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Army Medical University

23 ARTICLES PUBLISHED IN JoVE

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Neuroscience

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees (Apis mellifera)
Ying Wang 1, Nicholas Baker 1, Gro V. Amdam 1,2
1School of Life Sciences, Arizona State University , 2Department of Chemistry, Biotechnology, and Food Science, Norwegian University of Life Sciences

In this protocol, we describe two strategies that simultaneously suppress two genes (double gene knockdown) in honey bees. Then we present how to use the proboscis extension response (PER) assay to study the effect of double gene knockdown on honey bee gustatory perception.

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Biology

Analysis of Circadian Photoresponses in Drosophila Using Locomotor Activity
Yong Zhang 1, Xueyan Pang 1, Zhenxing Liu 1
1Department of Biology, University of Nevada Reno

Drosophila locomotor activity is a robust and quantitative measurement of circadian photo-responses. We describe protocols for designing behavior experiments for circadian photo-responses and analyzing the data. Studying the circadian photo-responses is important for dissecting the neuronal and molecular mechanisms of light entrainment.

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Immunology and Infection

In Vivo Investigation of Antimicrobial Blue Light Therapy for Multidrug-resistant Acinetobacter baumannii Burn Infections Using Bioluminescence Imaging
Yucheng Wang 1,2,3, Olivia D. Harrington 1, Ying Wang 1, Clinton K. Murray 4, Michael R. Hamblin 1, Tianhong Dai 1
1Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School, 2Department of Medical Oncology, Beijing Institute of Translational Medicine, Chinese Academy of Sciences, 3Cancer Center, Aviation General Hospital, Beijing, 4Infectious Disease Service, Brooke Army Medical Center

Infections caused by multidrug-resistant (MDR) bacterial strains have emerged as a serious threat to public health, necessitating the development of alternative therapeutics. We present a protocol to evaluate the effectiveness of antimicrobial blue light (aBL) therapy for MDR Acinetobacter baumannii infections in mouse burns by using bioluminescence imaging.

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Cancer Research

A Comprehensive Procedure to Evaluate the In Vitro Performance of the Putative Hemangioblastoma Neovascularization Using the Spheroid Sprouting Assay
Ying Wang 1, DanQi Chen 1, MingYu Chen 1, KaiYuan Ji 1, DeXuan Ma 1, LiangFu Zhou 1
1Department of Neurosurgery, Huashan Hospital, Fudan University

This paper presents a comprehensive procedure to evaluate in vitro whether classic tumor angiogenesis exists in hemangioblastomas (HBs) and its role in HBs. The results highlight the complexity of HB-neovascularization and suggest that this common form of angiogenesis is only a complementary mechanism in the HB-neovascularization.

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Environment

Isolation and Analysis of Microbial Communities in Soil, Rhizosphere, and Roots in Perennial Grass Experiments
Morgan R. McPherson 1, Peng Wang 1, Ellen L. Marsh 1, Robert B. Mitchell 2, Daniel P. Schachtman 1
1Department of Agronomy and Horticulture and Center for Plant Science Innovation, University of Nebraska-Lincoln, 2USDA-ARS, Wheat, Sorghum and Forage Research Unit, University of Nebraska-Lincoln East Campus

Excavation of plant roots from the field as well as processing of samples into endosphere, rhizosphere, and soil are described in detail, including DNA extraction and data analysis methods. This paper is designed to enable other laboratories to use these techniques for the study of soil, endosphere, and rhizosphere microbiomes.

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Engineering

Radio Frequency Magnetron Sputtering of GdBa2Cu3O7δ/ La0.67Sr0.33MnO3 Quasi-bilayer Films on SrTiO3 (STO) Single-crystal Substrates
Ying Wang 1,2, Zhen Li 1,2, YongSheng Liu 1, Yijie Li 2, Linfei Liu 2, Da Xu 2, Xiaojing Luo 1, Tian Gao 1,3, Yanyan Zhu 1,4, Luozeng Zhou 3, Jianming Xu 3
1Department of Physics, Mathematics, Shanghai Key Laboratory of Materials Protection and Advanced Materials in Electric Power, Shanghai University of Electric Power, 2Key Laboratory of Artificial Structure and Quantum Control, Ministry of Education, Department of Physics, Shanghai Jiao Tong University, 3Shanghai Institute of Space Power-sources, 4Shanghai Key Laboratory of High Temperature Superconductors, Shanghai University

Here, we present a protocol to grow LSMO nanoparticles and (Gd) BCO films on (001) SrTiO3 (STO) single-crystal substrates by radio frequency (RF)-sputtering.

