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Tianjin University

26 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

DNA Fingerprinting of Mycobacterium leprae Strains Using Variable Number Tandem Repeat (VNTR) - Fragment Length Analysis (FLA)
Ronald W. Jensen 1, Jason Rivest 1, Wei Li 1, Varalakshmi Vissa 1
1Department of Microbiology, Immunology and Pathology, Colorado State University

Leprosy, caused by Mycobacterium leprae, is still endemic in many places. In order to learn about the spread and mode of transmission of leprosy, it is important to determine which strain of M. leprae has infected a patient. Variable numbers of tandem repeats (VNTR) typing is one such method.

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Medicine

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies
Wei Li 1, Shu Zhu 1, Yusong Zhang 1, Jianhua Li 1, Andrew E. Sama 1, Ping Wang 1, Haichao Wang 1
1The Feinstein Institute for Medical Research, North Shore – LIJ Health System

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

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Engineering

Iridium Oxide-reduced Graphene Oxide Nanohybrid Thin Film Modified Screen-printed Electrodes as Disposable Electrochemical Paper Microfluidic pH Sensors
Jiang Yang 1,2, Tae-Joon Kwak 3, Xiaodong Zhang 4, Robert McClain 5, Woo-Jin Chang *3,6, Sundaram Gunasekaran *1
1Department of Biological Systems Engineering, University of Wisconsin-Madison, 2Environment, Energy and Natural Resources Center, Department of Environmental Science and Engineering, Fudan University, 3Department of Mechanical Engineering, University of Wisconsin-Milwaukee, 4Department of Physics, School of Science, Tianjin University, 5Department of Chemistry, University of Wisconsin-Madison, 6School of Freshwater Sciences, University of Wisconsin-Milwaukee

The study demonstrates the growth of iridium oxide-reduced graphene oxide (IrO2-RGO) nanohybrid thin films on irregular and rough screen-printed carbon substrate through a green electrochemical synthesis, and their implementation as a pH sensor with a patterned paper-fluidic platform.

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JoVE Core

SSVEP-based Experimental Procedure for Brain-Robot Interaction with Humanoid Robots
Jing Zhao 1, Wei Li 1,2, Xiaoqian Mao 1, Mengfan Li 1
1School of Electrical Engineering and Automation, Tianjin University, 2Department of Computer & Electrical Engineering and Computer Science, California State University

The overall goal of this method is to establish an SSVEP-based experimental procedure by integrating multiple software programs to enable the study of brain-robot interaction with humanoid robots, which is prospective in assisting the sick and elderly as well as performing unsanitary or dangerous jobs.

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Medicine

Ferric Chloride-induced Murine Thrombosis Models
Wei Li 1,2, Marvin Nieman 3, Anirban Sen Gupta 4
1Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, 2Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, 3Department of Pharmacology, Case Western Reserve University, 4Department of Biomedical Engineering, Case Western Reserve University

We report a refined procedure of the ferric chloride (FeCl3)-induced thrombosis models on carotid and mesenteric artery as well as vein, characterized efficiently using intravital microscopy to monitor time to occlusive thrombi formation.

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Bioengineering

Expanding Nanopatterned Substrates Using Stitch Technique for Nanotopographical Modulation of Cell Behavior
Kai Wang 1, Kam W. Leong 2, Yong Yang 1
1Department of Chemical and Biomedical Engineering, West Virginia University, 2Department of Biomedical Engineering, Columbia University

A protocol for producing a large area of nanopatterned substrate from small nanopatterned molds for study of nanotopographical modulation of cell behavior is presented.

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Neuroscience

Mitochondrial Ca2+ Retention Capacity Assay and Ca2+-triggered Mitochondrial Swelling Assay
Wei Li 1,2, Chen Zhang 1, Xiulian Sun 3
1Department of Neurology, Qilu Hospital of Shandong University, 2Department of Neurology, Qingdao Municipal Hospital, 3Brain Research Institute, Qilu Hospital of Shandong University

This protocol aims to describe a method to examine the Ca2+ retention capacity and Ca2+- triggered mitochondrial swelling of isolated mitochondria of SH-SY5Y cells step-by-step.

