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An optimized high-throughput SARS-CoV-2 dual reporter trans-complementation system for antiviral screening in vitro and in vivo

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Virologica Sinica 2024年第3期39卷 447-458页

作者： yingjian li Xue Tan Jikai Deng Xuemei liu Qianyun liu Zhen Zhang Xiaoya Huang Chao Shen Ke Xu li Zhou Yu Chen State Key Laboratory of Virology RNA InstituteCollege of Life Sciences and Frontier Science Center for Immunology and MetabolismWuhan UniversityWuhan430072China Institute for Vaccine Research at Animal Bio-safety Level III Laboratory Wuhan University School of MedicineWuhan430071China

The severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)is still epidemic around the *** manipulation of SARS-CoV-2 is restricted to biosafety level 3 laboratories(BSL-3).In this study,we developed a SARS-CoV-2ΔN-GFP-HiBiT replicon delivery particles(RDPs)encoding a dual reporter gene,GFP-HiBiT,capable of producing both GFP signal and luciferase *** optimal selection of the reporter gene,GFP-HiBiT demonstrated superior stability and convenience for antiviral ***,we established a RDP infection mouse model by delivering the N gene into K18-hACE2 KI mouse through *** mouse model supports RDP replication and can be utilized for in vivo antiviral *** summary,the RDP system serves as a valuable tool for efficient antiviral screening and studying the gene function of ***,this system can be manipulated in BSL-2 laboratories,decreasing the threshold of experimental requirements.

关键词： SARS-CoV-2 RDP Antiviral evaluation Mouse model BSL-2

Structural and functional insights into the 20-O-methyltransferase of SARS-CoV-2

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Virologica Sinica 2024年第4期39卷 619-631页

作者： Jikai Deng Feiyu Gong yingjian li Xue Tan Xuemei liu Shimin Yang Xianying Chen Hongyun Wang Qianyun liu Chao Shen li Zhou Yu Chen State Key Laboratory of Virology RNA InstituteCollege of Life Sciences and Frontier Science Center for Immunology and MetabolismWuhan UniversityWuhan430072China Animal Bio-Safety Level Ⅲ Laboratory/Institute for Vaccine Research Wuhan University School of MedicineWuhan430071China

A unique feature of coronaviruses is their utilization of self-encoded nonstructural protein 16(nsp16),20-Omethyltransferase(20-O-MTase),to cap their RNAs through ribose 20-O-methylation *** process is crucial for maintaining viral genome stability,facilitating efficient translation,and enabling immune *** considerable advances in the ultrastructure of SARS-CoV-2 nsp16/nsp10,insights into its molecular mechanism have so far been *** this study,we systematically characterized the 20-O-MTase activity of nsp16 in SARS-CoV-2,focusing on its dependence on nsp10 *** observed cross-reactivity between nsp16 and nsp10 in various coronaviruses due to a conserved interaction ***,a single residue substitution(K58T)in SARS-CoV-2 nsp10 restricted the functional activation of MERS-CoV ***,the cofactor nsp10 effectively enhanced the binding of nsp16 to the substrate RNA and the methyl donor Sadenosyl-L-methionine(SAM).Mechanistically,His-80,Lys-93,and Gly-94 of nsp10 interacted with Asp-102,Ser-105,and Asp-106 of nsp16,respectively,thereby effectively stabilizing the SAM binding ***-43 of nsp10 interacted with Lys-38 and Gly-39 of nsp16 to dynamically regulate the RNA binding pocket and facilitate precise binding of RNA to the nsp16/nsp10 *** assessing the conformational epitopes of nsp16/nsp10 complex,we further determined the critical residues involved in 20-O-MTase ***,we utilized an in vitro biochemical platform to screen potential inhibitors targeting 20-O-MTase ***,our results significantly enhance the understanding of viral 20-O methylation process and mechanism,providing valuable targets for antiviral drug development.