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Immunology and Infection

Human Serum Anti-aquaporin-4 Immunoglobulin G Detection by Cell-based Assay
Caiyun Liu 1, Mingqin Zhu 1, Ying Wang 1
1Department of Neurology and Neuroscience Center, The First Hospital of Jilin University

Cell-based assay is a widely used method to detect serum anti-aquaporin-4 immunoglobulin G. This method could be applied to clinical diagnosis and scientific researches of neuromyelitis optical spectrum disorders.

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Immunology and Infection

Design of Cecal Ligation and Puncture and Intranasal Infection Dual Model of Sepsis-Induced Immunosuppression
Zhihan Wang *1,2, Qinqin Pu *1,2, Ping Lin *2,3, Changlong Li 1, Jianxin Jiang 3, Min Wu 2
1West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, 2Department of Biomedical Sciences, School of Medicine and Health Sciences, University of North Dakota, 3State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Surgery Research, Daping Hospital, Army Medical University

This protocol describes techniques to measure infectious outcomes underlying secondary hospital-acquired infections in the immunosuppressive condition, first by establishing cecal ligation/puncture mice then challenging them with intranasal infection to create a clinically relevant model of immunosuppression sepsis.

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Genetics

A Bioinformatics Pipeline to Accurately and Efficiently Analyze the MicroRNA Transcriptomes in Plants
Ying Wang *1,2, Zheng Kuang *1,2, Lei Li 2, Xiaozeng Yang 1
1Beijing Key Laboratory of Agricultural Genetic Resources and Biotechnology, Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, 2State Key Laboratory of Protein and Plant Gene Research, Peking-Tsinghua Center for Life Sciences, School of Advanced Agricultural Sciences and School of Life Sciences, Peking University

A bioinformatics pipeline, namely miRDeep-P2 (miRDP2 for short), with updated plant miRNA criteria and an overhauled algorithm, could accurately and efficiently analyze microRNA transcriptomes in plants, especially for species with complex and large genomes.

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Immunology and Infection

Lentiviral CRISPR/Cas9-Mediated Genome Editing for the Study of Hematopoietic Cells in Disease Models
Soichi Sano 1, Ying Wang 1, Megan A. Evans 1, Yoshimitsu Yura 1, Miho Sano 1, Hayato Ogawa 1, Keita Horitani 1, Heather Doviak 1, Kenneth Walsh 1
1Hematovascular Biology Center, Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine

Described are protocols for the highly efficient genome editing of murine hematopoietic stem and progenitor cells (HSPC) by the CRISPR/Cas9 system to rapidly develop mouse model systems with hematopoietic system-specific gene modifications.

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JoVE Core

Optimized Sealing Process and Real-Time Monitoring of Glass-to-Metal Seal Structures
Zhichun Fan 1, Kangjia Hu 2, Zhiyong Huang 1, Yong Zhang 2, He Yan 1
1Institute of Nuclear and New Energy Technology, Key Laboratory of Advanced Reactor Engineering and Safety of Ministry of Education, Collaborative Innovation Center for Advanced Nuclear Energy Technology, Tsinghua University, 2Institute of Nuclear and New Energy Technology, Beijing Key Laboratory of Fine Ceramics, Tsinghua University

Key procedures to optimize the sealing process and achieve real-time monitoring of the metal-to-glass seal (MTGS) structure are described in detail. The embedded fiber Bragg grating (FBG) sensor is designed to achieve online monitoring of temperature and high-level residual stress in the MTGS with simultaneous environmental pressure monitoring.

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Biology

Detecting Establishment of Shared Blood Supply in Parabiotic Mice by Caudal Vein Glucose Injection
Xin Liu 1, Xue Bai 1, Mingqi Li 1, Huimin Li 1, Yong Zhang 1,2, Baofeng Yang 1,3
1Department of Pharmacology, The State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Research, Ministry of Education, College of Pharmacy, Harbin Medical University, 2Institute of Metabolic Disease, Heilongjiang Academy of Medical Science, 3Department of Pharmacology and Therapeutics, Melbourne School of Biomedical Sciences, Faculty of Medicine, Dentistry and Health Sciences University of Melbourne

Here we describe a new method of detecting successful establishment of shared blood circulation of two parabionts through a caudal vein injection of glucose, which causes minimal damage and is not fatal to the parabionts.

