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Chinese Academy of Medical Sciences & Peking Union Medical College

20 ARTICLES PUBLISHED IN JoVE

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Chemistry

Towards Biomimicking Wood: Fabricated Free-standing Films of Nanocellulose, Lignin, and a Synthetic Polycation
Karthik Pillai 1,2,3, Fernando Navarro Arzate 2,4, Wei Zhang 2,5, Scott Renneckar 1,2,5,6
1Institute for Critical Technology and Applied Science, Virginia Tech, 2Macromolecules and Interfaces Institute, Virginia Tech, 3Institute for Food Safety and Health, Illinois Institute of Technology- Moffett Campus, 4Wood, Cellulose, and Paper Research Department, University of Guadalajara, 5Department of Sustainable Biomaterials, Virginia Tech, 6Sustainable Nanotechnology Interdisciplinary Graduate Education Program, Virginia Tech

The objective of this research was to form synthetic plant cell wall tissue using layer-by-layer assembly of nanocellulose fibrils and isolated lignin assembled from dilute aqueous suspensions.  Surface measurement techniques of quartz crystal microbalance and atomic force microscopy were used to monitor the formation of the polymer-polymer nanocomposite material.

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Medicine

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery
Wei Zhang 1, Mohamed Trebak 1
1Nanobioscience, State University of New York College of Nanoscale Science and Engineering (SUNY CNSE)

This protocol uses a balloon catheter to cause an intraluminal injury on the rat carotid artery and henceforth elicit neointimal hyperplasia. This is a well-established model for studying the mechanisms of vascular remodeling in response to injury. It is also widely used to determine the validity of potential therapeutic approaches.

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Bioengineering

Methods for Characterizing the Co-development of Biofilm and Habitat Heterogeneity
Xiaobao Li 1, Jisun L. Song 2, Alessandro Culotti 1, Wei Zhang 1, David L. Chopp 3, Nanxi Lu 1, Aaron I. Packman 1
1Department of Civil and Environmental Engineering, Northwestern University, 2Department of Chemical and Biological Engineeering, Northwestern University, 3Department of Applied Mathematics and Engineering Sciences, Northwestern University

Biofilms have complex interactions with their surrounding environment. To comprehensively investigate biofilm-environment interactions, we present here a series of methods to create heterogeneous chemical environment for biofilm development, to quantify local flow velocity, and to analyze mass transport in and around biofilm colonies.

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Biochemistry

Sheathless Capillary Electrophoresis–Mass Spectrometry for Metabolic Profiling of Biological Samples
Wei Zhang 1, M. Can Gulersonmez 1, Thomas Hankemeier 1, Rawi Ramautar 1
1Division of Analytical Biosciences, Leiden Academic Center for Drug Research, Leiden University

A protocol for metabolic profiling of biological samples by capillary electrophoresis–mass spectrometry using a sheathless porous tip interface design is presented.

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Developmental Biology

Dissection of Larval Zebrafish Gonadal Tissue
Xinjian Wang *1, Sijie Chen *1, Wei Zhang 1, Yiyuan Ren 1, Quan Zhang 1, Gang Peng 1
1Institutes of Brain Science, State Key Laboratory of Medical Neurobiology and Collaborative Innovation Center for Brain Science, Fudan University

Here, we present a protocol for isolating gonadal tissue of larval zebrafish, which will facilitate investigations of zebrafish sex differentiation and maintenance.

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Medicine

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo
Mingming Liu 1, Xiaoyan Zhang 1, Bingwei Li 1, Bing Wang 1, Qingbin Wu 1, Fei Shang 1, Ailing Li 1, Hongwei Li 1, Ruijuan Xiu 1
1Institute of Microcirculation, Chinese Academy of Medical Sciences & Peking Union Medical College

Pancreatic islet microvascular vasomotion regulates islet blood distribution and maintains the physiological function of islet β cells. This protocol describes using a laser Doppler monitor to determine the functional status of pancreatic islet microvascular vasomotion in vivo and to assess the contributions of pancreatic islet microcirculation to pancreatic-related diseases.

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Engineering

Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes
Chao Tian 1, Wei Zhang 2,3, Van Phuc Nguyen 1, Xueding Wang 2,4, Yannis M. Paulus 1,2
1Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, 2Department of Biomedical Engineering, University of Michigan, 3Institute of Biomedical Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, 4Department of Radiology, University of Michigan

This manuscript describes the novel setup and operating procedure of a photoacoustic microscopy and optical coherence tomography dual-modality system for noninvasive, label-free chorioretinal imaging of larger animals, such as rabbits.

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Neuroscience

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method
Weiyue Zhang *1, Wei Zhang *1, Ning Dai *1, Chenxia Han 2, Fengzhi Wu 1, Xu Wang 1, Libo Tan 1, Jie Li 1, Feng Li 1, Qingjia Ren 3
1School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, 2Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University, 3Institute of Tibetan Medicine, Tibetan Traditional Medical College

Here, we present a protocol to introduce a rat model of central fatigue using the modified multiple platform method (MMPM).

