Shanghai Jiao Tong University School of Medicine 22 articles published in JoVE Medicine Early Weight-Bearing Rehabilitation Protocol After Anterior Cruciate Ligament Reconstruction Renren Du*1, Wenjiang Sun*2, Fei He2, Liming Jiang3, Wengao Cheng4, Bo Yu2, Jiwu Chen5 1Rehabilitation center, Shanghai Second Rehabilitation Hospital, 2Department of Rehabilitation Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 3 Based on the safety and feasibility, this article presents an early weight-bearing rehabilitation protocol after anterior cruciate ligament reconstruction. The protocol is clear and easy to operate, which is helpful to promote its use in clinical practice and accelerate the functional recovery of patients. Bioengineering IDG-SW3 Cell Culture in a Three-Dimensional Extracellular Matrix Kaizhe Chen*1, Haoyi Chen*1, Lianfu Deng1, Kai Yang1, Jin Qi1 1Department of Orthopedics, Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint Diseases, Shanghai Institute of Traumatology and Orthopaedics, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine Here, we present a protocol for culturing IDG-SW3 cells in a three-dimensional (3D) extracellular matrix. Biology A 3-D Visualization Technique for Bone Remodeling in a Suture Expansion Mouse Model Ziduan Ding1,2, Ruomei Li1,2, Yufeng Duan1,2, Zhenxia Li1,2, Bing Fang1,2, Dian Jing1,2 1Department of Orthodontics, China Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, 2National Center for Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology, Shanghai Jiao Tong University This protocol presents a standardized suture expansion mouse model and a 3-D visualization method to study the mechanobiological changes of the suture and bone remodeling under tensile force loading. Biology Isolation, Culture, and Adipogenic Induction of Stromal Vascular Fraction-derived Preadipocytes from Mouse Periaortic Adipose Tissue Min Liang*1, Yijie Huang*1, Yangjing Jiang1, Yunwen Hu1, Zhaohua Cai1, Ben He1 1Department of Cardiology, Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine Here, we describe the isolation, culture, and adipogenic induction of stromal vascular fraction-derived preadipocytes from mouse periaortic adipose tissue, allowing for the study of perivascular adipose tissue function and its relationship with vascular cells. Medicine Using Inducible Osteoblastic Lineage-Specific Stat3 Knockout Mice to Study Alveolar Bone Remodeling During Orthodontic Tooth Movement Yuanqi Liu*1,2,3,4,5, Siyuan Sun*1,2,3,4,5, Ziyi Jiang*6, Xinyi Gong1,2,3,4,5, Yiling Yang1,2,3,4,5, Yanfei Zhu1,2,3,4,5, Hongyuan Xu1,2,3,4,5, Anting Jin1,2,3,4,5, Xiangru Huang1,2,3,4,5, Xin Gao1,2,3,4,5, Tingwei Lu1,2,3,4,5, Jingyi Liu1,2,3,4,5, Xinyu Wang1,2,3,4,5, Qinggang Dai2,3,4,5,7, Lingyong Jiang1,2,3,4,5 1Center of Craniofacial Orthodontics, Department of Oral and Cranio-maxillofacial Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, 2College of Stomatology, Shanghai Jiao Tong University, 3National Center for Stomatology, 4National Clinical Research Center for Oral Diseases, 5Shanghai Key Laboratory of Stomatology, 6Shanghai Starriver Bilingual School, 7The 2nd Dental Center, Ninth People’s Hospital, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine This study provides a protocol for using inducible osteoblast lineage-specific Stat3 knockout mice to study bone remodeling under orthodontic force and describes methods for analyzing alveolar bone remodeling during orthodontic tooth movement, thus shedding light on skeletal mechanical biology. Medicine Establishment and Confirmation of a Postnatal Right Ventricular Volume Overload Mouse Model Sijuan Sun*1, Hongbin Zhu*2, Shoubao Wang3, Xiuxia Xu4, Lincai Ye2,5,6 1Department of Pediatric Intensive Care Unit, Shanghai Children’s Medical Center, School of Medicine, Shanghai Jiaotong University, 2Department of Thoracic and Cardiovascular Surgery, Shanghai Children’s Hospital, School of Medicine, Shanghai Jiaotong University, 3Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, 4Department of Radiology, Huangpu Branch, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, 5Institute of Pediatric Translational Medicine, Shanghai Children’s