关键词： SARS-CoV-2 nsp16/nsp10 20-O-Methyltransferase 3-Dimensional structure Molecular mechanism Inhibitors

Natural evidence of coronaviral 2′-O-methyltransferase activity affecting viral pathogenesis via improved substrate RNA binding

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Signal Transduction and Targeted Therapy 2024年第6期9卷 2819-2834页

作者： Jikai Deng Shimin Yang yingjian li Xue Tan Jiejie liu Yanying Yu Qiang Ding Chengpeng Fan Hongyun Wang Xianyin Chen Qianyun liu Xiao Guo Feiyu Gong li Zhou Yu Chen State Key Laboratory of Virology RNA InstituteCollege of Life Sciences and Frontier Science Center for Immunology and MetabolismWuhan UniversityWuhanChina School of Medicine Tsinghua UniversityBeijingChina School of Basic Medical Sciences Wuhan UniversityWuhanChina Animal Bio-Safety Level Ⅲ Laboratory/Institute for Vaccine Research Wuhan University School of MedicineWuhanChina

Previous studies through targeted mutagenesis of K-D-K-E motif have demonstrated that 2′-O-MTase activity is essential for efficient viral replication and immune ***,the K-D-K-E catalytic motif of 2′-O-MTase is highly conserved across numerous viruses,including flaviviruses,vaccinia viruses,coronaviruses,and extends even to ***,we observed a stronger 2′-O-MTase activity in SARS-CoV-2 compared to SARS-CoV,despite the presence of a consistently active catalytic *** further identified critical residues(Leu-36,Asn-138 and Ile-153)which served as determinants of discrepancy in 2′-O-MTase activity between SARS-CoV-2 and *** residues significantly enhanced the RNA binding affinity of 2′-O-MTase and boosted its versatility toward RNA *** interest,a triple substitution(Leu^(36)→Ile^(36),Asn^(138)→His^(138),Ile^(153)→Leu^(153),from SARS-CoV-2 to SARS-CoV)within nsp16 resulted in a proportional reduction in viral 2′-O-methylation and impaired viral ***,it led to a significant upregulation of type I interferon(IFN-I)and proinflammatory cytokines both in vitro and vivo,relying on the cooperative sensing of melanoma differentiation-associated protein 5(MDA5)and laboratory of genetics and physiology 2(LGP2).In conclusion,our findings demonstrated that alterations in residues other than K-D-K-E of 2′-O-MTase may affect viral replication and subsequently influence *** changes in nsp16 residues is crucial as it may aid in identifying and assessing future alteration in viral pathogenicity resulting from natural mutations occurring in nsp16.

关键词： pathogenesis impaired alterations

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氨纶包芯纱蠕变性能测试与分析

纺织学报 2007年第6期28卷 48-51页

作者：肖丰李营建河南纺织高等专科学校河南郑州450007

对大豆蛋白复合纤维、棉和涤纶纤维纺制的氨纶包芯纱的蠕变性能进行测试,基于纺织材料的黏弹性理论,使用四元素力学模型分析氨纶包芯纱的拉伸变形规律,并利用DPS数据处理系统,建立了氨纶包芯纱蠕变伸长的拟合方程,从本质上研究大豆蛋白... 详细信息

对大豆蛋白复合纤维、棉和涤纶纤维纺制的氨纶包芯纱的蠕变性能进行测试,基于纺织材料的黏弹性理论,使用四元素力学模型分析氨纶包芯纱的拉伸变形规律,并利用DPS数据处理系统,建立了氨纶包芯纱蠕变伸长的拟合方程,从本质上研究大豆蛋白/氨、棉/氨、涤/氨包芯纱的蠕变性能特征。结果表明:氨纶包芯纱外包纤维种类不同,其蠕变性能亦不同;四元素模型的理论分析结果比较接近实际测试值,即应用四元素模型可以较好地描述各种氨纶包芯纱的蠕变规律。

关键词：氨纶包芯纱蠕变性能力学模型拟合方程特征参数

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总能系统(IES)的动力驱动方式研究

石油与天然气化工 2003年第2期32卷 75-78页

作者：李迎建深圳大学理学院

动力和发电装置是IES的一个重要组成部分,选择动力驱动方式与整个系统的组合,直接关系到联产系统的效率、成本和环境影响。现试图对已经或即将投入商业化运行的三种动力驱动装置的特点作一分析和研究。

关键词：总能系统 IES 动力驱动方式研究动力装置性能能源利用

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生态纤维素纤维喷气纱针织物的性能研究

针织工业 2014年第3期 25-28页

作者：肖丰李营建河南工程学院纺织学院河南郑州450007

以天丝、莫代尔、竹黏等生态纤维素纤维为原料,采用不同的混纺比进行混纺,开发了提花、双珠地、罗纹等几种喷气纱针织物。重点测试了针织面料的透气性、透湿性、芯吸效应、硬挺度、悬垂性、光泽性、顶破强力、耐磨性、色牢度等性能,并... 详细信息

