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Key Laboratory of Theory and Therapy of Muscles and Bones, Ministry of Education

9 ARTICLES PUBLISHED IN JoVE

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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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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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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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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

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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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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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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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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