Medical Center, School of Medicine, Shanghai Jiaotong University, 6Shanghai Institute for Pediatric Congenital Heart Disease, Shanghai Children’s Medical Center, School of Medicine, Shanghai Jiaotong University This protocol presents the establishment and confirmation of a postnatal right ventricular volume overload (VO) model in mice with abdominal arteriovenous fistula (AVF), which can be applied to investigate how VO contributes to postnatal heart development. Medicine Isolation of Murine Intestinal Mesenchyme Resulting in a High Yield of Telocytes Marco Canella*1, Jianmei Tan*1,2, Bing Su2, Michal Shoshkes-Carmel1 1Department of Developmental Biology and Cancer Research, The Institute for Medical Research Israel-Canada, The Hebrew University - Hadassah Medical School, 2Shanghai Institute of Immunology, Department of Immunology and Microbiology, Shanghai Jiao Tong University School of Medicine Here, we present a protocol to isolate murine intestinal mesenchyme, including telocytes. These can be used for several applications, such as co-culture with mouse or human-derived organoids, to support growth and better reflect the situation in the original tissue. Biology Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis Chi-Ju Kim*1,2, Morgan D. Kuczler*1, Liang Dong1,3, Junyoung Kim2,4, Sarah R. Amend1, Yoon-Kyoung Cho2,4, Kenneth J. Pienta1 1The Brady Urological Institute, Johns Hopkins University School of Medicine, 2Department of Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), 3Department of Urology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, 4Center for Soft and Living Matter, Institute for Basic Science (IBS) Extracellular vesicles (EVs) contribute to cellular biology and intercellular communications. There is a need for practical assays to visualize and quantify EVs uptake by the cells. The current protocol proposes the EV uptake assay by utilizing three-dimensional fluorescence imaging via confocal microscopy, following EV isolation by a nano-filtration-based microfluidic device. Medicine Real-Time Dynamic Navigation System for the Precise Quad-Zygomatic Implant Placement in a Patient with a Severely Atrophic Maxilla Yihan Shen*1, Qinggang Dai*1, Baoxin Tao*1, Wei Huang2, Feng Wang2, Kengliang Lan1, Yuanyuan Sun1, Xiaowan Ling1, Lijun Yan1, Yueping Wang1, Yiqun Wu1 1The 2nd Dental Center, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, College of Stomatology, Shanghai Jiao Tong University, National Center for Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology, 2Department of Dental Implantology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, College of Stomatology, Shanghai Jiao Tong University, National Center for Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology Here, we present a protocol to achieve precise quad-zygomatic implant placement in patients with severely atrophic maxilla using a real-time dynamic navigation system. Immunology and Infection Flow Cytometric Analysis of Lymphocyte Infiltration in Central Nervous System during Experimental Autoimmune Encephalomyelitis Zhe Ji1,2, Chenghua Zhou1, Hongshen Niu3, Jinshen Wang1, Lei Shen3 1Translational Medicine Research Center, Ruijin Hospital North, Shanghai Jiao Tong University School of Medicine, 2Department of Microbiology and Immunology, Tongji University School of Medicine, 3Shanghai Institute of Immunology, Shanghai Key Laboratory for Tumor Microenvironment and Inflammation, Shanghai Jiao Tong University School of Medicine This manuscript presents a protocol to induce active experimental autoimmune encephalomyelitis (EAE) in mice. A method for the isolation and characterization of the infiltrated lymphocytes in the central nervous system (CNS) is also presented to show how lymphocytes are involved in the development of CNS autoimmune disease. Developmental Biology Isolation, Culture, and Adipogenic Induction of Neural Crest Original Adipose-Derived Stem Cells from Periaortic Adipose Tissue Yiding Qi*1, Xiang Miao*2, Lian Xu3, Mengxia Fu3, Shi Peng4, Kailei Shi5, Jun Li4, Maoqing Ye5, Ruogu Li1 1Department of Cardiology, Shanghai Chest Hospital, Shanghai Jiao Tong University, 2Shanghai Institute of Nutrition and