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26 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Rapid Acquisition of 3D Images Using High-resolution Episcopic Microscopy
Haochuan Zhang *1,2,3, JunGang Huang *2,3,4, Xin Liu 2,3, Ping Zhu 4, Zhongrong Li 1, Xue Li 2,3
1Department of Pediatric Surgery, The Second Affiliated Hospital & Yuying Children's Hospital, Wenzhou Medical University, 2Departments of Urology, Boston Children's Hospital, 3Department of Surgery, Harvard Medical School, 4Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences

We describe a detailed protocol using high-resolution episcopic microscopy to acquire three-dimensional (3D) images of mouse embryos. This improved protocol utilizes a modified tissue preparation method to enhance penetration of the fluorescent dye, thereby permitting morphometric analysis of both small and large-sized specimens.

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Neuroscience

Assessment of Neuronal Viability Using Fluorescein Diacetate-Propidium Iodide Double Staining in Cerebellar Granule Neuron Culture
Lin Jiajia *1, Mak Shinghung *2, Zheng Jiacheng 1, Wang Jialing 1, Xu Dilin 1, Hu Shengquan 2, Zhang Zaijun 3, Wang Qinwen 1, Han Yifan 2,4, Cui Wei 1,2
1Ningbo Key Laboratory of Behavioral Neuroscience, Zhejiang Provincial Key Laboratory of Pathophysiology, School of Medicine, Ningbo University, 2Department of Applied Biology and Chemistry Technology, Institute of Modern Chinese Medicine, The Hong Kong Polytechnic University, 3Institute of New Drug Research, Guangdong Provincial Key Laboratory of Pharmacodynamic, Constituents of Traditional Chinese Medicine & New Drug Research, College of Pharmacy, Jinan University, 4International Joint Laboratory (SYSU-PolyU HK) of Novel Anti-Dementia Drugs of Guangdong

This protocol describes how to accurately measure neuronal viability using Fluorescein diacetate (FDA) and Propidium Iodide (PI) double staining in cultured cerebellar granule neurons, a primary neuronal culture used as an in vitro model in neuroscience and neuropharmacology research.

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JoVE Journal

Screening Bioactive Nanoparticles in Phagocytic Immune Cells for Inhibitors of Toll-like Receptor Signaling
Hong Yang 1, Shan Yu Fung 2, Aihua Bao 1, Qiang Li 1, Stuart E. Turvey 2
1Department of Respiratory Medicine, Shanghai First People's Hospital, Shanghai Jiaotong University School of Medicine, 2Department of Pediatrics, BC Children's Hospital and University of British Columbia

Toll-like receptor (TLR) signaling plays an important role in the pathophysiology of many human inflammatory diseases, and regulating TLR responses by bioactive nanoparticles is anticipated to be beneficial in many inflammatory conditions. THP-1 cell-based reporter cells provide a versatile and robust screening platform for identifying novel inhibitors of TLR signaling.

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Environment

Composition and Properties of Aquafaba: Water Recovered from Commercially Canned Chickpeas
Youn Young Shim 1,2,3, Rana Mustafa 1, Jianheng Shen 1, Kornsulee Ratanapariyanuch 1, Martin J. T. Reaney 1,2,3
1Department of Plant Sciences, University of Saskatchewan, 2Prairie Tide Chemicals Inc., 3Guangdong Saskatchewan Oilseed Joint Laboratory (GUSTO), Department of Food Science and Engineering, Jinan University

Aquafaba is a viscous juice from canned chickpea that, when stirred vigorously, produces a relatively stable white froth or foam. The primary research goal is to identify the components of aquafaba that contribute viscosifying/thickening properties using nuclear magnetic resonance (NMR), ultrafiltration, electrophoresis, and peptide mass fingerprinting.

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Chemistry

Real-time Breath Analysis by Using Secondary Nanoelectrospray Ionization Coupled to High Resolution Mass Spectrometry
Xue Li *1,2, Dan D. Huang *3, Rui Du 1,2, Zhi J. Zhang 1, Chak K. Chan 3, Zheng X. Huang 1,2, Zhen Zhou 1,2
1Institute of Mass Spectrometer and Atmospheric Environment, Jinan University, 2Guangdong Provincial Engineering Research Center for On-line Source Apportionment System of Air Pollution, 3School of Energy and Environment, City University of Hong Kong

A protocol for characterizing chemical composition of exhaled breath in real time by using secondary nanoelectrospray ionization coupled to high resolution mass spectrometry is demonstrated.

