Chinese Academy of Medical Sciences and Peking Union Medical College 12 articles published in JoVE Biology Neutrophil Lifespan Extension with CLON-G and an In Vitro Spontaneous Death Assay Yuping Fan*1,2, Yan Teng*3, Fu tong Liu*4, Fengxia Ma1,2, Alan Y. Hsu5, Sizhou Feng1,2, Hongbo R. Luo5 1State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, 2Tianjin Institutes of Health Science, 3Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, 4Haihe Laboratory of Cell Ecosystem, Tianjin Medical University, 5Joint Program in Transfusion Medicine, Department of Pathology, Harvard Medical School; Division of Blood Bank, Department of Laboratory Medicine, The Stem Cell Program, Boston Children's Hospital, Dana-Farber /Harvard Cancer Center This protocol details the preparation of CLON-G to extend the neutrophil lifespan to greater than 5 days and provides a reliable procedure for evaluating neutrophil death with flow cytometry and confocal fluorescence microscopy. 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 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. Behavior Assessment of Sexual Behavior of Male Mice Zi-Wei Liu1, Ning Jiang2, Xue Tao2, Xiao-Ping Wang1, Xin-Min Liu2, Shui-Yuan Xiao3 1Department of Psychiatry, The Second Xiangya Hospital, Central South University, National Clinical Research Center for Mental Disorders and National Technology Institute on Mental Disorders, Hunan Key Laboratory of Psychiatry and Mental Health, 2Research Center for Pharmacology & Toxicology, Institute of Medicinal Plant Development (IMPLAD), Chinese Academy of Medical Sciences and Peking Union Medical College, 3Department of Social Medicine and Health Management, Xiangya School of Public Health, Central South University This article describes how to perform sexual behavior tests in male mice. Environment Sample Preparation Method of Scanning and Transmission Electron Microscope for the Appendages of Woodboring Beetle Yan-Ru Zhang1,2, Hai-Li Qiao3, Li-Li Ren1, Rong Wang4, Peng-Fei Lu1 1The Key Laboratory for Silviculture and Conservation of the Ministry of Education, School of Forestry, Beijing Forestry University, 2School of Forestry, Inner Mongolia Agricultural University, 3Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, 4School of Forestry, Fujian Agriculture and Forestry University In order to observe ultrastructure of insect sensilla, scanning and transmission electron microscopy (SEM and TEM, respectively) sample preparation protocol were presented in the study. Tween 20 was added into the fixative to avoid sample deformation in SEM. Fluorescence microscopy was helpful for improving slicing accuracy in TEM. Medicine Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice Fuliang Luo1, Xin Wang1, Xiaokang Luo1, Bo Li1, Desheng Zhu2, Hansong Sun3, Yue Tang1 1Beijing Key Laboratory of Pre-clinical Research and Evaluation for Cardiovascular Implant Materials, State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Centre for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, 2Laboratory Animal Center, Peking University, 3Center of Cardiac Surgery for Adults, State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Centre for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery approach. Biochemistry An In Vivo Estrogen Deficiency Mouse Model for Screening Exogenous Estrogen Treatments of Cardiovascular Dysfunction After Menopause Bing Sun1, Yue-zhang Yin2, Jing Xiao1 1Institute of Medicinal Plant Development (IMPLAD), Chinese Academy of Medical Sciences & Peking Union Medical College, 2Shandong University of Traditional Chinese Medicine Clinically, estrogen deficiency in menopausal women may aggravate the incidence of lipid disruption and atherosclerosis. We established an in vivo estrogen deficiency model by bilateral ovariectomy via a double dorsal-lateral incision in apoE-/- mice. The mouse model is applicable for screening exogenous estrogen treatments of cardiovascular dysfunction after menopause. Cancer Research Digital Analysis of Immunostaining of ZW10 Interacting Protein in Human Lung Tissues Yuan Wen1, Xie Song-ping2, Liu Pan3, Liu Xiao-yan1, Pan Shan4, Yin Qian4, Sun Meng5, Huang Xiao-xing6, Xiao Rui-jing4, Xiong Jie4, Zhang Qiu-ping4, Shao Liang1 1Department of Hematology, Zhongnan Hospital of Wuhan University, 2Department of Thoracic Surgery, Renmin Hospital of Wuhan University, 3Department of Human Anatomy, Histology and Embryology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, 4Department of Immunology, School of Basic Medical Sciences, Wuhan University, 5Department of Gastroenterology, Central Hospital of Wuhan, 6Department of Transfusion, Zhongnan Hospital of Wuhan University ZW10 interacting protein (ZWINT) participates in the mitotic spindle checkpoint and the pathogenesis of carcinoma. Here, we introduce a methodology of the immunostaining of ZWINT in human lung cancer tissues, followed by the digital scanning of whole slides and image analysis. This methodology can provide high-quality digital images and reliable results. Medicine A Flow Cytometry-based Assay for Measuring Mitochondrial Membrane Potential in Cardiac Myocytes After Hypoxia/Reoxygenation Guihao Chen1, Yuejin Yang1, Chuansheng Xu2, Shuo Gao2 1State Key Laboratory of Cardiovascular Disease, Department of Cardiology, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, 2State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College Here, we present a protocol that uses JC-1 dye to assess the mitochondrial membrane potential of cells after being exposed to hypoxia/reoxygenation with or without a protective agent. Bioengineering Surface Engineering of Pancreatic Islets with a Heparinized StarPEG Nanocoating Jingyi Yang1, Shaofeng Lou1, Deling Kong1, Chen Li1 1Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Chinese Academy of Medical Science & Peking Union Medical College This protocol aims to achieve surface engineering of pancreatic islets using a heparin-incorporated starPEG nanocoating via pseudo-bioorthogonal chemistry between the N-hydroxysuccinimide groups of the nanocoating and the amine groups of islet cell membrane. Engineering Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes Chao Tian1, Wei Zhang2,3, Van Phuc Nguyen1, Xueding Wang2,4, Yannis M. Paulus1,2 1Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, 2Department of Biomedical Engineering, University of Michigan, 3Institute of Biomedical Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, 4Department of Radiology, University of Michigan This manuscript describes the novel setup and operating procedure of a photoacoustic microscopy and optical coherence tomography dual-modality system for noninvasive, label-free chorioretinal imaging of larger animals, such as rabbits. Developmental Biology Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors Wei Wen1, Jian-Ping Zhang1, Wanqiu Chen2, Cameron Arakaki2, Xiaolan Li1,2, David Baylink2, Gary D. Botimer3, Jing Xu1, Weiping Yuan1, Tao Cheng1,4,5,6,7, Xiao-Bing Zhang1,2 1State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 2Division of Regenerative Medicine, Department of Medicine, Loma Linda University, 3Department of Orthopaedic Surgery, Loma Linda University, 4Center for Stem Cell Medicine, Chinese Academy of Medical Sciences, 5Department of Stem Cell & Regenerative Medicine, Peking Union Medical College, 6Collaborative Innovation Center for Cancer Medicine, 7Tianjin Key Laboratory of Blood Cell Therapy and Technology This protocol describes a detailed method for efficient generation of integration-free iPSCs from human adult peripheral blood cells. With the use of four oriP/EBNA-based episomal vectors to express the reprogramming factors, KLF4, MYC, BCL-XL, or OCT4 and SOX2, thousands of iPSC colonies can be obtained from 1 mL of peripheral blood.
