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Zhejiang University

47 ARTICLES PUBLISHED IN JoVE

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Biology

Measuring the Bending Stiffness of Bacterial Cells Using an Optical Trap
Siyuan Wang 1, Hugo Arellano-Santoyo 2, Peter A. Combs 2, Joshua W. Shaevitz 2
1Department of Molecular Biology, Lewis-Sigler Institute for Integrative Genomics, Princeton University, 2Department of Physics, Lewis-Sigler Institute for Integrative Genomics, Princeton University

We present a protocol for bending filamentous bacterial cells attached to a cover-slip surface with an optical trap to measure the cellular bending stiffness.

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Bioengineering

Elastomeric PGS Scaffolds in Arterial Tissue Engineering
Kee-Won Lee 1, Yadong Wang 1,2
1Department of Bioengineering, University of Pittsburgh, 2McGowan Institute for Regenerative Medicine, University of Pittsburgh

Elastomeric PGS scaffolds with vascular smooth muscle cells cultured in a pulsatile flow bioreactor may lead to promising small-diameter arterial constructs with native ECM production in a relatively short culture period.

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Biology

Measuring Spatial and Temporal Ca2+ Signals in Arabidopsis Plants
Xiaohong Zhu 1, Aaron Taylor 2, Shenyu Zhang 1, Dayong Zhang 1,3, Ying Feng 1,4, Gaimei Liang 1,5, Jian-Kang Zhu 1,6
1Department of Horticulture and Landscape Architecture, Purdue University, 2Bindley Bioscience Center, Purdue University, 3Institute of Biotechnology, Jiangsu Academy of Agricultural Sciences, 4College of Environmental & Resource Science, Zhejiang University, 5Dryland Agriculture Research Centre, Shanxi Academy of Agricultural Sciences, 6Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences

Ca2+ signaling regulates diverse biological processes in plants. Here we present approaches for monitoring abiotic stress induced spatial and temporal Ca2+ signals in Arabidopsis cells and tissues using the genetically encoded Ca2+ indicators Aequorin or Case12.

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Bioengineering

Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell
Yunfeng Chen *1, Baoyu Liu *2, Lining Ju *3, Jinsung Hong *1, Qinghua Ji 4,5, Wei Chen 6, Cheng Zhu 1
1Woodruff School of Mechanical Engineering, Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, 2Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 3Charles Perkins Centre, The University of Sydney, 4Institute of Biophysics, Laboratory of RNA Biology, Chinese Academy of Sciences, 5University of Chinese Academy of Sciences, 6School of Medicine and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University

We describe a technique for concurrently measuring force-regulated single receptor-ligand binding kinetics and real-time imaging of calcium signaling in a single T lymphocyte.

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Biology

Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy
Jingwen Yang 1, Yunjun Yan 1, Xiaowei Xiang 1, Yuchao Xu 1, Naiming Zhou 2, Tianming Wang 1
1National Engineering Research Center of Marine Facilities Aquaculture, Marine Science College, Zhejiang Ocean University, 2Institute of Biochemistry, College of Life Sciences, Zhejiang University

This protocol describes confocal microscopy detection of G protein-coupled receptor (GPCR) internalization in mammalian cells. It includes the basic cell culture, transfection, and confocal microscopy procedure and provides an efficient and easily interpretable method to detect the subcellular localization and internalization of fusion-expressed GPCR.

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Biology

Methods for the Extraction of Endosymbionts from the Whitefly Bemisia tabaci
Dan-Tong Zhu 1, Xin-Ru Wang 1, Fei-Xue Ban 1, Chi Zou 1, Shu-Sheng Liu 1, Xiao-Wei Wang 1
1Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogen and Insects, Institute of Insect Sciences, Zhejiang University

Here, we present a protocol to isolate endosymbionts from the whitefly Bemisia tabaci through dissection and filtration. After amplification, the DNA samples are suitable for subsequent sequencing and study of the mutualism between endosymbionts and whitefly.

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Neuroscience

Experimental Models to Study the Neuroprotection of Acidic Postconditioning Against Cerebral Ischemia
Yanrong Zheng *1, Zhe Shen *1, Xiaoli Wu 1, Lei Jiang 1, Weiwei Hu 1, Zhong Chen 1, Xiangnan Zhang 1
1Institute of Pharmacology & Toxicology, College of Pharmaceutical Sciences, Department of Pharmacology, Key Laboratory of Medical Neurobiology of The Ministry of Health of China, Zhejiang Province Key Laboratory of Neurobiology, Zhejiang University

Acidic postconditioning protects against cerebral ischemia. Here we present two models to execute APC. They are achieved respectively by transferring corticostriatal slices to acidic buffer after oxygen-glucose deprivation in vitro and by inhaling 20% CO2 after middle cerebral artery occlusion in vivo.

