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Chengdu University of Traditional Chinese Medicine

63 ARTICLES PUBLISHED IN JoVE

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Biology

Drosophila Pupal Abdomen Immunohistochemistry
Wei Wang 1, John H. Yoder 1
1Department of Biological Sciences, University of Alabama

Antibody staining of the Drosophila pupae can enhance genetic analyses of adult abdominal developmental genetics. We present our protocol for dissection, fixation and antibody staining of staged Drosophila pupal abdomen.

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Medicine

Analytical Techniques for Assaying Nitric Oxide Bioactivity
Hong Jiang 1, Deepa Parthasarathy 1, Ashley C. Torregrossa 1, Asad Mian 2, Nathan S. Bryan 1
1Texas Therapeutics Institute, University of Texas Health Science Center at Houston , 2Deptartment of Pediatrics, Baylor College of Medicine

The endogenous production of nitric oxide (NO) regulates a wide variety of biological functions. It is becoming increasingly clear that disruption or dysregulation of NO based signaling is involved in many human diseases. Methods to quantify relevant NO metabolites may provide novel diagnostic or prognostic biomarkers for human disease.

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Medicine

Imaging Glioma Initiation In Vivo Through a Polished and Reinforced Thin-skull Cranial Window
Lifeng Zhang *1, Andree Lapierre *1, Brittany Roy 1, Maili Lim 1, Jennifer Zhu 1, Wei Wang 1, Stephen B. Sampson 1, Kyuson Yun 1, Bonnie Lyons 1, Yun Li 1, Da-Ting Lin 1
1The Jackson Laboratory

By combining a polished and reinforced thin-skull (PoRTS) cranial window and glioblastoma (GBM) cell injection, we can observe glioma initiation and growth from injected GBM cells in the brain of a live mouse longitudinally.

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Neuroscience

Imaging pHluorin-tagged Receptor Insertion to the Plasma Membrane in Primary Cultured Mouse Neurons
Yun Li 1, Brittany D. Roy 1, Wei Wang 1, Lifeng Zhang 1, Stephen B. Sampson 1, Da-Ting Lin 1
1The Jackson Laboratory

By tagging the extracellular domains of membrane receptors with superecliptic pHluorin, and by imaging these fusion receptors in cultured mouse neurons, we can directly visualize individual vesicular insertion events of the receptors to the plasma membrane. This technique will be instrumental in elucidating the molecular mechanisms governing receptor insertion to the plasma membrane.

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Chemistry

Facile and Efficient Preparation of Tri-component Fluorescent Glycopolymers via RAFT-controlled Polymerization
Wei Wang 1, John M. Lester 1, Anthony E. Amorosa 2, Deborah L. Chance 3, Valeri V. Mossine 1, Thomas P. Mawhinney 1,4
1Department of Biochemistry, University of Missouri, 2Department of Biological Sciences, University of Missouri, 3Department of Molecular Microbiology & Immunology, University of Missouri, 4Department of Child Health, University of Missouri

An efficient, three-step synthesis of RAFT-based fluorescent glycopolymers, consisting of glycomonomer preparation, copolymerization, and post-modification, is demonstrated. This protocol can be used to prepare RAFT-based statistical glycopolymers with desired structures.

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Medicine

Working with Human Tissues for Translational Cancer Research
Alexandre Reuben *1, Vancheswaran Gopalakrishnan *1, Heidi E. Wagner 3, Christine N. Spencer 2, Jacob Austin-Breneman 1, Hong Jiang 1, Zachary A. Cooper 1, Jennifer A. Wargo 1
1Department of Surgical Oncology, University of Texas MD Anderson Cancer Center, 2Department of Genomic Medicine, University of Texas MD Anderson Cancer Center, 3Department of Pathology and Institutional Tissue Bank, University of Texas MD Anderson Cancer Center

Translational cancer research is dependent on extraction of human tissues. Much work has gone into optimizing extraction methods for ex vivo analysis. Here, we describe tissue processing methods allowing for maximal data output from limited samples.

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Chemistry

Laboratory Production of Biofuels and Biochemicals from a Rapeseed Oil through Catalytic Cracking Conversion
Siauw H. Ng 1, Yu Shi 1, Nicole E. Heshka 1, Yi Zhang 1, Edward Little 1
1CanmetENERGY, Natural Resources Canada

This paper presents an experimental method to produce biofuels and biochemicals from canola oil mixed with a fossil-based feed in the presence of a catalyst at mild temperatures. Gaseous, liquid, and solid products from a reaction unit are quantified and characterized. Conversion and individual product yields are calculated and reported.

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Genetics

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format
Muhammad Khairul Ramlee 1, Jing Wang 1, Alice M. S. Cheung 1, Shang Li 1
1Cancer & Stem Cell Biology Programme, Duke-NUS Medical School

The genotyping technique described here, which couples fluorescent polymerase chain reaction (PCR) to capillary gel electrophoresis, allows for high-throughput genotyping of nuclease-mediated knockout clones. It circumvents limitations faced by other genotyping techniques and is more cost effective than sequencing methods.

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Neuroscience

Assessment of Hippocampal Dendritic Complexity in Aged Mice Using the Golgi-Cox Method
Thomas R. Groves 1,2,3, Jing Wang 1,2, Marjan Boerma 1,2, Antiño R. Allen 1,2,3
1Division of Radiation Health, University of Arkansas for Medical Sciences, 2Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences, 3Neurobiology & Developmental Sciences, University of Arkansas for Medical Sciences

Here we present a Golgi-Cox protocol in extensive detail. This reliable tissue stain method allows for a high-quality assessment of the cytoarchitecture in the hippocampus, and throughout the entire brain, with minimal troubleshooting.

