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Shanghai University of Traditional Chinese Medicine

30 ARTICLES PUBLISHED IN JoVE

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Medicine

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD
Min Jia 1, Fei Meng 2, Stanley E. Smerin 1, Guoqiang Xing 1, Lei Zhang 1, David M. Su 2, David Benedek 1, Robert Ursano 1, Yan A. Su 2, He Li 1
1Department of Psychiatry, Center for the Study of Traumatic Stress, Uniformed Services University of the Health Sciences, Bethesda, Maryland, 2Department of Gene and Protein Biomarkers, GenProMarkers, Inc.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

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Medicine

Parabiosis in Mice: A Detailed Protocol
Paniz Kamran 1,2, Konstantina-Ioanna Sereti 1,2, Peng Zhao 1,2, Shah R. Ali 3, Irving L. Weissman 3, Reza Ardehali 1,2
1Department of Medicine-Division of Cardiology, University of California, Los Angeles, 2Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, 3Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine

Parabiotic joining of two organisms leads to the development of a shared circulatory system. In this protocol, we describe the surgical steps to form a parabiotic connection between a wild-type mouse and a constitutive GFP-expressing mouse.

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Biology

Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists
Hao Li *1, Wei Dou *2, Emil Padikkala 1, Sridhar Mani 1
1Department of Genetics, Albert Einstein College of Medicine , 2Shanghai Key Laboratory of Complex Prescription and MOE Key Laboratory for Standardization of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine

Ketoconazole binds to and antagonizes Pregnane X Receptor (PXR) activation. Yeast high throughput screens of PXR mutants define a unique region for ketoconazole binding. This yeast-based genetic method discovers novel nuclear receptor interactions with ligands that associate with surface binding sites.

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Medicine

Combined In vivo Optical and µCT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice
Nicholas M. Bernthal 1, Brad N. Taylor 2, Jeffrey A. Meganck 2, Yu Wang 3, Jonathan H. Shahbazian 3, Jared A. Niska 1, Kevin P. Francis 2, Lloyd S. Miller 3,4
1Orthopaedic Hospital Research Center, Orthopaedic Hospital Department of Orthopaedic Surgery, David Geffen School of Medicine at University of California, Los Angeles (UCLA), 2PerkinElmer, 3Department of Dermatology, Johns Hopkins University School of Medicine, 4Department of Medicine, Division of Infectious Diseases, Department of Orthopaedic Surgery, Johns Hopkins University School of Medicine

Combined optical and μCT imaging in a mouse model of orthopaedic implant infection, utilizing a bioluminescent engineered strain of Staphylococcus aureus, provided the capability to noninvasively and longitudinally monitor the dynamics of the bacterial infection, as well as the corresponding inflammatory response and anatomical changes in the bone.

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Developmental Biology

Defined and Scalable Generation of Hepatocyte-like Cells from Human Pluripotent Stem Cells
Yu Wang 1, Sharmin Alhaque 1, Kate Cameron 1, Jose Meseguer-Ripolles 1, Baltasar Lucendo-Villarin 1, Hassan Rashidi 1, David C. Hay 1
1MRC Centre for Regenerative Medicine, University of Edinburgh

The method presented here describes a scalable and good manufacturing practice (GMP)-ready differentiation system to generate human hepatocyte-like cells from pluripotent stem cells. It serves as a cost-effective and standardized system to generate human hepatocyte-like cells for basic and applied human liver research.

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Medicine

Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments
Shufen Liu 1, Yueli Sun 1, Ji Li 1, Jingcheng Dong 2, Qin Bian 1,2
1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2The Academy of Integrative Medicine of Fudan University

Here, we present the preparation of an er-xian decoction (EXD) in four steps—soaking, decoction, filtration, and concentration—and demonstrate the administration of a prepared EXD-containing serum to rats. These methods are applicable to the in vivo and in vitro study of herbal decoctions such as traditional Chinese medicines.

