基因-病毒治疗系统AdHA对血管内皮细胞的抑制作用

目的：研究基因－病毒治疗系统AdHA对血管内皮细胞增殖的抑制作用，意在为卵巢癌抗血管生成治疗提供实验依据。方法：采用直接感染台盼蓝染色排除法、MTF实验检测血管抑素K1-5对血管内皮细胞增殖的抑制作用。结果：直接感染台盼蓝染色排除法及MTT法检测表明，携带血管抑素K1-5基因的腺病毒能显著抑制血管内皮细胞增生，并对其具有杀伤力。结论：血管抑素K1—5能明显抑制血管内皮细胞增殖，为进一步行体内实验，研究其对血管形成的抑制作用及卵巢癌抗血管生成基因治疗奠定了基础。     

rAAV+HO-1基因对慢性脑缺血大鼠认知能力和氧化应激的影响

目的:研究从重组腺相关病毒介导血红素氧合酶-1(rAAV+HO-1)基因对大鼠慢性脑缺血认知能力改变和抗氧化应激作用。方法:成年雄性Wistar大鼠45只,每组各15只,随机分为假手术组(SH)、缺血2月组(2-VO),缺血2月+HO-1治疗组(2-VOH)。各组在脑立体定位仪下定位大鼠海马部位,2-VOH组缓慢注射rAAV+HO-1基因2μl,余组注入等量生理盐水。7 d后,采用永久节扎双侧颈总动脉方法制备慢性脑缺血动物模型。大鼠慢性缺血8周后,Morris水迷宫检测大鼠空间学习能力,Western blotting检测大鼠海马组织HO-1蛋白表达水平的变化,酶学法检测大鼠海马SOD活性、MDA含量、NO水平改变。结果:2-VO组大鼠认知水平下降与SH组比较,差异有统计学意义(P<0.01),2-VOH组比2-VO组逃避潜伏期明显减少,空间探索试验穿过平台次数增多,差异具有统计学意义(P<0.01),2-VOH组HO-1蛋白水平表达与2-VO组对比有统计学差异(P<0.01),2-VOH组与2-VO组对比SOD活性增多、MDA含量和NO水平下降,有统计学差异(P<0.05),2-VOH组与SH组SOD活性、MDA和NO含量无统计学差异(P>0.05)。结论:海马注射重组腺相关病毒介导HO-1基因,能够使慢性脑缺血大鼠海马组织HO-1蛋白水平表达升高,增加SOD活性,降低MDA、NO含量,减少氧化应激损伤,一定程度上改善大鼠空间认知学习能力。     

奥运会期间病媒生物控制保障面临的挑战及其防制策略

目的通过分析奥运会期间病媒生物控制保障所面临的风险与挑战,制定并实施有效保障策略和控制措施,确保奥运会期间不受病媒生物危害的影响。方法通过对近年来北京市病媒生物种类、密度消长特点以及危害程度的分析,提出奥运会期间媒介生物性传染病发生的危险性以及病媒生物可能造成的骚扰与不利影响;根据2005-2008年奥运会场馆及城区病媒生物密度、病原与抗药性监测结果,以及安全、环保、有效控制和管理措施的筛选,制定奥运会期间病媒生物控制保障策略。结果北京市存在媒介生物性传染病发生和病媒生物造成骚扰的危险;健全的管理机制、全面的密度监测、科学合理的用药、统一的灭虫除害、应急队伍的组建等措施成为奥运会期间病媒生物控制的主要策略。结论实施奥运会期间病媒生物监测与控制,开展防制技术应用研究,制定各类技术方案,为2008年北京奥运会期间病媒生物控制提供了有力的技术支撑,实现了北京奥运会无重大病媒生物危害事件发生的控制目标。     

2008年北京奥运会期间病媒生物控制效果分析与经验启示

目的通过对2008年北京奥运会期间病媒生物控制工作和控制效果的分析，总结奥运会期间病媒生物控制保障的成功经验与遗产。方法对奥运核心区、全市奥运场馆、场馆外围进行全面系统的病媒生物密度监测，以奥运场馆为重点，对周边2km及全市范围开展大规模的病媒生物控制活动。结果降低了奥运场馆和全市病媒生物密度，完全实现了“有效控制城市奥运涉及区的病媒生物种类及数量”的奥运公共卫生保障目标。结论围绕奥运会所采取的一些措施和做法是行之有效的，所形成的一些独特经验、技术措施和控制机制已成为奥运会的一份宝贵遗产，对今后北京市和我国的病媒生物控制工作都具有重要的指导意义。     

Control of Japanese encephalitis in India: A reality

Japanese encephalitis is considered as a serious disease due to the complexity of the disease and lack of specific treatment. A secular trend towards declining of JE has been brought in China, Korea, and Japan with widespread use of JE vaccine. In India, the actual JE burden could be estimated only by strengthening diagnostic facilities for JE confirmation in hospitals. However, the available records at present indicate a rising trend in JE-occurrence and expansion of the disease into JE non-endemic areas, which cannot be ignored. JE control through vector control methods has limitations owing to sustainability and cost effectiveness of the programs. Under these circumstances, feasibility of JE vaccination in India has to be considered as a preventive measure, for which identification of risk areas, target populations to be immunized, cost-evaluation of immunization is emphasized. Since, JE vaccine is produced in India, extension of the availability of this vaccine into routine JE-immunization programs is not remote. China has proved that countries with limited sources can produce safe and effective JE vaccines.     

