Ministry of Education

10 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Generation of Induced Neural Stem Cells from Peripheral Mononuclear Cells and Differentiation Toward Dopaminergic Neuron Precursors for Transplantation Studies

Wei Zheng^1,2, Zhiguo Chen^1,2

¹Cell Therapy Center, Beijing Institute of Geriatrics, Xuanwu Hospital, Capital Medical University, and Key Laboratory of Neurodegeneration, Ministry of Education, ²Center of Neural Injury and Repair, Beijing Institute for Brain Disorders

The protocol presents the reprogramming of peripheral blood mononuclear cells to induce neural stem cells by Sendai virus infection, differentiation of iNSCs into dopaminergic neurons, transplantation of DA precursors into the unilaterally-lesioned Parkinson's disease mouse models, and evaluation of the safety and efficacy of iNSC-derived DA precursors for PD treatment.

Medicine

The Miniature Pig: A Large Animal Model for Cochlear Implant Research

Xiaojun Ji^*1,2,3,4, Yi Luo^*1,2,3,4, Weiwei Guo^1,2,3,4, Fei Ji^1,2,3,4, Shuolong Yuan^1,2,3,4, Liangwei Xu^1,2,3,4, Wei Chen^*1,2,3,4

¹College of Otolaryngology Head and Neck Surgery, Chinese PLA General Hospital, ²National Clinical Research Center for Otolaryngologic Diseases, ³Key Lab of Hearing Science, Ministry of Education, ⁴Beijing Key Lab of Hearing Impairment for Prevention and Treatment

Miniature pigs (mini-pigs) are an ideal large animal model for research into cochlear implants. Cochlear implantation surgery in mini-pigs can be utilized to provide initial evidence of the safety and potential performance of novel electrode arrays and surgical approaches in a living system similar to human beings.

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.

Biology

Efficient PAM-Less Base Editing for Zebrafish Modeling of Human Genetic Disease with zSpRY-ABE8e

Shaohui Zheng^1,2, Fang Liang³, Yu Zhang^1,2, Ji-Feng Fei⁴, Wei Qin^1,2, Yanmei Liu^1,2

¹Key Laboratory of Brain, Cognition and Education Science, Ministry of Education, ²Institute for Brain Research and Rehabilitation, Guangdong Key Laboratory of Mental Health and Cognitive Science, South China Normal University, ³Institute of Modern Aquaculture Science and Engineering, Guangdong Provincial Engineering Technology Research Center for Environmentally Friendly Aquaculture, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Sciences, South China Normal University, ⁴Department of Pathology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences

This protocol describes how to perform efficient adenine base editing without PAM limitation to construct a precise zebrafish disease model using zSpRY-ABE8e.

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.

Medicine

Clinical Efficacy of Small Needle Knife Therapy on Stage I-II Frozen Shoulder

Suxia Guo¹, Dan Liu¹, Yanping Yang^1,2, Zhijun Hu¹

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.

Medicine

2D-HPLC-MS Technology Combined with Molecular Network for the Identification of Components in Tibetan Medicine Aconitum pendulum

Jing Ma^*1, Hanyu Lu^*1, Jia Liu¹, Ting Wang¹, Xing Fu², Xinmei Xu², Yi Zhang¹, Jing Zhang¹, Xiaolong Xie¹, Yingzhuang Chen^3,4, Jinsong Su¹

¹Ethnic Medicine Academic Heritage Innovation Research Center, Meishan Hospital, Chengdu University of Traditional Chinese Medicine, ²State Key Laboratory of Southwestern Chinese Medicine Resources, Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, ³Key Laboratory of Chemical Biology & Traditional Chinese Medicine Research, Ministry of Education, ⁴Key Laboratory of Phytochemical R&D of Hunan Province, Hunan Normal University

This study utilizes two-dimensional high-performance liquid chromatography-mass spectrometry (2D-HPLC-MS) technology in conjunction with molecular networking to unravel the intricate chemical composition of the Tibetan medicinal plant Aconitum pendulum Busch (APB). The article provides a detailed protocol for the systematic exploration and identification of complex chemical components of herbal medicines.

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.

Medicine

Endoscopic Balloon Dilatation of the Eustachian Tube via the Soft Palate Approach in Miniature Pigs

Hui Song^*1,2,3,4, Jianping Jia^*1,2,3,4, Danheng Zhao^*1,2,3,4, Mengyuan Guo^1,2,3,4, Fangyuan Wang^1,2,3,4, Kun Hou^1,2,3,4, Nan Jiang⁵, Zhenhao Fu^1,2,3,4, Jingcheng Zhou^1,2,3,4, Haoze Zhang^1,2,3,4, Yulin Ding^1,2,3,4, Yuting Yang⁵, Chen Chen⁵, Xinyuan Jia^1,2,3,4, Zhaohui Hou^1,2,3,4, Shiming Yang^1,2,3,4

¹Senior Department of Otolaryngology Head and Neck Surgery, Chinese PLA General Hospital, Chinese PLA Medical School, ²National Clinical Research Center for Otolaryngologic Diseases, ³Key Laboratory of Hearing Science, Ministry of Education, ⁴Beijing Key Laboratory of Hearing Impairment for Prevention and Treatment, ⁵Liaoning University of Traditional Chinese Medicine

In this study, we present a protocol to explore an eustachian tube surgery approach via the soft palate in miniature pigs. The surgical procedure is simple, with short surgical time, and wound healing is rapid; it is thus a good choice for procedures such as balloon dilation of the eustachian tube.

Biology

Enhancing Density Maps by Removing the Majority of Particles in Single Particle Cryogenic Electron Microscopy Final Stacks

Mengjia Cai^*1, Jianying Zhu^*2, Qi Zhang^*3,4,5,6, Yu Xu⁷, Zuoqiang Shi^2,8, Chenglong Bao^2,8,9, Mingxu Hu^1,5,6

¹Institute of Bio-Architecture and Bio-Interactions (IBABI), Shenzhen Medical Academy of Research and Translation, ²Yau Mathematical Sciences Center, ³Key Laboratory of Protein Science (Tsinghua University), Ministry of Education, ⁴School of Life Science, Tsinghua University, ⁵Beijing Advanced Innovation Center for Structural Biology, ⁶Beijing Frontier Research Center for Biological Structure, ⁷Department of Mathematical Sciences, Tsinghua University, ⁸Yanqi Lake Beijing Institute of Mathematical Sciences and Applications, ⁹State Key Laboratory of Membrane Biology, School of Life Science

An advanced particle selection method for cryo-EM, namely CryoSieve, improves density map resolution by removing a majority of particles in final stacks, as demonstrated through its application on a real-world dataset.