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Cancer Research

Detection of Rare Mutations in CtDNA Using Next Generation Sequencing
Xiaoxing Lv 1, Meiru Zhao 1, Yuting Yi 1, Lucheng Zhang 1, Yanfang Guan 1, Tao Liu 1, Ling Yang 1, Rongrong Chen 1, Jianhui Ma 1, Xin Yi 1
1Geneplus-Beijing Institute

This manuscript describes a technique for detecting mutations of low frequency in ctDNA, ER-Seq. This method is differentiated by its unique use of two-directional error correction, a special background filter, and efficient molecular acquirement.

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Chemistry

Co-expression of Multiple Chimeric Fluorescent Fusion Proteins in an Efficient Way in Plants
Xiaomin Peng *1, Guitao Zhong *1, Hao Wang 1
1College of Life Sciences, South China Agricultural University

We have developed a novel method for co-expressing multiple chimeric fluorescent fusion proteins in plants to overcome the difficulties of conventional methods. It takes advantage of using a single expression plasmid that contains multiple functionally independent protein expressing cassettes to achieve protein co-expression.

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Neuroscience

Preparation of Acute Spinal Cord Slices for Whole-cell Patch-clamp Recording in Substantia Gelatinosa Neurons
Mengye Zhu 1, Daying Zhang 1, Sicong Peng 2, Nana Liu 2, Jing Wu 2, Haixia Kuang 2, Tao Liu 2,3
1Department of Pain, First Affiliated Hospital of Nanchang University, 2Department of Pediatrics, First Affiliated Hospital of Nanchang University, 3Center for Laboratory Medicine, First Affiliated Hospital of Nanchang University

Here, we describe the essential steps for whole-cell patch-clamp recordings made from substantia gelatinosa (SG) neurons in the in vitro spinal cord slice. This method allows the intrinsic membrane properties, synaptic transmission and morphological characterization of SG neurons to be studied.

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Immunology and Infection

Effects of Taste Signaling Protein Abolishment on Gut Inflammation in an Inflammatory Bowel Disease Mouse Model
Ya-Wen Du *1, Qun Liu *1, Xiao-Cui Luo 1, Dong-Xiao Zhao 1, Jian-Bo Xue 1, Pu Feng 2, Robert F. Margolskee 2, Hong Wang 2, Liquan Huang 1,2
1College of Life Sciences, Zhejiang University, 2Monell Chemical Senses Center

Here we present a protocol to investigate the effect of the nullification of gustation-related genes on immune responses in a dextran sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) mouse model.

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Neuroscience

Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice
Liang Wen *1, Wendong You *1, Yadong Wang *1, Yuanrun Zhu 1, Hao Wang 1, Xiaofeng Yang 1
1Emergency and Trauma Center, The International Medical Center, The First Affiliated Hospital, School of Medicine, Zhejiang University

Presented here is a protocol to induce diffuse traumatic brain injury using a lateral fluid percussion device followed by the collection of the caecum content for gut microbiome analysis.

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Environment

Visualization of Flow Field Around a Vibrating Pipeline Within an Equilibrium Scour Hole
Dawei Guan 1,2, Yee-Meng Chiew 2, Maoxing Wei 3, Shih-Chun Hsieh 4
1Key Laboratory of Coastal Disaster and Defense, Ministry of Education, Hohai University, 2School of Civil and Environmental Engineering, Nanyang Technological University, 3Ocean College, Zhejiang University, 4AXESEA Information Technology Co. LTD

The goal of the protocol is to enable visualization of the detailed flow fields and determination of the near-boundary shear and normal stresses within an equilibrium scour hole induced by a vibrating pipeline.