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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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Behavior

Use of a Video Scoring Anchor for Rapid Serial Assessment of Social Communication in Toddlers
Natasha Marrus 1, Stefanie Kennon-McGill 1, Brooke Harris 2, Yi Zhang 1, Anne L. Glowinski 1, John N. Constantino 1
1Department of Psychiatry, Washington University School of Medicine, 2Veteran's Administration Palo Alto Health Care System

Here we present a protocol for administration of the video-referenced rating of reciprocal social behavior (vrRSB) using a paper or online version. This survey quantifies RSB, a prerequisite for social competence, in toddlers through "video-referenced" items whereby a subject is compared to a reference child shown in a three-minute video.

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JoVE Core

Measuring the Carotid to Femoral Pulse Wave Velocity (Cf-PWV) to Evaluate Arterial Stiffness
Hongwei Ji 1, Jing Xiong 1, Shikai Yu 1, Chen Chi 1, Bin Bai 1, Jiadela Teliewubai 1, Yuyan Lu 1, Yi Zhang 1, Yawei Xu 1
1Department of Cardiology, Shanghai Tenth People's Hospital, Tongji University School of Medicine

This protocol describes a method to standardize the measurements of carotid-femoral pulse wave velocity to evaluate arterial stiffness.

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Medicine

A Metadata Extraction Approach for Clinical Case Reports to Enable Advanced Understanding of Biomedical Concepts
John Harry Caufield 1,2, David A. Liem 1,2,3, Anders O. Garlid 1,2, Yijiang Zhou 4, Karol Watson 1,3, Alex A. T. Bui 1,5,6,7, Wei Wang 1,7,8,9, Peipei Ping 1,2,3,7,8
1The NIH BD2K Center of Excellence in Biomedical Computing, University of California, Los Angeles, 2Department of Physiology, University of California, Los Angeles, 3Department of Medicine/Cardiology, University of California, Los Angeles, 4Department of Cardiology, First Affiliated Hospital, Zhejiang University School of Medicine, 5Department of Radiological Sciences, University of California, Los Angeles, 6Department of Bioengineering, University of California, Los Angeles, 7Scalable Analytics Institute (ScAi), University of California, Los Angeles, 8Department of Bioinformatics, University of California, Los Angeles, 9Department of Computer Science, University of California, Los Angeles

We present a protocol and associated metadata template for the extraction of text describing biomedical concepts in clinical case reports. The structured text values produced through this protocol can support deep analysis of thousands of clinical narratives.

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JoVE Core

Intracranial Pharmacotherapy and Pain Assays in Rodents
Erik Martinez 1, Haocheng Zhou 1, Jing Wang 1,2
1Department of Anesthesiology, Perioperative Care and Pain Medicine, New York University School of Medicine, 2Department of Neuroscience and Physiology, New York University School of Medicine

Here we present a protocol to perform intracranial pharmacological experiments followed by pain behavior assays in rodents. This protocol allows researchers to deliver molecular and cellular targets in the brain, for pharmacologic agents in the treatment of pain.

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Cancer Research

Orthotopic Injection of Breast Cancer Cells into the Mice Mammary Fat Pad
Gan-Lin Zhang 1, Yi Zhang 1, Ke-Xin Cao 1, Xiao-Min Wang 1
1Oncology Department, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University

Here, we present a protocol to implant breast cancer cells into the mammary fat pad in a simple, less invasive, and easy-to-handle way, and this mouse orthotopic breast cancer model with a proper mammary fat pad environment can be used to investigate various aspects of cancer.

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Chemistry

Reactive Inkjet Printing and Propulsion Analysis of Silk-based Self-propelled Micro-stirrers
David A. Gregory 1, Piyush Kumar 1, Ana Jimenez-Franco 1, Yi Zhang 1, Yu Zhang  2, Stephen J. Ebbens 1, Xiubo Zhao 1
1Department of Chemical and Biological Engineering, University of Sheffield, 2School of Engineering, University of Liverpool

This protocol demonstrates the ability to utilize reactive inkjet printing to print self-motile biocompatible and environmentally friendly micro-stirrers for use in biomedical and environmental applications.

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Medicine

Cloud-Based Phrase Mining and Analysis of User-Defined Phrase-Category Association in Biomedical Publications
Dibakar Sigdel *1,2, Vincent Kyi *1,2, Aiden Zhang *1, Shaun P. Setty 3, David A. Liem 1,2,4, Yu Shi 5, Xuan Wang 5, Jiaming Shen 5, Wei Wang 1,6,7, JiaWei Han 5, Peipei Ping 1,2,4,6
1The NIH BD2K Center of Excellence in Biomedical Computing, University of California, Los Angeles, 2Department of Physiology, University of California, Los Angeles, 3Department of Pediatric and Adult Congenital Heart Surgery, Miller Children's and Women's Hospital and Long Beach Memorial Hospital, 4Department of Medicine/Cardiology, University of California, Los Angeles, 5NIH BD2K Program Centers of Excellence for Big Data Computing -- KnowEng Center, Department of Computer Science, University of Illinois at Urbana-Champaign (UIUC), 6Scalable Analytics Institute (ScAi), University of California, Los Angeles, 7Department of Computer Science, University of California, Los Angeles

We present a protocol and associated programming code as well as metadata samples to support a cloud-based automated identification of phrases-category association representing unique concepts in user selected knowledge domain in biomedical literature. The phrase-category association quantified by this protocol can facilitate in depth analysis in the selected knowledge domain.

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Medicine

Orthotopic Rat Kidney Transplantation: A Novel and Simplified Surgical Approach
Ali R. Ahmadi 1, Le Qi 1, Kenichi Iwasaki 1, Wei Wang 1, Russell N. Wesson 1, Andrew M. Cameron 1, Zhaoli Sun 1
1Department of Surgery, Johns Hopkins University School of Medicine

The purpose of this manuscript and protocol is to explain and demonstrate in detail the surgical procedure of orthotopic kidney transplantation in rats. This method is simplified to achieve the correct perfusion of the donor kidney and shorten the reperfusion time by using the venous and ureteral cuff anastomosis technique.

