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Asian Pacific Journal of Tropical Medicine 2016年第2期9卷 109-114页

作者： Drishya Diwaker Mishra K P Ganju L Singh S B Immunomodulation Lab Defence Institute of Physiology & Allied Sciences

Objective: To study protein-protein interaction between heterogeneous nuclear ribonucleoprotein H(hn RNP H) and Dengue virus(DENV) proteins. Methods: DENV proteins were screened against the host hn RNP H protein, in order to identify the host-viral protein-protein interactions in DENV infected THP-1 cells by co-immunoprecipitation. The co-localization of the interacting proteins was further confirmed by immunofluorescence microscopy. Results: The host protein hn RNP H was found to interact with DENV nonstructural 1 protein and help the virus to multiply in the cell. Conclusions: The non-structural 1 glycoprotein is a key modulator of host immune response and is also involved in viral replication. Therefore, disruption of this key interaction between hn RNP H and DENV nonstructural 1 could be an important therapeutic strategy for management of DENV infection.

关键词： Dengue virus non-structural 1 protein Heterogeneous nuclear ribonucleoprotein H Protein interactions

ACTIVITIES OF THE ESCAP/WMO TYPHOON COMMITTEE REGARDING SEDIMENT-RELATED DISASTER PREVENTION USING non-structural MEASURES,BASED ON JAPANESE SABO TECHNOLOGY

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Tropical Cyclone Research and Review 2013年第1期2卷 55-64页

作者： SHIN-ICHIRO HAYASHI HIDEAKI MIZUNO ATSUSHI OKAMOTO MASAKI HIRUMA REIJI KONDO National Institute for Land and Infrastructure Management Japan SABO and Landslide Technical Center Japan

In the project activities of the ESCAP/WMO Typhoon Committee(TC), the National Institute for Land and Infrastructure Management(NILIM), the SABO(Erosion and Sediment Control) Department of the Ministry of Land, Infrastructure, Transport and Tourism(MLIT), and the SABO and Landslide Technical Center(STC) implemented two projects: “Sediment-related Disaster Forecasting Warning System Project”(2002–2008) and “Project on Hazard Mapping for Sediment-related Disasters”(2009–2012). The aim of these projects was to distribute Japanese SABO technology, particularly non-structural measures, among TC members. This report presents the concepts and methods related to non-structural measures for sediment-related disaster prevention based on Japanese SABO technology, the contents of the two projects’ activities, the preparation of early warning information, the creation of a hazard map, and the preparation of an integrated system for resident evacuation.

关键词： sediment-related disaster prevention non-structural measures early warning hazard map

Inhibition of Hepatitis C Virus Genotype 1a non-structural Proteins by Small Interference RNA in Human Hepatoma Cell Lines

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Pharmacology & Pharmacy 2015年第11期6卷 502-517页

作者： Imran Shahid Waleed Hassan AlMalki Shaia Saleh R. Almalki Ismail Muhammad AlTurkestany Hassan Ali AlGhamdi Saleh Ali AlMenshawi Applied and Functional Genomics Laboratory Center of Excellence in Molecular Biology University of the Punjab Lahore Pakistan Department of Pharmacology and Toxicology College of Pharmacy Umm Al Qura University Makkah Saudi Arabia Laboratory Medicine Faculty of Applied Medical Sciences Albaha University Albaha Saudi Arabia Infection Control Pilgrims City Hospital Madina Saudi Arabia Toxicology Center Jeddah Saudi Arabia Toxiology Research Center Ministry of Interior Jeddah Saudi Arabia