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Behavior

Design and Use of an Apparatus for Presenting Graspable Objects in 3D Workspace
Kedi Xu 1,2,3, Junjun Chen 1,2, Guanghao Sun 1,2, Yaoyao Hao 1,3, Shaomin Zhang 1,2,3, Xingchen Ran 1,2, Weidong Chen 1,2, Xiaoxiang Zheng 1,2,3
1Qiushi Academy for Advanced Studies, Zhejiang University, 2Department of Biomedical Engineering, Zhejiang University, 3Key Laboratory of Biomedical Engineering of Ministry of Education, Zhejiang University

Presented here is a protocol to build an automatic apparatus that guides a monkey to perform the flexible reach-to-grasp task. The apparatus combines a 3D translational device and turning table to present multiple objects in an arbitrary position in 3D space.

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Neuroscience

The Detection of 5-Hydroxymethylcytosine in Neural Stem Cells and Brains of Mice
Yingliang Zhuang 1,2, Junchen Chen 1,2, Weize Xu 1, Qiang Shu 1, Xuekun Li 1,2
1The Children's Hospital, School of Medicine, Zhejiang University, 2The Institute of Translational Medicine, School of Medicine, Zhejiang University

Here, we present a protocol to detect 5-hydroxymethylcytosine in cells and brain tissues, utilizing immunofluorescence staining and DNA dot-blot methods.

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Genetics

Identifying Mutations by High Resolution Melting in a TILLING Population of Rice
Shan Li 1, Ying-Ping Yu 1, Song-Mei Liu 1, Hao-Wei Fu 2, Jian-Zhong Huang 3, Qing-Yao Shu 1, Yuan-Yuan Tan 4
1National Key Laboratory of Rice Biology, Zhejiang Provincial Key Laboratory of Crop Genetic Resources, Institute of Crop Science, Zhejiang University, 2Jiaxing Academy of Agricultural Sciences, 3Institute of Nuclear Agricultural Sciences, Zhejiang University, 4The New Countryside Development Institute at Zhejiang University

In this article, we present the protocol that is described as high-resolution melting analysis (HRM)-based Target Induced Local Lesions In Genomes (TILLING). This method utilizes fluorescence changes during the melting of the DNA duplex and is suitable for high-throughput screening of both insertion/deletion (Indel) and single base substition (SBS).

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Biology

Light Spot-Based Assay for Analysis of Drosophila Larval Phototaxis
Yixuan Sun 1, Peipei Zhou 1, Qianhui Zhao 1, Zhefeng Gong 1
1Department of Neurobiology, Zhejiang University

This protocol introduces a light-spot assay to investigate Drosophila larval phototactic behavior. In this assay, a light spot is generated as light stimulation, and the process of larval light avoidance is recorded by an infrared light-based imaging system.

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Bioengineering

Protocols of 3D Bioprinting of Gelatin Methacryloyl Hydrogel Based Bioinks
Mingjun Xie 1,2, Kang Yu 1,2, Yuan Sun 1,2, Lei Shao 1,2, Jing Nie 1,2, Qing Gao 1,2, Jingjiang Qiu 3, Jianzhong Fu 1,2, Zichen Chen 1,2, Yong He 1,2
1State Key Laboratory of Fluid Power and Mechatronic Systems, School of Mechanical Engineering, Zhejiang University, 2Key Laboratory of 3D Printing Process and Equipment of Zhejiang Province, School of Mechanical Engineering, Zhejiang University, 3School of Mechanics and Safety Engineering, Zhengzhou University

Presented here is a method for the 3D bioprinting of gelatin methacryloyl.

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Cancer Research

Establishing a Competing Risk Regression Nomogram Model for Survival Data
Lunpo Wu 1,2, Chenyang Ge 3, Hongjuan Zheng 4, Haiping Lin 5, Wei Fu 6, Jianfei Fu 4
1Department of Gastroenterology, Second Affiliated Hospital of Zhejiang University School of Medicine, 2Institute of Gastroenterology, Zhejiang University, 3Department of Colorectal Surgery, Afiliated Jinhua Hospital, Zhejiang University School of Medicine, 4Department of Medical Oncology, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, 5Department of Hepatobiliary Pancreatic Surgery, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, 6Division of Oncology, Johns Hopkins University School of Medicine

Presented here is a protocol to build nomograms based on the Cox proportional hazards regression model and competing risk regression model. The competing method is a more rational method to apply when competing events are present in the survival analysis.