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Biology

Laser Capture Microdissection of Highly Pure Trabecular Meshwork from Mouse Eyes for Gene Expression Analysis
Caleb Sutherland *1, Yu Wang *2, Robert V. Brown *1, Julie Foley 2, Beth Mahler 2, Kyathanahalli S. Janardhan 2,3, Ramesh C. Kovi 2,4, Anton M. Jetten 1
1Immunity, Inflammation, and Disease Laboratory, Division of Intramural Research, National Institute of Environmental Health Sciences, NIH, 2Cellular and Molecular Pathology Branch, Division of the National Toxicology Program, National Institute of Environmental Health Sciences, NIH, 3Integrated Laboratory Systems Inc., 4Experimental Pathology Laboratories Inc.

Here, we describe a protocol for a reproducible laser capture microdissection (LCM) for isolating trabecular meshwork (TM) for downstream RNA analysis. The ability to analyze changes in gene expression in the TM will help in understanding the underlying molecular mechanisms of TM-related ocular diseases.

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Developmental Biology

Semi-automated Production of Hepatocyte Like Cells from Pluripotent Stem Cells
Jose Meseguer-Ripolles 1, Baltasar Lucendo-Villarin 1, Yu Wang 1, David C. Hay 1
1MRC Centre for Regenerative Medicine, University of Edinburgh

This protocol describes a semi-automated approach to produce hepatocyte-like cells from human pluripotent stem cells in a 96 well plate format. This process is rapid and cost-effective, allowing the production of quality assured batches of hepatocyte-like cells for basic and applied human research.

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Medicine

Preparation Of Gushukang (GSK) Granules for In Vivo and In Vitro Experiments
Yongjian Zhao 1,2,3, Qiang Wang 1,2,3, Shufen Liu 1,2,3, Yongjun Wang 1,2,3,4, Bing Shu 1,2,3, Dongfeng Zhao 1,2,3
1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Spine Institute, Shanghai University of Traditional Chinese Medicine, 3Key Laboratory of Theory and Therapy of Muscles and Bones, Ministry of Education, 4School of Rehabilitation Science, Shanghai University of Traditional Chinese Medicine

This article provides a detailed protocol for preparing a working solution of Gushukang granules for animal studies and GSK granule containing serum for in vitro experiments. This protocol can be applied to pharmacological investigations of herbal medicines as well as prescriptions for both in vivo and in vitro experiments.

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Biology

Assessment of Vascular Tone Responsiveness using Isolated Mesenteric Arteries with a Focus on Modulation by Perivascular Adipose Tissues
Daniels Konja 1, Cuiting Luo 1, Wai Yan Sun 1, Kangmin Yang 1, Andy W.C. Man 1, Aimin Xu 1, Paul M. Vanhoutte 1, Yu Wang 1
1The State Key Laboratory of Pharmaceutical Biotechnology and the Department of Pharmacology and Pharmacy, University of Hong Kong

The protocol describes the use of wire myography to evaluate the transmural isometric tension of mesenteric arteries isolated from mice, with special consideration of the modulation by factors released from endothelial cells and perivascular adipose tissues.

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JoVE Core

Home-Based Prescribed Pulmonary Exercise in Patients with Stable Chronic Obstructive Pulmonary Disease
Xiaodan Liu 1,2, Peijun Li 3, Jian Li 3, Lu Xiao 1, Ning Li 3, Yufan Lu 3, Zhengrong Wang 3, Jianqing Su 3, Zhenwei Wang 4, Chunlei Shan 1,2, Weibing Wu 3
1School of Rehabilitation Science, Shanghai University of Traditional Chinese Medicine, 2Institute of Rehabilitation Medicine, Shanghai Academy of Traditional Chinese Medicine, 3Department of Sports Medicine, Shanghai University of Sport, 4Department of Respiratory Medicine, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine

Presented here is a protocol to investigate the effects of home-based prescribed pulmonary exercise in stable chronic obstructive pulmonary disease (COPD) patients, which is modified based on traditional Chinese exercises according to dyspnea and limited exercise capacity observed in COPD patients.

