Shanghai Jiaotong University 20 articles published in JoVE 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. Biology Real-Time Detection of Reactive Oxygen Species Production in Immune Response in Rice with a Chemiluminescence Assay Ying Wang1,2, Zhuo An1,2, Zhifang Zhao1,2, Can Li3, Jiangbo Fan1,2 1Shanghai Collaborative Innovation Center of Agri-Seeds, School of Agriculture and Biology, Shanghai Jiao Tong University, 2Joint Center for Single Cell Biology, School of Agriculture and Biology, Shanghai Jiao Tong University, 3Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education), Shanghai Jiao Tong University Here, we describe a method for the real-time detection of apoplastic reactive oxygen species (ROS) production in rice tissues in pathogen-associated molecular pattern-triggered immune response. This method is simple, standardized, and generates highly reproducible results under controlled conditions. Biology Agrobacterium-Mediated Genetic Transformation, Transgenic Production, and Its Application for the Study of Male Reproductive Development in Rice Dawei Xu*1,2, Palash Chandra Mondol*1, Muhammad Uzair1, Matthew R. Tucker3, Dabing Zhang1,3 1Joint International Research Laboratory of Metabolic and Developmental Sciences, Shanghai Jiao Tong University-University of Adelaide Joint Centre for Agriculture and Health, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 2College of Agronomy, Anhui Agricultural University, Heifei, China, 3School of Agriculture, Food and Wine, University of Adelaide This work describes the use of CRISPR-Cas9 genome editing technology to knockout endogenous gene OsABCG15 followed by a modified Agrobacterium-mediated transformation protocol to produce a stable male-sterile line in rice. Genetics Semiconductor Sequencing for Preimplantation Genetic Testing for Aneuploidy Baoheng Gui*1,2,3, Yingxin Zhang*4, Bo Liang5, Yvonne Ka Yin Kwok4, Wai Ting Lui4, Queenie Sum Yee Yeung4, Lingyin Kong6, Liming Xuan6, Jacqueline Pui Wah Chung4, Kwong Wai Choy3,4 1Department of Genetics and Metabolism, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, 2Birth Defects Prevention and Control Institute of Guangxi Zhuang Autonomous Region, 3Shenzhen Research Institute, The Chinese University of Hong Kong, 4Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, 5State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 6Basecare Medical Device Co., Ltd Here, we introduce a semiconductor sequencing method for preimplantation genetic testing for aneuploidy (PGT-A) with the advantages of short turnaround time, low cost, and high throughput. 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. Engineering Radio Frequency Magnetron Sputtering of GdBa2Cu3O7−δ/ La0.67Sr0.33MnO3 Quasi-bilayer Films on SrTiO3 (STO) Single-crystal Substrates Ying Wang1,2, Zhen Li1,2, YongSheng Liu1, Yijie Li2, Linfei Liu2, Da Xu2, Xiaojing Luo1, Tian Gao1,3, Yanyan Zhu1,4, Luozeng Zhou3, Jianming Xu3 1Department of Physics, Mathematics, Shanghai Key Laboratory of Materials Protection and Advanced Materials in Electric Power, Shanghai University of Electric Power, 2Key Laboratory of Artificial Structure and Quantum Control, Ministry of Education, Department of Physics, Shanghai Jiao Tong University, 3Shanghai Institute of Space Power-sources, 4Shanghai Key Laboratory of High Temperature Superconductors, Shanghai University Here, we present a protocol to grow LSMO nanoparticles and (Gd) BCO films on (001) SrTiO3 (STO) single-crystal substrates by radio frequency (RF)-sputtering. Genetics Purification and Transplantation of Myogenic Progenitor Cell Derived Exosomes to Improve Cardiac Function in Duchenne Muscular Dystrophic Mice Xuan Su*1,2, Yan Shen*1, Yue Jin*1,2, Meng Jiang2, Neal Weintraub1, Yaoliang Tang1 1Vascular Biology Center, Medical College of Georgia, Augusta University, 2Renji Hospital, School of Medicine, Shanghai