Preliminary study on the Oncolytic Activity of Newcastle Disease Virus strain F48E9

恶性肿瘤已成为当前影响人类健康的头号杀手。虽然手术、放化疗等治疗方法已经显著提高了肿瘤患者的生存率，但仍存在副作用大，对中晚期、远端转移或放化疗不敏感的肿瘤患者疗效差等不足，因此有必要探寻新的更有效的肿瘤治疗手段。近年来，溶瘤病毒因其具有杀伤效率高、特异性好、安全性高、副作用小等特点，已逐渐成为极具潜力的恶性肿瘤治疗手段。 新城疫病毒（Newcastlediseasevirus，NDV）是一种对禽类高致病、高致死性的单股负链RNA病毒，属副黏病毒科、副黏病毒亚科中的禽副黏病毒属。自20世纪50年代首次发现NDV能够抑制晚期胃癌转移以来，许多研究已经证实NDV具有特异性杀伤肿瘤细胞的潜力，同时...Cancer has been the first leading cause of death in the world. The overall survival rate of cancer patients has been significantly improved by surgery, radiotherapy and chemotherapy. However, for patients with advanced cancer and distant metastasis, there is still lack of effective treatment measures. Therefore, it is necessary to explore more effective methods for tumor therapy. Oncolytic virus i...学位：理学硕士院系专业：生命科学学院_生物化学与分子生物学学号：2162014115256

Impeded Nedd4-1-Mediated Ras Degradation Underlies Ras-Driven Tumorigenesis

RAS genes are among the most frequently mutated proto-oncogenes in cancer. However, how Ras stability is regulated remains largely unknown. Here, we report a regulatory loop involving the E3 ligase Nedd4-1, Ras, and PTEN. We found that Ras signaling stimulates the expression of Nedd4-1, which in turn acts as an E3 ubiquitin ligase that regulates Ras levels. Importantly, Ras activation, either by oncogenic mutations or by epidermal growth factor (EGF) signaling, prevents Nedd4-1-mediated Ras ubiquitination. This leads to Ras-induced Nedd4-1 overexpression, and subsequent degradation of the tumor suppressor PTEN in both human cancer samples and cancer cells. Our study thus unravels the molecular mechanisms underlying the interplay of Ras, Nedd4-1, and PTEN and suggests a basis for the high prevalence of Ras-activating mutations and EGF hypersignaling in cancer. © 2014 The Authors

ATR/Chk1 signaling induces autophagy through sumoylated RhoB-mediated lysosomal translocation of TSC2 after DNA damage

RhoB作为抑癌蛋白通过诱导肿瘤细胞的凋亡在抑制肿瘤的发生发展中发挥着重要作用，并且与肿瘤耐药性密切相关，但关于RhoB如何促进细胞死亡的分子机理的研究仍不清楚。在本研究中，该团队发现在DNA单链损伤情况下，ATR-Chk1信号通路的激活，会使RhoB被Chk1磷酸化，该磷酸化修饰会使RhoB从细胞质膜解离下来进入细胞质中，进而被SUMO化修饰。SUMO化修饰后的RhoB会与TSC2形成复合物，并将TSC2复合物带到溶酶体上，引起细胞自噬的发生。 该文共同第一作者为刘明冬、曾涛玲和张新，通讯作者为王洪睿教授和赵同金教授。【Abstract】DNA damage can induce autophagy; however, the underlying mechanism remains largely unknown. Here we report that DNA damage leads to autophagy through ATR/Chk1/RhoB-mediated lysosomal recruitment of TSC complex and subsequent mTORC1 inhibition. DNA damage caused by ultraviolet light (UV) or alkylating agent methyl methanesulphonate (MMS) results in phosphorylation of small GTPase RhoB by Chk1. Phosphorylation of RhoB enhances its interaction with the TSC2, and promotes its sumoylation by PIAS1, which is required for RhoB/TSC complex to translocate to lysosomes. As a result, mTORC1 is inhibited, and autophagy is activated. Knockout of RhoB severely attenuates lysosomal translocation of TSC complex and the DNA damage-induced autophagy. Reintroducing wild-type but not sumoylation-resistant RhoB into RhoB−/− cells restores the onset of autophagy. Hence, our study identifies a molecular mechanism for translocation of TSC complex to lysosomes in response to DNA damage, which depends on ATR/Chk1-mediated RhoB phosphorylation and sumoylation.This work was supported by the National Natural Science Foundation of China (U1605222, 81472459, 31671223), the National Key Research and Development Project of China (2016YFC1302400, 2016YFA0502003), the Fundamental Research Funds for the Central Universities (20720140550, 20720160070), the National Science Foundation for Fostering Talents in Basic Research of the National Natural Science Foundation of China (J1310027), the Project 111 sponsored by the State Bureau of Foreign Experts and Ministry of Education (B12001), the National Natural Science Foundation of China (31601132) to T.Z., the National Natural Science Foundation of China (81402290) to Q.L., and the National Natural Science Foundation of China (U1405223) to X.D

中国超大城市空气污染和人体健康 (APHH-Beijing) 中英联合研究计划 最终报告

Chinese versio

Development of monoclonal antibodies against nucleoprotein of Newcastle disease virus and establishment of a quantitative double-antibody sandwich ELISA for NDV antigen