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Environment

A Uniaxial Compression Experiment with CO2-Bearing Coal Using a Visualized and Constant-Volume Gas-Solid Coupling Test System
Weitao Hou 1, Hanpeng Wang 1, Wei Wang 1, Zhongzhong Liu 1, Qingchuan Li 1
1Research Center of Geotechnical and Structural Engineering, Shandong University

This protocol demonstrates how to prepare a briquette sample and conduct a uniaxial compression experiment with a briquette in different CO2 pressures using a visualized and constant-volume gas-solid coupling test system. It also aims to investigate changes in terms of coal’s physical and mechanical properties induced by CO2 adsorption.

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Medicine

Immunoglobulin G N-Glycan Analysis by Ultra-Performance Liquid Chromatography
Di Liu *1, Xizhu Xu *2, Yuejin Li 2, Jie Zhang 1, Xiaoyu Zhang 1, Qihuan Li 1, Haifeng Hou 2, Dong Li 2, Wei Wang 1,2,3, Youxin Wang 1
1Beijing Key Laboratory of Clinical Epidemiology, School of Public Health, Capital Medical University, 2School of Public Health, Shandong First Medical University & Shandong Academy of Medical Sciences, 3School of Medical and Health Sciences, Edith Cowan University

Immunoglobulin G (IgG) N-glycan is characterized using hydrophilic interaction chromatography UPLC. In addition, the structure of IgG N-glycan is clearly separated. Presented here is an introduction to this experimental method so that it can be widely used in research settings.

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Biology

Quantification of Proliferative and Dead Cells in Enteroids
Hua-Shan Li *1, Shao-Fang Xu *1, Jian-Ying Sheng *1, Zhi-Hui Jiang 1, Jing Wang 1, Ning Ding 1, Tao Wang 1, Matthew A. Odenwald 2, Jerrold R. Turner 2,3, Wei-Qi He 1, Hong Xu 1, Juan-Min Zha 1
1Jiangsu Key Laboratory of Neuropsychiatric Diseases and Cambridge-Suda (CAM-SU) Genomic Resource Center, Medical College of Soochow University, Department of Oncology, The First Affiliated Hospital of Soochow University, 2Department of Pathology, University of Chicago, 3Department of Pathology, Brigham and Women's Hospital–Harvard Medical School

The presented protocol uses flow cytometry to quantify the number of proliferating and dead cells in cultured mouse enteroids. This method is helpful to evaluate the effects of drug treatment on organoid proliferation and survival.

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Chemistry

A Femtoliter Droplet Array for Massively Parallel Protein Synthesis from Single DNA Molecules
Yi Zhang 1, Kanako Kurosawa 1, Daisuke Nishiura 2, Mika Tei 1, Mikiko Tsudome 3
1SUGAR Program, X-star, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2Center for Mathematical Science and Advanced Technology, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 3Deep-Sea Nanoscience Research Group, Japan Agency for Marine-Earth Science and Technology (JAMSTEC)

The overall goal of the protocol is to prepare over one million ordered, uniform, stable, and biocompatible femtoliter droplets on a 1 cm2 planar substrate that can be used for cell-free protein synthesis.

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Developmental Biology

In Situ Detection of Ribonucleoprotein Complex Assembly in the C. elegans Germline using Proximity Ligation Assay
Nicholas J. Day 1, Xiaobo Wang 1, Ekaterina Voronina 1
1Division of Biological Sciences, University of Montana

This protocol demonstrates use of the proximity ligation assay to probe for protein-protein interactions in situ in the C. elegans germline.

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Behavior

A Chronic Sleep Fragmentation Model using Vibrating Orbital Rotor to Induce Cognitive Deficit and Anxiety-Like Behavior in Young Wild-Type Mice
Yi Xie 1, Sai-Yue Deng 1, Si-Miao Chen 1, Xue-Jiao Chen 1, Wen-Wen Lai 1, Li-Fang Huang 1, Li Ba 1, Wei Wang 1,2, Feng-Fei Ding 1,3
1Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430000, China, 2Key Laboratory of Neurological Diseases of Chinese Ministry of Education, The School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430000,China, 3Department of Pharmacology, School of Basic Medical Science; State Key Laboratory of Medical Neurobiology, Institutes of Brain Science and Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China

Presented here is a protocol for chronic sleep fragmentation (CSF) model achieved by an electrically controlled orbital rotor, which could induce confirmed cognitive deficit and anxiety-like behavior in young wild-type mice. This model can be applied to explore the pathogenesis of chronic sleep disturbance and related disorders.

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Neuroscience

Establishment of a Rat Model of Superior Sagittal-Sinus Occlusion via a Thread-Embolism Method
Weiyan Jiang *1, Congcong Jin *2, Weiming Xu *3, Yingxian Li 4, Yinghong Lin 5, Shengxiang Liang 6,7, Wei Wang 8
1Department of Clinical Laboratory, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, 2Rehabilitation Department, Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine, 3Department of Neurosurgery, the Hospital of Changle, 4Pediatrics Ward 2, Shouguang Hospital of Traditional Chinese Medicine, 5Fujian University of Traditional Chinese Medicine, 6National-Local Joint Engineering Research Center of Rehabilitation Medicine Technology, Fujian University of Traditional Chinese Medicine, 7Rehabilitation Industry Institute, Fujian University of Traditional Chinese Medicine, 8Department of Neurosurgery, the First Affiliated Hospital of Wenzhou Medical University

Here, we establish a novel Sprague-Dawley (SD) rat model of superior sagittal sinus (SSS) thrombosis via a thread-embolization method, and the stability and reliability of the model were verified.

