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China Academy of Chinese Medical Sciences

18 ARTICLES PUBLISHED IN JoVE

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Medicine

Evaluation of Stem Cell Properties in Human Ovarian Carcinoma Cells Using Multi and Single Cell-based Spheres Assays
Hui Wang 1,2, Anna Paczulla 1, Claudia Lengerke 1,2
1Department of Biomedicine, University Hospital Basel, 2Department of Internal Medicine II, University Hospital Tübingen

In vitro spheres assays are commonly used to identify cancer stem cells. Here we compare single with multi cell-based spheres assays. The more laborious single cell-based assays or methylcellulose supplementation give more accurate results while multi cell-based assays performed in liquid medium can be highly influenced by cell density.

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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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Neuroscience

Studying Brain Function in Children Using Magnetoencephalography
Hannah Rapaport 1,2, Robert A. Seymour 1,2,3, Paul F. Sowman 1,2, Nick Benikos 1,2, Elisabeth Stylianou 1,2, Blake W. Johnson 1,2, Stephen Crain 1,4, Wei He 1,2
1ARC Centre of Excellence in Cognition and its Disorders, Macquarie University, 2Department of Cognitive Science, Macquarie University, 3Aston Brain Centre, School of Life and Health Sciences, Aston University, 4Department of Linguistics, Macquarie University

This article introduces a child-friendly research protocol designed to improve data quality by reducing head movement during pediatric magnetoencephalography (MEG). We familiarize families with the MEG environment, train children to remain still using an MEG simulator, and correct for residual head movement artefacts using a real-time head movement detection system.

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Cancer Research

Three-Dimensional Bone Extracellular Matrix Model for Osteosarcoma
Yu Zhang 1, Yupeng Yao 1, Yan Zhang 1
1MOE Key Laboratory of Gene Function and Regulation, School of Life Sciences, Sun Yat-sen University

The bone extracellular matrix (BEM) model for osteosarcoma (OS) is well established and shown here. It can be used as a suitable scaffold for mimicking primary tumor growth in vitro and providing an ideal model for studying the histologic and cytogenic heterogeneity of OS.

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JoVE Journal

Thermocapillary Convection Space Experiment on the SJ-10 Recoverable Satellite
Li Duan *1,2, Yongli Yin *3, Jia Wang *1, Qi Kang 1,2, Di Wu 1, Huan Jiang 1, Pu Zhang 1, Liang Hu 1
1National Microgravity Laboratory, Institute of Mechanics, Chinese Academy of Sciences, 2School of Engineering Sciences, University of Chinese Academy of Sciences, 3China Astronaut Research and Training Center

A protocol for the space payload design, the space experiment on thermocapillary convection, and analyses of experimental data and images are presented in this paper.

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Behavior

Analysis of Learning and Memory Ability in an Alzheimer's Disease Mouse Model using the Morris Water Maze
Huiling Tian 1, Ning Ding 2, Mengwei Guo 1, Shun Wang 1, Zidong Wang 1, Hao Liu 1, Jiayi Yang 1, Yujie Li 3, Jingyu Ren 1, Jing Jiang 4, Zhigang Li 1
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.

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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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Behavior

Fully Automated Leg Tracking in Freely Moving Insects using Feature Learning Leg Segmentation and Tracking (FLLIT)
Animesh Banerjee 1, Shuang Wu 2, Li Cheng 3, Sherry Shiying Aw 1
1Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, 2Bioinformatics Institute, Agency for Science, Technology and Research, 3Department of Electrical and Computer Engineering, University of Alberta

We describe detailed protocols for using FLLIT, a fully automated machine learning method for leg claw movement tracking in freely moving Drosophila melanogaster and other insects. These protocols can be used to quantitatively measure subtle walking gait movements in wild type flies, mutant flies and fly models of neurodegeneration.