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Bioengineering

Microhoneycomb Monoliths Prepared by the Unidirectional Freeze-drying of Cellulose Nanofiber Based Sols: Method and Extensions
Zheng-Ze Pan 1,2, Hirotomo Nishihara 3, Wei Lv 1, Cong Wang 1,2, Yi Luo 1,2, Liubing Dong 1,2, Houfu Song 1,4, Wenjie Zhang 2, Feiyu Kang 1,2,4, Takashi Kyotani 3, Quan-Hong Yang 1,4,5
1Engineering Laboratory for Functionalized Carbon Materials and Shenzhen Key Laboratory for Graphene-based Materials, Graduate School at Shenzhen, Tsinghua University, 2School of Materials Science and Engineering, Tsinghua University, 3Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 4Tsinghua-Berkeley Shenzhen Institute (TBSI), Tsinghua University, 5School of Chemical Engineering and Technology, Tianjin University

Here, we present a general protocol to prepare a variety of microhoneycomb monoliths (MHMs) in which fluid can pass through with an extremely low pressure drop. MHMs obtained are expected to be used as filters, catalyst supports, flow-type electrodes, sensors and scaffolds for biomaterials.

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JoVE Journal

A Random-displacement Measurement by Combining a Magnetic Scale and Two Fiber Bragg Gratings
Lianqing Zhu 1,2, Lidan Lu 3,4, Wei Zhuang 2,5, Zhoumo Zeng 3,4, Mingli Dong 1,2
1School of Instrument Science and Opto-electronics Engineering, Beijing Information Science and Technology University, 2Beijing Engineering Research Center of Optoelectronic Information and Instruments, Beijing Key Laboratory for Optoelectronics Measurement Technology, 3School of Precision Instrument & Opto-electronics Engineering, Tianjin University of Science and Technology, 4School of Precision Instrument and Opto-electronics Engineering, State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University, 5School of Instrument and Opto-electronics Engineering, Hefei University of Technology

A protocol to create a full-range linear displacement sensor, combining two packaged fiber Bragg grating detectors with a magnetic scale, is presented.

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Engineering

Measurement of Dynamic Force Acted on Water Strider Leg Jumping Upward by the PVDF Film Sensor
Lele Zhang 1, Meirong Zhao 1, Zhiyi Wang 1, Yunxi Li 1, Yinguo Huang 1, Yelong Zheng 1
1State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University

The protocol here is dedicated to investigating the free and quick maneuvering of water strider on water surface. The protocol includes observing the microstructure of legs and measuring the adhesion force when departing from water surface at different speeds.

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Biochemistry

Crystal Structure of the N-terminal Domain of Ryanodine Receptor from Plutella xylostella
Bidhan Chandra Nayak *1, Jie Wang *1, Lianyun Lin *1,2,3,4, Weiyi He 2,3,4, Minsheng You 2,3,4, Zhiguang Yuchi 1,2
1Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency, Collaborative Innovation Center of Chemical Science and Engineering, School of Pharmaceutical Science and Technology, Tianjin University, 2State Key Laboratory of Ecological Pest Control for Fujian/Taiwan Crops and Institute of Applied Ecology, Fujian Agriculture and Forestry University, 3Joint International Research Laboratory of Ecological Pest Control, Ministry of Education, Fuzhou, 4Fujian-Taiwan Joint Centre for Ecological Control of Crop Pests, Fujian Agriculture and Forestry University

In this article, we describe the protocols of protein expression, purification, crystallization and structure determination of the N-terminal domain of ryanodine receptor from diamondback moth (Plutella xylostella).

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Education

Measuring the Interaction Force Between a Droplet and a Super-hydrophobic Substrate by the Optical Lever Method
Shuya Zhuang 1, Meirong Zhao 1, Zhiyi Wang 1, Lele Zhang 1, Yinguo Huang 1, Yelong Zheng 1
1State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University

The protocol aims to investigate the interaction between droplets and super-hydrophobic substrates in the air. This includes calibrating the measurement system and measuring the interaction force at super-hydrophobic substrates with different grid fractions.

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Biology

Photostimulation by Femtosecond Laser Activates Extracellular-signal-regulated Kinase (ERK) Signaling or Mitochondrial Events in Target Cells
Shaoyang Wang 1,2, Minglie Hu 2, Tao Ren 1, Hao He 3
1Department of Respiratory Medicine, Shanghai Sixth People's Hospital, School of Medicine, Shanghai Jiao Tong University, 2Ultrafast Laser Laboratory, College of Precision Instrument and Optoelectronics Engineering, Tianjin University, 3School of Biomedical Engineering, Shanghai Jiao Tong University

Tightly-focused femtosecond laser can deliver precise stimulation to cells by being coupled into a confocal microscopy enabling the real-time observation and photostimulation. The photostimulation can activate cell molecular events including ERK signaling pathway and mitochondrial flashes of reactive oxygen species.