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Medicine

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction
Lifei Yang *1,2, Jianwen Lu *1,2,3, Yue Wang 1,2,3, Mei Zhang 1,2,3, Yuan Shi 4, Shasha Wei 1,2, Peng Liu 1,2,3, Zheng Wu 3, Yi Lv 1,2,3, Rongqian Wu 1,2
1Shaanxi Provincial Center for Regenerative Medicine and Surgical Engineering, First Affiliated Hospital, Xi'an Jiaotong University, 2Institute of Advanced Surgical Technology and Engineering, First Affiliated Hospital, Xi'an Jiaotong University, 3Department of Hepatobiliary Surgery, First Affiliated Hospital, Xi'an Jiaotong University, 4Organ Transplantation Center, Tianjin First Center Hospital

The reconstruction of the suprahepatic vena cava (SHVC) remains a difficult step in rat orthotopic liver transplantation. In this article, we show a step-by-step protocol for SHVC reconstruction in rats using a novel magnetic anastomosis technique.

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JoVE Core

Evaluating the Anti-depression Effect of Xiaoyaosan on Chronically-stressed Mice
Zhi-Yi Yan *1, Xiao-Juan Li *1,2, Xiu-Fang Ding 1, Yue-Yun Liu 1, Jia-xu Chen 1,2
1School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, 2Formula-pattern Research Center, School of Traditional Chinese Medicine, Jinan University

Here, we present a protocol to establish the mouse depressive model, observe the behavior changes associated with chronic unpredictable mild stress (CUMS), and evaluate the anti-depression effect of Xiaoyaosan.

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Bioengineering

Construction of a Multilayered Mesenchymal Stem Cell Sheet with a 3D Dynamic Culture System
Yingwei Wang *1,2, Cheng Lu *1,2, Chengzhi He 3, Baoxin Chen 1,2, Youling Zheng 1,2, Junming Zheng 1,2, Jianhua Zhang 4, Zheng Wu 1,2
1Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, 2Department of Developmental and Regenerative Biology, Jinan University, 3Nansha College Preparatory Academy, 4Department of Cardiology, First Affiliated Hospital of Jinan University

This article provides an efficient and feasible method for constructing multilayered stem cell sheets with favorable stem cell property.

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Immunology and Infection

Renal Subcapsular Transplantation of 2'-Deoxyguanosine-Treated Murine Embryonic Thymus in Nude Mice
Jianxue Wang 1,3,4, Guolin Chen 1,3,4, Qianwen Cui 2,3,4, Erfei Song 1,3,4, Weihua Tao 1, Wanqun Chen 2, Cunchuan Wang 1,3,4, Shiqi Jia 1,3,4
1The First Affiliated Hospital, Jinan University, 2Department of Biochemistry and Molecular Biology, School of Basic Medical Science, Jinan University, 3The Institute of Clinical Medicine, Jinan University, 4Joint institute of Metabolic Medicine Between Jinan University and University of Hong Kong

We provide a simple and efficient method to transplant 2’-deoxyguanosine treated E18.5 thymus into the renal capsule of a nude mouse. This method should aide in the study of both thymic epithelial cells function and T cells maturation.

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Medicine

The Lower Body Positive Pressure Treadmill for Knee Osteoarthritis Rehabilitation
Junjie Liang *1,2, Yuanyuan Guo *1,2, Yuxin Zheng 1,2, Shijuan Lang 1, Hongxin Chen 1,2, Yaoyao You 1,2, Bryan O’Young 3,4, Haining Ou *1,2, Qiang Lin *1,2
1Department of Rehabilitation, The Fifth Affiliated Hospital of Guangzhou Medical University, 2Experiment Education Model Center of Rehabilitation Medicine, Guangzhou Medical University, 3Department of Physical Medicine and Rehabilitation, Geisinger Health System, 4Department of Rehabilitation Medicine, New York University School of Medicine

Here, based on a clinician’s point-of-view, we propose a two-model lower body positive pressure (LBPP) protocol (walking and squatting models) in addition to a clinical, functional assessment methodology, including details for further encouragement of the development of non-drug surgical intervention strategies in knee osteoarthritis patients. However, we only present the effect of LBPP training in improvement of pain and knee function in one patient through three-dimensional gait analysis. The exact, long-term effects of this approach should be explored in future studies.