The Ozobranchus leech is a candidate mechanical vector for the fibropapilloma-associated turtle herpesvirus found latently infecting skin tumors on Hawaiian green turtles (Chelonia mydas)

Fibropapillomatosis (FP) of marine turtles is a neoplastic disease of ecological concern. A fibropapilloma-associated turtle herpesvirus (FPTHV) is consistently present, usually at loads exceeding one virus copy per tumor cell. DNA from an array of parasites of green turtles (Chelonia mydas) was examined with quantitative PCR (qPCR) to determine whether any carried viral loads are sufficient to implicate them as vectors for FPTHV. Marine leeches (Ozobranchus spp.) were found to carry high viral DNA loads; some samples approached 10 million copies per leech. Isopycnic sucrose density gradient/qPCR analysis confirmed that some of these copies were associated with particles of the density of enveloped viruses. The data implicate the marine leech Ozobranchus as a mechanical vector for FPTHV. Quantitative RT-PCR analysis of FPTHV gene expression indicated that most of the FPTHV copies in a fibropapilloma have restricted DNA polymerase expression, suggestive of latent infection.     

Selection of Rabbit Single-chain Fv Fragments against the Herbicide Atrazine using a New Phage Display System

A convenient new bacteriophage display vector, pSD3, has been constructed and used to generate rabbit monoclonal anti-pesticide antibody fragments. Following amplification of immunoglobulin light chain, and heavy chain variable region gene libraries, restriction enzymes Sfi I and PflM I are used to assemble scFv libraries in pSD3. This allows the number of stages involving the polymerase chain reaction and restriction enzyme digestion to be minimized to optimize maintenance of the original diversity of the variable region genes in the libraries. The vector also incorporates an amber codon, a 6xHis tag and a c-myc epitope to facilitate soluble single-chain Fv production detection and purification. Using the pSD3 system two anti-atrazine single-chain Fvs were isolated from a library derived from the spleen cells of a rabbit immunized with bovine serum albumin-atrazine conjugate. Characterization of single-chain Fvs by competition and equilibrium ELISA indicated good specificity and affinity to atrazine.     

髓样相关蛋白14真核表达载体的构建及其细胞内定位研究

目的:构建小鼠髓样相关蛋白14(MRP14)的真核表达载体,观察其在NIH3T3细胞中的表达定位情况。方法:提取BALB/c小鼠肝脏组织的总RNA,通过逆转录-聚合酶链反应(RT-PCR)扩增得到MRP14编码序列。然后将该编码序列克隆到带有血凝素(HA)标记的载体pcDNA3-HA上,随后转染NIH3T3细胞,在荧光显微镜下观察结果。结果:重组质粒经聚合酶链反应(PCR)、酶切和测序鉴定证明构建正确。转染实验发现,该质粒能够在NIH3T3细胞中表达,表达产物主要定位在细胞质中。结论:成功构建带HA标签的MRP14真核表达载体。该载体能在哺乳动物细胞中有效表达并正确定位,为下一步深入研究MRP14作用细胞的信号通路提供了一个重要工具。     

Influential observations in cointegrated VAR models: Danish money demand 1973–2003

Summary This paper suggests a set of simple diagnostic tools for assessing the influence of a patch of κ observations in a cointegrated vector autoregressive model. The diagnostics are based on the leave‐κ‐out principle ( Bruce and Martin, 1989 Journal of the Royal Statistical Society, Series B, 51, 363–424) and the influence is measured by the likelihood displacement ( Cook and Weisberg, 1982 Residuals and Influence in Regression. London: Chapman and Hall). An application to Danish money demand 1973–2003 suggests that the observations for real money in 1999 are affected by institutional factors related to the definition of broad money, and that the dynamic adjustment following the international oil‐price shock in 1973 is very influential for the long‐run parameters.     

E2F1的siRNA真核表达载体转染对胆管癌细胞的作用

目的:探讨E2F1蛋白信号的转导通路对胆管癌细胞凋亡的影响中的作用。方法:根据E2F1基因cDNA序列，设计针对目的基因的4个siRNA靶序列，将其插入U6启动子下游，克隆到真核表达载体pGsensil中，通过酶切鉴定和DNA测序鉴定。采用脂质体介导法将构建好的4个siRNA表达载体分别转染于QBC939细胞中；以RT-PCR检测E2F1基因在转染前后的表达，流式细胞术检测转染后对细胞凋亡率的影响。结果:重组质粒酶切后呈线性化，酶切鉴定及DNA测序结果显示插入片断正确。转染48h后，4个siRNA表达载体均抑制了E2F1 mRNA的表达，沉默E2F1基因后细胞的凋亡率显著增加。结论:E2F1的siRNA真核表达载体转染QBC939细胞后，该表达载体具有抑制E2F1基因表达、促进细胞凋亡的功能，可以用于后续胆管癌的实验研究。