Nanjing Medical University 16 articles published in JoVE Cancer Research Glioblastoma Relapse Post-Resection Model for Therapeutic Hydrogel Investigations Shuo Sun*1, Dongyan Shi*1, Jian Liu1, JinYing Lu1, Pingnan Dou1, Zewei Zhou1, Yun Chen1,2 1Department of Immunology, Key Laboratory of Human Functional Genomics of Jiangsu Province, Gusu School, Nanjing Medical University, 2Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center for Cancer Personalized Medicine, Nanjing Medical University The present protocol establishes a glioblastoma (GBM) relapse post-resection model using microscopy to investigate the therapeutic effect of an injectable, bioresponsive hydrogel in vivo. Cancer Research Development of a 68Gallium-Labeled D-Peptide PET Tracer for Imaging Programmed Death-Ligand 1 Expression Lulu Zhang*1, Siqi Zhang*2, Wenyu Wu1, Xingkai Wang2, Jieting Shen2, Dongyuan Wang4, Kuan Hu*2,3, Ming-Rong Zhang*3, Feng Wang*1, Rui Wang*2 1Department of Nuclear Medicine, Nanjing First Hospital, Nanjing Medical University, 2State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, 3Department of Advanced Nuclear Medicine Sciences, Institute for Quantum Medical Science, National Institutes for Quantum Science and Technology, 4Department of Pharmacy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology This study developed a noninvasive and real-time method to evaluate the distribution of programmed death-ligand 1 in the whole body, based on positron emission tomographic imaging of [68Ga] D-dodecapeptide antagonist. This technique has advantages over conventional immunohistochemistry and improves the efficiency of identifying appropriate patients who will benefit from immune checkpoint blockade therapy. Developmental Biology Generation of a Mouse Artificial Decidualization Model with Ovariectomy for Endometrial Decidualization Research Yang Zhang*1,2,3, Zhe Zhang*1,2,3, Nannan Kang1,2,3, Xiaoqiang Sheng1,2,3 1Center for Reproductive Medicine and Obstetrics and Gynecology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, 2State Key Laboratory of Reproductive Medicine, Nanjing Medical University, 3Center for Molecular Reproductive Medicine, Nanjing University Here, we describe the method of generating an artificial decidualization model using the ovariectomized mouse, a classic endometrial decidualization experiment in the research field of endometrial decidualization. Cancer Research A Rapid Screening Workflow to Identify Potential Combination Therapy for GBM using Patient-Derived Glioma Stem Cells Ziyi Hu*1, Tingting Zhou*1, Fangrong Wu1, Fan Lin1,2 1Department of Cell Biology, School of Basic Medical Sciences, Nanjing Medical University, 2Institute for Brain Tumors & Key Laboratory of Rare Metabolic Diseases, Nanjing Medical University; Nanjing Medical University Affiliated Cancer Hospital; Key Laboratory of Human Functional Genomics of Jiangsu Province The glioma stem cells (GSCs) are a small fraction of cancer cells which play essential roles in tumor initiation, angiogenesis, and drug resistance in glioblastoma (GBM), the most prevalent and devastating primary brain tumor. The presence of GSCs makes the GBM very refractory to most of individual targeted agents, so high-throughput screening methods are required to identify potential effective combination therapeutics. The protocol describes a simple workflow to enable rapid screening for potential combination therapy with synergistic interaction. The general steps of this workflow consist of establishing luciferase-tagged GSCs, preparing matrigel coated plates, combination drug screening, analyzing, and validating the results. Biology Quantification of Circulating Pig-Specific DNA in the Blood of a Xenotransplantation Model Yangyang Deng*1,2, Ming Zhou*1,3, Ying Lu1, Jiao Chen1, Zuhui Pu4, Dongjing Yu1,5, Yifan Dai6, Yongqiang Zhan7, Lisha Mou1 1 In this protocol, porcine specific primers were designed, plasmids-containing porcine specific DNA fragments were constructed, and standard curves for