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Genetics

A Robust Polymerase Chain Reaction-based Assay for Quantifying Cytosine-guanine-guanine Trinucleotide Repeats in Fragile X Mental Retardation-1 Gene
Huilin Wang *1, Xiaofan Zhu *2,3, Baoheng Gui *3,4, Wan Chee Cheung 2, Mengmeng Shi 2,3, Zhenjun Yang 2,3, Ka Yin Kwok 2, Ricky Lim 5, Sanna Pietilä 5, Yuanfang Zhu 6,7, Kwong Wai Choy 2,3
1Central Laboratory, Bao'an Maternity and Child Health Hospital, Jinan University, 2Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, 3Shenzhen Research Institute, The Chinese University of Hong Kong, 4Department of Genetics and Metabolism, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, 5PerkinElmer Diagnostics, 6Department of Obstetrics and Gynecology, Bao'an Maternity and Child Health Hospital, Jinan University, 7Maternal-Fetal Medicine Institute, Bao'an Maternity and Child Health Hospital, Jinan University

An accurate and robust polymerase chain reaction-based assay for quantifying cytosine-guanine-guanine trinucleotide repeats in the Fragile X mental retardation-1 gene facilitates molecular diagnosis and screening of Fragile X syndrome and Fragile X-related disorders with shorter turn-around time and investment in equipment.

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Behavior

Loneliness Assuaged: Eye-Tracking an Audience Watching Barrage Videos
Guangyao Chen 1, Shuhua Zhou 2
1School of Journalism & Communication/National Media & Experimental Teaching Center, Jinan University, 2School of Journalism, University of Missouri

The study proposes an activation-match model to study how loneliness is mitigated when a lonely audience watches barrage videos of rational and emotional appeals. The protocol uses eye tracking to document duration and fixation, accounting for the degree of satisfaction when emotional needs are appeased by content and barrage.

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Biochemistry

An Integrated Workflow of Identification and Quantification on FDR Control-Based Untargeted Metabolome
Dehua Li *1, Junze Liang *1, Yongjian Zhang 2, Gong Zhang 1,2
1Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes and MOE Key Laboratory of Tumor Molecular Biology, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University, 2Chi-Biotech Co. Ltd.

We constructed an untargeted metabolomic workflow that integrated XY-Meta and metaX together. In this protocol, we displayed how to use XY-Meta to generate a decoy spectral library from open access spectra reference, and then performed FDR control and used the metaX to quantitate the metabolites after identifying the metabolomics spectra.

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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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Cancer Research

Ex Vivo Organoid Model of Adenovirus-Cre Mediated Gene Deletions in Mouse Urothelial Cells
Dongbo Xu 1, Li Wang 1, Kyle Wieczorek 1, Yanqing Wang 2, Xiaojing Zhang 2, David W. Goodrich 2, Qiang Li 1,2
1Department of Urology, Roswell Park Comprehensive Cancer Center, 2Department of Pharmacology and Therapeutics, Roswell Park Comprehensive Cancer Center

This protocol describes the process of the generation and characterization of mouse urothelial organoids harboring deletions in genes of interest. The methods include harvesting mouse urothelial cells, ex vivo transduction with adenovirus driving Cre expression with a CMV promoter, and in vitro as well as in vivo characterization.

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Medicine

Establishment and Evaluation of a Porcine Vein Graft Disease Model
Xiaohui Li *1, Jinlin Hu *1, Weijiang Tan *2, Zhaoming Lin *1, Caiyi Zhu 2, Chengfeng Huang 1, Jiawen Huang 3, Yintian Liu 1, Qiuying Liao 1, Hua Lu 1, Xiaoshen Zhang 1
1Department of Cardiovascular Surgery, the First Affiliated Hospital of Jinan University, 2Guangdong Provincial Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, 3The First Affiliated Hospital of Jinan University

In this protocol, novel pig vein bypass grafting was performed through a small incision in the left chest wall without cardiopulmonary bypass. A postoperative pathology study was done, which showed intimal thickening.

