Preliminary study of novel live attenuated varicella-zoster virus of ORF7 knock out mutation (rOka△7) and virulence critical factor of ORF7

水痘是一种急性、高传染性的儿童常见呼吸道传播疾病，由水痘-带状疱疹病毒（Varicella-zostervirus，VZV）引起。同时VZV也是导致带状疱疹（HZ）的病原体，具有严重的危害性。现今使用的VZV减毒活疫苗株（vOka）是唯一获WHO批准使用的水痘/带状疱疹疫苗株，然而有研究显示，vOka为混合毒株且仍具备感染和潜伏神经系统的能力，可能存在较复杂的安全性隐患。因此，迫切需要发展新型的可消除长期隐患的水痘疫苗株，而VZV关键毒力因子的发现和验证是开展该研究的核心基础。本研究在前期发现VZVORF7抗原缺失可影响病毒在体外人皮肤中感染的工作基础上通过建立包括人皮肤、胸腺、背根神经节移植...Varicella is an acute, high contagious viral disease, and the causative agent is Varicella-zoster virus(VZV), VZV is also the causative agent of Herpes zoster(HZ) So,it is a hamful virus.VZV live attenuated vaccine strain (vOka) used nowadays is the only licenced varicella zoster vaccine by WHO. But research shows that this vaccine is a mixed strain remain neurovirulence and the capacity of establ...学位：理学硕士院系专业：生命科学学院生物学系_生物化学与分子生物学学号：2172009115209

Development of aquantitative ELISA detection method for Varicella Zoster Virus(VZV) antigen

目的:建立水痘-带状疱疹病毒(VzV)抗原的双抗体夹心ElISA定量检测方法,用于质控VzV灭活疫苗研发和生产中抗原含量。方法:以VzV中和单抗5f6C8为包被抗体、8H5d1为酶标抗体,构建定量检测VzV抗原的双抗体夹心ElISA方法,并对本方法的特异性、灵敏度、准确性、线性和稳定性等性能进行分析。结果:建立的双抗体夹心定量检测VzV抗原的ElISA方法,线性范围为0.4μg~13μg/Ml,相关系数为r2=0.994,定量限度为0.4μg/Ml;变异系数CV80%。与VzV以外的相关病毒样本没有交叉反应。结论:构建的VzV抗原ElISA定量检测方法的各项性能符合定量检测需要,可用于VzV灭活疫苗的研发和生产过程的抗原含量检测。Objective:To develop a quantitative enzyme linked immunosorbent assay(Q-ELISA) to determine the concentration of Varicella Zoster Virus(VZV) antigen.This method was used to determine VZV antigen content at each stage of VZV inactived vaccine developing and manufacturing process.Methods: A double antibody sandwich Q-ELISA was developed to determine concentration of VZV antigen,which was based on the the high-affinity neutralizing monoclonal antibodies 5F6C8 as capture antibodies,and 8H5D1 as HRP-labeled antibody.The performance of reagent were evaluated.Results: The Q-ELISA for VZV antigen content was successfully developed.The reagent had good performance.The quantitation scope was 0.4 μg~13 μg/ml,The coefficient correlation was 0.994,the limit of detection was 0.4 μg /ml,the recovery was between 87.5% and 111.6%.The stability was up to 80% after reagent was heated for 6 days at 37℃.The variation coefficient was lower than 15%,and the reagent was no reaction with other sample except VZV antigen.Conclusion: The Q-ELISA for VZV antigen was developed with good specificity,accuracy and stability.The method can be used to determine VZV antigen content during development and production of VZV inactived vaccine

Construction and Identification of siRNA Targeting to Multidrug Resistance Gene 1

通信作者:[email protected][中文文摘]mdr1基因及其表达产物P-gp是引起肿瘤细胞多药耐药(MDR)的主要原因,抑制mdr1基因的表达可用于逆转MDR.RNAi可用于特异抑制靶基因的表达,本研究的目的是构建获得可特异有效靶向mdr1基因的siRNA元件.应用siRNA设计软件与mRNA结构分析软件设计构建了3个分别靶向mdr1基因mRNA环结构和茎结构的siRNA元件,同时构建了携带mdr1基因序列的luc报告质粒,通过siRNA表达质粒与携带靶序列的报告质粒的共转染抑制实验检测不同siRNA的抑制效率,结果显示靶向环结构siMDR1B具有较好的抑制效率和特异性.进一步将siMDR1B表达载体与mdr1基因表达载体共转染细胞,应用免疫流式细胞术检测显示,相比对照细胞,siMDR1B可显著抑制其转染后mdr1基因产物P-gp蛋白的表达活性.同时采用CCK-8细胞活性检测试剂评价了siMDR1B对细胞活性的影响,结果显示siMDR1B不会影响细胞活性,具有良好的特异性.本研究获得的可有效靶向mdr1基因的siRNA元件可为进一步开展逆转MDR研究提供重要基础.[英文文摘]The over-expression of mdr1 gene is the main reason of multidrug resistance(MDR) of tumor cells.Inhibiting the expression of mdr1 gene can be used to reverse the MDR.RNA interfence(RNAi) is a sequence-specific intracellular mechanism of gene silence.In this study,the construction of small interfering RNAs(siRNAs) element against mdr1 gene effectively and specifically were explored.Three siRNA sequences targeting to mRNA stem or loop stucture separately were selected based on the sequence of mdr1 gene.Using various reporter plasmids contaiining different target sequences,the inhibition efficiencies and specificities of the constructs against mdrlgene were determined and siMDR1Btargeting to the loop structure was selected from the candidates.siMDR1B expression vector was then co-transfected with the mdrigene expression vector in cells,and the expression level of P-gp protein encoded by mdr1gene was analysed by immune flow cytometry.Results showed that P-gp protein expression can be effectively inhibited by siMDR1Bin transformed cells comparing to control cells.CCK-8assay was used for cell viablity analsis in this study,and results showed that cell viability was not affected by siMDR1Bexpression in transformed cells.In conclusion,siMDR1Bis a promising candidate for the next research on MDR reverse.福建省卫生厅青年科研课题计划项目(2008-1-53); 厦门市卫生局基金项目(3502z20089012

Determination of the number of ψ(3686) events taken at BESIII

The number of ψ(3686) events collected by the BESIII detector during the 2021 run period is determined to be (2259.3±11.1)×106 by counting inclusive ψ(3686) hadronic events. The uncertainty is systematic and the statistical uncertainty is negligible. Meanwhile, the numbers of ψ(3686) events collected during the 2009 and 2012 run periods are updated to be (107.7±0.6)×106 and (345.4±2.6)×106, respectively. Both numbers are consistent with the previous measurements within one standard deviation. The total number of ψ(3686) events in the three data samples is (2712.4±14.3)×10^