以天丝、莫代尔、竹黏等生态纤维素纤维为原料,采用不同的混纺比进行混纺,开发了提花、双珠地、罗纹等几种喷气纱针织物。重点测试了针织面料的透气性、透湿性、芯吸效应、硬挺度、悬垂性、光泽性、顶破强力、耐磨性、色牢度等性能,并与环锭纱针织物进行对比分析。结果表明:纤维素纤维喷气纱针织物透气透湿、芯吸效应明显;光泽明亮,色牢度好;织物耐磨性好,顶破强力接近环锭纱织物;织物相对较硬,有身骨,悬垂性比环锭纱针织物略差,立体感强。该类针织物适合于针织T恤和内衣面料的生产。

关键词：生态纤维素纤维喷气纱针织物环锭纱针织物性能测试

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集聚赛络纱及其织物性能分析

棉纺织技术 2015年第2期43卷 57-60页

作者：肖丰李营建河南工程学院河南郑州450007

探讨集聚赛络纱及其织物的性能特点。为了推广应用开发集聚赛络纱织物,测试了集聚赛络纱的结构、毛羽、捻度、条干、强伸性及其织物的透气性、芯吸、悬垂性、拉伸强力、耐磨性能,并与环锭纱织物进行对比分析。结果表明:集聚赛络纱具有... 详细信息

探讨集聚赛络纱及其织物的性能特点。为了推广应用开发集聚赛络纱织物,测试了集聚赛络纱的结构、毛羽、捻度、条干、强伸性及其织物的透气性、芯吸、悬垂性、拉伸强力、耐磨性能,并与环锭纱织物进行对比分析。结果表明:集聚赛络纱具有股线结构特征,成纱紧密,表面光洁,毛羽少,强力高;集聚赛络纱织物的透气性好,芯吸效应明显,坚牢耐磨,悬垂性比环锭纱织物稍差。认为:用集聚赛络纺纱织物适合开发男式衬衫面料和床上用品面料。

关键词：集聚纺赛络纺成纱结构纱线性能织物性能

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天丝/棉/涤纶单丝包芯纱的开发

上海纺织科技 2017年第1期45卷 31-33,40页

作者：李营建肖丰河南工程学院河南郑州450007 纺织新产品开发河南省工程实验室河南郑州451191

根据天丝、棉纤维及涤纶单丝的特性,对生产涤纶单丝包芯纱的纺纱工艺进行研究。探讨了在环锭细纱机上纺制涤纶单丝包芯纱的主要技术难点。通过对细纱机进行适当改造,利用天丝纤维和棉纤维混纺作皮层,30 D涤纶单丝作芯纱,纺制55/45 18.4 ... 详细信息

根据天丝、棉纤维及涤纶单丝的特性,对生产涤纶单丝包芯纱的纺纱工艺进行研究。探讨了在环锭细纱机上纺制涤纶单丝包芯纱的主要技术难点。通过对细纱机进行适当改造,利用天丝纤维和棉纤维混纺作皮层,30 D涤纶单丝作芯纱,纺制55/45 18.4 tex天丝/棉(33.3 dtex)包芯纱,成纱质量达到了较高水平。

关键词：包芯纱涤纶单丝工艺设计产品开发天丝纤维

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信息自助时代高校图书馆微知识服务模式探析

图书情报导刊 2021年第2期6卷 33-36页

作者：李英剑江苏海洋大学图书馆江苏连云港222005

信息自助时代,需要高质、多元、个性、便捷的知识获取方式,微知识服务应运而生。从高校图书馆开展微知识服务的优势和特征入手,探讨了微知识服务的创新模式:包括通过整合图书馆优势资源,建立微知识库和检索平台;应用数据挖掘技术,分析... 详细信息

信息自助时代,需要高质、多元、个性、便捷的知识获取方式,微知识服务应运而生。从高校图书馆开展微知识服务的优势和特征入手,探讨了微知识服务的创新模式:包括通过整合图书馆优势资源,建立微知识库和检索平台;应用数据挖掘技术,分析用户需求并进行有针对性的微知识服务;建立奖励机制,促进隐性知识的交流与转移;不断通过效果评价、用户反馈等措施调整服务内容与策略,促进微知识服务模式的良性、动态发展。

关键词：微知识服务高校图书馆个性化信息服务