草鱼免疫相关基因Prkrip1的克隆和表达分析

随着草鱼养殖规模的扩大, 草鱼的病毒性疾病极大地影响着草鱼的产量。开展鱼类病毒免疫反应相关功能基因的研究意义重大。研究首先通过同源克隆的方法从草鱼中克隆到了一段Prkrip1基因的EST序列, 进一步通过RACE、长片段PCR和Genome walking的方法获得了该基因的全长cDNA序列、基因组DNA序列和启动子区序列。氨基酸序列分析显示, Prkrip1含有3个核定位信号和一个双链RNA结合区, 并具有与PKR结合的保守N端区; 荧光报告基因的表达证实我们所克隆到的启动子区是有活性的, 可用于后续该基因的转录调控分析; Real-time PCR分析发现, Prkrip1 基因在草鱼的肝和血中表达量最高, GCRV感染后在大部分免疫组织中均上调表达, 说明该基因确实与病毒感染相关。研究结果为Prkrip1基因在硬骨鱼类的功能研究提供了线索, 也为鱼类天然免疫反应中调控PKR信号通路的系统研究提供了理论依据。       

The parasitoid factor in the virulence and spread of lepidopteran baculoviruses

Insect parasitoids and baculoviruses play important roles in the natural and strategic biological control of insects. The two parasites are frequent competitors within common hosts and much research has focused on the negative impact that baculoviral host infections have on parasitoids. This review summarizes the impacts that parasitoids may have on the virulence and spread of lepidopteran baculoviruses. By changing host behavior and development, parasitoids have been shown to decrease baculovirus virulence and productivity within parasitized baculovirus-susceptible hosts; however, studies of the tools used by hymenopteran parasitoids to overcome their hosts' immune systems, suggest that parasitoids may, in some cases, facilitate baculoviral infections in less susceptible hosts. Laboratory and field research have demonstrated that parasitoids can mechanically transmit bacuioviruses between insects, and in this way, increase the efficacy of the viruses. Instances of new, more virulent isolates of baculoviruses have been recorded from specifically parasitoid-targeted hosts suggesting other possible benefits from the transmission or activation of baculoviruses by parasitoids.     

非溶细胞性抗病毒作用机制的研究进展

本文综述了非溶细胞性作用控制病毒感染的研究进展,并着重指出了细胞因子在这一过程中的地位,即直接激活抗病毒作用以及通过调节免疫反应上调病毒抗原决定簇在感染细胞表面的呈递和表达,从而间接控制病毒感染。     

应用抑制消减杂交和cDNA芯片技术筛选流行性感冒病毒感染相关基因

病毒基因组有限的编码能力和以病毒蛋白为靶的抗病毒药物易出现耐药性,使从病毒感染宿主筛选病毒感染相关生物大分子作为抗病毒药靶和诊断标志物成为新的研究方向。为了筛选流行性感冒（流感）病毒感染相关基因,采用抑制消减杂交（suppression subtractive hybridization,SSH)技术,以流感病毒A／鲁防／93－9（H3N2）感染的MDCK细胞及正常MDCK细胞为材料,构建病毒感染特异性差减cDNA文库。从文库中随机挑取约800个克隆,PCR扩增其中插入片段,经纯化、紫外定量后,用基因芯片自动点样仪点在氨基片上,制备cDNA芯片。将流感病毒感染的MDCK细胞和正常MDCK细胞的总RNA分别用Cy3、Cy5反转录荧光标记后,与cDNA芯片杂交,用芯片扫描仪扫描获得芯片杂交信号,经阳性对照校正和归一化处理后,以如下条件作为判定基因差异表达的标准;（a)Cy3与Cy5的信号比值大于1．5（正常细胞用Cy5标记）或小于0．67（正常细胞用Cy3标记）;（b)Cy3和Cy5信号值之一必须大于1000。经cDNA芯片筛选获得了18个流感病毒感染特异性克隆,经测序和生物信息学分析发现均为流感病毒感染相关新基因EST。流感病毒感染相关基因cDNA片段的获得,为新型病毒药靶诊断标志物发现和功能研究提供了基础。     

趋化因子在病毒感染中的作用及意义

趋化因子是近年发现的一大类结构相似、功能多样的新型分子。其不仅与淋巴细胞迁移、炎症反应、新生血管生成、造血、肿瘤有关,越来越多的证据表明趋化因子在病毒感染的发生、发展中扮演着重要角色。病毒感染宿主细胞后可导致组成性和诱导性趋化因子表达谱的改变,这些表达水平改变了趋化因子通过直接和间接的方式参与了病毒的致病过程;同时某些病毒可编码趋化因子样或趋化因子受体样分子,从而干扰趋化因子网络功能。研究趋化因子在病毒感染中的作用及意义,将有利于阐明在病毒性疾病的发病机制和抗病毒感染所策略的制定,具有重要的理论和应用价值。     

