER1 and PTEN genes. Dampened DICER1 expression supported expansion of MDSCs , while decreased PTEN led to activation of the PI3K pathway, resulting in increased ARG1 expression. Furthemore, gastric cancer-derived miR-107 TDEs, when dosed intravenously into mice, were also capable of inducing expansion of CD11b+,Gr1+/high ,MDSCs in mouse peripheral blood and altering expression of DICER1, PTEN, ARG1, and NOS2 in the MDSCs.Conclusions: Our findings demonstrate for the first time that gastric cancer-secreted exosomes are able to deliver miR-107 to the host MDSCs where they induce their expansion and activition by targeting DICER1 and PTEN genes, thereby may provide novel cancer therapeutics target for gastric cancer.Keywords: exosome, miRNA-107, myeloid-derived suppressor cell, arginase 1, gastric cancer

Yushan Liang,1,2 Guofei Feng,1,2 Liang Wu,3 Suhua Zhong,1,2 Xiaoyu Gao,1,2 Yan Tong,1,2 Wanmeng Cui,1 Yongying Qin,1 WenQing Xu,1 Xue Xiao,1,2 Zhe Zhang,1,2 Guangwu Huang,1,2 Xiaoying Zhou1,41Key laboratory of High-Incidence-Tumor Prevention & Treatment, Ministry of Education, Guangxi Medical University, Nanning, People’s Republic of China; 2Department of Otolaryngology Head & Neck Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning, People’s Republic of China; 3Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing, People’s Republic of China; 4Life Science Institute, Guangxi Medical University, Nanning, People’s Republic of ChinaPurpose: Caffeic acid phenethyl ester (CAPE) is the main polyphenol extracted from honeybee propolis, which inhibits the growth of several kinds of tumor. This study aimed to assess the inhibitory effect of CAPE in nasopharyngeal carcinoma (NPC), evaluate the synergistic action of CAPE in radiotherapy sensitivity of NPC cell lines and further elucidate the possible molecular mechanism involved.Materials and methods: CCK-8 assay was used to analyze cell proliferation ability. Colony formation assay was used to evaluate the clonogenic ability and radio-sensitiveness of NPC cells by CAPE treatment. Wound-healing and transwell assay were used to assess the motility of cells. The expression of key molecules of the epithelial–mesenchymal transition (EMT) was determined by western blot analysis and changes in radiation sensitivity were measured by colony-formation assay. cDNA microarray analysis was used to determine differentially expressed genes with and without CAPE treatment, with Gene Ontology enrichment of gene function and KEGG pathways determined. Cell cycle and apoptosis were detected by flow cytometry and western blot analysis.Results: CAPE suppressed the viability of NPC cell lines time- and dose-dependently. It induced apoptosis in NPC cells along with decreased expression of Bcl-XL and increased cleavage of PARP and expression of Bax. G1 phase arrest was induced by CAPE with ower expression of CDK4, CDK6, Rb and p-Rb. The migratory and invasive ability of NPC cells was decreased by the EMT pathway. The irradiation sensitivity of NPC cells was enhanced with CAPE treatment. CAPE specifically inhibited nuclear factor κB (NF-κB) signaling pathway by suppressing p65 subunit translocation from cytoplasm to nucleus. CAPE treatment was synergistic with chemotherapy and radiotherapy.Conclusion: CAPE may inhibit the proliferation and metastasis of NPC cells but enhance radiosensitivity in NPC therapy by inhibiting the NF-κB pathway. CAPE could be a potential therapeutic compound for NPC therapy.Keywords: nasopharyngeal carcinoma, CAPE, proliferation, metastasis, NF-κB pathway