Health, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, 3Shanghai Key Laboratory of Hypertension, Shanghai Institute of Hypertension, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, 4Department of Cardiology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 5Department of Cardiology, Shanghai Key Laboratory of Clinical Geriatric Medicine, Huadong Hospital Affiliated to Fudan University We present a protocol for the isolation, culture, and adipogenic induction of neural crest derived adipose-derived stem cells (NCADSCs) from the periaortic adipose tissue of Wnt-1 Cre+/-;Rosa26RFP/+ mice. The NCADSCs can be an easily accessible source of ADSCs for modeling adipogenesis or lipogenesis in vitro. Biochemistry A Colorimetric Assay of Citrate Synthase Activity in Drosophila Melanogaster Ping Wei*1, Qihong Liu*2, Wen Xue2, Jiwu Wang2 1 We present a protocol for a colorimetric assay of citrate synthase activity for quantification of intact mitochondrial mass in Drosophila tissue homogenates. Biology Using En Face Immunofluorescence Staining to Observe Vascular Endothelial Cells Directly Chang Li*1,2, Zhu Hui Liu*1,2, Jia Wei Chen1,2, Xin Yi Shu1,2, Ying Shen1, Feng Hua Ding1, Rui Yan Zhang1, Wei Feng Shen1, Lin Lu1, Xiao Qun Wang1 1Department of Cardiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, 2Institute of Cardiovascular Diseases, Shanghai Jiao Tong University Here, we present a protocol for immunofluorescence staining to observe the endothelial cells of the mouse aorta directly. This technique is useful when studying the cellular and molecular phenotype of endothelial cells in different flow patterns and in the development of atherosclerosis. Biology Photostimulation by Femtosecond Laser Activates Extracellular-signal-regulated Kinase (ERK) Signaling or Mitochondrial Events in Target Cells Shaoyang Wang1,2, Minglie Hu2, Tao Ren1, Hao He3 1Department of Respiratory Medicine, Shanghai Sixth People's Hospital, School of Medicine, Shanghai Jiao Tong University, 2Ultrafast Laser Laboratory, College of Precision Instrument and Optoelectronics Engineering, Tianjin University, 3School of Biomedical Engineering, Shanghai Jiao Tong University Tightly-focused femtosecond laser can deliver precise stimulation to cells by being coupled into a confocal microscopy enabling the real-time observation and photostimulation. The photostimulation can activate cell molecular events including ERK signaling pathway and mitochondrial flashes of reactive oxygen species. Immunology and Infection Laminar Flow-based Assays to Investigate Leukocyte Recruitment on Cultured Vascular Cells and Adherent Platelets Tanja Vajen1, Alexandra C.A. Heinzmann1, Annemiek Dickhout1, Zhen Zhao2, Magdolna Nagy1, Johan W.M. Heemskerk1, Rory R. Koenen1 1Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, 2Department of Vascular Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine Leukocytes avidly interact with vascular cells and platelets after vessel wall injury or during inflammation. Here, we describe a straightforward laminar flow-based assay to characterize the molecular mechanisms that underlie the interactions between leukocytes and their cellular partners. Neuroscience An Improved Method for Collection of Cerebrospinal Fluid from Anesthetized Mice Nastasia K-H Lim1,2, Visse Moestrup3,4, Xiao Zhang1, Wen-An Wang5, Arne Møller3,4,6, Fu-De Huang1,4 1Shanghai Advanced Research Institute, University of Chinese Academy of Sciences, Chinese Academy of Science, 2Shanghai Institute of Materia Medica, University of Chinese Academy of Sciences, Chinese Academy of Science, 3Center of Functionally Integrative Neuroscience, Department of Clinical Medicine, Aarhus University, 4Sino-Danish Center for Education and Research (SDC), 5Department of Neurology, Xinhua Hospital Chongming Branch Affiliated to Shanghai Jiao Tong University School of Medicine, 6Department of Nuclear Medicine and PET-centre, Aarhus University Hospital This protocol describes an improved technique for the abundant collection of cerebrospinal fluid (CSF) with no contamination from blood. With greater sample collection and purity, more analyses can be performed using CSF to further