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Behavior

Moxibustion Strategies for Mice
Xiao-qin Dai *1,2, Peng Lv *3, Rui-zhen Huang *4, Han-xiao Zhang 3, Si-rui Lin 5, Gang Shi 4, Cheng-shun Zhang 3, Chuan-yi Zuo 6
1Department of Traditional Chinese Medicine, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, 2Chinese Academy of Sciences Sichuan Translational Medicine Research Hospital, 3Acupuncture and Tuina School, Chengdu University of Traditional Chinese Medicine, 4Department of Pharmacy Service, Hospital of Chengdu University of Traditional Chinese Medicine, 5Department of Acupuncture, Southwest Medical University Affiliated Traditional Chinese Medicine Hospital, 6Department of Acupuncture, Chongqing Traditional Chinese Medicine Hospital

The present protocol describes a moxibustion application method for mice, with the benefits of reducing mouse fearfulness, guaranteeing their welfare, and improving efficiency for researchers.

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Medicine

Standardized Rat Coronary Ring Preparation and Real-Time Recording of Dynamic Tension Changes Along Vessel Diameter
Pengmei Guo *1, Wenqiao An *2, Ying Guo 1, Zhihao Sun 2, Xiaobo Wang 1,3, Sanyin Zhang 1,3
1Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 2School of Basic Medicine, Chengdu University of Traditional Chinese Medicine, 3Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine

The present protocol describes the wire myograph technique for measuring vascular reactivity of the rat coronary artery.

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Neuroscience

Real-Time Dynamic Collection of Hippocampal Extracellular Fluid from Conscious Rats Using a Microdialysis System
Xiaobo Wang 1,2, Na Xie 3, Yi Zhang 4, Xianli Meng 1,2,3, Ya Hou 3, Sanyin Zhang 1,2
1Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine, 3School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 4School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine

The protocol here provides a detailed real-time dynamic sampling of extracellular fluid from the hippocampus of awake rats using a microdialysis system.

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Medicine

Dynamic Continuous Blood Extraction from Rat Heart via Noninvasive Microdialysis Technique
Ya Hou 1, Jinrong Bai 1,2, Yi Zhang 3, Xianli Meng 1,2,4, Sanyin Zhang 4, Xiaobo Wang 2
1School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine, 3School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, 4Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine

The present protocol describes a simple and efficient method for the real-time and dynamic collection of rat heart blood using the microdialysis technique.

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JoVE Core

Meta-Analysis of the Effectiveness and Safety of Shugan Jieyu Capsules for the Treatment of Insomnia
Jing Wang *1, Peipei Du *2, Linda LD Zhong 2, Yaxuan Ren 1, Li Feng 1, Shu Yang 1, Haibei Song 1
1School of Intelligent Medicine, Chengdu University of Traditional Chinese Medicine, 2School of Chinese Medicine, Hong Kong Baptist University

The presented protocol describes an investigation strategy and systematic review to evaluate the clinical effectiveness and safety of Shugan Jieyu capsules for treating insomnia.

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JoVE Core

An In Vitro Dissolution Determination of Multi-Index Components in Tibetan Medicine Rhodiola Granules
Qinyun Du 1, Qingxiu He 2, Fengming Zhang 3, Jiao Mi 3, Yuying Li 3, Shaohui Wang 1, Yi Zhang 1
1School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, 3Tibet Rhodiola Pharmaceutical Holding Co. Ltd

Here, we test the dissolution of Rhodiola granules (RG) in vitro, draw dissolution curves of salidroside, gallic acid, and ethyl gallate in ultrapure water, and fit the curves to different mathematical models. This protocol provides information and guidance for in vivo bioequivalence and in vivo-in vitro correlation studies of RG.

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Bioengineering

Synthesis of Strong Adhesive Hydrogel, Gelatin O-Nitrosobenzaldehyde
Yuelong Liang *1,2, Zhengze Huang *1, Yi Zhang *3,4, Yi Hong 3,4, Qijiang Mao 1,2,5, Xu Feng 1
1Department of General Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 2Zhejiang Province Medical Research Center of Minimally Invasive Diagnosis and Treatment of Abdominal Diseases, 3Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cells and Regenerative Medicine, Department of Orthopedic Surgery of the Second Affiliated Hospital, Zhejiang University School of Medicine, 4Zhejiang University-University of Edinburgh Institute, Zhejiang University School of Medicine, Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, Zhejiang University School of Medicine, 5Zhejiang Provincial Key Laboratory of Laparoscopic Technology

The protocol presented here shows the synthesis of a strong adhesive hydrogel gelatin o-nitrosobenzaldehyde (gelatin-NB). Gelatin-NB has rapid and efficient tissue adhesion ability, which can form a strong physical barrier to protect wound surfaces, so it is expected to be applied to the field of injury repair biotechnology.

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Medicine

Voltage-Dependent Potassium Current Recording on H9c2 Cardiomyocytes via the Whole-Cell Patch-Clamp Technique
Hong Jiang *1, Yating Zhang *1, Ya Hou 1, Lijun Li 2, Sanyin Zhang 3,4, Yi Zhang 5, Xianli Meng 1,3,4, Xiaobo Wang 4
1State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Chengdu Techman Instrument Co., Ltd., 3Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 4Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine, 5School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine

The present protocol describes an efficient method for the real-time and dynamic acquisition of voltage-gated potassium (Kv) channel currents in H9c2 cardiomyocytes using the whole-cell patch-clamp technique.

