Rift Valley fever virus(Bunyaviridae: Phlebovirus): an update on pathogenesis, molecular epidemiology, vectors, diagnostics and prevention

Rift Valley fever(RVF) virus is an arbovirus in the Bunyaviridae family that, from phylogenetic analysis, appears to have first emerged in the mid-19th century and was only identified at the beginning of the 1930's in the Rift Valley region of Kenya. Despite being an arbovirus with a relatively simple but temporally and geographically stable genome, this zoonotic virus has already demonstrated a real capacity for emerging in new territories, as exemplified by the outbreaks in Egypt (1977), Western Africa (1988) and the Arabian Peninsula (2000), or for re-emerging after long periods of silence as observed very recently in Kenya and South Africa. The presence of competent vectors in countries previously free of RVF, the high viral titres in viraemic animals and the global changes in climate, travel and trade all contribute to make this virus a threat that must not be neglected as the consequences of RVF are dramatic, both for human and animal health. In this review, we present the latest advances in RVF virus research. In spite of this renewed interest, aspects of the epidemiology of RVF virus are still not fully understood and safe, effective vaccines are still not freely available for protecting humans and livestock against the dramatic consequences of this virus.     

中内毒素（LPS）诱导后家蝇幼虫抗菌肽CecMd基因的克隆、生物信息学分析及其表达载体的构建

从脂多糖（LPS）诱导或非诱导的家蝇幼虫体内抽提总RNA,根据家蝇Cecropin基因（GenBank：DQ 232774）设计特异性引物,应用RT—PCR技术扩增出编码CecMd开放阅读框（ORF）的eDNA序列,将其克隆入pMD18-T载体,经PCR、双酶切和测序鉴定正确后,将CecMd亚克隆基因与线性化的pET32a（＋）连接,构建重组表达载体pET32a（＋）／CecMid,并对此基因及其推导氨基酸序列进行系统的生物信息学分析。结果表明从脂多糖（LPS）诱导的家蝇幼虫体内成功的扩增出家蝇抗菌肽Cecropin基因,命名为CecMd,但是从未诱导的虫体内未能获得此基因;CecMd开放阅读框的cDNA全长为192bp,编码由63个氨基酸残基组成的多肽;此多肽最可能的裂解位点在氨基酸残基A23和G24之间,提示1～23氨基酸残基组成信号肽,24～63氨基酸残基构成成熟肽;CecMd的全长肽包含4个螺旋,成熟肽（不包括信号肽）含有2个螺旋,两者均不合二硫键;CecMd与果蝇Cecropins的同源性显著高于其他昆虫,达到81．0％以上;经PCR,限制酶切和DNA测序鉴定表明,成功构建了重组表达载体pET32a（＋）／Cec Md 。重组表达载体pET32a（＋）／Cec Md的构建及其CecMd基因的生物信息学分析对其生物学功能研究和高效表达工程菌的构建具有重要意义。     

人β2微球蛋白成熟肽基因克隆及其原核表达载体的构建

根据GenBank基因库中人β2微球蛋白(β2m)成熟肽基因序列设计2对引物,使用RT-PCR法从健康人血液中扩增人β2m成熟肽基因,扩增产物进行T-A克隆和测序.结果表明,获得的人β2m成熟肽基因为297 bp,与模板序列的同源性为100%.利用基因重组技术,将β2m成熟肽基因亚克隆入pET-28a(+)载体中,经PCR、酶切和测序鉴定,证实所获重组表达质粒pET-28/Humβ2m中含有目的片段,且连接、构建正确,表明成功构建了重组表达质粒pET-28/Ham β2m.     

马铃薯ARF基因RNAi载体的遗传转化及对其试管薯生理特性的影响

用含有ADP核糖基化因子(ARF)基因的干扰载体的农杆菌转化马铃薯栽培品种“甘农薯2号”,获得转化植株48株,经卡那霉素抗性筛选和PCR检测证明ARF基因已成功整合到马铃薯基因组中。用实时荧光定量PCR检测表明,转化植株中ARF基因的表达较对照均有不同程度的下降,在转基因株系Z-11和Z-12中,ARF基因表达的干扰程度分别高达92.53%和93.22%。对转基因植株诱导的试管薯的淀粉、可溶性糖、蛋白质、蔗糖和葡萄糖含量测定结果表明,与未转基因的对照相比,转基因植株的淀粉含量提高了4.29%~34.27%,蛋白质含量提高了1.47%~7.35%,可溶性糖含量提高了1.44%~17.39%;蔗糖含量提高了11.53%~53.84%,而葡萄糖含量降低了6.06%~21.21%。     