新现与再现的病毒及朊毒子

近年出现的新现或再现病毒包括禽流感病毒,Nipah病毒,Ebola病毒,人中间肺为病毒及朊毒子,这些病原体可致人,畜（禽）共患病,由昆虫或节肢动物媒物媒介传播,随国际交往频繁及工作习性变化而致感染,引起的感染必须作病原学诊断以识别,重新新现与再现病原体,对医学微生物学的理论与应用研究将开辟新源头。     

职业献血员中庚型肝炎病毒感染状况的分析

为了解职业献血员中庚型肝炎病毒（HGV）的感染状况,应用ELISA法和RT-PCR法进行检测,在10069名职业献血员中有516例抗-HGV阳性,总感染率为5.12%,其中男性感染率5.79%。女性感染率4.45%,甘肃省感染率4.96%,宁夏区感染率5.70%,青海感染率5.67%以及陕西感染率4.91%,年龄18-25岁的感染率4.48%,26-35岁感染率4.87%;36-45岁感染率6.90%;将抗-HGV阳性作RT-PCR,HBsAg,抗HCV检测,其中HGV-RNA阳性362例,HBsAg阳性25例,抗HCV阳性37例,HBsAg和抗-HCV阳性8例,以上说明,在职业献血员中,HGV的感染与性别,地域及年龄无关,与HBV,HCV有重叠感染现象。     

MicroRNA在埃博拉病毒感染中的作用研究进展

MicroRNA是一类通过影响RNA表达、抑制蛋白质翻译来调节基因表达的内源性非编码小分子RNA。自从2014年首次发现苏丹型和扎伊尔型埃博拉病毒能够编码microRNA以来,不断有研究发现新的病毒microRNA以及其在埃博拉病毒复制、扩散、免疫逃逸中发挥的作用。宿主microRNA则参与到抗病毒防御机制中,但也面临着病毒蛋白对其调控的阻力包括相应宿主microRNA水平的变化以及RNA沉默抑制子的编码。深入研究埃博拉病毒与宿主在microRNA水平上的动态相互作用,有利于进一步了解埃博拉病毒的致病机制以及开发新的诊断、治疗策略。本文就microRNA在埃博拉病毒感染中的作用做一简要综述。     

The host type I interferon response to viral and bacterial infections

Type I interferons (IFN) are well studied cytokines with anti-viral and immune-modulating functions. Type I IFNs are produced following viral infections, but until recently, the mechanisms of viral recognition leading to IFN production were largely unknown. Toll like receptors (TLRs) have emerged as key transducers of type I IFN during viral infections by recognizing various viral components. Furthermore, much progress has been made in defining the signaling pathways downstream of TLRs for type I IFN production. TLR7 and TLR9 have become apparent as universally important in inducing type I IFN during infection with most viruses, particularly by plasmacytoid dendritic cells. New intracellular viral pattern recognition receptors leading to type I IFN production have been identified. Many bacteria can also induce the up-regulation of these cytokines. Interestingly, recent studies have found a detrimental effect on host cells if type Ⅰ IFN is produced during infection with the intracellular gram-positive bacterial pathogen, Listeria monocytogenes. This review will discuss the recent advances made in defining the signaling pathways leading to type I IFN production.     

Viral infections and cell cycle G2／M regulation

Progression of cells from G2 phase of the cell cycle to mitosis is a tightly regulated cellular process that requires activation of the Cdc2 kinase, which determines onset of mitosis in all eukaryotic cells. In both human and fission yeast(Schizosaccharomyces pombe) cells, the activity of Cdc2 is regulated in part by the phosphorylation status of tyrosine 15 (Tyrl5) on Cdc2, which is phosphorylated by Weel kinase during late G2 and is rapidly dephosphorylated by the Cdc25 tyrosine phosphatase to trigger entry into mitosis. These Cdc2 regulators are the downstream targets of two wellcharacterized G2/M checkpoint pathways which prevent cells from entering mitosis when cellular DNA is damaged or when DNA replication is inhibited. Increasing evidence suggests that Cdc2 is also commonly targeted by viral proteins,which modulate host cell cycle machinery to benefit viral survival or replication. In this review, we describe the effect of viral protein R (Vpr) encoded by human immunodeficiency virus type 1 (HIV-Ⅰ) on cell cycle G2/M regulation. Based on our current knowledge about this viral effect, we hypothesize that Vpr induces cell cycle G2 arrest through a mechanism that is to some extent different from the classic G2/M checkpoints. One the unique features distinguishing Vpr-induced G2 arrest from the classic checkpoints is the role of phosphatase 2A (PP2A) in Vpr-induced G2 arrest.Interestingly, PP2A is targeted by a number of other viral proteins including SV40 small T antigen, polyomavirus T antigen, HTLV Tax and adenovirus E4orf4. Thus an in-depth understanding of the molecular mechanisms underlying Vpr-induced G2 arrest will provide additional insights into the basic biology of cell cycle G2/M regulation and into the biological significance of this effect during host-pathogen interactions.