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Immunology and Infection

Localization and Quantification of Begomoviruses in Whitefly Tissues by Immunofluorescence and Quantitative PCR
Fei-Xue Ban 1, Tian-Yan Yin 1, Qi Guo 1, Li-Long Pan 1, Yin-Quan Liu 1, Xiao-Wei Wang 1
1Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogen and Insects, Institute of Insect Sciences, Zhejiang University

We describe an immunofluorescence and quantitative PCR method for the localization and quantification of begomoviruses in insect tissues. The immunofluorescence protocol can be used to colocalize viral and vector proteins. The quantitative PCR protocol can be extended to quantify viruses in whole whitefly bodies and virus-infected plants.

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Medicine

OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery
Tao Liu *1,2,3,4,5, Xueling Song *1,2,3,4,5, Xiaoying Zheng *1,2,3,4,5, Liying Yan 1,2,3,4,5, Caihong Ma 1,2,3,5, Rong Li 1,2,3,4,5, Jie Yan 1,2,3,4,5, Jie Qiao 1,2,3,4,5
1Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, 2National Clinical Research Center for Obstetrics and Gynecology, 3Key Laboratory of Assisted Reproduction, Ministry of Education, 4Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction, 5Research Units of Comprehensive Diagnosis and Treatment of Oocyte Maturation Arrest, Chinese Academy of Medical Sciences

In vitro maturation (IVM) before gynecological operation (OP-IVM) combines IVM following oocyte retrieval with routine gynecological surgery and serves as an extension of conventional IVM applications for fertility preservation.

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Biology

High-Throughput Metabolic Profiling for Model Refinements of Microalgae
Amnah Alzahmi 1,2, Sarah Daakour 1, Diana Charles El Assal 3, Bushra S. Dohai 3, Amphun Chaiboonchoe 3, Weiqi Fu 3,4, David R. Nelson 1, Alexandra Mystikou 1, Kourosh Salehi-Ashtiani 1,3
1Center for Genomics and Systems Biology (CGSB), New York University Abu Dhabi Research Institute, 2Department of Biology, United Arab Emirates University, 3Laboratory of Algal, Systems, and Synthetic Biology (LASSB), Division of Science and Math, New York University Abu Dhabi, 4Department of Marine Science, Ocean College, Zhejiang University

This protocol demonstrates the use of a phenotype microarray (PM) technology platform to define metabolic requirements of Chlamydomonas reinhardtii, a green microalga, and refine an existing metabolic network model.

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Biology

Profiling of H3K4me3 Modification in Plants using Cleavage under Targets and Tagmentation
Xiaoyuan Tao *1,2, Mengtao Gao *3, Siyuan Wang *1, Xueying Guan 1,4
1College of Agriculture and Biotechnology, Zhejiang University, 2Central Laboratory, State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Zhejiang Academy of Agricultural Sciences, 3State Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Hybrid R & D Engineering Center (the Ministry of Education), College of Agriculture, Nanjing Agricultural University, 4Hainan Institute of Zhejiang University

Cleavage under targets and tagmentation (CUT&Tag) is an efficient chromatin epigenomic profiling strategy. This protocol presents a refined CUT&Tag strategy for the profiling of histone modifications in plants.

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Environment

Isolation of Quartz Grains for Optically Stimulated Luminescence (OSL) Dating of Quaternary Sediments for Paleoenvironmental Research
Liliana C. Marin 1, Steven L. Forman 1, Victoria T. Todd 1, Connor Mayhack 1, Ashley Gonzalez 1, Peng Liang 2
1Geoluminescence Dating Research Laboratory, Dept. of Geosciences, Baylor University, 2School of Earth Sciences, Zhejiang University

This protocol is for the isolation of quartz grains by size for luminescence dating of sediments. Outlined are physical cleansing and chemical digestions by soaking sequentially in H2O2, HCl, HF, and HCl again to isolate quartz grains. The quartz purity is quantified with microscopic assessment, Raman spectroscopy, and IR depletion ratio.

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Developmental Biology

An Explant System for Time-Lapse Imaging Studies of Olfactory Circuit Assembly in Drosophila
Tongchao Li 1, Liqun Luo 1
1Department of Biology, Howard Hughes Medical Institute, Department of Biology, Stanford University

This protocol describes the dissection procedure, culture condition, and live imaging of an antennae-brain explant system for the study of the olfactory circuit assembly.