目的制备新城疫病毒(NDV)核蛋白(NP)的单克隆抗体(单抗),并用于建立一种可定量检测NDV病毒含量的双抗体夹心酶联免疫吸附试验检测方法(ND; V NP ELISA)。方法基于NDV病毒株F48E9获得NP基因,经原核表达方式制备出重组抗原rNP;免疫小鼠制备NDV; NP特异性单抗;单抗经HRP标记和配对筛选,建立NDV NP; ELISA,分析其特异性、灵敏度、精密度、准确度和检测线性,并分析本方法定量检测的NP含量与PFU病毒滴度的定量相关性。结果建立基于单抗3C10; 和4E7的NDV NP ELISA,其定量检测NDV; rNP的最佳线性范围为0.015~0.250mug/ml(R2=0.9974),回收率在88.4%~106.01%,变异系数小于3.4%;该方法; 具有良好特异性;该方法定量检测NDV抗原含量与PFU病毒感染滴度有较好的相关性(R2=0.9209)。结论建立NDV NP; ELISA,可准确定量检测NDV病毒中的NP抗原含量,为NDV病毒含量的测定提供一种可靠简便的分析方法。We developed the monoclonal antibodies against nucleoprotein(NP)of; Newcastle disease virus(NDV),and established a double antibody sandwich; ELISA method for quantitative determination of NP antigen of NDV(NDV NP; ELISA). The recombination NP protein derived from strain F48E9of NDV; were prepared and used to immunize BLAB/c mice.The mouse splenic cells; from immunized mice were fused with SP2/0cells to generate monoclonal; antibodies(mAb).The NDV NP specific mAbs were paired to establish a; double antibody sandwich ELISA method.The performance of the NDV NP; ELISA was evaluated,including specificity,sensitivity,precision,accuracy; and linearity.The correlation between the ELISA and PFU virus titer was; analyzed by regression analysis method.Two monoclonal antibodies 3C10and; 4E7were selected to establish double antibody sandwich ELISA for NP; antigen of NDV.The linearity and performance of the NDV NP ELISA was; characterized. The detection linearity fell in the range of; 0.015-0.250mug/mL(R2=0.997 4).The detection limit of the assay was; 0.015mug/mL.The recovery was between 88.4%and 106.01%;the variation; coefficient was below 3.4%.In testing of 50 NDV virus samples,this assay; performed well and correlated comparably with PFU virus titer(R2=0.920; 9).The NDV NP ELISA for quantitative detection of NDV is a reliable; quantifiable assay for detection of NDV NP protein;it provides a new; approach for rapid and quantitative detection of Newcastle disease; virus.国家自然科学基

知母中Officinalisinin Ⅰ降糖机制及中枢神经保护研究

目的:观察知母中OfficinalisininⅠ对糖尿病的降糖效果,并探讨其降糖机制以及对糖尿病引起的神经损伤的保护作用。方法:利用四氧嘧啶小鼠造模后灌胃药品测定糖耐量;糖苷酶抑制效果体外检测;糖尿病小鼠禁食后给药检测胰高血糖素样肽-1(GLP-1);糖尿病小鼠给药20d后利用酶联免疫法对小鼠血清中脑源性神经营养因子(BDNF)和谷胱甘肽过氧化物酶(GSH-PX)进行检测。结果:OfficinalisininⅠ具有降血糖并显著提高糖尿病小鼠糖耐量的功能,实验排除糖苷酶抑制剂途径,并且通过促进GLP-1以及BDND的产生以及提高GSH-PX活力起到降糖及神经保护作用。结论:OfficinalisininⅠ可以有效降低糖尿病小鼠血糖,对神经损伤具有保护作用

communication protocol reverse engineering of malware using dynamic taint analysis

对恶意代码通信协议的逆向分析是多种网络安全应用的重要基础.针对现有方法在协议语法结构划分的完整性和准确性方面存在不足,对协议字段的语义理解尤为薄弱,提出了一种基于动态污点分析的协议逆向分析方法,通过构建恶意进程指令级和函数级行为的扩展污点传播流图(Extended Taint Propagation Graph,ETPG),完成对协议数据的语法划分和语义理解.通过实现原型系统并使用恶意代码样本进行测试,结果表明本方法可以实现有效的语法和语义分析,具有较高的准确性和可靠性.Communication protocol reverse engineering of malwares is significant base for various network security applications. However, recent works have limited accuracy and integrity in identifying protocol fields and are especially weak in understanding fields' semantics. This paper proposed a method for communication protocol reverse engineering based on dynamic taint analysis. By building an extended taint propagation graph (ETPG) recording both instruction and function level behaviors of a malicious process, dividing the protocol data into different syntax fields and inducing the semantic information of individual fields were achieved. A prototype system was implemented and evaluated with malware samples. The results show that this method can divide the syntax fields and extract semantic information accurately and effectively

颗粒孔结构的积木分形模型

构造了立方和四面体两种积木分形体,得到一般积木分形体模型,导出关联表面积和体积增量的3个分形表达式,并分析了表面分数维的几何意义. 实验结果表明,利用该模型的表面积与体积增量分形表达式可以从压汞和BET的实验数据计算表面分数维,相关系数较高. 对同一种颗粒,两种实验方法可以得到相同的分数维. 讨论了体积增量的计算方法