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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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Chemistry

Optimization of Radiochemical Reactions using Droplet Arrays
Alejandra Rios 1,2, Travis S. Holloway 2,3, Jia Wang 2,4, R. Michael van Dam 1,2,3,4
1Physics and Biology in Medicine Interdepartmental Graduate Program, University of California Los Angeles (UCLA), 2Crump Institute of Molecular Imaging, UCLA, 3Department of Molecular & Medical Pharmacology, David Geffen School of Medicine, 4Department of Bioengineering, UCLA

This method describes the use of a novel high-throughput methodology, based on droplet chemical reactions, for the rapid and economical optimization of radiopharmaceuticals using nanomole amounts of reagents.

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Bioengineering

3D Printing of In Vitro Hydrogel Microcarriers by Alternating Viscous-Inertial Force Jetting
Tiankun Liu *1,2, Yongchun Shao *1,2, Zitong Wang 1,2, Yuqiu Chen 1,2, Yuan Pang 1,2, Ding Weng 1,2, Wei Sun 1,2,3
1Biomanufacturing Center, Dept. of Mechanical Engineering, Tsinghua University, 2Biomanufacturing and Rapid Forming Technology Key Laboratory of Beijing, 3Department of Mechanical Engineering, Drexel University

Presented here is a mild 3D printing technique driven by alternating viscous-inertial forces to enable the construction of hydrogel microcarriers. Homemade nozzles offer flexibility, allowing easy replacement for different materials and diameters. Cell binding microcarriers with a diameter of 50-500 µm can be obtained and collected for further culturing.

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Bioengineering

Automated Microbial Cultivation and Adaptive Evolution using Microbial Microdroplet Culture System (MMC)
Xingjin Jian *1,2, Xiaojie Guo *1,2,3, Jia Wang 4, Zheng Lin Tan 1,2,6, Xin-hui Xing 1,2,5, Liyan Wang 3, Chong Zhang 1,2,5
1Department of Chemical Engineering, Institute of Biochemical Engineering, Tsinghua University, 2Key Laboratory of Industrial Biocatalysis, Ministry of Education, Tsinghua University, 3Luoyang TMAXTREE Biotechnology Co., Ltd., 4Biochemical Engineering Research Group, School of Chemical Engineering and Technology, Xi’an Jiaotong University, 5Center for Synthetic & Systems Biology, Tsinghua University, 6School of Life Science and Technology, Tokyo Institute of Technology

This protocol describes how to use the Microbial Microdroplet Culture system (MMC) to conduct automated microbial cultivation and adaptive evolution. MMC can cultivate and sub-cultivate microorganisms automatically and continuously and monitor online their growth with relatively high throughput and good parallelization, reducing labor and reagent consumption.

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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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Medicine

Binocular Dynamic Visual Acuity in Eyeglass-Corrected Myopic Patients
Yuexin Wang *1, Yining Guo *1, Shanshan Wei 2, Yifei Yuan 1, Tingyi Wu 1, Yu Zhang 1, Yueguo Chen 1, Xuemin Li 1
1Department of Ophthalmology, Beijing Key Laboratory of Restoration of Damaged Ocular Nerves, Peking University Third Hospital, 2Beijing Tongren Eye Center, Beijing Tongren Hospital

The present research demonstrates a method to accurately examine dynamic visual acuity (DVA) in myopic subjects with eyeglass correction. Further analysis indicated that the closer the refraction state to emmetropia, the better the eyeglass-corrected binocular DVA is at both 40 and 80 degrees per second.

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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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Neuroscience

Dissecting Cell-Autonomous Function of Fragile X Mental Retardation Protein in an Auditory Circuit by In Ovo Electroporation
Qiwei Fan *1, Xiaotan Zhang *2, Yuan Wang 3, Xiaoyu Wang 1
1Division of Histology & Embryology, Key Laboratory for Regenerative Medicine of the Ministry of Education, College of Medicine, Jinan University, 2Department of Clinical Pathology, First Affiliated Hospital of Jinan University, 3Program in Neuroscience, Department of Biomedical Sciences, College of Medicine, Florida State University

Using in ovo electroporation, we devised a method to selectively transfect the auditory inner ear and cochlear nucleus in chicken embryos to achieve a cell-group-specific knockdown of fragile X mental retardation protein during discrete periods of circuit assembly.

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