Jiaotong University Here, we present a protocol to transiently improve cardiac function in Duchenne muscular dystrophy mice by transplanting exosomes derived from normal myogenic progenitor cells. Bioengineering Protein Kinase C-delta Inhibitor Peptide Formulation using Gold Nanoparticles Hisato Konoeda1, Hong Yang2, Chengliang Yang1, Annette Gower1, Chun Xu1, Wei Zhang1, Mingyao Liu1,3 1Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, 2Respiratory Medicine Research Laboratory, Institute of Translation Medicine, Shanghai General Hospital, Shanghai Jiaotong University, 3Institute of Medical Science, Faculty of Medicine, University of Toronto We have previously used a gold nanoparticle peptide hybrid to intravenously deliver a synthetic peptide, protein kinase C-delta inhibitor, which reduced ischemia-reperfusion-induced acute lung injury. Here we show the detailed protocol of the drug formulation. Other intracellular peptides can be formulated similarly. Engineering Evaluating Targeting Accuracy in the Focal Plane for an Ultrasound-guided High-intensity Focused Ultrasound Phased-array System Ke Li1,2, Jingfeng Bai1,2, Yazhu Chen1,2, Xiang Ji1,2 1Biomedical Instrument Institute, School of Biomedical Engineering, Shanghai Jiao Tong University, 2Shanghai Med-X Engineering Center for Medical Equipment and Technology, Shanghai Jiao Tong University This study describes a protocol to evaluate the targeting accuracy in the focal plane of an ultrasound-guided high-intensity focused ultrasound phased-array system. Cancer Research Dual Effects of Melanoma Cell-derived Factors on Bone Marrow Adipocytes Differentiation Juan Wang1, Jin Wen2, Xiao-Xiang Chen1, Guang-Liang Chen3 1Department of Rheumatology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 2Department of Nephrology and Rheumatology, Yongchuan Hospital of Chongqing Medical University, 3Department of Medical Oncology, Fudan University Shanghai Cancer Center; Department of Oncology, Shanghai Medical College, Fudan University Here, we present a reliable and straightforward two-dimensional (2D) coculture system for studying the interaction between tumor cells and bone marrow adipocytes, which reveals a dual effect of melanoma cell-derived factors on the bone marrow adipocytes differentiation and also poses a classic method for the mechanistic study of bone metastasis. Engineering Fabrication of Soft Pneumatic Network Actuators with Oblique Chambers Lisen Ge*1,2, Tianyu Wang*1,3, Ningbin Zhang1,2, Guoying Gu1,2 1State Key Laboratory of Mechanical System and Vibration, Shanghai Jiao Tong University, 2Robotics Institute, School of Mechanical Engineering, Shanghai Jiao Tong University, 3University of Michigan-Shanghai Jiao Tong University Joint Institute, Shanghai Jiao Tong University Here we present a fabrication method of soft pneumatic network actuators with oblique chambers. The actuators are capable of generating coupled bending and twisting motions, which broadens their application in soft robotics. Genetics Canalostomy As a Surgical Approach to Local Drug Delivery into the Inner Ears of Adult and Neonatal Mice Jing-Ying Guo1, Lu He1, Teng-Fei Qu1, Yu-Ying Liu2, Ke Liu1, Guo-Peng Wang1, Shu-Sheng Gong1 1Department of Otolaryngology-Head and Neck Surgery, Beijing Friendship Hospital, Capital Medical University, 2Department of Otolaryngology-Head and Neck Surgery, Shanghai First People's Hospital, Shanghai Jiao Tong University Here we describe canalostomy procedure which allows local drug delivery into the inner ears of adult and neonatal mice through the semicircular canal with minimum damage to hearing and vestibular function. This method can be used to inoculate viral vectors, pharmaceuticals, and small molecules into the mouse inner ear. Biology A Simple and Efficient Method for In Vivo Cardiac-specific Gene Manipulation by Intramyocardial Injection in