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Medicine

Indocyanine Green-Guided Intraoperative Imaging to Facilitate Video-Assisted Retroperitoneal Debridement for Treating Acute Necrotizing Pancreatitis
Li Huang 1, Wei Chen 1, Jiancong Chen 1, Dong Chen 1, Kunsong Zhang 1, Jianpeng Cai 1, Hong Peng 1, Chensong Huang 1, Guangyan Zeng 1, Mingjian Ma 1, Jiahua Liang 1, Borui Xu 1, Xiaoyu Yin 1, Jiaming Lai 1, Lijian Liang 1
1Department of Pancreatobiliary Surgery, The First Affiliated Hospital, Sun Yat-sen University

This protocol presents Indocyanine Green-guided video-assisted retroperitoneal debridement (ICG-guided VARD) for treating severe acute necrotizing pancreatitis.

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Medicine

The Miniature Pig: A Large Animal Model for Cochlear Implant Research
Xiaojun Ji *1,2,3,4, Yi Luo *1,2,3,4, Weiwei Guo 1,2,3,4, Fei Ji 1,2,3,4, Shuolong Yuan 1,2,3,4, Liangwei Xu 1,2,3,4, Wei Chen *1,2,3,4
1College of Otolaryngology Head and Neck Surgery, Chinese PLA General Hospital, 2National Clinical Research Center for Otolaryngologic Diseases, 3Key Lab of Hearing Science, Ministry of Education, 4Beijing Key Lab of Hearing Impairment for Prevention and Treatment

Miniature pigs (mini-pigs) are an ideal large animal model for research into cochlear implants. Cochlear implantation surgery in mini-pigs can be utilized to provide initial evidence of the safety and potential performance of novel electrode arrays and surgical approaches in a living system similar to human beings.

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Biology

Analyzing Starvation-Induced Autophagy in the Drosophila melanogaster Larval Fat Body
Kexin Shi 1, Chao Tong 1
1Ministry of Education Key Laboratory of Biosystems Homeostasis and Protection and Innovation Center for Cell Signaling Network, Life Sciences Institute, Zhejiang University

The present protocol describes the induction of autophagy in the Drosophila melanogaster larval fat body via nutrient depletion and analyzes changes in autophagy using transgenic fly strains.

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Medicine

Laminectomy for the Removal of Thoracic Ossification of the Ligamentum Flavum (TOLF) Using Ultrasonic and Conventional Osteotomes
Junhui Liu 1,2, Zhi Shan 1,2, Yilei Chen 1,2, Fengdong Zhao 1,2
1Department of Orthopedics, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, 2Key Laboratory of Musculoskeletal System Degeneration and Regeneration Translational Research of Zhejiang Province

We present a methodology for using an ultrasonic osteotome combined with a conventional osteotome for laminectomy in thoracic ossification of the ligamentum flavum. This is a relatively safe and easy-to-learn method that avoids the perioperative complications associated with the conventional method.

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Biology

An Electroporation Method to Transform Rickettsia spp. with a Fluorescent Protein-Expressing Shuttle Vector in Tick Cell Lines
Xin-Ru Wang 1, Nicole Y. Burkhardt 1, Lisa D. Price 1, Ulrike G. Munderloh 1
1Department of Entomology, University of Minnesota

Electroporation is a rapid, broadly adopted method for introducing exogenous DNA into the genus Rickettsia. This protocol provides a useful electroporation method for the transformation of obligate intracellular bacteria in the genus Rickettsia.

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Methods And Protocols For Live Imaging In Development
Tongchao Li 1,2,3
1Department of Neurobiology and Department of Neurosurgery of Second Affiliated Hospital, Zhejiang University School of Medicine, 2Liangzhu Laboratory, MOE Frontier Science Center for Brain Science and Brain-machine Integration, State Key Laboratory of Brain-machine Intelligence, Zhejiang University, 3NHC and CAMS Key Laboratory of Medical Neurobiology, Zhejiang University

Methods And Protocols For Live Imaging In Development

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Behavior

Usability Evaluation of Augmented Reality: A Neuro-Information-Systems Study
Jun Wu *1, Di Zhang *1, Tao Liu 2,3,4, Helen Hong Yang 5, Yi Wang 1, Huili Yao 1, Shinan Zhao 1
1Economics and Management School, Jiangsu University of Science and Technology, 2School of Health, Fujian Medical University, 3School of Management, Shanghai University, 4School of Management, Zhejiang University, 5Business School, La Trobe University

This study presents an experimental paradigm for a usability test combining subjective and objective evaluations. The objective evaluation adopted Neuro-Information-Systems (NeuroIS) methods, and the subjective evaluation adopted a usability questionnaire and a NASA-Task Load Index (NASA-TLX) scale.

