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Zhongkai University of Agriculture and Engineering

7 ARTICLES PUBLISHED IN JoVE

Biology

Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System

Gang Tian¹, Qing Lu², Li Zhang², Susanne E. Kohalmi¹, Yuhai Cui²

¹Department of Biology, University of Western Ontario, ²Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada

We have developed a technique to test protein-protein interactions in plant. A yellow fluorescent protein (YFP) is split into two non-overlapping fragments. Each fragment is cloned in-frame to a gene of interest via Gateway system, enabling expression of fusion proteins. Reconstitution of YFP signal only occurs when the inquest proteins interact.

Medicine

Staining Protocols for Human Pancreatic Islets

Martha L. Campbell-Thompson¹, Tiffany Heiple¹, Emily Montgomery¹, Li Zhang¹, Lynda Schneider¹

¹Department of Pathology, Immunology, and Laboratory Medicine, University of Florida

This video demonstrates procedures for characterization of human pancreatic islets using hematoxylin and eosin (H&E) and immunohistochemistry (IHC). Pancreatic sections from head, body, and tail regions are stained by both H&E and IHC to determine islet endocrine composition (insulin, glucagon, and pancreatic polypeptide), cell replication (Ki67), and inflammatory infiltrates (H&E, CD3). The uncinate region is localized using IHC for pancreatic polypeptide.

Biology

Measurement of Heme Synthesis Levels in Mammalian Cells

Jagmohan Hooda¹, Maksudul Alam¹, Li Zhang¹

¹Department of Molecular and Cell Biology, Center for Systems Biology, University of Texas at Dallas

Altered intracellular heme levels are associated with common diseases such as cancer. Thus, there is a need to measure heme biosynthesis levels in diverse cells. The goal of this protocol is to provide a fast and sensitive method to measure and compare the levels of heme synthesis in different cells.

Immunology and Infection

High-Efficiency Generation of Antigen-Specific Primary Mouse Cytotoxic T Cells for Functional Testing in an Autoimmune Diabetes Model

Howard W. Davidson¹, Joseph Ray Cepeda², Nitin S. Sekhar², Junying Han², Ling Gao³, Tomasz Sosinowski¹, Li Zhang²

¹Barbara Davis Center for Childhood Diabetes, University of Colorado Denver, ²Department of Medicine, Endocrinology, Diabetes & Metabolism, Baylor College of Medicine, ³Scientific Center, Shandong Provincial Hospital affiliated to Shandong University

This article describes a protocol for the generation of antigen-specific CD8 T cells, and their expansion in vitro, with the aim of yielding high numbers of functional T cells for use in vitro and in vivo.

JoVE Core

International Expert Consensus and Recommendations for Neonatal Pneumothorax Ultrasound Diagnosis and Ultrasound-guided Thoracentesis Procedure

Jing Liu^1,2, Dalibor Kurepa³, Francesco Feletti^4,5, Almudena Alonso-Ojembarrena⁶, Jovan Lovrenski⁷, Roberto Copetti⁸, Erich Sorantin⁹, Javier Rodriguez-Fanjul¹⁰, Karishma Katti³, Andrea Aliverti⁴, Huayan Zhang^11,12, Misun Hwang¹³, Tsu F. Yeh¹⁴, Cai-Bao Hu¹⁵, Xing Feng¹⁶, Ru-Xin Qiu^1,2, Jing-Han Chi¹⁷, Li-Li Shang¹⁸, Guo-Rong Lyu¹⁹, Shao-Zheng He²⁰, Yan-Fen Chai²¹, Zhan-Jun Qiu²², Hai-Ying Cao^2,23, Yue-Qiao Gao^1,2, Xiao-Ling Ren^1,2, Guo Guo^1,24, Li Zhang^1,2, Ying Liu^1,2, Wei Fu^1,2, Zu-Lin Lu^1,2, Hong-Lei Li^1,2