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Neuroscience

Single Sensillum Recordings for Locust Palp Sensilla Basiconica
Hongwei Li 1, Yinwei You 2, Long Zhang 1
1Department of Entomology, China Agricultural University, 2Bio-tech Research Center, Shandong Academy of Agricultural Sciences

This paper describes a detailed and highly effective protocol for single sensillum recordings from the sensilla basiconica on the palps of insect mouthparts.

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Medicine

Introduction of Intracapsular Rotary-cut Procedures (IRCP): A Modified Hysteromyomectomy Procedures Facilitating Fertility Preservation
Liping Zeng 1,2, Weixia Wei 1,2, Huiru Tang 1,2, Fang He 1,2, Shilin Zhong 1,2, Wei Zhang 1,2, Yuting Xiang 1, Xinfeng Qu 3, Ruifang Wu 1,2
1Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital, China, 2Shenzhen Key Laboratory of Gynecological Diagnostic Technology Research, China, 3Specialist of SZSM Project, Peking University Shenzhen Hospital, China

Here, we present a protocol for performing an intracapsular rotary-cut procedure (IRCP), a modified laparoscopic intracapsular myomectomy that promotes fertility preservation.

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Chemistry

Methionine Functionalized Biocompatible Block Copolymers for Targeted Plasmid DNA Delivery
Yang Wu *1, Wei Zhang *2, Jian Zhang 2, Zhi-Xiang Mao 3, Li Ding 2, Hao Li 3, Rong Ma 1, Jin-Hai Tang 2
1Research Center of Clinical Oncology, Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research & Nanjing Medical University Affiliated Cancer Hospital, 2Department of General Surgery, The First Affiliated Hospital with Nanjing Medical University, 3School of Clinical Medicine, Xuzhou Medical University

This work presents the preparation of methionine functionalized biocompatible block copolymers (mBG) via the reversible addition-fragmentation chain transfer (RAFT) method. The plasmid DNA complexing ability of the obtained mBG and their transfection efficiency were also investigated. The RAFT method is very beneficial for polymerizing monomers containing special functional groups.

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Bioengineering

Protein Kinase C-delta Inhibitor Peptide Formulation using Gold Nanoparticles
Hisato Konoeda 1, Hong Yang 2, Chengliang Yang 1, Annette Gower 1, Chun Xu 1, Wei Zhang 1, Mingyao Liu 1,3
1Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, 2Respiratory Medicine Research Laboratory, Institute of Translation Medicine, Shanghai General Hospital, Shanghai Jiaotong University, 3Institute of Medical Science, Faculty of Medicine, University of Toronto

We have previously used a gold nanoparticle peptide hybrid to intravenously deliver a synthetic peptide, protein kinase C-delta inhibitor, which reduced ischemia-reperfusion-induced acute lung injury. Here we show the detailed protocol of the drug formulation. Other intracellular peptides can be formulated similarly.

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Biochemistry

An In Vivo Estrogen Deficiency Mouse Model for Screening Exogenous Estrogen Treatments of Cardiovascular Dysfunction After Menopause
Bing Sun 1, Yue-zhang Yin 2, Jing Xiao 1
1Institute of Medicinal Plant Development (IMPLAD), Chinese Academy of Medical Sciences & Peking Union Medical College, 2Shandong University of Traditional Chinese Medicine

Clinically, estrogen deficiency in menopausal women may aggravate the incidence of lipid disruption and atherosclerosis. We established an in vivo estrogen deficiency model by bilateral ovariectomy via a double dorsal-lateral incision in apoE-/- mice. The mouse model is applicable for screening exogenous estrogen treatments of cardiovascular dysfunction after menopause.

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Medicine

A Training and Testing System for Performing Vascular Reconstruction In Vitro
Yue Wang 1,2, Lijun Mu 3, Wei Zhang 1,2, Huan Chen 1,2, Qingshan Li 1,2, Aihua Shi 1, Bo Tang 2, Xiaogang Zhang 1,2, Dinghui Dong 1,2, Yi Lv 1,2
1National Local Joint Engineering Research Center for Precision Surgery and Regenerative Medicine, First Affiliated Hospital of Xi'an Jiaotong University, 2Department of Hepatobiliary Surgery, First Affiliated Hospital of Xi'an Jiaotong University, 3Department of General Surgery, First Affiliated Hospital of Xi'an Jiaotong University

Here we present a training and testing system where a trainee can complete manual vascular reconstruction in vitro individually using a magnetic anchoring technique. The system can also be used to test the quality of reconstruction.