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Genetics

Investigating the Pathogenesis of MYH7 Mutation Gly823Glu in Familial Hypertrophic Cardiomyopathy using a Mouse Model
Yu Xia *1,2, Jinlin Hu *3, Xiang Li 4, Shuang Zheng 4, Ge Wang 1,2, Songtao Tan 1,2, Zengxiao Zou 1,2, Qiong Ling 2,5, Fenghua Yang 4, Xiaoping Fan 1,2
1Department of Cardiovascular Surgery, Guangdong Provincial Hospital of Traditional Chinese Medicine, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine, 2The Second Clinical College of Guangzhou University of Chinese Medicine, 3Department of Cardiovascular Surgery, The First Affiliated Hospital, Jinan University, 4Guangdong Provincial Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals Monitoring Institute, 5Department of Anesthesiology, Guangdong Provincial Hospital of Chinese Medicine, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine

Based on the familial hereditary cardiomyopathy family found in our clinical work, we created a C57BL/6N mouse model with a point mutation (G823E) at the mouse MYH7 locus through CRISPR/Cas9-mediated genome engineering to verify this mutation.

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Neuroscience

Dual-Task Stroop Paradigm for Detecting Cognitive Deficits in High-Functioning Stroke Patients
Songjun Lin 1,2, Qiang Lin 3,4,5, Biyi Zhao 3,4,6, Yongchun Jiang 3,4, Wanqi Zhuang 3,4, Delong Chen 3,4, Yajie Zhang 3,4, Aijia Chen 3,4, Qianrong Zhang 7, Yuxin Zheng 3,4, Jianjun Wang 1, Fangqiu Xu 8, Xi Qin 3,4,9, Yefeng Cai 10
1Department of Neurology and Psychology, the Fourth Clinical Medical College, Guangzhou University of Chinese Medicine, Shenzhen Traditional Chinese Medicine Hospital, 2Guangzhou University of Chinese Medicine, 3Department of Rehabilitation Medicine, The Fifth Affiliated Hospital of Guangzhou Medical University, 4Department of Rehabilitation Medicine, Guangzhou Medical University, 5Key Laboratory of Biological Targeting Diagnosis, Therapy and Rehabilitation of Guangdong Higher Education Institutes, The Fifth Affiliated Hospital of Guangzhou Medical University, 6School of Traditional Chinese Medicine, Jinan University, 7Health College of Guangdong Pharmaceutical University, 8Department of Clinical Medicine, Guangzhou Medical University, 9Department of Rehabilitation Medicine, Panzhihua Central Hospital, 10Department of Neurology and Psychology, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine

Clinical assessment scales are notsensitive enough to cognitive dysfunction in high-functioning stroke patients. The dual-task paradigm presents advantages and potential in the assessment and cognitive training of cognitive dysfunction. The study here proposes a dual-task Stroop paradigm to identify cognitive dysfunction in high-functioning stroke patients.

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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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Medicine

An Apical Resection Model in the Adult Xenopus tropicalis Heart
Si-Yi He *1, Yi-Min Zhou *1, Na Wen 1, Ke Meng 1, Dong-Qing Cai 1, Xu-Feng Qi 1
1Key Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, Jinan University

Xenopus tropicalis is an ideal model for regenerative research as many of its organs possess a remarkable regenerative capacity. Here, we present a method for constructing a heart injury model in X. tropicalis via apex resection.

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Medicine

Laparoscopic Common Bile Duct Exploration in Patients with a Previous History of Biliary Tract Surgery
Junjie Liang *1, Jiexing Li *1, Yuchuan Jiang 1, Qiang Li 1, Zhilong Liu 1, Leyang Xiang 1, Youzhu Hu 1,2, Mingrong Cao 1
1Department of Hepatobiliary Surgery, The First Affiliated Hospital, Jinan University, 2Department of General Surgery, The Affiliated Shunde Hospital, Jinan University

Programmed surgery based on surgical methods and anatomical markers helps shorten the operation time, reduce complications, and improve the safety of the surgery. This study investigated and summarized the surgical methods and anatomic markers of laparoscopic common bile duct exploration in patients with a history of biliary tract surgery.

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Neuroscience

Motor Imagery Brain-Computer Interface in Rehabilitation of Upper Limb Motor Dysfunction After Stroke
Yongchun Jiang *1,2,3, Junxiao Yin *4, Biyi Zhao 1,3,5, Yajie Zhang 1,3, Tingting Peng 1,3, Wanqi Zhuang 1,3, Siqing Wang 1,3, Siqi Huang 1,3, Meilian Zhong 1,3, Yanni Zhang 1,3, Guibing Tang 1,3, Bingchi Shen 6, Haining Ou 1,3, Yuxin Zheng *2,3, Qiang Lin *2,3
1Guangzhou Medical University, 2Department of Rehabilitation Medicine, The Seventh Affiliated Hospital of Sun Yat-sen University, 3Department of Rehabilitation Medicine, The Fifth Affiliated Hospital of Guangzhou Medical University, 4Clinical Medical College of Acupuncture and Rehabilitation, Guangzhou University of Traditional Chinese Medicine, 5School of Traditional Chinese Medicine, Jinan University, 6Department of Stomatology, Second Clinical Medical College, Dongguan Campus of Guangdong Medical University

The purpose of this study is to provide an important reference for the standard clinical operation of motor imagery brain-computer interface (MI-BCI) for upper limb motor dysfunction after stroke.

