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China Academy Of Chinese Medical Sciences

13 ARTICLES PUBLISHED IN JoVE

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Medicine

Assessment of Sarcoplasmic Reticulum Calcium Reserve and Intracellular Diastolic Calcium Removal in Isolated Ventricular Cardiomyocytes
Jing Gao *1, Xiaolu Shi *2, Hong He 1, Juhong Zhang 1, Ding Lin 3, Guosheng Fu 1, Dongwu Lai 1
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.

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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 Wang 1, Zhiyun Zhang 1, Chen She 1, Wanzhu Bai 1
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.

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Genetics

Inducing Hairy Roots by Agrobacterium rhizogenes-Mediated Transformation in Tartary Buckwheat (Fagopyrum tataricum)
Yaolei Mi 1,2, Zhihui Zhu 3, Guangtao Qian 3, Yu Li 4, Xiangxiao Meng 1, Jianping Xue 3, Qingfu Chen 5, Wei Sun 1, Yuhua Shi 1,2
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.

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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 Cui 1, Chen She 1, Hui Wang 1, Shuang Wu 1, Ling Zou 1, Jianliang Zhang 1, Wanzhu Bai 1
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.

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Medicine

Aggravation of Myocardial Ischemia upon Particulate Matter Exposure in Atherosclerosis Animal Model
Yuanmin Yang *1, Shuoqiu Deng *1, Shuiqing Qu 1, Yu Zhang 1, Zhongyuan Zheng 1, Lina Chen 1, Yujie Li 1
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.

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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 Zhang 1, Zhengyang Qu 1, Dongsheng Xu 1, Hongye Wan 1, Yangshuai Su 1, Wei He 1, Xianghong Jing 1, Wanzhu Bai 1
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.

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Neuroscience

Visualizing the Morphological Characteristics of Neuromuscular Junction in Rat Medial Gastrocnemius Muscle
Jingjing Cui *1, Shuang Wu *1, Jia Wang 1, Yuqing Wang 1, Yuxin Su 1, Dongsheng Xu 1, Yihan Liu 1, Junhong Gao 1, Xianghong Jing 1, Wanzhu Bai 1
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.

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Medicine

Dioscin Mediated IgA Nephropathy Alleviation by Inhibiting B Cell Activation In Vivo and Decreasing Galactose-Deficient IgA1 Production In Vitro
Lan Lin *1, JiaChen Shen *2, XiaoCong Wu 1, Yun You 3, Shen Li 1
1Department of Nephrology, Guanganmen Hospital, 2Department of Nephrology, 906 Hospital of Joint Logistic Support Force of People's Liberation Army (PLA), 3Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences

This study provides experimental data for treating immunoglobulin A nephropathy (IgAN) with Dioscin (DIO), the active ingredient of Dioscoreae Nipponicae Rhizoma (DNR), and a paradigm for studying herbal medicine's effects and underlying mechanisms in vivo and in vitro.

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Neuroscience

In Vivo Calcium Imaging of Dorsal Root Ganglia Neurons' Response to Somatic and Visceral Stimuli
Kun Liu *1, Yun Liu *1, Xia Li 1, Shuya Wang 1, Xiaoxi Wang 1, Zhiyun Zhang 1, Xinyan Gao 1
1Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences

The present protocol outlines in vivo calcium imaging for measuring the responses of ensembles of lumbar-6 DRG neurons to somatic and visceral stimuli. Thorough comparisons can be made among neurons responding to different stimuli. This protocol is valuable for investigating mechanisms of visceral pain and somatic stimulation, such as acupuncture.

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Neuroscience

In Vivo Thoracic Dorsal Root Ganglia (DRG) Calcium Imaging and ECG Recording for Studying Peripheral Nerve Stimulation
Xia Li 1, Yun Liu 1, Kun Liu 1, Longhua Du 1, Tao Lv 1, Bing Zhu 1, Xinyan Gao 1
1Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences

This study presents a surgical manipulation to expose the T-DRG in anesthetized mice for in vivo calcium imaging, along with synchronous ECG recordings. This method represents a cutting-edge tool for studying peripheral electric nerve stimulation and thoracic visceral organ inputs, as well as their interactions at the primary sensory level.

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Medicine

Primary Sjogren's Syndrome Associated with Lung Adenocarcinoma: Probing the Potential Common Pathogenic Mechanisms and Experimental Verification
Ying Li 1, Xue-lei Chu 2, Peng Xue 2, Peng Wang 3, Ting-ting Zhou 4, Shi-jie Zhu 2
1Graduate School, Beijing University of Chinese Medicine, 2Oncology Department, Wangjing Hospital, China Academy of Chinese Medical Sciences, 3The Second Department of Healthcare, China-Japan Friendship Hospital, 4Rheumatology Department, Shangqiu Hospital of Traditional Chinese Medicine

This manuscript describes the operational procedures and precautions to probe the potential common pathogenic mechanisms linking primary Sjogren's syndrome and lung adenocarcinoma through bioinformatics analysis and experimental verification.

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Biochemistry

3D Visualization of Retinal Vascular Pericytes in Mice by Immunostaining
Shuang Zhang *1, Yuqing Wang *2, Zheng Wang 1, Yihan Liu 2, Jingjing Cui 2, Yuxin Su 2, Jingyan Han 1, Jia Wang 2, Wanzhu Bai 2
1Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, 2Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences

The pericytes in retinal vasculature were examined by immunofluorescent staining with platelet-derived growth factor receptor β after retro-orbital injection of fluorescent tomato lectin. The labeled retina was further treated with the tissue-clearing method and whole mounted for visualizing the three-dimensional views of pericytes surrounding retinal vasculature under a confocal microscope.

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