Lining Shi,1,* Jiao Feng,2,* Zhe Zhan,2 Yuzong Zhao,2,3 Haijian Zhou,4 Haifeng Mao,5 Yingjie Gao,6 Ying Zhang,7 Zhe Yin,2 Bo Gao,2 Yigang Tong,2 Yanping Luo,7 Defu Zhang,2,3 Dongsheng Zhou2 1Institute of Medical Laboratory Sciences, Jinling Hospital, School of Medicine, Nanjing University, Nanjing 210002, China; 2State Key Laboratory of Pathogen & Biosecurity, Beijing Institute of Microbiology & Epidemiology, Beijing 100071, China; 3College of Food Science & Project Engineering, Bohai University, Jinzhou 121013, China; 4National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China; 5Department of Laboratory Medicine, The First People’s Hospital of Lianyungang, Lianyungang 222002, China; 6College of Veterinary Medicine, Jilin University, Changchun 130062, China; 7Medical Laboratory Center, Chinese People’s Liberation Army General Hospital, Beijing 100085, China *These authors contributed equally to this work Background: We recently reported the complete sequence of a blaKPC-2- and rmtB-carrying IncFII-family plasmid p675920-1 with the pKPC-LK30/pHN7A8 hybrid structure. Comparative genomics of additional sequenced plasmids with similar hybrid structures and their prevalence in blaKPC-carrying Klebsiella pneumoniae strains from China were investigated in this follow-up study. Methods: A total of 51 blaKPC-carrying K. pneumoniae strains were isolated from 2012 to 2016 from five Chinese hospitals and genotyped by multilocus sequence typing. The blaKPC-carrying plasmids from four representative strains were sequenced and compared with p675920-1 and pCT-KPC. Plasmid transfer, carbapenemase activity determination, and bacterial antimicrobial susceptibility test were performed to characterize resistance phenotypes mediated by these plasmids. The prevalence of pCT-KPC-like plasmids in these blaKPC-carrying K. pneumoniae strains was screened by PCR. Result: The six KPC-encoding plasmids p1068-KPC, p20049-KPC, p12139-KPC and p64917-KPC (sequenced in this study) and p675920-1 and pCT-KPC slightly differed from one another due to deletion and acquisition of various backbone and accessory regions. Two major accessory resistance regions, which included the blaKPC-2 region harboring blaKPC-2 (carbapenem resistance) and blaSHV-12 (β-lactam resistance), and the MDR region carrying rmtB (aminoglycoside resistance), fosA3 (fosfomycin resistance), blaTEM-1B (β-lactam resistance) and blaCTX-M-65 (β-lactam resistance), were found in each of these six plasmids and exhibited several parallel evolution routes. The pCT-KPC-like plasmids were present in all the 51 K. pneumoniae isolates, all of which belonged to CG258. Conclusion: There was clonal dissemination of K. pneumoniae CG258 strains, harboring blaKPC-2- and rmtB-carrying IncFII-family pKPC-LK30/pHN7A8 hybrid plasmids, among multiple Chinese hospitals. Keywords: Klebsiella pneumoniae, multidrug resistance, carbapenemase, plasmids

Sha Yang,* Yun Yang,* Sixin Cui, Ziqi Feng, Yuzhi Du, Zhen Song, Yanan Tong, Liuyang Yang, Zelin Wang, Hao Zeng, Quanming Zou, Hongwu Sun National Engineering Research Center of Immunological Products & Department of Microbiology and Biochemical Pharmacy, College of Pharmacy, Third Military Medical University of Chinese PLA, Chongqing, 400038, People’s Republic of China *These authors contributed equally to this work Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most predominant and fatal pathogens at wound infection sites. MRSA is difficult to treat because of its antibiotic resistance and ability to form biofilms at the wound site. Methods: In this study, a novel nanoscale liquid film-forming system (LFFS) loaded with benzalkonium bromide was produced based on polyvinyl alcohol and chitosan. Results: This LFFS showed a faster and more potent effect against MRSA252 than benzalkonium bromide aqueous solution both in vitro and in vivo. Additionally, the LFFS had a stronger ability to destroy biofilms (5 mg/mL) and inhibit their formation (1.33 µg/mL). The LFFS inflicted obvious damage to the structure and integrity of MRSA cell membranes and caused increases in the release of alkaline phosphate and lactate dehydrogenase in the relative electrical conductivity and in K+ and Mg2+ concentrations due to changes in the MRSA cell membrane permeability. Conclusion: The novel LFFS is promising as an effective system for disinfectant delivery and for application in the treatment of MRSA wound infections. Keywords: liquid film-forming system, methicillin-resistant Staphylococcus aureus, wound healing, antibiofilm, antibacterial