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Medicine

Enema of Traditional Chinese Medicine for Patients with Severe Acute Pancreatitis
Shi Wang 1,2, Tong Zhou 1,2, Kunlan Long 1, Jun Chen 1, Xiujuan Zhou 1, Peiyang Gao 1
1Department of Critical Care Medicine, Hospital of Chengdu University of Traditional Chinese Medicine, 2Clinical Medicine Department, Chengdu University of Traditional Chinese Medicine

An enema is a type of traditional Chinese medicine (TCM) therapy used to treat severe acute pancreatitis (SAP). The therapy involves instilling a TCM liquid into the rectum and colon through the anus to keep the liquid in the intestine, absorbing through the intestinal mucosa. This study aims to standardize the enema procedure for SAP patients.

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Medicine

Acupoint Needle-Embedding Combined with Ironing Therapy for Postoperative Pain After Anal Surgery
Xiao Tang 1,2, Jian Kang 1, Ya Liu 1
1Hospital of Chengdu University of Traditional Chinese Medicine, 2Chengdu University of Traditional Chinese Medicine

The high incidence of pain after anal surgery is due to the physiological structure and pathological status of the anus. At present, the use of painkillers cannot provide satisfactory pain relief, which makes complementary therapy important. This paper proposes acupoint needle-embedding combined with ironing therapy to relieve postoperative pain.

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Medicine

Visualization of Metabolites Identified in the Spatial Metabolome of Traditional Chinese Medicine Using DESI-MSI
Binjie Xu 1,4, Lirun Chen 3, Fengqi Lv 3, Yuan Pan 1, Xing Fu 1, Zhaoqing Pei 1,2
1Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Tianfu Chinese Medicine Innovation Harbour, Chengdu University of Traditional Chinese Medicine, 3School of Medical Technology, Chengdu University of Traditional Chinese Medicine, 4China Resources Sanjiu (Ya’an) Pharmaceutical Co., Ltd.

In this study, a series of methods are presented to prepare DESI-MSI samples from plants, and a procedure of DESI assembly installation, MSI data acquisition, and processing is described in detail. This protocol can be applied in several conditions for acquiring spatial metabolome information in plants.

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Medicine

A Protocol for Constructing a Rat Wound Model of Type 1 Diabetes
Wei Wang *1, Dingxi Bai *1, Chenxi Wu *1, Hang Li 1, Xuemei Xie 1, Wenting Ji 1, Jing Gao 1
1College of Nursing, Chengdu University of Traditional Chinese Medicine

The streptozotocin-induced diabetic wound model in male SD rats is currently the most widely used model for studying wound healing in type I diabetes mellitus. This protocol describes the methods used to construct this model. It also presents and addresses potential challenges and examines the progression and angiogenic characteristics of diabetic wounds.

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Medicine

Fast Inspection of Quality of Indigo Naturalis by Multiple Light Scattering
Xuemei Liu 1, Hui Ye 1, Haozhou Huang 1, Ming Yang 2, Li Han 1, Runchun Xu 1, Dingkun Zhang 1
1School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2State Key Laboratory of Innovation Drug and Efficient Energy-saving Pharmaceutical Equipment, Jiangxi University of Traditional Chinese Medicine

Here, we present a quality evaluation method based on a multiple light scattering technique to evaluate the quality of Indigo Naturalis.

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Chemistry

Standardized Identification of Compound Structure in Tibetan Medicine Using Ion Trap Mass Spectrometry and Multiple-Stage Fragmentation Analysis
Xing Fu 1, Yuan Pan 1, Yili Wang 1, Zhaoqing Pei 1, Binjie Xu 1, Jing Zhang 2, Jinsong Su 1
1State Key Laboratory of Southwestern Chinese Medicine Resources, Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 2School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine

Here, we describe a general protocol and design that could be applied to identify trace amounts and minor constituents in the complex natural product formulations (matrixes) in Tibetan medicine.

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JoVE Core

Network Pharmacology Prediction and Metabolomics Validation of the Mechanism of Fructus Phyllanthi against Hyperlipidemia
Baihan Zeng *1,2,3, Luming Qi *1,2,3, Sha Wu *1,2,3, Nannan Liu 1,2,3, Jie Wang 1,2,3, Kaidi Nie 1,2,3, Lina Xia 1,2,3, Shuguang Yu 2,3
1School of Health Preservation and Rehabilitation, Chengdu University of Traditional Chinese Medicine, 2State Administration of Traditional Chinese Medicine Key Laboratory of Traditional Chinese Medicine, Regimen and Health, Chengdu University of Traditional Chinese Medicine, 3Key Laboratory of Traditional Chinese Medicine Regimen and Health of Sichuan Province

The present protocol describes an integrated strategy for exploring the key targets and mechanisms of Fructus Phyllanthi against hyperlipidemia based on network pharmacology prediction and metabolomics verification.

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Medicine

Using Multiple Light Scattering to Examine the Stability of Phyllanthus emblica L. Extracts Obtained with Different Extraction Methods
Haozhou Huang *1,2, Mengqi Li *3, Chuanhong Luo 4, Sanhu Fan 5, Taigang Mo 5, Li Han 4, Dingkun Zhang 4, Junzhi Lin 6
1State Key Laboratory of Southwestern Chinese Medicine Resources, Innovative Institute of Chinese Medicine and Pharmacy/Academy for Interdiscipline, Chengdu University of Traditional Chinese Medicine, 2Meishan Hospital of Chengdu University of Traditional Chinese Medicine, 3Sichuan Nursing Vocational College, 4State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 5Sanajon Pharmaceutical Group, 6TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Hospital of Chengdu University of Traditional Chinese Medicine

Here, we introduce a stability evaluation method based on multiple light scattering technology to evaluate the stability of traditional Chinese medicine extracts.

