舰艇病媒生物本底调查研究

目的开展大范围的舰艇环境病媒生物本底调查研究,为相关防治工作提供科学依据和指导性建议。方法在北海、东海及南海2个区域设调查点,监测当地气温在20～25℃之间时舰艇上的病媒生物侵害情况。结果水面舰艇主要病媒生物种类为蟑螂与鼠类,侵害率分别高达74.45%与66.15%;其次为蚊、蝇,侵害率分别为27.54%和57.73%;鼠类体表寄生蚤与寄生革螨侵害率分别为43.1%和27.06%;舱室尘螨孳生较为普遍,达到83.2%。潜艇环境只捕获到蟑螂与尘螨,侵害率分别为11.11%和50.39%。舰艇病媒生物侵害有显著的地域性,呈从北到南增强的趋势,而且随服役年限增加,鼠类与蟑螂侵害呈明显上升趋势。结论水面舰艇病媒生物侵害严重,特别是鼠类体表寄生虫及其传播的疾病应引起相关管理部门的重视。     

“Bird biting” mosquitoes and human disease: A review of the role of Culex pipiens complex mosquitoes in epidemiology

The transmission of vector-borne pathogens is greatly influenced by the ecology of their vector, which is in turn shaped by genetic ancestry, the environment, and the hosts that are fed on. One group of vectors, the mosquitoes in the Culex pipiens complex, play key roles in the transmission of a range of pathogens including several viruses such as West Nile and St. Louis encephalitis viruses, avian malaria (Plasmodium spp.), and filarial worms. The Cx. pipiens complex includes Culex pipiens pipiens with two forms, pipiens and molestus, Culex pipiens pallens, Culex quinquefasciatus, Culex australicus, and Culex globocoxitus. While several members of the complex have limited geographic distributions, Cx. pipienspipiens and Cx. quinquefasciatus are found in all known urban and sub-urban temperate and tropical regions, respectively, across the world, where they are often principal disease vectors. In addition, hybrids are common in areas of overlap. Although gaps in our knowledge still remain, the advent of genetic tools has greatly enhanced our understanding of the history of speciation, domestication, dispersal, and hybridization. We review the taxonomy, genetics, evolution, behavior, and ecology of members of the Cx. pipiens complex and their role in the transmission of medically important pathogens. The adaptation of Cx. pipiens complex mosquitoes to human-altered environments led to their global distribution through dispersal via humans and, combined with their mixed feeding patterns on birds and mammals (including humans), increased the transmission of several avian pathogens to humans. We highlight several unanswered questions that will increase our ability to control diseases transmitted by these mosquitoes.     

HSV 2多功能蛋白ICP27真核表达载体构建及其在Vero细胞中的表达

目的构建单纯疱疹病毒2型（HSV 2） 多功能蛋白感染细胞多肽27（ICP27）真核表达载体pCDNA3.0 ICP27,并检测其在非洲绿猴肾细胞（Vero）中的表达。方法提取病毒株HSV 2333 DNA,用高保真DNA聚合酶对多功能蛋白ICP27基因进行高保真扩增,用双酶切连接至真核表达载体pCDNA3.0中。pCDNA3.0 ICP27经双酶切、测序验证,并通过脂质体介导质粒瞬时转染Vero 细胞,经RT PCR和Western blot检测ICP27蛋白的表达。结果pCDNA3.0 ICP27经双酶切可切出目的片段,测序结果经比对,与基因库中的序列完全一致。转染后经RT PCR 和Western blot检测,证实转染重组质粒组有ICP27蛋白表达,而转染空质粒组及未转染组没有检测到ICP27的表达。结论成功构建了HSV 2 多功能蛋白ICP27真核表达载体pCDNA3.0 ICP27,并能在Vero 细胞中表达。     