Hepatitis C virus (HCV) infection and associated liver diseases are still challenging and represent a significant health care burden around the world. Although, the treatment strategies have been improved by the development of novel direct-acting antivirals, but such therapeutic options are still expensive and beyond the financial range of the most infected individuals in developing or even in resource replete countries. It demands an urgent need to search novel and improved alternate treatment strategies to treat the infection. The present study was aimed to develop an in vitro stable cell culture system, persistently expressing HCV genotype 1a non-structural genes and to characterize the inhibitory effects of synthetic siRNAs (short interference RNA) directed against the most conserved regions of nonstructural genes in an in vitro cell culture model. The continuous expression of nonstructural genes for more than 30 days post transfection was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis in stable human hepatoma cell line (Huh-7). The gene expression studies revealed significantly reduced gene expression of HCV nonstructural genes (i.e., NS2, NS4A and NS5A) both at mRNA and protein levels when treated against genome specific synthetic siRNAs in stable cell lines (51%, 47% and 54% respectively, p < 0.05). Similarly, a vivid decrease in HCV viral titer was exhibited by synthetic siRNAs in an in vitro viral replicate cell culture model (58%, 48% and 50%, respectively, p < 0.05) determined by quantitative Real-Time PCR (qPCR). Our data indicate that siRNA mediated gene silencing may be considered a promising alternate treatment strategy against HCV in combination with other effective therapeutic regimens in future.

关键词： Hepatitis C Virus non-structural Proteins Stable Cell Line Anti-HCV Drugs Short Interference RNA

The cloning of non-structural-1 (NS1) gene of H9N2 subtype of avian influenza virus in pGEX-4T-1 and pMAL-c2X plasmids and expression in <i>Escherichia coli</i>DH5<i>α</i>strain

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Advances in Bioscience and Biotechnology 2012年第3期3卷 283-289页

作者： Ramin Soleimani Mehdi Vasfi Marandi Mohammad Bagher Hashemi-Soteh Farhid Hemmatzadeh Department of Poultry Diseases Faculty of Veterinary Medicine University of Tehran-Iran Tehran Iran Medical Faculty and Cellular and Molecular Research Center Mazandaran University of Medical Sciences Sari Iran School of Animal and Veterinary Sciences The University of Adelaide Adelaide Australia

Avian influenza is a viral contagious disease that affects poultry industry and human health. Vaccination has been considered as a preventive tool in the eradication of AI, but it causes some limitations including trade embargoes and interfering with serologic surveillance in differentiation between infected and vaccinated animals (DIVA strategy). Several distinct DIVA strategies have been presented to conquer these limitations. In this study, the open reading frame of NS1 gene of a H9N2 subtype of AI virus was amplified by polymerase chain reaction. After extraction and purification of NS1 gene from agarose gel, it was inserted into two different pGEX-4T-1 and pMAL-c2X plasmids and transferred in DH5α strain of Escherichia coli by using electroporation procedure. The E. coli colonies possessing recombinant NS1 gene were screened using PCR, restriction mapping and sequencing analysis. The expressed rNS1 protein was purified using affinity chromatography based on MBP (pMAL- c2X) and GST (pGEX-4T-1). The MBP-NS1 and GST- NS1 proteins on SDS-PAGE had bands with molecular weight of 68 and 52 kDa respectively. Western blotting with MBP-NS1 protein showed positive reaction using antisera obtained from chickens challenged with a H9N2 subtype strain. But, the most sera prepared from H9N2 vaccinated chickens were negative in WB. These findings indicated that the MBP-rNS1 protein of 26 kDa expressed by pMAL-c2X plasmid can be used in a DIVA for differentiation of AI infected and vaccinated chickens.

关键词： Avian Influenza Virus H9N2 Subtype Recombinant Protein non-structural Protein 1 DIVA

Study on the Changes of non-structural Carbohydrates and Nitrogen Contents of Quercus aquifolioides Scrub along Different Elevation Gradient

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Agricultural Science & Technology 2011年第4期12卷 576-580页

作者：黄亚洲吴杰孟玉山吕俊王三根朱万泽西南大学农学与生物科技学院重庆400716 中国科学院成都山地灾害与环境研究所四川成都610041

[Objective] The aim was to compare the content changes between the non-structural carbohydrates（NSC）and the total nitrogen in various growing seasons,and to explore the response relationship between altitude and the contents.[Method] Taking Quercus aquifolioides scrub which widely distributed in Zheduoshan in the west of Sichuan as the experimental objects,the changes between NSC and the toal nitrogen in various growing seasons at different altitude were studied.[Result] The results showed that the content of NSC in Quercus aquifolioides underground increased with the lift of elevation in the dormancy,but decreased in the early germination,growing period and growth *** content of NSC in the ground tissue changed non-linearly with increasing *** addition,the total nitrogen of Quercus aquifolioides organizations was decreasing with increasing elevation in the dormant period,which did not change significantly in the other *** result implied that the content of NSC in Quercus aquifolioides underground was more sensitive to temperature.[Conclusion] The experiment laid basis for the exploration of the physical and ecological mechanism of underground plants adaptability to highland environment,their response to global climate changes and adjustment to high altitude ecological system.