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Medicine

Optimization of Processing Technology for Tiebangchui with Zanba Based on CRITIC Combined with Box-Behnken Response Surface Method
Shiling Li 1, Liqiong Yu 1, Congying Li 2, Naiyu Wang 1, Xianrong Lai 2, Yue Liu 2, Yi Zhang 2
1State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine

The present protocol describes an efficient and standard detoxification processing method for Zanba-stir-fried Tiebangchui using CRITIC combined with the Box-Behnken response surface method.

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Medicine

Objectification of Tongue Diagnosis in Traditional Medicine, Data Analysis, and Study Application
Li Feng *1, Wenke Xiao *1, Chuanbiao Wen 1, Qiaoling Deng 1, Jinhong Guo 1, Haibei Song 1
1School of Intelligent Medicine, Chengdu University of Traditional Chinese Medicine

The present study employed U-Net and other deep learning algorithms to segment a tongue image and compared the segmentation results to investigate the objectification of tongue diagnosis.

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Biology

Noninvasive Intratracheal Lipopolysaccharide Instillation in Mice
Peng Yu *1, Bo Lin *2, Jingyang Li 1, Yanqing Luo 1, Di Zhang 1, Jiayi Sun 1, Xianli Meng 1, Yingfan Hu 3, Li Xiang 1
1State Key Laboratory of Southwestern Chinese Medicine Resources, College of Pharmacy, Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 2Department of Pharmacy, The Second Affiliated Hospital of Hainan Medical University, 3The School of Preclinical Medicine, Chengdu University

Here, we propose a protocol for intratracheal lipopolysaccharide (LPS) delivery via noninvasive oropharyngeal endotracheal intubation. This method minimizes the trauma of the surgical procedure for the animal and accurately delivers LPS to the trachea and then to the lungs.

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Medicine

Optimization of Processing of Tiebangchui with Highland Barley Wine Based on the Box-Behnken Design Combined with the Entropy Method
Liqiong Yu 1, Shiling Li 1, Xiaoyan Tan 1, Chengping Wang 2, Xianrong Lai 1,2, Yue Liu 2, Yi Zhang 2
1State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 2State Key Laboratory of Southwestern Chinese Medicine Resources, School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine

The present protocol describes an efficient method for optimization of the processing technology of Tiebangchui processed with highland barley wine based on a Box-Behnken design response surface combined with the entropy method.

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Chemistry

An Improved Technique for Trimethylamine Detection in Animal-Derived Medicine by Headspace Gas Chromatography-Tandem Quadrupole Mass Spectrometry
Hui Ye 1, Xuemei Liu 1, JiaBao Liao 2, Haozhou Huang 3, Lin Huang 1, Yang Bao 1, Hongyan Ma 1, Junzhi Lin 4, Xiaoming Bao 5, Dingkun Zhang 1, Runchun Xu 1
1State Key Laboratory of Southwestern Chinese Medicine Resources, Pharmacy School, Chengdu University of Traditional Chinese Medicine, 2China Resources Sanjiu Modern Chinese Medicine Pharmaceutical Co., Ltd, 3Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 4TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Hospital of Chengdu University of Traditional Chinese Medicine, 5Shimadzu (China) Co., Ltd

Here, a headspace gas chromatography-tandem quadrupole mass spectrometry (HS-GC-MS/MS) method suitable for the determination of trimethylamine (TMA) in animal-derived medicines is described. The protocol includes sample pretreatment, headspace treatment, analysis conditions, methodological validation, and the determination of TMA in animal-derived medicines.

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Medicine

Operational and Intervention Effects of Targeted Tuina in Lumbar Intervertebral Disc Degeneration Model Rabbits
Jiang-nan Ye 1,2, Chang Liu 1,2, Yun-ru Guo 3, Zhi-jun Ma 1,2, Xiang-yue Li 1, Kun-lin Kuang 1, Xiao-xia Zheng 3, Wen-xi Sun 3, Ding-kun Shen 1, Jun Zhu 1,2, Cheng-guo Su 1
1Tuina Teaching and Research Department, School of Acupuncture-Moxibustion and Tuina, Chengdu University of Traditional Chinese Medicine, 2Acupuncture and Massage Rehabilitation Center, The Third Affiliated Hospital of Chengdu University of Traditional Chinese Medicine, 3Graduate School, Guangzhou University of Chinese Medicine

The finger tactile measurement system quantifies tactile pressure on the finger using a highly sensitive electrosurgical pressure sensor, with software displaying and accurately recording the pressure data and video in real time. Two analysis modules unify the data processing for thumb tracking and conditioning during Tuina.

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Medicine

Real-Time Detection of Ferulic Acid Effects on Rat Left Ventricle Using Pressure-Volume Conductivity Catheter
Zhihao Sun 1,2, Wenqiao An 2, Taiping He 2, Qingqing Liu 2, Ziming Wang 2, Pengmei Guo 2, Sanyin Zhang 2,3
1School of Basic Medical Sciences, Chengdu University of Traditional Chinese Medicine, 2Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 3Research Institute of Integrated TCM & Western Medicine, Chengdu University of Traditional Chinese Medicine

This protocol describes a method for measuring left ventricular pressure and volume using the pressure-volume conductance technique. This method enables continuous real-time monitoring of the effects of drugs on the heart.

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Neuroscience

Barrier Functional Integrity Recording on bEnd.3 Vascular Endothelial Cells via Transendothelial Electrical Resistance Detection
Fuhan Fan 1, Hong Jiang 1, Ya Hou 1, Yi Zhang 1,2,3, Qipeng Zhao 4, Yong Zeng 1,2, Xianli Meng 1,2,5, Xiaobo Wang 2
1School of Pharmacy/School of Modern Chinese Medicine Industry, Chengdu University of Traditional Chinese Medicine, 2Meishan Hospital of Chengdu University of Traditional Chinese Medicine, 3School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, 4School of Pharmacy, Ningxia Medical University, 5Innovative Institute of Chinese Medicine and Pharmacy/Academy for Interdiscipline, Chengdu University of Traditional Chinese Medicine

This protocol describes a dependable and efficient in vitro model of the brain blood barrier. The method uses mouse cerebral vascular endothelial cells bEnd.3 and measures transmembrane electrical resistance.