our understanding of diseases that affect the brain and spinal cord. Biochemistry Capturing the Interaction Kinetics of an Ion Channel Protein with Small Molecules by the Bio-layer Interferometry Assay Bo Han1, Man Zhang2, Peng Sun1, Shangwei Hou3 1Department of General Surgery, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, 2Key Laboratory of Systems Biomedicine (Ministry of Education), Institute of Systems Biomedicine, Shanghai Jiao Tong University, 3Hongqiao International Institute of Medicine, Tongren Hospital, Shanghai Jiao Tong University School of Medicine The protocol here describes the interactions of purified hEAG1 ion channel protein with the small molecule lipid ligand phosphatidylinositol 4, 5-bisphosphate (PIP2). The measurement demonstrates that BLI could be a potential method for novel small-molecule ion channel ligand screening. Medicine Gene Regulation and Targeted Therapy in Gastric Cancer Peritoneal Metastasis: Radiological Findings from Dual Energy CT and PET/CT Bowen Shi1, Huimin Lin1, Miao Zhang2, Wei Lu3, Ying Qu4, Huan Zhang1 1Department of Radiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, 2Department of Nuclear Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, 3GE Healthcare China, 4Department of Surgery, Cedars-Sinai Medical Center This protocol describes the value of dual energy CT and PET/CT imaging methods in tumor imaging and efficacy evaluation. This article demonstrates the research methods and results acquired by dual energy CT and PET/CT to evaluate the gene regulation and targeted treatment of gastric cancer peritoneal metastasis. Neuroscience Live-cell Measurement of Odorant Receptor Activation Using a Real-time cAMP Assay Yuetian Zhang*1, Yi Pan*1, Hiroaki Matsunami2,3, Hanyi Zhuang1,4 1Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education, Shanghai Jiao Tong University School of Medicine, 2Department of Molecular Genetics and Microbiology, Duke University Medical Center, 3Department of Neurobiology, Duke Institute for Brain Sciences, Duke University Medical Center, 4Institute of Health Science, Chinese Academy of Science/Shanghai Jiao Tong University School of Medicine Characterizing the function of odorant receptors serves an indispensable part in the deorphanization process. We describe a method to measure the activation of odorant receptors in real time using a cAMP assay. Bioengineering Microfluidic Bioprinting for Engineering Vascularized Tissues and Organoids Yu Shrike Zhang*1, Qingmeng Pi*1,2, Anne Metje van Genderen1,3 1Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, 2Department of Plastic and Reconstructive Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, 3Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Utrecht University We provide a generalized protocol based on a microfluidic bioprinting strategy for engineering a microfibrous vascular bed, where a secondary cell type could be further seeded into the interstitial space of this microfibrous structure to generate vascularized tissues and organoids. Medicine Screening Bioactive Nanoparticles in Phagocytic Immune Cells for Inhibitors of Toll-like Receptor Signaling Hong Yang1, Shan Yu Fung2, Aihua Bao1, Qiang Li1, Stuart E. Turvey2 1Department of Respiratory Medicine, Shanghai First People's Hospital, Shanghai Jiaotong University School of Medicine, 2 Toll-like receptor (TLR) signaling plays an important role in the pathophysiology of many human inflammatory diseases, and regulating TLR responses by bioactive nanoparticles is anticipated to be beneficial in many inflammatory conditions. THP-1 cell-based reporter cells provide a versatile and robust screening platform for identifying novel inhibitors of TLR signaling. Developmental Biology A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro Zhaofeng Jia1,2, Yujie Liang3, Bin Ma4,5, Xiao Xu2,6, Jianyi Xiong2, Li Duan2, Daping Wang1,2 1Guangzhou Medical University, 2 We present a method to quantify DNA methylation based on the 5-methylcytosine (5-mC) dot blot. We determined the 5-mC levels during chondrocyte dedifferentiation. This simple technique could be used to quickly determine the chondrocyte phenotype in ACI treatment.