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JoVE Core

Bone Marrow Transplantation Procedures in Mice to Study Clonal Hematopoiesis
Eunbee Park 1, Megan A. Evans 4, Heather Doviak 4, Keita Horitani 4, Hayato Ogawa 4, Yoshimitsu Yura 4, Ying Wang 2, Soichi Sano 3,4, Kenneth Walsh 1,4
1Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, 2Department of Cardiology, Xinqiao Hospital, Army Medical University, 3Department of Cardiology, Osaka City University Graduate School of Medicine, 4Hematovascular Biology Center, Robert M. Berne Cardiovascular Research Center, University of Virginia School of Medicine

We describe three methods of bone marrow transplantation (BMT): BMT with total-body irradiation, BMT with shielded irradiation, and BMT method with no pre-conditioning (adoptive BMT) for the study of clonal hematopoiesis in mouse models.

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Neuroscience

A Pilot Study on the Repetitive Transcranial Magnetic Stimulation of Aβ and Tau Levels in Rhesus Monkey Cerebrospinal Fluid
Ling-Yi Liao 1,2,3, Ying-Qian Zhang 4, Benson Wui-Man Lau 5, Qiong Wu 6, Zi-Yan Fan 1,3, Qiang Gao 1,3, Zhi-Hui Zhong 6,7
1Department of Rehabilitation Medicine, West China Hospital, Sichuan University, 2Department of Rehabilitation Medicine, Daping Hospital, Army Medical University, 3Key Laboratory of Rehabilitation Medicine in Sichuan Province, West China Hospital, Sichuan University, 4Department of physiology, Southwest Medical University, 5Department of Rehabilitation Sciences, The Hong Kong Polytechnic University, 6Laboratory of Nonhuman Primate Disease Modeling Research, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, 7Sichuan Kangcheng Biotech Co., Inc.

Here, we describe the procedure for a pilot study to explore the effect of repetitive transcranial magnetic stimulation with different frequencies (1 Hz/20 Hz/40 Hz) on Aβ and tau metabolism in rhesus monkey cerebrospinal fluid.

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Medicine

A Rat Orthotopic Renal Transplantation Model for Renal Allograft Rejection
Hang You 1,2, Xin Mao 2, Chenyang Wang 2, Gang Huang 3, Marcus Groettrup 4,5, Jun Li 2
1School of Medicine, Chongqing University, 2Department of Urological Oncology Surgery, Chongqing University Cancer Hospital, 3Department of Biochemistry and Molecular Biology, College of Basic Medical Science, Army Medical University, 4Division of Immunology, Department of Biology, University of Konstanz, 5Biotechnology Institute Thurgau, University of Konstanz

The rat orthotopic renal transplantation model contributes to investigating the mechanism of renal allograft rejection. The current model increases the recipients' survival without interference with blood supply and venous reflux of the lower body using an end-to-end anastomosis of kidney implantation and an end-to-side "tunnel" method of ureter-bladder anastomosis.

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Medicine

A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model
Xin Mao *1, Peng Xian *1, Hang You 1,2, Gang Huang 3, Jun Li 1
1Department of Urological Oncology Surgery, Chongqing University Cancer Hospital, 2School of Medicine, Chongqing University, 3Department of Biochemistry and Molecular Biology, College of Basic Medical Science, Army Medical University

In the present protocol, a mouse heart transplantation model is used for investigating the mechanism of cardiac allograft rejection. In this heterotopic heart transplantation model, operation efficiency is improved, and the survival of cardiac grafts is ensured by a cervical end-to-end anastomosis of heart implantation using a modified Cuff technique.

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Medicine

Microscopic Replantation of Penile Glans Amputation Due to Circumcision
Da-Chun Jin *1, Bo Zhou *1, Jun Li 2, Cong-Cong Bao 1, Yong Luo 1, Yong Zhang 1, Peng Wang 1, Gang Bi 1, Yan-Feng Li 1
1Department of Urology, Daping Hospital, Army Medical University, 2Department of Urology, Chongqing 13th People's Hospital

The present protocol describes the emergency management of microscopic replantation of penile glans amputation due to circumcision.

