基于立即早期蛋白IE62建立水痘-带状疱疹病毒感染性滴度检测方法

目的:基于水痘-带状疱疹病毒(VZV)立即早期蛋白IE62与酶联免疫斑点检测技术(ELISpot)建立一种新型的VZV感染性滴度的快速检测方法。方法:应用生物信息学方法设计并合成VZV-IE62蛋白的多肽抗原,牛血清白蛋白偶联后免疫BALB/c小鼠,筛选和制备抗VZV-IE62单克隆抗体,应用Western blot和免疫荧光法等开展抗体性能评价,继而应用酶联免疫斑点技术(ELISpot)结合生物素-亲和素放大法建立新型的VZV感染性滴度检测方法,并与经典空斑计数法进行比较。结果:获得抗VZV-IE62的单克隆抗体1B7,应用于ELISpot方法可特异识别VZV感染后的细胞,以之为基础建立了新型的VZV感染性滴度检测方法。相比经典空斑计数法,该方法可显著缩短检测时间(从5至7 d缩短至32 h)检测结果具有较好的一致性。结论:本研究建立了一种新型的基于VZV立即早期蛋白IE62与酶联免疫斑点技术的VZV感染性滴度检测方法(VZV-IE62 ELISpot),具有潜在的转化应用前景,可为VZV的防治研究提供支持。国家自然科学基金项目(81601762)资

什么是好的教育政策研究

为进一步加强教育政策研究,更好服务新时代中国特色社会主义教育事业发展,在教育部-上海市人民政府共建的教育经济宏观政策研究院和华东师范大学国家宏观教育政策研究院的支持下,我们特别邀请全国知名教育政策研究专家围绕\"什么是好的教育政策研究\"展开笔谈。笔谈按作者姓氏笔画排序。华东师范大学国家宏观教育政策研究院委托项目“学界有影响力研究成果评判

Preparation and Application of Soluble Human Squamous Cell Carcinoma Antigen Expressed by Escherichia coli

旨在建立基于大肠杆菌表达系统的高效可溶性表达人鳞状上皮细胞癌抗原(SCCAg)方法,获得具有较好活性的重组SCCAg抗原并应用于建立抗原检测方法; 。基于pGEX-6P-l载体和大肠杆菌E. coli; ER2566菌株开展重组SCCAg抗原可溶性表达纯化方法研究,评价纯化抗原活性,筛选特异性单克隆抗体,初步建立并评价SCCAg抗原检测方法。结果; 显示,pGEX-6P-l载体和E coli; ER2566菌株可用于建立较高效的可溶性表达和纯化SCCAg抗原的方法,获得了具有较高纯度和活性的重组SCCAg抗原,筛选获得特异性单克隆抗体并; 初步建立了 SCCAg管式化学发光检测方法。建立了有效的基于大肠杆菌表达系统的可溶性表达和纯化SCCAg的方法。The aims of this study are to establish a method for efficient soluble; expression of human squamous cell carcinoma antigen(SCCAg ) based on; Escherichia coli expression system and obtain the recombinant SCCAg; antigen in fine activity, then apply it in the detection method; establishment of antigen. The study on the method of soluble expression; and purification of recombinant SCCAg antigen was conducted based on; pGEX-6P-l vector and E. coli ER2566 strain. The activity of the purified; antigen was evaluated by Abbott Kit and the specific monoclonal antibody; was screened by indirect ELISA. It was proved that PGEX-6P-1 vector and; E. coli strain ER2566 could be used to establish efficient soluble; expression and purification method for recombinant SCCAg antigen.; Moreover, the recombinant SCCAg antigen was proved to be in high purity; and activity. Thus,the SCCAg detection method of chemical luminous tube; was established with the specific monoclonal antibodies. In conclusion,; an effective method for the expression and purification of SCCAg, which; is based on the E. coli expression system, is established.国家高技术研究发展计划(863计划

水痘-带状疱疹病毒主要衣壳蛋白ORF40单克隆抗体的制备及初步应用

目的:制备抗水痘-带状疱疹病毒(VZV)主要衣壳蛋白ORF40的单克隆抗体,对该蛋白在VZV感染细胞内的表达、分布以及在VZV病毒颗粒中的包含情况进行初步探究。方法:设计并合成VZV ORF40多肽片段,偶联...国家自然科学基金项目(81601762)资

Near-atomic cryo-electron microscopy structures of varicella-zoster virus capsids