quantitation were established. Using species-specific primers, cpsDNA was quantified by qPCR in pig-to-mouse cell transplantation models and pig-to-monkey artery patch transplantation models. Environment Experimental Protocol for Detecting Mitochondrial Function in Hepatocytes Exposed to Organochlorine Pesticides Qian Liu1,2,3, Zhaoyan Jiang3, Aihua Gu1,2 1State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, 2Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, 3Center of Gallbladder Disease, Shanghai East Hospital, Institute of Gallstone Disease, Tongji University School of Medicine Understanding the influence of environmental organochlorine pesticides (OCPs) on mitochondrial function in hepatocytes is important in exploring the mechanism of OCPs causing metabolic disorders. This paper presents detailed methods on detecting hepatic mitochondrial function. Developmental Biology Whole-Mount In Situ Hybridization in Zebrafish Embryos and Tube Formation Assay in iPSC-ECs to Study the Role of Endoglin in Vascular Development Yong Wang*1, Ding Zhang*2, Fang Zhou2, Meijun Zhou2, Qiujie Li1, Jingyu Chen3, Jun Yang1,2 1Department of Physiology and Department of Cardiology of the Second Affiliated Hospital, Zhejiang University School of Medicine, 2Department of Cell Biology, State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, 3Wuxi Lung Transplant Center, Wuxi People's Hospital affiliated to Nanjing Medical University Presented here is a protocol for whole-mount in situ RNA hybridization analysis in zebrafish and tube formation assay in patient-derived induced pluripotent stem cell-derived endothelial cells to study the role of endoglin in vascular formation. Medicine Inguinal Subcutaneous White Adipose Tissue (ISWAT) Transplantation Model of Murine Islets Yuanzheng Peng*1, Zhicheng Zou*1, Jiao Chen1, Hancheng Zhang1, Ying Lu1, Rito Bittino2, Hongxing Fu3, David K. C. Cooper4, Shan Lin1, Mengtao Cao1, Yifan Dai5, Zhiming Cai1, Lisha Mou1 1Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Health Science Center, 2Institute for Cellular Therapeutics, Allegheny-Singer Research Institute, 3School of Pharmaceutical Sciences, Wenzhou Medical University, 4Xenotransplantation Program/Department of Surgery, University of Alabama at Birmingham, 5Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University In this protocol, a method of murine islet isolation and transplantation into the inguinal subcutaneous white adipose tissue is described. Isolated syngeneic murine islets are transplanted into a murine recipient using a basement membrane hydrogel. The blood glucose level of the recipients is monitored, and histology analysis of the islet grafts is performed. Medicine Murine Appendectomy Model of Chronic Colitis Associated Colorectal Cancer by Precise Localization of Caecal Patch Yaguang Li1, Junhong Liu2,3, Gaixia Liu1, Zhenhong Pan2,3, Mingxia Zhang1, Yao Ma4, Qingxia Wei1,5, Hongping Xia1,6, Rui Xue Zhang1,2, Junjun She1 1 The presented protocol describes a facile surgical removal of the appendix (caecal patch) in a mouse followed by the induction of inflammatory bowel disease-associated colorectal cancer. This murine appendectomy model enables investigation of the biological role of the appendix in the pathogenesis of human gastrointestinal disease. Chemistry Methionine Functionalized Biocompatible Block Copolymers for Targeted Plasmid DNA Delivery Yang Wu*1, Wei Zhang*2, Jian Zhang2, Zhi-Xiang Mao3, Li Ding2, Hao Li3, Rong Ma1, Jin-Hai Tang2 1Research Center of Clinical Oncology, Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research & Nanjing Medical University Affiliated Cancer Hospital, 2Department of General Surgery, The First Affiliated Hospital with Nanjing Medical University, 3School of Clinical Medicine, Xuzhou Medical University This work presents the preparation of methionine functionalized biocompatible block copolymers (mBG) via the reversible addition-fragmentation chain transfer (RAFT) method. The plasmid DNA complexing ability