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Peking Union Medical College

9 ARTICLES PUBLISHED IN JoVE

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Medicine

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model
Jing Feng 1, Yvonne Fitz 1, Yan Li 1, Melinda Fernandez 1, Irene Cortes Puch 1, Dong Wang 1, Stephanie Pazniokas 2, Brandon Bucher 3, Xizhong Cui 1, Steven B. Solomon 1
1Critical Care Medicine Department, Clinical Center, National Institutes of Health, 2Harvard Apparatus, 3ADInstruments

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

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Engineering

Using Synchrotron Radiation Microtomography to Investigate Multi-scale Three-dimensional Microelectronic Packages
Holly D. Carlton 1, John W. Elmer 1, Yan Li 2, Mario Pacheco 2, Deepak Goyal 2, Dilworth Y. Parkinson 3, Alastair A. MacDowell 3
1Materials Engineering Division, Lawrence Livermore National Laboratory, 2Assembly Test and Technology Development Failure Analysis Labs, Intel Corporation, 3Advanced Light Source, Lawrence Berkeley National Laboratory

For this study synchrotron radiation micro-tomography, a non-destructive three-dimensional imaging technique, is employed to investigate an entire microelectronic package with a cross-sectional area of 16 x 16 mm. Due to the synchrotron's high flux and brightness the sample was imaged in just 3 min with an 8.7 µm spatial resolution.

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Developmental Biology

Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors
Wei Wen 1, Jian-Ping Zhang 1, Wanqiu Chen 2, Cameron Arakaki 2, Xiaolan Li 1,2, David Baylink 2, Gary D. Botimer 3, Jing Xu 1, Weiping Yuan 1, Tao Cheng 1,4,5,6,7, Xiao-Bing Zhang 1,2
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.

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Bioengineering

Probing the Roles of Physical Forces in Early Chick Embryonic Morphogenesis
Yan Li *1, Hannah Grover *1, Eric Dai 2, Kevin Yang 1, Zi Chen 1
1Thayer School of Engineering, Dartmouth College, 2Department of Bioengineering, University of Pennsylvania

Here, we present a protocol introducing a set of new ex-ovo experiments and physical modeling approaches for studying the mechanics of morphogenesis during early chick embryonic brain torsion.

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Cancer Research

Digital Analysis of Immunostaining of ZW10 Interacting Protein in Human Lung Tissues
Yuan Wen 1, Xie Song-ping 2, Liu Pan 3, Liu Xiao-yan 1, Pan Shan 4, Yin Qian 4, Sun Meng 5, Huang Xiao-xing 6, Xiao Rui-jing 4, Xiong Jie 4, Zhang Qiu-ping 4, Shao Liang 1
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.

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Immunology and Infection

Functionalization of Atomic Force Microscope Cantilevers with Single-T Cells or Single-Particle for Immunological Single-Cell Force Spectroscopy
Jiahuan Chen 1, Ying Xu 2, Yan Shi 2, Tie Xia 2
1State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 2Institute for Immunology, School of Medicine, Tsinghua University

We present a protocol to functionalize atomic force microscope (AFM) cantilevers with a single T cell and bead particle for immunological studies. Procedures to probe single-pair T cell-dendritic cell binding by AFM and to monitor the real-time cellular response of macrophages to a single solid particle by AFM with fluorescence imaging are shown.

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Neuroscience

Purification of Fibroblasts and Schwann Cells from Sensory and Motor Nerves in Vitro
Qianru He 1, Fanhui Yu 1, Yan Li 1, Junjie Sun 1, Fei Ding 1
1Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-Innovation Center of Neuroregeneration, Jiangsu Clinical Medicine Center of Tissue Engineering and Nerve Injury Repair, Nantong University

Here, we present a method to purify fibroblasts and Schwann cells from sensory and motor nerves in vitro.

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Biochemistry

Transplantation of Neonatal Mouse Cardiac Macrophages into Adult Mice
Yandong Li 1, Jie Feng 1, Yan Li 1, Jianqiu Pei 1, Shengshou Hu 1, Yu Nie 1,2
1Fuwai Hospital & Chinese Academy of Medical Sciences, State Key Laboratory of Cardiovascular Disease, Cardiovascular Institute, National Center for Cardiovascular Diseases, Peking Union Medical College, 2National Health Commission Key Laboratory of Cardiovascular Regenerative Medicine, Fuwai Central-China Hospital, Central-China Subcenter of National Center for Cardiovascular Diseases

We provide a protocol for neonatal cardiac macrophage separation and transplantation into an adult mouse heart, which could be a promising way to promote cardiac repair.

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Immunology and Infection

Establishment of Deep Hypothermic Circulatory Arrest in Rats
Weidong Yan 1, Bingyang Ji 1
1Department of Cardiopulmonary Bypass, National Center for Cardiovascular Disease and Fuwai Hospital, State Key Laboratory of Cardiovascular Medicine, Chinese Academy of Medical Science, Peking Union Medical College

This protocol presents the establishment of deep hypothermic circulatory arrest in rats, which can be applied to investigate systemic inflammatory response syndrome, ischemia/reperfusion injury, oxidative stress, neuroinflammation, etc.

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