Mice Yanan Fu*1,2, Wenlong Jiang*2, Yichao Zhao*2, Yuli Huang1, Heng Zhang1, Hongju Wang1, Jun Pu1,2 1Department of Cardiology, The First Affiliated Hospital of Bengbu Medical College, 2Department of Cardiology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University Here we present a protocol for cardiac-specific gene manipulation in mice. Under anesthesia, the mouse hearts were externalized through the fourth intercostal space. Subsequently, adenoviruses encoding specific genes were injected with a syringe into the myocardium, followed by protein expression measurement via in vivo imaging and Western blot analysis. 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. Bioengineering Visual Detection of Multiple Nucleic Acids in a Capillary Array Jianwei Chen*1,2,3, Ning Shao*1,2,3, Jiaying Hu*4, Rong Li4, Yuanshou Zhu1,2,3, Dabing Zhang4,5, Shujuan Guo1,2,3, Junhou Hui6, Peng Liu6, Litao Yang4, Sheng-Ce Tao1,2,3 1Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, 2State Key Laboratory of Oncogenes and Related Genes, 3School of Biomedical Engineering, Shanghai Jiao Tong University, 4Collaborative Innovation Center for Biosafety of GMOs, National Center for the Molecular Characterization of Genetically Modified Organisms, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 5Key Laboratory of Crop Marker-Assisted Breeding of Huaian Municipality, Jiangsu Collaborative Innovation Center of Regional Modern Agriculture and Environmental Protection, 6Department of Biomedical Engineering, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, School of Medicine, Tsinghua University This protocol describes the fabrication of a small, ready-to-use cassette that can be applied for visual detection of multiple nucleic acids in a single, test that is easy to operate. In this approach, a capillary array was used for multiplex and highly efficient detection of GMO targets. Medicine Generation of a Chronic Obstructive Pulmonary Disease Model in Mice by Repeated Ozone Exposure Zhongwei Sun1,2, Feng Li3, Xin Zhou3, Wen Wang1,2 1Cellular Biomedicine Group, Shanghai, 2Cellular Biomedicine Group, Cupertino, 3Department of Respiratory Medicine, Shanghai General Hospital, Shanghai Jiaotong University This study describes the successful generation of a new chronic obstructive pulmonary disease (COPD) animal model by repeatedly exposing mice to high concentrations of ozone. 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. Neuroscience Real-time Imaging of Axonal Transport of Quantum Dot-labeled BDNF in Primary Neurons Xiaobei Zhao1, Yue Zhou2, April M. Weissmiller1, Matthew L. Pearn3,4, William C. Mobley1, Chengbiao Wu1 1Department of Neurosciences, University of California, San Diego, 2School of Biomedical Engineering and Med-X Research Institute, Shanghai Jiao Tong University, 3Department of Anesthesiology, University of California, San Diego, 4VA San Diego Healthcare System Axonal transport of BDNF, a neurotrophic factor, is critical for the survival and function of several neuronal populations. Some degenerative disorders are marked by disruption of axonal structure and function. We demonstrated the techniques used to examine live trafficking of QD-BDNF in microfluidic chambers using primary neurons. Medicine Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging Xiaobing Liu*1,2, Toya Terry*1, Su Pan1, Zhongwei Yang1, James T. Willerson1, Richard A. F. Dixon1, Qi Liu1 1The Texas Heart Institute at St. Luke's Episcopal Hospital, 2Department of Vascular Surgery, The Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University We show here the in vivo insertion of an osmotic pump for constant local drug delivery and the creation of hindlimb ischemia in a mouse model. Moreover, the hindlimb vasculature is perfused with Microfil, a silicone radiopaque agent, to prepare for micro-computed tomography (micro-CT) imaging.