of the obtained mBG and their transfection efficiency were also investigated. The RAFT method is very beneficial for polymerizing monomers containing special functional groups. Genetics In Vitro Biochemical Assays using Biotin Labels to Study Protein-Nucleic Acid Interactions Lina Yu*1, Wenxiu He*1, Jie Xie*1, Rui Guo*1, Juan Ni2, Xia Zhang3, Quishi Xu1, Caifeng Wang1, Qiuling Yue1, Fangfang Li1, Mengcheng Luo4, Bo Sun3, Lan Ye1, Ke Zheng1 1State Key Laboratory of Reproductive Medicine, Nanjing Medical University, 2The Affiliated Hospital of Hangzhou Normal University, 3School of Life Science and Technology, ShanghaiTech University, 4Department of Tissue and Embryology, School of Basic Medical Sciences, Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan University Presented here are protocols for in vitro biochemical assays using biotin labels that may be widely applicable for studying protein-nucleic acid interactions. Biology Enhanced Crosslinking Immunoprecipitation (eCLIP) Method for Efficient Identification of Protein-bound RNA in Mouse Testis Qiushi Xu*1, Caifeng Wang*1, Li Ling*1, Qiuling Yue*1, Mengrou Liu1, Shuya Zhang1, Kaiqiang Fu1, Lan Ye1, Ke Zheng1 1State Key Laboratory of Reproductive Medicine, Nanjing Medical University Here, we present an eCLIP protocol to determine major RNA targets of RBP candidates in testis. Behavior A Behavioral Test Battery for the Repeated Assessment of Motor Skills, Mood, and Cognition in Mice Ran You1,2,3, Yan Liu3, Raymond Chuen-Chung Chang3 1 A comprehensive behavioral test battery of motor skills, mood—including social interaction, depression, and anxiety—and cognition is designed for the repeated assessment of neurodegeneration-related behavioral changes in mice. Developmental Biology An In Vivo Method to Study Mouse Blood-Testis Barrier Integrity Mengrou Liu*1, Chunsen Zhu*1, Shun Bai*2, Xin Li1, Kaiqiang Fu1, Lan Ye1, Ke Zheng1 1State Key Laboratory of Reproductive Medicine, Nanjing Medical University, 2Center for Reproductive Medicine, Department of Obstetrics and Gynecology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China Here, we present a protocol to assess the blood-testis barrier integrity by injecting inulin-FITC into testes. This is an efficient in vivo method to study blood-testis barrier integrity that can be compromised by genetic and environmental elements. Biochemistry Regeneration of Arrayed Gold Microelectrodes Equipped for a Real-Time Cell Analyzer Zhihui Xu*1, Yiyan Song*1, Huijun Jiang2, Yan Kong3, Xiaoming Li3, Jin Chen1, Yuan Wu4 1School of Public Health, Nanjing Medical University, 2School of Pharmacy, Nanjing Medical University, 3State Key Laboratory of Materials-Oriented Chemical Engineering, College of Chemistry and Chemical Engineering, Nanjing Tech University, 4Department of Medical Oncology, Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, The Affiliated Cancer Hospital of Nanjing Medical University This protocol describes a general strategy to regenerate commercial arrayed gold microelectrodes equipped for a label-free cell analyzer aimed at saving on the high running costs ofmicrochip-based assays. The regeneration process includes trypsin digestion, rinsing with ethanol and water, and a spinning step, which enables repeated usage of microchips. Medicine Construction and Evaluation of a Murine Calvarial Osteolysis Model by Exposure to CoCrMo Particles in Aseptic Loosening Hui Jiang*1, Yicun Wang*1, Zhantao Deng*1,2,3, Jiewen Jin2,3, Jia Meng1, Shuo Chen1, Jun Wang1, Yang Qiu1, Ting Guo1, Jianning Zhao1 1Department of Orthopedics, Jinling Hospital, School of Medicine, Nanjing University, 2Center for Translational Medicine, Nanjing University Medical School, 3Jiangsu Key Laboratory for Molecular Medicine, Nanjing University Medical School This manuscript describes a murine calvarial osteolysis model by exposure to CoCrMo particles, which constitutes an ideal animal model for assessing the interactions between wear particles and various cells in aseptic loosening.