Jing Ouyang,1,2,* Fengjun Sun,1,* Dongsheng Zhou,3 Jiao Feng,3 Zhe Zhan,3 Zhijian Xiong,4 Bo Yang,1 Zhirui Liu,1 Tingbo Li,1 Yigang Tong,3 Peiyuan Xia1 1Department of Pharmacy, Southwest Hospital, The Third Military Medical University, Chongqing 400038, China; 2Department of Pharmacy, Chongqing Public Health Medical Center, Chongqing 400038, China; 3State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 10007, China; 4Outpatient Department of August First Film Studio, Beijing 100161, China *These authors contributed equally to this work Background: Plasmid-mediated multi-drug resistance (MDR) has been widely found in Citrobacter freundii. C. freundii P10159 was isolated from a human case of postoperative urinary tract infection in a Chinese teaching hospital.Methods: The complete nucleotide sequences of five resistance plasmids pP10159-1, pP10159-2, pP10159-3, pP10159-4 and pP10159-5 from C. freundii P10159 were determined through high-throughput genome sequencing, and then compared with related plasmids sequences. Plasmid transfer, CarbaNP test of carbapenemase activity, and bacterial antimicrobial susceptibility test were performed to characterize resistance phenotypes mediated by these plasmids.Results: pP10159-1 carrying blaNDM-1 and pP10159-2 harboring blaIMP-4 plus qnrS1 were almost identical to IncX3 plasmid pNDM-HN380 and IncN1 plasmid pP378-IMP, respectively. The blaKPC-2-carrying plasmids pP10159-3, pHS062105-3 and pECN49-KPC were highly similar to each other, and constituted a novel group of plasmids belonging to an unknown incomparability group. The MDR plasmids pP10159-4 and pP10159-5 had the backbones highly similar to IncHI4 plasmid pNDM-CIT and type 2 IncC plasmid pR55, respectively, but their accessory resistance regions differed from pNDM-CIT and pR55, respectively. The five plasmids from the P10159 isolate contained a total of 24 different genes or gene loci, which contributed to resistance to 13 distinct antibiotic molecules or toxic compounds.Conclusion: This is the first report of co-occurrence of five different resistance plasmids, with determination of their complete sequences. Data presented here provide a deeper insight into co-selection and maintenance of multiple plasmids and an extremely large number of resistance genes in a single bacterial isolate. Keywords: Citrobacter freundii, multi-drug resistance, plasmids, mobile elements

Lian Zhang,1 Sungkyoung Kim,1 Wenping Ding,1 Yingying Tong,1 Xiuli Zhang,1 Minggui Pan,2 Siyu Chen1 1Department of Oncology, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 2Department of Oncology and Hematology, Kaiser Permanente Medical Center, Santa Clara, CA, USA Background: We previously showed that arsenic sulfide (As4S4) induced cell cycle arrest and apoptosis in several human solid tumor cell lines, including those of gastric cancer. In this study, we investigated the effect of As4S4 on the migration and invasion of gastric cancer cells both in vitro and in vivo.Methods: The human gastric cancer cell lines AGS and MGC803 were selected as in vitro models. Wound-healing migration assay and Transwell invasion assay were carried out to determine the effects of As4S4 on cell migration and invasion. The expressions of E-cadherin, β-catenin, Sp1, KLF4, and VEGF were measured by Western blotting analysis. The activities of matrix metalloproteinase (MMP)-2 and MMP-9 in MGC803 cells were demonstrated by zymography assay. A mouse xenograft model was established by inoculation with MGC803 cells, then intraperitoneal injected with As4S4 for 3 weeks and monitored for body weight and tumor changes. Finally, the inhibition rate of tumor growth was calculated, and the expression of proteins and genes associated with tumor invasion and metastasis in tumor tissues were measured by immunohistochemistry, Western blotting, and real-time polymerase chain reaction assay.Results: As4S4 significantly inhibited the migration and invasion of gastric cancer cell lines. The expression of E-cadherin and KLF4 was upregulated, while the expressions of β-catenin, VEGF, and Sp1 were downregulated following treatment with As4S4. Moreover, the protease activities of MMP-2 and MMP-9 were suppressed by As4S4 in MGC803 cells. Meanwhile, As4S4 effectively suppressed the abilities of tumor growth and invasion in the xenograft tumor model. We found that As4S4 upregulated the expression of E-cadherin and downregulated the expression of β-catenin, Sp1, VEGF, and CD34 in mouse tumor tissues, consistent with the results in vitro.Conclusion: As4S4 inhibited the migration and invasion of gastric cancer cells by blocking tumor cell adhesion, decreasing the ability of tumor cells to destroy the basement membrane, and therefore suppressing their angiogenesis. Keywords: As4S4, xenograft, realgar, E-cadherin, MMPs, VEGF

Lian Zhang,1 Yingying Tong,1 Xiuli Zhang,1 Minggui Pan,2,3 Siyu Chen1 1Department of Oncology, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 2Department of Oncology and Hematology, Kaiser Permanente Medical Center, Santa Clara, 3Division of Research, Kaiser Permanente, Oakland, CA, USA Background: Arsenic compounds have modest cytotoxic activity in solid tumors. We investigated if arsenic sulfide (As4S4) in combination with other distinct agents could enhance its cytotoxic activity. Methods: We used gastric and colon cancer cell lines to study the synergistic effect of As4S4 in combination with BRD4 inhibitor JQ1, or with chemotherapy drug cisplatin and irinotecan or with COX2 inhibitor celecoxib. We investigated the mechanism of the cytotoxic effect of these novel combinations. Results: We found that when As4S4 was combined with JQ1, cisplatin, irinotecan or celecoxib, its cytotoxic activity was dramatically enhanced in both gastric and colon cancer cell lines. As4S4 and JQ1 inhibited BRD4 and c-Myc while activating p53 expression synergistically. As4S4 inhibited COX2 and cyclin D1 expression. When As4S4 was combined with chemotherapy drug cisplatin or COX2 inhibitor celecoxib, its inhibition of COX2, BCL2, and p38 expression was enhanced. As4S4 and cisplatin synergistically stimulated p53, phosphor-p38 (p-p38), and increased cleaved caspase 3 (c-caspase 3). Conclusion: As4S4 in combination with JQ1, cisplatin, irinotecan or celecoxib showed enhanced cytotoxic effect on gastric and colon cancer cells, indicating the potential application of these novel drug combinations as part of treatment strategy that warrants further investigation. As4S4 and JQ1 demonstrate synergistic activation of p53 and inhibition of c-Myc. As4S4 and cisplatin and celecoxib activated multiple apoptosis pathways. Keywords: As4S4, JQ1, cisplatin, irinotecan, celecoxib, gastric cancer, colon cancer