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Engineering

Sensitivity Enhancement of Soft Capacitive Pressure Sensors Using a Solvent Evaporation-Based Porosity Control Technique
Ziying Zhu 1, Yunqi Cao 1, Haozhen Chi 1, Xiaowei Wang 1, Dibo Hou 1
1State Key Laboratory of Industrial Control Technology, College of Control Science and Engineering, Zhejiang University

A simple and cost-efficient fabrication method based on the solvent evaporation technique is presented to optimize the performance of a soft capacitive pressure sensor, which is enabled by porosity control in the dielectric layer using different mass ratios of the molding PDMS/toluene solution.

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Neuroscience

Modeling the Functional Network for Spatial Navigation in the Human Brain
Fengxiang Zhang *1, Chenghui Zhang *1, Yi Pu 2, Xiang-Zhen Kong 1,3
1Department of Psychology and Behavioral Sciences, Zhejiang University, 2Department of Neuroscience, Max Planck Institute for Empirical Aesthetics, 3Department of Psychiatry of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine

This paper presents an integrative approach to investigating the functional network for spatial navigation in the human brain. This approach incorporates a large-scale neuroimaging meta-analytic database, resting-state functional magnetic resonance imaging, and network modeling and graph-theoretical techniques.

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Engineering

Microfluidic Channel-Based Soft Electrodes and Their Application in Capacitive Pressure Sensing
Xiaowei Wang 1, Peijun Shangguan 1, Pingjie Huang 1, Dibo Hou 1
1State Key Laboratory of Industrial Control Technology, College of Control Science and Engineering, Zhejiang University

Flexible electrodes have a wide range of applications in soft robotics and wearable electronics. The present protocol demonstrates a new strategy to fabricate highly stretchable electrodes with high resolution via lithographically defined microfluidic channels, which paves the way for future high-performance soft pressure sensors.

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Bioengineering

Quantitative Characterization of Liquid Photosensitive Bioink Properties for Continuous Digital Light Processing Based Printing
Yang Li *1,2,3, Yue Wang *4, Jun Yin 3, Jin Qian 1,2
1School of Aeronautics and Astronautics, Zhejiang University, 2Department of Engineering Mechanics, Key Laboratory of Soft Machines and Smart Devices of Zhejiang Province, Zhejiang University, 3The State Key Laboratory of Fluid Power and Mechatronic Systems, School of Mechanical Engineering, Zhejiang University, 4Institute of Artificial Intelligence, School of Future Technology, Shanghai University

This study uses temperature and material composition to control the yield stress properties of yield stress fluids. The solid-like state of the ink can protect the printing structure, and the liquid-like state can continuously fill the printing position, realizing the digital light processing 3D printing of extremely soft bioinks.

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Biology

Hand-Rearing Method for Infant Marmosets
Hao Sun 1,2, Rui Li 1, Yingxu Lin 1,2, Xinyuan Cao 1, Lingzhu Fan 1, Guanglong Sun 1, Min Xie 1, Lin Zhu 1, Chencen Yu 1, Ruolan Cai 1, Chenfei Lyu 1, Xiaohui Wang 1,2, Yuanqing Zhang 1,2, Siyi Bai 1,2, Runze Qi 1, Binliang Tang 1,3, Guoqiang Jia 1,4, Xinjian Li 1,2,4, Lixia Gao 1,2,4
1Department of Neurology of the Second Affiliated Hospital and Interdisciplinary Institute of Neuroscience and Technology, School of Medicine, Zhejiang University, 2Key Laboratory of Biomedical Engineering of Ministry of Education, College of Biomedical Engineering and Instrument Science, Zhejiang University, 3Rehabilitation Medicine Center, Rehabilitation & Sports Medicine Research Institute of Zhejiang Province, Department of Rehabilitation Medicine, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital, Hangzhou Medical College, 4NHC and CAMS Key Laboratory of Medical Neurobiology, MOE Frontier Science Center for Brain Science and Brain-Machine Integration, School of Brain Science and Brain Medicine, Zhejiang University

Here, we describe a hand-rearing method for raising infant marmosets in an animal incubator. This method greatly increases the survival rate of marmoset infants, which provides the opportunity to study the development of marmoset infants with similar genetic backgrounds raised in different postnatal environments.