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Neuroscience

Automated Impactor for Contusive Spinal Cord Injury Model in Mice
Maocong Wu *1, Jianxian Luo *1, Yujing Gao *1, Chen Peng 1, Tianjun Chen 1, Guowei Zhang 1, Hua Yang 1, Hongsheng Lin 1, Zhisheng Ji 1
1Department of Orthopedics, The First Affiliated Hospital, Jinan University

Presented here is a novel automated spinal cord injury contusion device for mice, which can accurately produce spinal cord injury contusion models with varying degrees.

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Medicine

A Rat Dry Eye Model with Lacrimal Gland Dysfunction Induced by Scopolamine
Shixu Li 1,2, Yu Xiao 3, Yan Tang 1,2, Yongxin Zhang 1,2, Yingge Ma 1,2, Ling Wang *1,2, Lin Ye *1,2
1Shenzhen Eye Hospital, Jinan University, 2Shenzhen Eye Institute, 3Shenzhen Research Institute, Wuhan University

Here, we establish a rat model of lacrimal gland dysfunction to provide a basis for the study of aqueous-deficient dry eye.

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Medicine

Y-90 Radioembolization and PD-1 Inhibitor as Neoadjuvant Treatment in Hepatocellular Carcinoma
Qiang Li *1, Xiaolan Wang *1,2, Deju Li *1, Yulong Liu *3, Kangshou Liu 1, Junjie Liang 1, Jian Sun 1, Quan Jiang 1, Jiexing Li 1, Zhilong Liu 1, Jian Gong 4, Leyang Xiang 1, Zhidong Jia 1, Zhiwei Chen 1, Mingrong Cao 1, Yuchuan Jiang 1,5
1Department of Hepatobiliary Surgery, The First Affiliated Hospital, Jinan University, 2Dongpu Branch, The First Affiliated Hospital, Jinan University, 3Department of Vascular Intervention, The First Affiliated Hospital, Jinan University, 4Department of Nuclear Medicine, The First Affiliated Hospital, Jinan University, 5Department of Gastroenterology, The Second Affiliated Hospital of Nanchang University

This study illustrates the methodological potential of combining Yttrium-90 Trans-Arterial Radioembolization (Y-90 TARE) with an anti-PD-1 monoclonal antibody as an effective neoadjuvant strategy leading to hepatectomy in hepatocellular carcinoma (HCC) patients with a high initial recurrence risk. It emphasizes the safety, feasibility, and step-by-step procedural guidance of this approach.

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Neuroscience

Scalp Acupuncture Synchronizing Dual Task Gait for Enhancing Prefrontal Cortex Response
Jun Su 1, Yuanyuan Guo 2, Biyi Zhao 3, Yongchun Jiang 2,4,5, Xi Qin 6, Meilian Zhong 2,4,5, Dathan Nevin Dychingco Leung 7, Junxiao Yin 8, Qiang Lin *4, Ke Chen *9
1Department of Orthopaedics, Anshun Hospital of Traditional Chinese Medicine, 2The Fifth Affiliated Hospital of Guangzhou Medical University, 3School of Traditional Chinese Medicine, Jinan University, 4Department of Rehabilitation Medicine, The Seventh Affiliated Hospital of Sun Yat-sen University, 5Guangzhou Medical University, 6Panzhihua Central Hospital, 7Department of Physical Therapy, College of Rehabilitation Sciences, University of Santo Tomas, 8Clinical Medical College of Acupuncture and Rehabilitation, Guangzhou University of Traditional Chinese Medicine, 9Department of Spine Surgery, Zhongshan Torch Development Zone People's Hospital

This protocol demonstrates the standardized procedure of scalp acupuncture synchronizing motor-cognitive dual task and motor-cognitive dual task. This can provide an important reference for the clinical exploration of the new and effective non-drug treatment of integrated Chinese and Western medicine.

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