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Developmental Biology

Serum Free Production of Three-dimensional Human Hepatospheres from Pluripotent Stem Cells
Balta Lucendo-Villarin 1, Hassan Rashidi 1,2, Sharmin Alhaque 1,3, Lena Fischer 1,4, Jose Meseguer-Ripolles 1, Yu Wang 1, Cliona O'Farrelly 4, Michael Themis 3, David C. Hay 1
1MRC Centre for Regenerative Medicine, University of Edinburgh, 2UCL Great Ormond Street Institute of Child Health, University College London, 3Division of Biosciences, Department of Life Sciences, College of Health and Life Sciences, Brunel University London, 4School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin

This protocol describes an approach to produce hepatospheres from human pluripotent stem cells using a defined culture system and cell self-assembly. This protocol is reproducible in a number of cell lines, cost effective and allows the production of stable human hepatospheres for biomedical application.

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Bioengineering

Screening and Identification of Small Peptides Targeting Fibroblast Growth Factor Receptor2 using a Phage Display Peptide Library
Ying Zhao *1, Qiang Wang *1, An Hong 1, Xiaojia Chen 1
1Institute of Biomedicine & Department of Cell Biology, Jinan University, National Engineering Research Center of Genetic Medicine, Guangdong Provincial Key Laboratory of Bioengineering Medicine

Herein, we present a detailed protocol for screening small peptides that bind to FGFR2 using a phage display peptide library. We further analyze the affinity of the selected peptides toward FGFR2 in vitro and its ability to suppress cell proliferation.

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Genetics

Introducing Point Mutations into Human Pluripotent Stem Cells Using Seamless Genome Editing
Yu Wang 1, Andrew J. H. Smith 1, David C. Hay 1
1MRC Centre for Regenerative Medicine, University of Edinburgh

Here, we describe a detailed method for seamless gene editing in human pluripotent stem cells using a piggyBac-based donor plasmid and the Cas9 nickase mutant. Two point mutations were introduced into exon 8 of the hepatocyte nuclear factor 4 alpha (HNF4α) locus in human embryonic stem cells (hESCs).

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Biology

Hepatic Progenitor Specification from Pluripotent Stem Cells using a Defined Differentiation System
Jose Meseguer-Ripolles 1, Yu Wang 1, Agnes Sorteberg 1, Aishwariya Sharma 2, Nan-Linda Ding 2, Baltasar Lucendo-Villarin 1, Philipp Kramer 2, Charis-Patricia Segeritz 2, David C. Hay 1
1MRC Centre for Regenerative Medicine, University of Edinburgh, 2Research & Development, STEMCELL Technologies Inc

The goal of this article is to provide a standardized approach to induce human hepatic progenitor differentiation from pluripotent stem cells. The development of this procedure with ready-to-use media formulations offer the user a facile system to generate human liver cells for biomedical research and translation.

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Developmental Biology

Differentiation and Characterization of Neural Progenitors and Neurons from Mouse Embryonic Stem Cells
Aflah Hanafiah 1, Zhuangzhuang Geng 1, Qiang Wang 1, Zhonghua Gao 1
1Departments of Biochemistry and Molecular Biology, Penn State College of Medicine, Penn State Hershey Cancer Institute

We describe the procedure for the in vitro differentiation of mouse embryonic stem cells into neuronal cells using the hanging drop method. Furthermore, we perform a comprehensive phenotypic analysis through RT-qPCR, immunofluorescence, RNA-seq, and flow cytometry.

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Biology

A Mouse Model of Lumbar Spine Instability
Shufen Liu 1, Yueli Sun 1, Jingcheng Dong 2, Qin Bian 2
1Spine Research Institute, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Department of Integrative Medicine, Huashan Hospital, Fudan University

We developed a lumbar intervertebral disc degeneration mouse model by resection of L3–L5 spinous processes along with supra- and inter-spinous ligaments and detachment of paraspinous muscles.

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Medicine

Acupuncture in a Rat Model of Asthma
Dong-Dong Zhou 1, Guang-Quan Zhang 1, Qing-Yi Zhao 1, Mi Cheng 1, Jin Lu 1, Yu Wang 1, Yu-Dong Xu 1, Yan-Jiao Chen 1, Yong-Qing Yang 1, Lei-Miao Yin 1,2
1Laboratory of Molecular Biology, Shanghai Research Institute of Acupuncture and Meridian, Shanghai University of Traditional Chinese Medicine, 2Shanghai Innovation Center of Traditional Chinese Medicine Health Service

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes methods for the fabrication of the high platform, establishes a rat model of asthma and measures changes in respiratory function using a noninvasive and real-time whole-body plethysmography (WBP) system.