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Medicine

Real-Time Detection of Dynamic Affinity between Biomolecules Using Surface Plasmon Resonance (SPR) Technology
Wenqiao An *1, Zhihao Sun *1, Pengmei Guo 2, Xiaobo Wang 2,3, Sanyin Zhang 2,3
1School of Basic Medical Sciences, Chengdu University of Traditional Chinese Medicine, 2Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 3Research Institute of Integrated Traditional Chinese Medicine and Western Medicine, Chengdu University of Traditional Chinese Medicine

The present study aims to elucidate the principle and methodology of surface plasmon resonance (SPR) technology, which finds versatile applications across multiple domains. This article describes SPR technology, its operational simplicity, and its remarkable efficacy, with the goal of fostering broader awareness and adoption of this technology among readers.

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Medicine

Field Collection and Laboratory Routine Identification of Rhodiola crenulata
Jiayi Wang *1, Na Xie *2,3, Mengjia Li 1, Jinsong Su 4, Ya Hou 3, Yi Zhang 1,2,3, Xiaobo Wang 2
1School of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, 2Meishan Hospital of Chengdu University of Traditional Chinese Medicine, 3School of Pharmacy, Chengdu University of Traditional Chinese Medicine, 4Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine

Here, we describe the identification of Rhodiola Crenulata from habitat, plant morphology, medicinal properties, microscopic features, and thin-layer chromatography.

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Medicine

2D-HPLC-MS Technology Combined with Molecular Network for the Identification of Components in Tibetan Medicine Aconitum pendulum
Jing Ma *1, Hanyu Lu *1, Jia Liu 1, Ting Wang 1, Xing Fu 2, Xinmei Xu 2, Yi Zhang 1, Jing Zhang 1, Xiaolong Xie 1, Yingzhuang Chen 3,4, Jinsong Su 1
1Ethnic Medicine Academic Heritage Innovation Research Center, Meishan Hospital, Chengdu University of Traditional Chinese Medicine, 2State Key Laboratory of Southwestern Chinese Medicine Resources, Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 3Key Laboratory of Chemical Biology & Traditional Chinese Medicine Research, Ministry of Education, 4Key Laboratory of Phytochemical R&D of Hunan Province, Hunan Normal University

This study utilizes two-dimensional high-performance liquid chromatography-mass spectrometry (2D-HPLC-MS) technology in conjunction with molecular networking to unravel the intricate chemical composition of the Tibetan medicinal plant Aconitum pendulum Busch (APB). The article provides a detailed protocol for the systematic exploration and identification of complex chemical components of herbal medicines.

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Medicine

Using An In Vitro Tissue Perfusion System to Detect the Functional Activities of Isolated Intestinal Tubes in Real Time
Pengmei Guo 1, Wenqiao An 2, Yili Wang 1, Yajing Ren 3, Sanyin Zhang 1,4
1Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 2School of Basic Medical College, Chengdu University of Traditional Chinese Medicine, 3School of Medical and Life Sciences, Chengdu University of Traditional Chinese Medicine, 4Chengdu Integrated TCM and Western Medicine Hospital

We present a method for isolating rat intestinal tubes and assessing the impact of drugs on their tension, frequency, and amplitude in vitro. This method offers a valuable approach for researchers investigating intestinal tubes.

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Neuroscience

Biomarker Identification for Gender Specificity of Alzheimer's Disease Based on the Glial Transcriptome Profiles
Hanjie Liu *1, Hui Yang *2, Shuqing Liu 1, Siyu Li 1, Wen Yang 3, Anwar Ayesha 1, Xin Tan 4, Cen Jiang 1, Yi Liu 5, Lushuang Xie 1
1Chengdu University of Traditional Chinese Medicine, 2Chengdu Shuangliu Hospital of Traditional Chinese Medicine, 3Department of Automation, Tsinghua University, 4Affiliated Reproductive & Women-Children Hospital, Chengdu University of Traditional Chinese Medicine, 5Department of Neurology, Dalian Municipal Central Hospital, Central Hospital of Dalian University of Technology

This study analyzed single-nuclei transcriptomes of thirty-three individuals with Alzheimer's disease (AD), revealing sex-specific DEGs in glial cells. Functional enrichment analysis highlighted synaptic, neural, and hormone-related pathways. Key genes, namely NLGN4Y and its regulators, were identified, and potential therapeutic candidates for gender-specific AD were proposed.

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Medicine

Protein Target Prediction and Validation of Small Molecule Compound
Liuling Luo 1, Jinrui Xiong 1, Yancai Tang 1, Xiaorui Chen 1, Hai Zhang 1
1State Key Laboratory of Southwestern Chinese Medicine Resources, School of Pharmacy/School of Modern Chinese Medicine Industry, Chengdu University of Traditional Chinese Medicine

The experiment used here shows a method of molecular docking combined with cellular thermal shift assay to predict and validate the interaction between small molecules and protein targets.

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Chemistry

13C6-Glucose Labeling Associated with LC-MS: Identification of Plant Primary Organs in Secondary Metabolite Synthesis
Siyu Chen 1,2, Fei Chang 1,2, Liting Lin 1,2, Yue Wang 3, Feiyan Wen 1,2, Tao Zhou 1,2, Jin Pei 1,2
1State Key Laboratory of Southwestern Chinese Medicine Resources, Chengdu University of Traditional Chinese Medicine, 2College of Pharmacy, Chengdu University of Traditional Chinese Medicine, 3Department of Pharmacy, Ya'an People's Hospital

The developed method of 13C6-Glucose labeling combined with liquid chromatography high-resolution mass spectrometry is versatile and lays the foundation for future studies on the primary organs and pathways involved in the synthesis of secondary metabolites in medicinal plants, as well as the comprehensive utilization of these secondary metabolites.