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Chemistry

Capillary Electrophoresis Mass Spectrometry Approaches for Characterization of the Protein and Metabolite Corona Acquired by Nanomaterials
Andrew J. Chetwynd *1, Wei Zhang *2, Klaus Faserl *3, James A. Thorn 4, Iseult Lynch 1, Rawi Ramautar 2, Herbert H. Lindner 3
1School of Geography Earth and Environmental Sciences, University of Birmingham, 2Biomedical Microscale Analytics, Leiden University, 3Institute of Medical Biochemistry, Medical University of Innsbruck, 4AB Sciex UK Ltd

Here we present a protocol to characterize the complete biomolecular corona, proteins, and metabolites, acquired by nanomaterials from biofluids using a capillary electrophoresis – mass spectrometry approach.

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Developmental Biology

A Protocol for Immunohistochemistry and RNA In-situ Distribution within Early Drosophila Embryo
Wei Zhang *1, Xinjuan Lei *1, Xin Zhou *2,3, Boling He 1, Liqin Xiao 1, Huimin Yue 1, Shulin Wang 1, Yuting Sun 1, Yajun Wu 1, Liyang Wang 1,4, George Ghartey-Kwansah 1, Odell D. Jones 5, Joseph L. Bryant 6, MengMeng Xu 7, Jianjie Ma 3, Xuehon Xu 1
1National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest of China/CGDB, Shaanxi Normal University College of Life Sciences, 2Shaanxi Key Laboratory of Ischemic Cardiovascular Disease, Shaanxi Key Laboratory of Brain disorders, Institute of Basic & Translational Medicine, Xi'an Medical University, 3Ohio State University College of Medicine, 4Hematology-Oncology Division, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 5University of Pennsylvania ULAR, 6University of Maryland School of Medicine, 7Columbia University Medical Center

Here, we describe a protocol for detection and localization of Drosophila embryo protein and RNA from collection to pre-embedding and embedding, immunostaining, and mRNA in situ hybridization.

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Behavior

Embryo Microinjection and Knockout Mutant Identification of CRISPR/Cas9 Genome-Edited Helicoverpa Armigera (Hübner)
Dong Ai 1,2, Bing Wang 2, Zhennan Fan 1, Yuyao Fu 1, Caihong Yu 1, Guirong Wang 2,3
1School of Chemical and Environmental Engineering, China University of Mining and Technology, 2State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 3Guangdong Laboratory of Lingnan Modern Agriculture, Shenzhen; Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences

Presented here is a protocol of Helicoverpa armigera (Hübner) embryo microinjection and knockout mutant identification created by CRISPR/Cas9 genome editing. Mutant insects enable further research of gene function and interaction among different genes in vivo.

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Biochemistry

Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases
Olivia Roscow 1, Wei Zhang 1
1Department of Molecular and Cellular Biology, College of Biological Science, University of Guelph

Ubiquitination is a critical protein post-translational modification, dysregulation of which has been implicated in numerous human diseases. This protocol details how phage display can be utilized to isolate novel ubiquitin variants that can bind and modulate the activity of E3 ligases that control the specificity, efficiency, and patterns of ubiquitination.

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Biology

Neutrophil Lifespan Extension with CLON-G and an In Vitro Spontaneous Death Assay
Yuping Fan *1,2, Yan Teng *3, Fu tong Liu *4, Fengxia Ma 1,2, Alan Y. Hsu 5, Sizhou Feng 1,2, Hongbo R. Luo 5
1State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, 2Tianjin Institutes of Health Science, 3Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, 4Haihe Laboratory of Cell Ecosystem, Tianjin Medical University, 5Joint Program in Transfusion Medicine, Department of Pathology, Harvard Medical School; Division of Blood Bank, Department of Laboratory Medicine, The Stem Cell Program, Boston Children's Hospital, Dana-Farber /Harvard Cancer Center

This protocol details the preparation of CLON-G to extend the neutrophil lifespan to greater than 5 days and provides a reliable procedure for evaluating neutrophil death with flow cytometry and confocal fluorescence microscopy.

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Bioengineering

Effects of Extracorporeal Membrane Oxygenation on the Coagulation System
Tianlong Wang *1, Weidong Yan *2, Mingru Zhang 3, Jing Wang 1, Han Zhang 1, Gang Liu 1, Jian Wang 1, Bingyang Ji 1
1Department of Cardiopulmonary Bypass, Fuwai Hospital, National Center for Cardiovascular Disease, State Key Laboratory of Cardiovascular Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College, 2Department of Anesthesiology, Beijing Chao-Yang Hospital, Capital Medical University, 3Department of Anesthesiology, Beijing Tongren Hospital, Capital Medical University

We present a protocol for establishing a long-term awake extracorporeal membrane oxygenation (ECMO) model in sheep. Special attention is given to the management and evaluation of the coagulation system during the ECMO model.

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