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Neuroscience

New Framework for Understanding Cross-Brain Coherence in Functional Near-Infrared Spectroscopy (fNIRS) Hyperscanning Studies
Hila Zahava Gvirts Provolovski 1, Mini Sharma 1, Itai Gutman 1, Anat Dahan 2, Yafeng Pan 3, Jacqueline Stotler 4, Teresa Wilcox 4
1The Department of Behavioral Sciences and Psychology, Ariel University, 2Braude College of Engineering, 3Department of Psychology and Behavioral Sciences, Zhejiang University, 4Department of Psychology, Florida Atlantic University

Wavelet transform coherence (WTC) is a common methodology for assessing the coupling between signals that is used in functional near-infrared spectroscopy (fNIRS) hyperscanning studies. A toolbox for assessing the directionality of the signal interaction is presented in this work.

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Medicine

Predictive Measurement for Windlass Change in Length and Selected Treatment Outcomes in Chronic Plantar Fasciitis
Sahar Ahmed Abdalbary 1, Ahmad Alhaskawi 2, Haiying Zhou 2, Yanzhao Dong 2, Tian Tu 3, Pengfei Li 3, Hui Lu 2
1Department of Orthopaedic Physical Therapy, Faculty of Physical Therapy, Nahda University in Beni Suef, 2Department of Orthopedics, The First Affiliated Hospital, Zhejiang University, 3Department of Plastic and Aesthetic Center, First Affiliated Hospital, College of Medicine, Zhejiang University

This protocol presents an objective, easy, and economical method to measure the change in length of the windlass in plantar fasciitis and assess the effectiveness of a selected treatment procedure employed in this study over a one-month period.

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Medicine

Oblique Lumbar Interbody Fusion at L5-S1 Segment Through an Approach Between the Psoas Muscle and the Great Vessels
Xigong Li *1, Yanzhao Dong *1, Haiying Zhou 1, Ahmad Alhaskawi 1, Zhongyang Gao 1, Yuzhu Zhang 1, Hui Lu 1
1Department of Orthopedics, The First Affiliated Hospital, Zhejiang University

The protocol presents an OLIF L5-S1 technique that offers a more feasible approach for lumbosacral fusion sharing a common surgical plane with OLIF L2-5. This facilitates reproducible multi-level interbody fusions extending from L2 to S1 through a retroperitoneal oblique corridor between the Psoas muscle and the great vessels.

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Biology

Comparison of Two Representative Methods for Differentiation of Human Induced Pluripotent Stem Cells into Mesenchymal Stromal Cells
Fangqin Wang 1, Langping Gao 2,3, Xudong Fu 4, Qintao Yan 2, Lidan Hu 1,2, Jianhua Mao 2,3
1National Clinical Research Center for Child Health, The Children's Hospital, Zhejiang University School of Medicine, 2Department of Nephrology, The Children's Hospital, Zhejiang University School of Medicine, 3Zhejiang University School of Medicine, 4Liangzhu Laboratory, Zhejiang University

This protocol describes and compares two representative methods for differentiating hiPSCs into mesenchymal stromal cells (MSCs). The monolayer method is characterized by lower cost, simpler operation, and easier osteogenic differentiation. The embryoid bodies (EBs) method is characterized by lower time consumption.

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Medicine

Metacarpal Small Incision for Carpal Tunnel Syndrome
Jingen Hu 1,2, Caibao Hu 3
1Department of Orthopedics, The First Affiliated Hospital, School of Medicine, Zhejiang University, 2Department of Orthopedics, Liangzhu Branch of the First Affiliated Hospital, School of Medicine, Zhejiang University (The First People's Hospital Of Yuhang District, Hangzhou), 3Intensive Care Unit, Zhejiang Hospital

Carpal tunnel decompression with transverse carpal ligament transection is a very successful treatment of carpal tunnel syndrome. The standard long-curved incision technique has become the best treatment for median nerve entrapment decompression. Here we introduce the metacarpal small incision carpal tunnel release (MSICTR) in the treatment of carpal tunnel syndrome (CTS).

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Medicine

Cognitive Function and Upper Limb Rehabilitation Training Post-Stroke Using a Digital Occupational Training System
Zhao Yao 1, Tianfang Zhang 2, Fenfen Chen 1,3, Weiyi Shi 2, Jinjin zheng 2, Ziwei Zhang 2, Zuobing Chen 1,2
1The Second Affiliated Hospital and Yuying Children's Hospital, Wenzhou Medical University, 2Department of Rehabilitation Medicine, The First Affiliated Hospital, School of Medicine, Zhejiang University, 3Department of Rehabilitation Medicine, Taizhou Hospital Affiliated to Wenzhou Medical University

The current protocol outlines how the VR-based digital occupational training system enhances the rehabilitation of patients with cognitive impairment and upper limb dysfunction following a stroke.