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Bioengineering

Fabricating Highly Open Porous Microspheres (HOPMs) via Microfluidic Technology
Sheng-Chang Luo 1,2, Ying Wang 3, Ranjith Kumar Kankala 1,2, Yu Shrike Zhang 4, Ai-Zheng Chen 1,2
1Institute of Biomaterials and Tissue Engineering, Huaqiao University, 2Fujian Provincial Key Laboratory of Biochemical Technology, Huaqiao University, 3Affiliated Dongguan Hospital, Southern Medical University, 4Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School

The present protocol describes the fabrication of poly(lactic-co-glycolic acid)-based highly open porous microspheres (HOPMs) via the single-emulsion formulation based facile microfluidic technology. These microspheres have potential applications in tissue engineering and drug screening.

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Neuroscience

Using a Bipolar Electrode to Create a Temporal Lobe Epilepsy Mouse Model by Electrical Kindling of the Amygdala
Yongchang Lu *1,2, Yang Dai *1,2, Siqi Ou 1,2, Yujing Miao 1, Ying Wang 1,2, Quanlei Liu 1,2, Yihe Wang 1,2, Penghu Wei 1,2, Yongzhi Shan 1,2, Guoguang Zhao 1,2,3
1Department of Neurosurgery, Xuanwu Hospital Capital Medical University, 2China Clinical Research Center for Epilepsy Capital Medical University, 3China Beijing Municipal Geriatric Medical Research Center

The amygdala plays a key role in temporal lobe epilepsy, which originates in and propagates from this structure. This article provides a detailed description of the fabrication of deep brain electrodes with both recording and stimulating functions. It introduces a model of medial temporal lobe epilepsy originating from the amygdala.

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Medicine

Indwelling Needle Puncture and Irrigation in the Conservative Treatment of Breast Abscess During Non-Lactation Period
Yanhong Hu *1, Yanping Li *2, Jiaqun Zou 3, Yan Xu 3
1Chongqing Wuyi Polytechnic, 2Chongqing Medical and Pharmaceutical College, 3Department of Breast and Thyroid surgery, Daping Hospital, Army Medical University

Here, we present a protocol detailing a minimally invasive treatment for breast abscesses during the non-lactation period. This method effectively treats single breast abscesses, and the indwelling needle can be used as an effective tool for the puncture and irrigation of single breast abscesses in a non-lactation period.

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Biology

Bulk Electroacupuncture Operation for Mice or Young Rats
Siqi Yao *1, Yun Wei *1, Yuping Zhang 1, Xiaolin Yang 2, Zhifeng Wu 1
1Department of Pediatrics, Second Affiliated Hospital, Army Medical University, 2Department of Neurosurgery, Second Affiliated Hospital, Army Medical University

Here, we present a protocol for efficient and rapid electroacupuncture (EA) in mice or young rats using a three-dimensional (3D) printed holder. This technique allows simultaneous operation on multiple animals, saving time and increasing experimental efficiency.

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Medicine

Microscopic Electric Rotary Grinding of Plaques Combined with Graft Repair in the Management of Peyronie's Disease
Da-Chun Jin 1, Yong Luo 1, Peng Wang 1, Yong Zhang 1, Gang Bi 1, Da-Li Tong 1, Yi-Ye Wang 1, Wan-Yi Zhou 1, Yan-Feng Li 1
1Department of Urology, Army Medical Center, Daping Hospital, Army Medical University

The presented protocol describes a microscopic surgical method for the treatment of Peyronie's disease by using electric rotary grinding of the plaques followed by using the tunica vaginalis or bovine pericardium as a graft for the repairing of the defects of the tunica albuginea.

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Medicine

Mouse Wound Models and Preparation of Single-Cell Suspensions
Yuanyuan Liu 1, Mingwang Zhang 2, Kai Luo 3, Yurui Zou 4, Rongya Yang 5
1Medical School of Chinese People's Liberation Army, 2Department of Dermatology, Southwest Hospital, Army Medical University, 3Biomedical Treatment Center, the Seventh Medical Center, Chinese PLA General Hospital, 4The 83th Affiliated Hospital, Xinxiang Medical University, 5Department of Dermatology, the Seventh Medical Center, Chinese PLA General Hospital

This study presents two crucial experimental techniques: the construction of a murine wound model for assessing wound closure and the preparation of single-cell suspensions from murine skin to preserve cellular viability.

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