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Medicine

Three-Dimensional Printing Guide Template Assisted Percutaneous Vertebroplasty (PVP)
PeiLun Hu *1, JiSheng Lin *1, JunChuan Xu 1, Hai Meng 1, Nan Su 1, Yong Yang *1, Qi Fei 1
1Department of Orthopedics, Beijing Friendship Hospital, Capital Medical University

Herein, we present a three-dimensional printing guide template for percutaneous vertebraplasty. A patient with a T11 vertebral compression fracture was selected as a case study.

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Bioengineering

Databases to Efficiently Manage Medium Sized, Low Velocity, Multidimensional Data in Tissue Engineering
Alexander R. Ochs 1,2, Mehrsa Mehrabi 1,2, Danielle Becker 1,2, Mira N. Asad 1,2, Jing Zhao 1,2, Michael V. Zaragoza 3,4, Anna Grosberg 1,2,5,6,7
1Department of Biomedical Engineering, University of California, Irvine, 2The Edwards Lifesciences Center for Advanced Cardiovascular Technology, University of California, Irvine, 3Pediatrics-Genetics & Genomics Division-School of Medicine, University of California, Irvine, 4Biological Chemistry-School of Medicine, University of California, Irvine, 5Department of Chemical and Biomolecular Engineering, University of California, Irvine, 6Center for Complex Biological Systems, University of California, Irvine, 7The NSF-Simons Center for Multiscale Cell Fate Research (CMCF), University of California, Irvine

Many researchers generate "medium-sized", low-velocity, and multi-dimensional data, which can be managed more efficiently with databases rather than spreadsheets. Here we provide a conceptual overview of databases including visualizing multi-dimensional data, linking tables in relational database structures, mapping semi-automated data pipelines, and using the database to elucidate data meaning.

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Environment

Studying Neurobehavioral Effects of Environmental Pollutants on Zebrafish Larvae
Bin Zhang 1,2, Xinyue Yang 2, Jing Zhao 3, Ting Xu 2,4, Daqiang Yin 2,4
1State Key Laboratory of Marine Geology, Tongji University, 2Key Laboratory of Yangtze River Water Environment, Ministry of Education, College of Environmental Science and Engineering, Tongji University, 3Shanghai Collaborative Innovation Centre for WEEE Recycling, WEEE Research Center of 10 Shanghai Polytechnic University, 4Shanghai Institute of Pollution Control and Ecological Security

A detailed experimental protocol is presented in this paper for the evaluation of neurobehavioral toxicity of environmental pollutants using a zebrafish larvae model, including the exposure process and tests for neurobehavioral indicators.

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Chemistry

Direct-Coupled Electroretinogram (DC-ERG) for Recording the Light-Evoked Electrical Responses of the Mouse Retinal Pigment Epithelium
Kiyoharu J. Miyagishima 1, Congxiao Zhang *2, Volha V. Malechka *3, Kapil Bharti 2, Wei Li 1
1Retinal Neurophysiology Section, National Eye Institute, National Institutes of Health, 2Ocular and Stem Cell Translational Research Section, National Eye Institute, National Institutes of Health, 3Human Visual Function Core, National Eye Institute, National Institutes of Health

Here, we present a method for recording light-evoked electrical responses of the retinal pigment epithelium (RPE) in mice using a technique known as DC-ERGs first described by Marmorstein, Peachey, and colleagues in the early 2000s.

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Engineering

Design and Development of a Three-Dimensionally Printed Microscope Mask Alignment Adapter for the Fabrication of Multilayer Microfluidic Devices
Celine R. Garcia *1, Zhenya Ding *1, Hilario C. Garza 1, Wei Li 1
1Department of Chemical Engineering, Texas Tech University

This project allows small laboratories to develop an easy-to-use platform for the fabrication of precise multilayer microfluidic devices. The platform consists of a three-dimensionally printed microscope mask alignment adapter using which multilayer microfluidic devices with alignment errors of <10 µm were achieved.

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Biology

Two-layered Membrane Sandwich Method for Laodelphax striatellus Saliva Collection
Jing Zhao 1,2,3, Jie Yang 1,2,3, Lili Zhang 1,2, Rongxiang Fang 1,2, Yan Huo 1,2
1State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, 2National Plant Gene Research Center, 3University of the Chinese Academy of Sciences

The present protocol describes a method to collect sufficient saliva from piercing-sucking insects using an artificial medium. This is a convenient method for collecting insect saliva and studying salivary function on insect feeding behavior and vector-borne virus transmission.