端粒逆转录酶基因修饰对绵羊骨髓间充质干细胞端粒酶活性影响的研究

骨髓间充质干细胞(BMSC)具有多向分化的特性,但其生命周期有限,为进一步探讨端粒逆转录酶(TERT)对端粒酶活性的影响,本试验利用将外源性TERT导入间充质干细胞技术,构建重组表达载体pcDNA3.1-EGFP-TERT,将其转染BMSC,通过筛选及鉴定,结果显示成功构建了重组表达载体pcDNA3.1-EGFP-TERT.将TERT-BMSCs在体外进行传代培养及生长曲线的测定,利用PCR技术对其原有干细胞因子的表达情况进行了鉴定.结果显示,TERT-BMSC具有快速扩增的能力,细胞形态良好,且仍具有干细胞特性.本试验结果揭示了外源性TERT基因的表达能激活绵羊BMSC的端粒酶活性.     

反转录病毒载体RCASBP介导的EGFP基因在DF-1细胞中的表达

为建立反转录病毒载体RCASBP介导的增强型绿色荧光蛋白(EGFP)基因在DF-1细胞中的表达体系,本研究将PCR获得的EGFP基因插入反转录病毒载体RCASBP,构建重组反转录病毒载体RCASBP-EGFP,之后将重组载体转染DF-1细胞;用基于禽白血病病毒(ALV) p27抗原的ELISA检测盲传至第4代的细胞上清,ELISA阳性结果说明重组病毒拯救成功;荧光显微镜观察发现80%以上DF-1细胞都有明显的绿色荧光信号,证明DF-1细胞表达绿色荧光蛋白;对DF-1细胞基因组进行特异性PCR检测,扩增出特异性条带说明重组反转录病毒载体RCASBP-EGFP携带的EGFP基因整合到DF-1细胞的基因组中。本研究建立的RCASBP介导反转录病毒表达体系为研究ALV基因的结构和功能奠定了基础。     

介体传播的草莓病毒研究进展

草莓病毒在世界各草莓栽培区广泛分布,可随无性繁殖材料和传毒介体传播扩散,严重影响草莓的产量和品质。目前已知传播草莓病毒的介体主要有蚜虫、蓟马、粉虱、线虫和真菌等,总共可传播19种病毒,其中2个为病毒暂定种。本文综述了不同介体传播的草莓病毒的分类地位、分子生物学和生物学特性等方面的研究进展,同时总结并展望了介体传播草莓病毒的检测技术和防治方法的新策略。     

Control methods against invasive Aedes mosquitoes in Europe: a review

Five species of invasive Aedes mosquitoes have recently become established in Europe: Ae. albopictus, Ae. aegypti, Ae. japonicus japonicus, Ae. koreicus and Ae. atropalpus. These mosquitoes are a serious nuisance for people and are also competent vectors for several exotic pathogens such as dengue and chikungunya viruses. As they are a growing public health concern, methods to control these mosquitoes need to be implemented to reduce their biting and their potential for disease transmission. There is a crucial need to evaluate methods as part of an integrated invasive mosquito species control strategy in different European countries, taking into account local Aedes infestations and European regulations. This review presents the control methods available or in development against invasive Aedes mosquitoes, with a particular focus on those that can be implemented in Europe. These control methods are divided into five categories: environmental (source reduction), mechanical (trapping), biological (e.g. copepods, Bacillus thuringiensis var. israelensis, Wolbachia), chemical (insect growth regulators, pyrethroids) and genetic (sterile insect technique and genetically modified mosquitoes). We discuss the effectiveness, ecological impact, sustainability and stage of development of each control method. © 2015 Society of Chemical Industry     

基于NURBS曲面的小麦叶片三维可视化研究与实现

作物三维可视化将为作物生长动态预测、栽培管理调控、作物株型设计等提供形象逼真的可视化工具,从而有力促进虚拟农业和信息农业的发展。针对小麦形态结构复杂、叶片三维可视化不易表达的问题,基于田间实测数据和NURBS曲面建模实现了小麦叶片的三维重建。首先,通过田间实验定期观测小麦形态数据(叶长、叶宽、茎叶夹角、叶片弯曲度、茎秆直径等),建立小麦叶片主脉控制点构造算法,将小麦形态数据转换为主脉控制点坐标;其次,确定所有控制点坐标和节点矢量,构建小麦叶片和叶鞘NURBS曲面;最后,借助Open GL图形库可实现小麦不同生育期的模拟模型,并实现了小麦叶片的扭曲形变。结果表明该方法选取的小麦形态数据具有代表性,重建的小麦叶片模型逼真度较高,接近实测模型,易于实现小麦生长过程的动态模拟。