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Neuroscience

Regulating Schwann Cell Growth by Nanosecond Pulsed Electric Field for Peripheral Nerve Regeneration In Vitro
Jiahang Han *1,2, Zewei Wang *1,2, Yanzhao Dong *1, Xiaodi Zou 3, Haoyu Wang 1,2, Yonggang Chen 1, Sahar Ahmed Abdalbary 4, Tian Tu 5, Hui Lu 1
1Department of Orthopedics, The First Affiliated Hospital, College of Medicine, Zhejiang University, 2College of Medicine, Zhejiang University, 3Department of Orthopedics, The Second Affiliated Hospital of Zhejiang Chinese Medical University, 4Department of Orthopedic Physical Therapy, Faculty of Physical Therapy, Nahda University in Beni Suef, 5Plastic & Cosmetic Center, The First Affiliated Hospital, College of Medicine, Zhejiang University

Here, we present a protocol for applying nanosecond pulse electric field (nsPEF) to stimulate Schwann cells in vitro. The synthesis and secretion ability of relevant factors and cell behavior changes validated the successful stimulation using nsPEF. The study gives a positive view of the peripheral nerve regeneration method.

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Medicine

Combination of Anatomical Plates and Rotator Cuff Reinforcement for Minimally Invasive Treatment of Greater Tuberosity Fractures of the Humerus
Xiao Lu *1, Xiaodi Zou *2, Ahmad Alhaskawi 3, Yanzhao Dong 3, Sohaib Hasan Abdullah Ezzi 4, Vishnu Goutham Kota 5, Mohamed Hasan Abdulla Hasan Abdulla 5, Sahar Ahmed Abdalbary 6, Hui Lu 3,7
1Department of Orthopedics, Sanmen People's Hospital, 2Department of Orthopedics, The Second Affiliated Hospital of Zhejiang Chinese Medical University, 3Department of Orthopedics, The First Affiliated Hospital, Zhejiang University, 4Department of Orthopedics, Third Xiangya Hospital, Central South University, 5Zhejiang University School of Medicine, 6Department of Orthopedic Physical Therapy, Faculty of Physical Therapy, Nahda University in Beni Suef, 7Alibaba-Zhejiang University Joint Research Center of Future Digital Healthcare, Zhejiang University

Here, we present a protocol to utilize anatomical plates alongside rotator cuff reinforcement in a minimally invasive surgery to efficiently treat humerus greater tuberosity fractures, promising quicker healing and improved stability.

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Medicine

Enhancing Upper Limb Function and Motor Skills Post-Stroke Through an Upper Limb Rehabilitation Robot
Tianfang Zhang 1, Zhao Yao 1,2, Fenfen Chen 2,3, Jie Wang 4, Weiyi Shi 1, Jinjin Zheng 1, Ziwei Zhang 1, Zuobing Chen 1,2,3
1Department of Rehabilitation Medicine, The First Affiliated Hospital, School of Medicine, Zhejiang University, 2The Second Affiliated Hospital and Yuying Children's Hospital, Wenzhou Medical University, 3Department of Rehabilitation Medicine, Taizhou Hospital Affiliated to Wenzhou Medical University, 4Affiliate Hospital of Shandong University of Traditional Chinese Medicine

This protocol describes an upper limb rehabilitation robot that provides intelligent feedback through four modes. These modes enhance upper limb function and flexibility, thereby improving patients' quality of life.

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Medicine

Improved Thoracoscopic-Assisted Surgery for the Treatment of Metastatic Thoracic Vertebral Tumors
Xiao Long *1, Xiaodi Zou *1,2, Xiang Tong 1, Dongdong Yu 1, Hui Li 1, Yuxin Chang 1, Bin Chen 1, Hui Lu 1
1Department of Orthopedics, The First Affiliated Hospital, Zhejiang University, 2Department of Orthopedics, The Second Affiliated Hospital, Zhejiang Chinese Medical University

Here, we present a protocol to demonstrate an improved thoracoscopic-assisted surgery for the treatment of metastatic thoracic vertebral tumors.

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