China Academy of Chinese Medical Sciences 10 articles published in JoVE Neuroscience Demonstrating Hairy and Glabrous Skin Innervation in a 3D Pattern Using Multiple Fluorescent Staining and Tissue Clearing Approaches Xiaoyu Wang*1, Wanying Cao*1, Jingtao Shi*1, Xiaoning Zhang1, Zhengyang Qu1, Dongsheng Xu1, Hongye Wan1, Yangshuai Su1, Wei He1, Xianghong Jing1, Wanzhu Bai1 1Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences The thickness of tissue sections limited the morphological study of the skin innervation. The present protocol describes a unique tissue clearing technique to visualize cutaneous nerve fibers in thick 300 µm tissue sections under confocal microscopy. Neuroscience Visualizing the Morphological Characteristics of Neuromuscular Junction in Rat Medial Gastrocnemius Muscle Jingjing Cui*1, Shuang Wu*1, Jia Wang1, Yuqing Wang1, Yuxin Su1, Dongsheng Xu1, Yihan Liu1, Junhong Gao1, Xianghong Jing1, Wanzhu Bai1 1Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences The protocol shows a method to examine spatial correlation among the pre-synaptic terminals, post-synaptic receptors, and peri-synaptic Schwann cells in the rat medial gastrocnemius muscle using fluorescent immunohistochemistry with different biomarkers, namely, neurofilament 200, vesicular acetylcholine transporter, alpha-bungarotoxin, and S100. Medicine Aggravation of Myocardial Ischemia upon Particulate Matter Exposure in Atherosclerosis Animal Model Yuanmin Yang*1, Shuoqiu Deng*1, Shuiqing Qu1, Yu Zhang1, Zhongyuan Zheng1, Lina Chen1, Yujie Li1 1China Academy of Chinese Medical Sciences This protocol describes a composite animal model with exposure to particulate matter (PM) that aggravates myocardial ischemia with atherosclerosis. Medicine OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery Tao Liu*1,2,3,4,5, Xueling Song*1,2,3,4,5, Xiaoying Zheng*1,2,3,4,5, Liying Yan1,2,3,4,5, Caihong Ma1,2,3,5, Rong Li1,2,3,4,5, Jie Yan1,2,3,4,5, Jie Qiao1,2,3,4,5 1Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, 2National Clinical Research Center for Obstetrics and Gynecology, 3Key Laboratory of Assisted Reproduction, Ministry of Education, 4Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction, 5Research Units of Comprehensive Diagnosis and Treatment of Oocyte Maturation Arrest, Chinese Academy of Medical Sciences In vitro maturation (IVM) before gynecological operation (OP-IVM) combines IVM following oocyte retrieval with routine gynecological surgery and serves as an extension of conventional IVM applications for fertility preservation. Neuroscience Visualizing the Calcitonin Gene-Related Peptide Immunoreactive Innervation of the Rat Cranial Dura Mater with Immunofluorescence and Neural Tracing Jia Wang*1, Dongsheng Xu*1, Jingjing Cui1, Chen She1, Hui Wang1, Shuang Wu1, Ling Zou1, Jianliang Zhang1, Wanzhu Bai1 1Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences Here we present a protocol to visualize spatial correlation of calcitonin gene-related peptide (CGRP)-immunoreactive nerve fibers and blood vessels in the cranial dura mater using immunofluorescence and fluorescent histochemistry with CGRP and phalloidin, respectively. In addition, the origin of these nerve fibers was retrograde traced with a fluorescent neural tracer. Medicine A High-Throughput Electrochemiluminescence 7-Plex Assay Simultaneously Screening for Type 1 Diabetes and Multiple Autoimmune Diseases Xiaofan Jia*1,2, Ling He*1,3, Yong Gu1, Hilary High1, Liping Yu1 1Barbara Davis Center for Diabetes, University of Colorado School of Medicine, 2Department of Endocrinology, Beijing Hospital, National Center of Gerontology; Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, 3Department of endocrinology, Guangzhou First People's Hospital, School of Medicine, South China University of Technology We model a simple multiplexed ECL assay that combines 7 autoantibody assays together. The assay is capable of screening for T1D and multiple other autoimmune diseases, simultaneously, including celiac disease, autoimmune thyroid disease, and autoimmune polyglandular syndrome 1. Genetics Inducing Hairy Roots by Agrobacterium rhizogenes-Mediated Transformation in Tartary Buckwheat (Fagopyrum tataricum) Yaolei Mi1,2, Zhihui Zhu3, Guangtao Qian3, Yu Li4, Xiangxiao Meng1, Jianping Xue3, Qingfu Chen5, Wei Sun1, Yuhua Shi1,2 1Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, 2Artemisinin Research Center, China Academy of Chinese Medical Sciences, 3College of Life Science, Huaibei Normal University, 4Economic Crop Research Institute Sichuan Academy of Agriculture Sciences, 5Research Center of Buckwheat Industry Technology, Guizhou Normal University We describe a method of inducing hairy roots by Agrobacterium rhizogenes-mediated transformation in Tartary buckwheat (Fagopyrum tataricum). This can be used to investigate gene functions and production of secondary metabolites in Tartary buckwheat, be adopted for any genetic transformation, or used for other medicinal plants after improvement. Behavior Analysis of Learning and Memory Ability in an Alzheimer's Disease Mouse Model using the Morris Water Maze Huiling Tian1, Ning Ding2, Mengwei Guo1, Shun Wang1, Zidong Wang1, Hao Liu1, Jiayi Yang1, Yujie Li3, Jingyu Ren1, Jing Jiang4, Zhigang Li1 1School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, 2Guang'anmen Hospital of China Academy of Chinese Medical Science, 3The Third Affiliated Hospital of Beijng University of Chinese Medicine, 4School of Nursing, Beijing University of Chinese Medicine Herein, a protocol to conduct the Morris water maze tests to evaluate the ability of learning and memory of Alzheimer’s Disease model mice and to assess the effect of manual acupuncture for treating them is described. Neuroscience Improving the Application of High Molecular Weight Biotinylated Dextran Amine for Thalamocortical Projection Tracing in the Rat Dongsheng Xu*1, Jingjing Cui*1, Jia Wang1, Zhiyun Zhang1, Chen She1, Wanzhu Bai1 1Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences Here, we present a refined protocol to effectively reveal biotinylated dextran amine (BDA) labeling with a fluorescent staining method through a reciprocal neural pathway. It is suitable for analyzing the fine structure of BDA labeling and distinguishing it from other neural elements under a confocal laser scanning microscope. Medicine Assessment of Sarcoplasmic Reticulum Calcium Reserve and Intracellular Diastolic Calcium Removal in Isolated Ventricular Cardiomyocytes Jing Gao*1, Xiaolu Shi*2, Hong He1, Juhong Zhang1, Ding Lin3, Guosheng Fu1, Dongwu Lai1 1Department of Cardiology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 2Experimental Research Center, China Academy of Chinese Medical Sciences, 3Department of Cardiology, The Third Hospital of Hangzhou City Intracellular calcium recycling plays a critical role in regulation of systolic and diastolic function in cardiomyocytes. Here, we describe a protocol to evaluate sarcoplasmic reticulum Ca2+ reserve and diastolic calcium removal function in cardiomyocytes by a calcium imaging system.