Lian Zhang,1,* Wei Tian,1,2,* Sungkyoung Kim,1 Wenping Ding,1 Yingying Tong,1 Siyu Chen1 1Department of Oncology, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 2Department of Oncology, Central Hospital of Zibo, Shandong, People’s Republic of China *These authors contributed equally to this work Background: Arsenic sulfide (As4S4), the main component of realgar, a traditional Chinese medicine, has shown antitumor efficacy in several tumor types, especially for acute promyelocytic leukemia. In this study, we aimed to explore the efficacy and mechanism of As4S4 in gastric cancer.Methods: The effect of As4S4 on cell proliferation and apoptosis of gastric cancer cells was investigated by MTT assay, 4',6-diamidino-2-phenylindole (DAPI) staining, and annexin V–fluorescein isothiocyanate/propidium iodide staining using gastric cancer cell lines AGS (harboring wild-type p53) and MGC803 (harboring mutant p53) in vitro. The expression of apoptosis-related proteins was measured by Western blotting, real-time polymerase chain reaction, and immunohistochemistry analysis. Mouse xenograft models were established by inoculation with MGC803 cells, and the morphology and the proportion of apoptotic cells in tumor tissues were detected by hematoxylin and eosin staining and TdT-mediated dUTP nick end labeling (TUNEL) assay, respectively. Results: As4S4 inhibited the proliferation and induced apoptosis of AGS and MGC803 cells in a time- and dose-dependent manner. As4S4 upregulated the expression of Bax and MDM2 while downregulated the expression of Bcl-2. The expression of p53 increased significantly in the AGS cells but did not readily increase in the MGC803 cells, which harbored mutant p53. Pifithrin-α, a p53 inhibitor, blocked the modulation of As4S4 on AGS cells, but not on MGC803 cells. Using xenograft as a model, we showed that As4S4 suppressed tumor growth and induced apoptosis in vivo and that the expression of p53 increased accordingly.Conclusion: As4S4 is a potent cytotoxic agent for gastric cancer cells, as it induced apoptosis both in vitro and in vivo through a p53-dependent pathway. Our data indicate that As4S4 may have therapeutic potential in gastric cancer. Keywords: As4S4, p53, realgar, antitumor, xenograft

Yingqi Li,1,2 Yaoli Tong,1 Ruixia Cao,1 Zhimei Tian,2 Binsheng Yang,2 Pin Yang2 1Department of Chemistry, College of Chemistry and Chemical Engineering, 2Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Molecular Science, Shanxi University, Taiyuan, People's Republic of China Background: This study investigated the use of nanodiamond particles (NDs) as a promising material for drug delivery in vivo and in vitro. Methods: HepG2 cells (a human hepatic carcinoma cell line) were used to determine the characteristics of a nanodiamond-doxorubicin complex (ND-DOX) when taken up by cells in vitro using laser scanning confocal microscopy and dialysis experiments. We also compared the survival rate and histopathology of tumor-bearing mice after treatment with NDs or ND-DOX in vivo. Results: In vitro investigation showed that ND-DOX has slow and sustained drug release characteristics compared with free doxorubicin. In vivo, the survival rate of tumor-bearing mice treated with ND-DOX was four times greater than that of mice treated with free doxorubicin. Interestingly, the survival rate in mice treated with NDs alone was close to that of mice treated with free doxorubicin. This indicates that treatment with ND-DOX can prolong the lifespan of tumor-bearing mice significantly compared with conventional doxorubicin and that NDs can have this effect as well. Histopathological analysis showed that neither the NDs nor ND-DOX were toxic to the kidney, liver, or spleen in contrast with the well-known toxic effects of free doxorubicin on the kidney and liver. Further, both the bare NDs and ND-DOX could suppress tumor growth effectively. Conclusion: NDs can potentially prolong survival, and ND-DOX may act as a nanodrug with promising chemotherapeutic efficacy and safety. Keywords: nanodiamond, drug delivery, sustained release, survival rate, cancer, treatment