VZV是一种广泛存在并且具有高度传染性的人类α-疱疹病毒。初次感染VZV可导致水痘，人群普遍易感（感染率约为61%～100%）。该病毒可在背根神经节潜伏感染，持续终生。夏宁邵教授团队长期开展VZV相关基础与新型疫苗研究，通过系统和精细探索建立了高效的VZV规模化培养和病毒颗粒纯化技术体系，成功获得高质量的VZV颗粒样品。首次揭示了疱疹病毒α家族的水痘-带状疱疹病毒（VZV）不同类型核衣壳的近原子分辨率结构，阐明了VZV核衣壳不同组成蛋白的相互作用网络与衣壳装配机制，可为进一步开展新型载体疫苗设计及抗病毒药物等研究提供重要支持。 我校博士后王玮、高级工程师郑清炳、博士生潘德全和俞海副教授为该论文共同第一作者，我校夏宁邵教授、程通副教授、李少伟教授以及美国罗格斯大学朱桦（Hua Zhu）教授、加利福尼亚大学洛杉矶分校周正洪（Z. Hong Zhou）教授为该论文的共同通讯作者。【Abstract】Varicella-zoster virus (VZV) is a medically important human herpesvirus that causes chickenpox and shingles, but its cell-associated nature has hindered structure studies. Here we report the cryo-electron microscopy structures of purified VZV A-capsid and C-capsid, as well as of the DNA-containing capsid inside the virion. Atomic models derived from these structures show that, despite enclosing a genome that is substantially smaller than those of other human herpesviruses, VZV has a similarly sized capsid, consisting of 955 major capsid protein (MCP), 900 small capsid protein (SCP), 640 triplex dimer (Tri2) and 320 triplex monomer (Tri1) subunits. The VZV capsid has high thermal stability, although with relatively fewer intra- and inter-capsid protein interactions and less stably associated tegument proteins compared with other human herpesviruses. Analysis with antibodies targeting the N and C termini of the VZV SCP indicates that the hexon-capping SCP—the largest among human herpesviruses—uses its N-terminal half to bridge hexon MCP subunits and possesses a C-terminal flexible half emanating from the inner rim of the upper hexon channel into the tegument layer. Correlation of these structural features and functional observations provide insights into VZV assembly and pathogenesis and should help efforts to engineer gene delivery and anticancer vectors based on the currently available VZV vaccine.This research was supported by grants from the National Science and Technology Major Projects for Major New Drugs Innovation and Development (no. 2018ZX09711003-005-003), the National Science and Technology Major Project of Infectious Diseases (no. 2017ZX10304402), the National Natural Science Foundation of China (no. 81871648, 81601762), the Research Unit of Frontier Technology of Structural Vaccinology of Chinese Academy of Medical Sciences (no. 2019RU022) and the US National Institutes of Health (DE025567/028583). 该研究获得了国家自然科学基金、新药创制国家科技重大专项和传染病防治国家科技重大专项等资助

Two grades optimal design of submain manifold for sprinkle irrigation

采用两级优化设计模型进行喷微灌支管设计,第一级优化采用拉格朗日法求解非线性规划模型,以年费用最低为目标函数,以连续管径为决策变量,获得一组最优管径组合。第二级采用枚举法进行优化设计,在第一次连续管径基础上,按照施工要求的变径数,选择出靠近连续管径的几个商用管径,以年费用最低为目标函数,以管径和管长作为决策变量,获得最优管径和管长组合,最后计算出各节点的水压力。该方法不仅使年费用最低,而且满足了施工要求,可直接用于喷微灌支管的施工设计

Experimental Study on Influencing Factors of Soil Wetted Body of Clay Loam under Bubbled Root Irrigation

滴头流量、灌水量及埋深与土壤湿润体的关系是涌泉根灌设计的依据。通过室内土槽试验,研究了涌泉根灌的滴头流量、灌水量及滴头埋深对均质粘壤土湿润体变化的影响。结果表明:涌泉根灌土壤入渗湿润体在垂直向下距离和水平扩散距离均与灌水时间成幂函数关系,相关系数达到0.98以上;滴头流量和灌水器埋深对土壤入渗湿润体影响较大,而灌水量对土壤入渗湿润体无显著影响

Study on the Emission Uniformity for the Emitter under the Condition of Farmland Micro-terrain Action

首先对滴灌系统中田面微地形偏差、水力偏差、制造偏差三因素独立作用及共同作用下产生的滴头流量偏差率、偏差系数、均匀度的计算公式进行分析,对符合正态分布的田面局部高、滴头制造系数进行计算机模拟,并随机分配给滴头;然后借助计算机模拟计算出上述三种因素独立作用及共同作用下流量偏差率、偏差系数、均匀度等指标;最后将上述四种计算结果的500组数据, 经过统计分析,确定了灌水均匀度计算公式,并建立了各均匀度参数之间的关系,分析得到的公式可直接用于滴灌系统的水力学计算