关键词： Quercus aquifolioides Total nitrogen non-structural Carbohydrates Elevation

Chikungunya infection:A potential re-emerging global threat

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Asian Pacific Journal of Tropical Medicine 2016年第10期9卷 910-915页

作者： Shanmugaraj Bala Murugan Ramalingam Sathishkumar Plant Genetic Engineering Laboratory Department of BiotechnologyBharathiar University

Infectious diseases are indeed a lifelong threat to everyone irrespective of age, sex, lifestyle and socio-economic status. The infectious diseases have persisted among the prominent causes of death globally. Recently, re-emergence of Chikungunya viral infection harmed many in Asian and African countries. Chikungunya was considered as a major threat in developing and underdeveloped countries; the recent epidemiological outbreak of Chikungunya in La Reunion urges the global researchers to develop effective vaccine against this viral disease. In this review, Chikungunya, pathogenesis and epidemiology were briefly described.

关键词： Aedes mosquitoes Alphavirus Arthropod-borne disease Chikungunya non-structural Proteins structural Proteins

Research on Cohesion and Coherence of Spoken Language—— A Case of Desperate Housewives

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海外英语 2019年第3期 233-235页

作者：罗宇晓电子科大学中山学院外国语学院

This paper mainly gives a research of cohesion and coherence in the spoken language. It takes one of the most popular teleplays—Desperate Housewives as its source of language data and tries to give a rather specific analysis of the application of thetheory of cohesion and coherence. The paper mainly focuses on the structural cohesion and non-structural cohesion as well as thecontext of situation, which includes thematic structure, information structure, cohesive devices, transitivity, turn-taking, adjacencypair, etc. In addition, it encourages more people to pay attention to this theory and its application so as to make a little contributionto the development of discourse analysis.

关键词： Cohesion Coherence structural Cohesion non-structural Cohesion

人星状病毒非结构蛋白nsP1a／1表达纯化及多克隆抗体的制备

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医学分子生物学杂志 2016年第3期13卷 158-162页

作者：苑荣亮崔成成井敏敏刘志成汪玉婷岳风先张成井申荣毛小琴昆明理工大学医学院昆明市650500 云南省第一人民医院、昆明理工大学附属医院昆明市 650032 云南省临床微生物分子研究中心昆明市650032

目的：表达纯化人星状体病毒（ human astrovirus， HAstV）非结构蛋白nsP1a／1，免疫动物制备多克隆抗体。方法利用PCR技术扩增nsP1a／1基因序列，构建到大肠埃希菌原核表达系统中表达重组nsP1a／1蛋白，使用镍柱亲和层析法对重组蛋白... 详细信息

目的：表达纯化人星状体病毒（ human astrovirus， HAstV）非结构蛋白nsP1a／1，免疫动物制备多克隆抗体。方法利用PCR技术扩增nsP1a／1基因序列，构建到大肠埃希菌原核表达系统中表达重组nsP1a／1蛋白，使用镍柱亲和层析法对重组蛋白进行纯化，十二烷基磺酸钠-聚丙烯酰胺凝胶电泳（ SDS-PAGE）和二噻啉甲酸（BCA）实验对重组蛋白的纯度与浓度进行分析，以重组的nsP1a／1蛋白为抗原，免疫雄性SPF级SD 大鼠获得多抗血清，用 ELISA 测定抗体效价、 Western 印迹检测抗体特异性。结果nsP1a／1-pET28a原核表达载体构建成功，将其转化至大肠埃希菌BL21（DE3）细菌中诱导表达了重组蛋白，免疫大鼠获得的多抗血清几何平均效价达到1∶406374。结论本实验成功地运用原核表达系统表达并鉴定了人星状体病毒非结构蛋白nsP1a／1，为进一步研究人星状病毒的复制及病毒感染的临床诊断奠定基础。

关键词：人星状病毒非结构蛋白nsP1a/1 原核表达多克隆抗体 non-structural proteins nsP1 a/1