大鼠PEDF基因RNA干扰慢病毒载体的构建与鉴定

目的 构建针对大鼠PEDF 基因的RNA干扰（RNAi）慢病毒表达载体并进行鉴定,探讨PEDF基因对缺血心肌血管新生的影响.方法 构建PEDF基因过表达质粒,测序鉴定.针对PEDF基因不同干扰靶点构建4种RNAi 病毒载体质粒pGCsi-U6/Neo/GFP/shRNA,测序鉴定.过表达质粒和RNAi质粒共转染293T细胞后应用Western blot方法进行外源筛靶确定PEDF基因RNAi 有效靶序列.有效RNAi 病毒质粒和其他3种辅助包装原件载体质粒通过Lipofectamine 2000共转染293T细胞,培养48 h后, 收集细胞培养上清液,将其浓缩后用孔稀释法测定病毒滴度.结果 过表达质粒及4种RNAi质粒构建成功.Western blot外源筛靶显示2个靶点能有效敲减目的基因的表达.PEDF shRNA慢病毒表达载体Serpinf1-RNAi-LV经293T细胞包装成功,收集293T细胞分泌的病毒上清测定浓缩病毒悬液的滴度为2×1012Tu/L.结论 成功构建了PEDF基因的RNAi慢病毒载体,为研究PEDF基因对缺血心肌血管新生的影响打下基础.     

猪APOBEC3F基因克隆及真核表达载体的构建与鉴定

目的 克隆猪载脂蛋白B mRNA编辑酶催化多肽样蛋白3F(apolipoprotein B mRNA editing enzyme,catalytic polypeptide 3F,APOBEC3F)基因,构建真核表达载体实现其在体外表达与鉴定。方法分离4头21周龄,体质量25～30kg的雌性健康五指山猪的外周血单个核...     

RPS14基因重组慢病毒载体的构建及其在MUTZ-8细胞中的表达

目的 构建携带人RPS14基因重组慢病毒载体获得稳定产毒的细胞,并观察其在人MDS细胞株MUTZ-8细胞中的表达.方法 通过反转录方法,获得目的基因RPS14;将目的基因克隆入慢病毒表达质粒pGC-FU Vector,构建含有RPS14基因的重组慢病毒载体pGC-FU RPS14,经PCR、酶切及测序鉴定其正确性;在...     

癌胚抗原相关细胞黏附分子6-shRNA及双自杀基因yCDglyTK的联合基因载体构建

目的 构建新型靶向联合双自杀基因载体pCDNA3.1-yCDglyTK-CEACAM6-shRNA.方法 根据已知的癌胚抗原相关细胞黏附分子6(Carcinoembryonic antigen associated cell adhesion molecule 6,CEA CAM6)序列构建含有hU6启动子和CEACAM6-shRNA表达框的重组质粒,并将CEACAM6-shRNA表达框(含hU6启动子)通过双酶切方式亚克隆至双自杀基因栽体pCDNA3.1-yCDglyTK,构建新型靶向联合双自杀基因载体pCDNA3.1-yCDglyTK-CEACAM6-shRNA.通过酶切、测序等鉴定重组质粒.结果 实验所构建的最终裁体酶切产物所得的片段大小为7.5 kb,与预期片段一致.测序证实最终载体与pCDNA3.1-ycDglyTK-CEACAM6-shRNA序列一致.结论 成功构建新型靶向联合双自杀基因载体pCDNA3.1-yCDglyTK-CEACAM6-shRNA.     

针对小鼠VEGFA基因的siRNA干扰载体的构建和慢病毒包装

目的 针对NM_001025250.3,NM_001025257.3,NM_009505.4,构建4个针对靶基因小鼠VEGFA(血管内皮生长因子A)的小干扰RNA(siRNA)干扰载体,并将其重组转成慢病毒载体.方法 根据靶基因设计并合成4对siRNA干抚序列,将siRNA干扰序列克隆到pcDNA6.2-GW/EmGF...     

临沂口岸医学媒介生物本底调查名录

目的 为全面了解临沂口岸医学媒介生物的种群组成及分类地位,为本口岸媒介生物防治及卫生口岸创建提供科学依据.方法 1998年以来,坚持对所辖区域媒介生物的监测和调查.结果 鉴定出媒介昆虫2纲9目21科51属83种.结论 通过调查与鉴定,建立了临沂口岸医学媒介生物本底名录,为今后实施口岸媒介生物监测和控制提供了便利.