¹Department of Neonatology and NICU, Beijing Chaoyang District Maternal and Child Healthcare Hospital, ²The National Neonatal Lung Ultrasound Training Base, ³Division of Neonatal-Perinatal Medicine, Cohen Children's Medical Center, ⁴Department of Electronics, Information and Bioengineering, Politecnico di Milano, ⁵Dipartimento di Diagnostica per Immagini, Ausl della Romagna, S. Maria delle Croci Hospital, ⁶Neonatal Intensive Care Unit, Puerta del Mar University Hospital, ⁷Faculty of Medicine, University of Novi Sad, Serbia, Institute for Children and Adolescents Health Care of Vojvodina, ⁸Emergency Department, University Hospital of Cattinara, ⁹Division of Pediatric Radiology, Department of Radiology, Medical University Graz, ¹⁰Pediatric Intensive Care Unit, Pediatric Service Hospital Joan XXIII Tarragona, University Rovira i Virgil, ¹¹Center for Newborn Care, Guangzhou Women and Children's Medical Center, ¹²Division of Neonatology, Children's Hospital of Philadelphia, ¹³Section of Neonatal Imaging, Department of Radiology, Children's Hospital of Philadelphia, ¹⁴Maternal Child Health Research institute, Taipei Medical University and China Medical University, ¹⁵Intensive Care Unit, Zhejiang Hospital, ¹⁶Department of Neonatology, Children's Hospital of Soochow University, ¹⁷Department of Neonatology and NICU, Bayi Children's Hospital Affiliated to the Seventh Medical Center of Chinese PLA General Hospital, ¹⁸Intensive Care Unit, Second Affiliated Hospital of Heilongjiang University of Chinese Medicine, ¹⁹Collaborative Innovation Center for Maternal and Infant Health Service Application Technology, Quanzhou Medical College, ²⁰Department of Ultrasound, Second Affiliated Hospital of Fujian Medical University, ²¹Department of Emergency Medicine, Tianjin Medical University General Hospital, ²²Department of Emergency and Critical Care Medicine, Affiliated Hospital of Traditional Chinese Medicine, ²³Department of Ultrasound, GE Healthcare, ²⁴The Neonatal Intensive Care Unit, Fifth Medical Center of Chinese PLA General Hospital

Pneumothorax is a common emergency and critical disease in newborn infants that needs rapid, clear diagnosis and timely treatment. Diagnosis and treatment based on chest X-rays are associated with delayed management and radiation damage. Lung ultrasound (US) provides useful guidance for rapid, accurate diagnosis and the precise thoracentesis of pneumothorax.

Biology

Isolation of Primary Rat Hepatocytes with Multiparameter Perfusion Control

Inn Chuan Ng^*1, Li Zhang^*2, Narelle Nichola Yi Ying Shen³, Yun Ting Soong⁴, Chan Way Ng⁵, Phoebe Kang Sheing Koh¹, Yan Zhou³, Hanry Yu^1,3,4,5,6

¹Department of Physiology & The Institute for Digital Medicine (WisDM), National University of Singapore, ²College of Agriculture and Biology, Zhongkai University of Agriculture and Engineering, ³Mechanobiology Institute, National University of Singapore, ⁴Institute of Bioengineering and Nanotechnology, A*STAR, ⁵NUS Graduate School for Integrative Sciences and Engineering, ⁶CAMP, Singapore-MIT Alliance for Research and Technology

This protocol details the use of a special intravenous catheter, standardized sterile disposable tubing, temperature control complemented by real-time monitoring, and an alarm system for two-step collagenase perfusion procedure to improve the consistency in the viability, yield, and functionality of isolated primary rat hepatocytes.

JoVE Journal

Genome-wide Analysis of Histone Modifications Distribution using the Chromatin Immunoprecipitation Sequencing Method in Magnaporthe oryzae

Zechi Wu^*1, Wanyu Sun^*1, Sida Zhou¹, Li Zhang¹, Xinyu Zhao¹, Yang Xu¹, Weixiang Wang¹

¹Beijing Key Laboratory of New Technology in Agricultural Application, National Demonstration Center for Experimental Plant Production Education, Department of Agronomy, Beijing University of Agriculture

Here, we present a protocol to analyze the genome-wide distribution of histone modifications, which can identify new target genes in the pathogenesis of M. oryzae and other filamentous fungi.