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Neuroscience

A Neuronal Apoptosis Model induced by Spinal Cord Compression in Rat
Yue-li Sun *1,2, Gan Li *1,2, Zhong Zheng *1,2, Min Yao 1,2, Jia-wen Cui, Shu-fen Liu 1,2, Long-Yun Zhou 1,2, Kim-Sia Sng 1,2, Xue-Jun Cui 1,2, Yong-jun Wang 1,2,3
1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Spine Research Institute, Shanghai University of Traditional Chinese Medicine, 3Shanghai University of Traditional Chinese Medicine

Here, we present a protocol to generate a rat spinal cord compression model, assess its behavioral score, and observe the compressed spinal cord region. The behavioral assessments showed decreased monitor motor disability. Hematoxylin and eosin staining and immunostaining revealed considerable neuronal apoptosis in the compressed region of the spinal cord.

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Medicine

Preparation of Naringenin Solution for In Vivo Application
Shufen Liu 1, Jingcheng Dong 2, Qin Bian 2
1Spine Research Institute, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Department of Integrative Medicine, Huashan Hospital, Fudan University

Here, the protocol presents the preparation of naringenin solution for in vivo intraperitoneal administration. Naringenin is fully dissolved in a mixture of dimethylsulfoxide, Tween 80, and saline. The antidiabetic osteoporotic effects of naringenin were assessed by blood glucose testing, tartrate-resistant acid phosphatase staining, and enzyme-linked immunosorbent assay.

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Medicine

Three-Dimensional Finger Motion Tracking during Needling: A Solution for the Kinematic Analysis of Acupuncture Manipulation
Liu-Liu Xu 1, Fan Wang 1, Hua-Yuan Yang 1, Wen-Chao Tang 1
1School of Acupuncture-Moxibustion and Tuina, Shanghai University of Traditional Chinese Medicine

This experimental method describes a solution for the kinematic analysis of acupuncture manipulation with three-dimensional finger motion tracking technology.

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Medicine

Robot-assisted Total Mesorectal Excision and Lateral Pelvic Lymph Node Dissection for Locally Advanced Middle-low Rectal Cancer
Chenhao Hu *1,2,3, Zhe Zhang *1,2,3, Lei Zhang 1,2,3, Ruihan Liu 2,3, Jun Yan 1,2,3, Qi Sun 1,2,3, Guanghui Wang 1, Junjun She 1,2,3
1Department of General Surgery, The First Affiliated Hospital of Xian Jiaotong University, 2Center for Gut Microbiome Research, Med-X Institute, The First Affiliated Hospital of Xian Jiao tong University, 3Department of High Talent, The First Affiliated Hospital of Xian Jiaotong University

The robotic technique described herein aims to detail a stepwise approach to robot-assisted total mesorectal excision and lateral pelvic lymph node dissection for locally advanced (T3/T4) rectal cancer located below the peritoneal reflection.

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Cancer Research

Intratibial Osteosarcoma Cell Injection to Generate Orthotopic Osteosarcoma and Lung Metastasis Mouse Models
Junli Chang *1,2, Fulai Zhao *1,2, Xingyuan Sun 1,2, Xiaoping Ma 1,2, Wenlan Zhi 1,2, Yanping Yang 1,2
1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Key Laboratory of theory and therapy of muscles and bones, Ministry of Education

The present protocol describes intratibia osteosarcoma cell injection to generate mouse models bearing orthotopic osteosarcoma and pulmonary metastasis lesions.