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Medicine

Detection of CD40 Protein-Umbelliferone Interaction via Differential Scanning Fluorescence
Nanjia Cairang *1, Yating Zhang *2, Hong Jiang *2, Tsering Sonan 1, Xiaobo Wang 2
1Department of Tibetan Medicine, University of Tibetan Medicine, 2Innovative Institute of Chinese Medicine and Pharmacy / Academy for Interdiscipline, Chengdu University of Traditional Chinese Medicine

This protocol describes a unique method for detecting the binding between compounds and protein molecules, offering the advantages of minimal protein sample loss and high data accuracy.

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Medicine

Press Needle: A Traditional Chinese Medicine Therapy for Myopia Patients with Dry Eye
Tianyao Gong *1,2, Qun Huang *3, Haiyan Zhang *1, Chunmeng Liu 1, Xiaojuan Su 1, Yiqi Yang 2, Xinglin Liu 2, Yanlin Zheng 3, Hejiang Ye 3
1Eye School of Chengdu University of Traditional Chinese Medicine, 2Chengdu University of Traditional Chinese Medicine, 3Hospital of Chengdu University of Traditional Chinese Medicine

Here, we present a protocol to standardize the operation of acupuncture in the treatment of myopia with dry eye.

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Medicine

Acupoint Application Combined with Ear Plaster Therapy for Treating Sleep Disorders with Acute Exacerbation of Chronic Obstructive Pulmonary Disease
Hualing Wang *1, Li Liu *2, Chunyi Wang 3, Fanyi Yuan 1, Yufei Liu 1, Feifei Yang 1, Cuifang He 1, Yuanyue Tang 4, Tianbao Wang 5
1Department of Respiratory Medicine, Hospital of Chengdu University of Traditional Chinese Medicine, 2Department of Pharmacy, Hospital of Chengdu University of Traditional Chinese Medicine, 3Clinical Medical College, Chengdu University of Traditional Chinese Medicine, 4Nursing Department, Hospital of Chengdu University of Traditional Chinese Medicine, 5Outpatient Department, Sichuan Cancer Hospital

The present protocol describes the specific process and efficacy of the acupoint application combined with ear plaster therapy for sleep disorders in a patient with acute exacerbation of chronic obstructive pulmonary disease, including medical devices, selection of acupoints, operating methods, and posttreatment care.

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Biology

LPS and ATP-induced Death of PMA-differentiated THP-1 Macrophages and its Validation
Huan Wang 1, Yuejia Lan 1, Cuiping Chen 1, Lu Yang 2, Jiayi Sun 2, Yong Zeng 1, Jiasi Wu 3
1State Key Laboratory of Southwestern Chinese Medicine Resources, Chengdu University of Traditional Chinese Medicine, 2Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, 3Acupuncture and Tuina School, Chengdu University of Traditional Chinese Medicine

This protocol is based on LPS and ATP-induced death of PMA-differentiated THP-1 macrophages. We use flow cytometry to analyze Annexin V and 7-AAD double staining to detect cell death, using the whole cell and employing scanning electron microscopy to observe the cell membrane morphology.

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Medicine

Acupoint Application Combined with Acupressure as an Adjunctive Therapy for Chemotherapy-Induced Nausea and Vomiting
Qiuyu Chen 1, Ma HongMei 2, Liu Yi 1
1Chengdu University of Traditional Chinese Medicine, 2Oncology Department of Chengdu First People's Hospital

Acupoint application combined with acupressure-assisted treatment has a good effect on chemotherapy-induced nausea and vomiting and is more convenient and cost-effective.

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Genetics

Application of DNA Barcoding to Identify Medicinal Plants
Shuai Yang 1,2, Shilin Chen 1,2, Jing Mo 2, Junfei Liu 2, Qi Pan 2, Chi Song 2,3
1School of Pharmacy, Guizhou University of Traditional Chinese Medicine, 2School of Pharmacy/School of Modern Chinese Medicine Industry, Chengdu University of Traditional Chinese Medicine, 3Institute of Herbgenomics, Chengdu University of Traditional Chinese Medicine

The present protocol describes the use of DNA barcoding technology to authenticate plant-based medicinal materials, and the medicinal plant Angelica sinensis (Oliv.) Diels was identified as an example.

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Medicine

Monitoring Pulmonary Nodule Progression Using Micro-Computed Tomography and Blood Sampling in a Mouse Model
Ying Guo *1,2, Fuhan Fan *2,3, Huan Xu 4, Ting Zhang 2, Peng Yang 5, Sanyin Zhang 2,3, Fengming You 6
1Affiliated Hospital of Chengdu University of Traditional Chinese Medicine, 2State Key Laboratory of Southwestern Chinese Medicine Resources, Innovative Institute of Chinese Medicine and Pharmacy/Academy for Interdisciplinary, Chengdu University of Traditional Chinese Medicine, 3Research Institute of Integrated Traditional Chinese Medicine and Western Medicine, Chengdu University of Traditional Chinese Medicine, 4Chengdu University of CTM, Basic Medical College, 5Chengdu Fifth People's Hospital, 6TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province, Affiliated Hospital of Chengdu University of Traditional Chinese Medicine

This protocol describes an efficient, simple, and minimally invasive method for studying pulmonary nodules. Submaxillary vein blood collection and micro-CT imaging are used as investigative techniques.

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