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Medicine

Laparoscopy-endoscopy Cooperative Surgery for the Treatment of Gastric Gastrointestinal Stromal Tumors
Dongjie Yang *1,2, Xun Hou *1, Wentong Lan *3, Minxuan Shen 1, Shaohua Yang 4, Lele Zhang 1, Qinbo Cai 1, Yan Qian 1, Yuan Lin 5, Xia Feng 6, Yulong He 1,2,4
1Center for Gastrointestinal Surgery, The First Affiliated Hospital, Sun Yat-sen University, 2Guangdong Provincial Key Laboratory of Digestive Cancer Research, 3Department of Endoscopy, The First Affiliated Hospital, Sun Yat-sen University, 4Digestive Medicine Center, The Seventh Affiliated Hospital, Sun Yat-sen University, 5Department of Pathology, The First Affiliated Hospital, Sun Yat-sen University, 6Department of Anesthesiology, The First Affiliated Hospital, Sun Yat-sen University

Laparoscopy-endoscopy cooperative surgery is appropriate for the treatment of gastric gastrointestinal stromal tumors less than 5 cm in size. It can achieve the respective advantages of endoscopic and laparoscopic surgeries while avoiding their drawbacks.

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Engineering

The Traceable Calibration of Thrust Stand by Electrostatic Force
Bin Wang 1,2, Yong Yang 3, Zhan Hu 1,2, Yelong Zheng 3
1Shanghai Engineering Center for Microsatellites, 2Innovation Academy for Microsatellites, Chinese Academy of Sciences, 3State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University

Traceability calibration of mechanical characteristics of thrust stand is an essential prerequisite to ensure traceability measurement of thrust. Here, we describe how to calibrate the thrust stand by the electrostatic force generated by the parallel plate capacitor.

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Biology

Primary Culture of Porcine Retinal Pigment Epithelial Cells
Feng Wen *1,2,3, Yanzi Wang *1,2,3, Danxue He 1,2,3, Chunyan Liao 1,2,3, Weijie Ouyang 1,2,3, Zuguo Liu 1,2,3, Wei Li 1,2,3, Yi Liao 1,2,3
1Eye Institute of Xiamen University, Fujian Provincial Key Laboratory of Ophthalmology and Visual Science, School of Medicine, Xiamen University, 2Department of Ophthalmology, Xiang'an Hospital of Xiamen University, School of Medicine, Xiamen University, 3Xiamen University Affiliated Xiamen Eye Center, School of Medicine, Xiamen University

Here, an easy-to-follow method to culture primary porcine retinal pigment epithelial cells in vitro is presented.

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JoVE Journal

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
Lifang Yu *1, Yadan Zhang *1, Mario Andrea Marchisio 1
1School of Pharmaceutical Science and Technology, Tianjin University

CRISPR-Cas systems and anti-CRISPR proteins were integrated into the scheme of Boolean gates in Saccharomyces cerevisiae. The new small logic circuits showed good performance and deepened the understanding of both dCas9/dCas12a-based transcription factors and the properties of anti-CRISPR proteins.

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Engineering

A Micro-Displacement Measurement Based on the Shadow Technique
Yong Yang 1, Meirong Zhao 1, Yinguo Huang 1, Yu Tian 2, Yelong Zheng 1
1State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University, 2State Key Laboratory of Tribology, Department of Mechanical Engineering, Tsinghua University

High-precision micro-displacement measurement is significant in the field of aerospace engineering, ultra-precision machining, and micro-assembly. The present protocol describes measuring micro displacements based on the shadow technique.

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Biology

A Magnetic Separation-Assisted High-Speed Homogenization Method for Large-Scale Production of Endosome-Derived Vesicles
Dujuan Wang 1,2, Shili Yao 2,3, Peng Guo 1,2,3,4
1Academy of Medical Engineering and Translational Medicine, Tianjin University, 2Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences, 3School of Materials Science and Engineering, Tianjin University, 4Zhejiang Cancer Hospital

Here, we describe a magnetic separation-assisted high-speed homogenization method for large-scale production of endosome-derived nanovesicles as a new type of exosome mimics (EMs) that share the same biological origin and similar structure, morphology, and protein composition of native extracellular vesicles (EVs).

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Bioengineering

Fabrication and Characterization of Microneedle Patches for Loading and Delivery of Exosomes
Sijia Mu 1,2, Hao Chang 2, Fengli Qu 2
1Medical School, Faculty of Medicine, Tianjin University, 2Hangzhou Institute of Medicine, Chinese Academy of Sciences

Exosomes possess significant clinical potential, but their practical application is limited due to easy in vivo clearance and poor stability. Microneedles present a solution by enabling localized delivery by puncturing physiological barriers and dry-state preservation, thereby addressing the limitations of exosome administration and expanding their clinical utility.

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