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Biochemistry

Alleviation of Diabetic Tendon Injury via Activation of Tendon Fibroblasts Autophagy under Berberine Treatment
Mei-Dong Zhu *1, Qi Li *2, Jia-Qin Yuan 1, Fu-Chen Song 1, Sheng-Hua Liu 2, Lei Zhang 1, Cheng Zhao 3
1Department of Vascular Surgery, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, 2Yueyang Clinical Medical College, Shanghai University of Traditional Chinese Medicine, 3Department of Vascular Disease, Shanghai TCM-Integrated Hospital, Shanghai University of Traditional Chinese Medicine

Alleviation of Diabetic Tendon Injury via Activation of Tendon Fibroblasts Autophagy under Berberine Treatment

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Cancer Research

Intra-Cardiac Injection of Human Prostate Cancer Cells to Create a Bone Metastasis Xenograft Mouse Model
Junli Chang *1,2, Xingyuan Sun *1,2, Xiaoping Ma 1,2, Peng Zhao 1,2, Binhao Shi 1,2, Yongjun Wang 1,2, Xianghui Han 1,3, Yanping Yang 1,2
1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Key Laboratory of Theory and Therapy of Muscles and Bones, Ministry of Education, 3Institute of Chinese Traditional Surgery, Longhua Hospital, Shanghai University of Traditional Chinese Medicine

Here, we present a protocol for the intra-cardiac injection of human prostate cancer cells to generate a mouse model with bone metastasis lesions.

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Cancer Research

RNA Fluorescence In Situ Hybridization for Long Non-Coding RNA Localization in Human Osteosarcoma Cells
Junli Chang *1,2, Xiaoping Ma *1,2, Xingyuan Sun 1,2, Chujie Zhou 1,2, Peng Zhao 1,2, Yongjun Wang 1,2, Yanping Yang 1,2
1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Key Laboratory of Theory and Therapy of Muscles and Bones, Ministry of Education

The present protocol describes a method of RNA fluorescence in situ hybridization to localize the lncRNAs in human osteosarcoma cells.

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Medicine

Tuina Intervention in Sodium Monoiodoacetate Injection-Induced Rat Model of Knee Osteoarthritis
Yifei Xu *1, Xuanying Zhu *2, Xinrong Li *3, Yuxiang Lu *4, Jiangwei Wu 3, Weimin Cai 3, Juanjuan Zheng 3, Zhiwei Wu 5, Guangxin Guo 3,6
1Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, 2Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, 3School of Acupuncture-Moxibustion and Tuina, Shanghai University of Traditional Chinese Medicine, 4School of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, 5Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, 6Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine

This protocol describes the methods of Tuina intervention in sodium monoiodoacetate injection-induced rat model of knee osteoarthritis (KOA), which provides a reference for the application of Tuina in KOA animal models. This protocol also studies the effective mechanism of Tuina for KOA, and the results will help promote its application.

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Medicine

Clinical Efficacy of Small Needle Knife Therapy on Stage I-II Frozen Shoulder
Suxia Guo 1, Dan Liu 1, Yanping Yang 1,2, Zhijun Hu 1
1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Key Laboratory of theory and therapy of muscles and bones, Ministry of Education

The protocol presented here describes the detailed operation method and proves the effectiveness of a small needle knife in the treatment of frozen shoulders.

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Medicine

Gavage Strategy for Decoction Formula of Traditional Chinese Medicine in Osteosarcoma Model Mice
Chujie Zhou 1,2, Xingyuan Sun 1,2, Junli Chang 1,2, Junjie Tong 1,2, Peng Zhao 1,2, Suxia Guo 1,2, Yanping Yang 1,2
1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Key Laboratory of Theory and Therapy of Muscles and Bones, Ministry of Education

The protocol provides a detailed standardization process of decoction formula and gavage technique with Yiqi Jiedu decoction in the osteosarcoma mouse model as an example. It describes animal protection and improves reliability of research data, providing effective strategies for investigating therapeutic efficacy and molecular mechanisms of decoction formulas in vivo.

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Cancer Research

Establishment of Patient-Derived Xenograft Mouse Model with Human Osteosarcoma Tissues
Xingyuan Sun *1,2, Junli Chang *1,2, Chujie Zhou 1,2, Peng Zhao 1,2, Suxia Guo 1,2, Junjie Tong 1,2, Yongjun Wang 1,2, Yanping Yang 1,2
1Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 2Key Laboratory of theory and therapy of muscles and bones, Ministry of Education

The present protocol describes the method for establishing a patient-derived xenograft (PDX) mouse model using human osteosarcoma tissue.

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