汉坦病毒感染者血清病毒RNA的临床意义

汉坦病毒感染啮齿类动物,但不引起疾病。人类可通过接触这些宿主动物的尿、粪等排泄物中携带的病毒而感染。汉坦病毒RNA检测技术已经由核酸分子杂交,发展为实时荧光定量PCR,敏感性、特异性和可重复性明显提高。汉坦病毒感染为急性感染性疾病,病程早期可以在患者血清中检出病毒RNA。汉坦病毒感染者血清中病毒RNA与凝血功能、炎症因子、疾病严重程度、疾病预后等密切相关。在肾综合征出血热(HFRS)患者的发热期和低血压休克期病毒载量较高,随着病程进展,病毒载量逐渐下降,直至消失。检测汉坦病毒感染者血清中病毒RNA载量有重要临床价值。     

抗汉坦病毒核蛋白细胞内抗体抗病毒作用的研究

本文报道在内质网和细胞浆内稳定表达抗汉坦病毒核蛋白细胞内抗体,研究抗汉坦病毒核蛋白细胞内抗体抗病毒作用及其抗病毒作用机理。在获得稳定表达抗汉坦病毒核蛋白细胞内抗体的细胞系的基础上,用MTT法检测细胞内表达抗汉坦病毒单链抗体对细胞增殖的影响,证实细胞内抗体的表达不会影响到细胞的生长。不同时间收集病毒感染毒后细胞培养培养上清和细胞裂解物,用ELISA方法检测细胞内外病毒结构蛋白含量的变化,结果表明定位于细胞内质肉和细胞浆内的抗汉坦病毒核蛋白细胞内抗体均能不同程度降低感染细胞上清中的核蛋白含量,但是不能或轻微抑制细胞内汉坦病毒核蛋白的合成。利用病毒微量滴定免疫荧光方法,对细胞内抗体与病毒的增殖关系进行了研究,证实无论在内质网还是在胞质,抗核蛋白细胞内抗体都能够抑制病毒的增殖,具有抗病毒活性。抗汉坦病毒核蛋白细胞内抗体不能抑制病毒结构蛋白的合成,其抗病毒效果是通过抑制病毒的包装而实现。     

汉坦病毒及所致疾病

汉坦病毒是引起多种人类疾病的病原体,为布尼亚病毒科的一个属.已发现至少有20个血清/基因型,每型均有其特定啮齿类动物宿主,病毒种系发生与宿主种系发生密切相关.不同病毒型别对人类致病性不同,其损伤器官和病情轻重各异.已发现2种主要疾病肾综合征出血热(HFRS)和汉坦病毒肺综合征(HPS).在过去的几十年里,对汉坦病毒及其所致疾病的认识有了很大进展病毒型别不断增加,病毒致病谱不断扩大.有证据表明,在英国,汉坦病毒也可引起人类疾病,但无明显特征.     

汉坦病毒的基因分型及其序列分析

为了探讨从核苷酸水平耐汉坦病毒进行分型,设计两对型特异性引物,采用反转录和聚合酶链式反应（ＲＴ－ＰＣＲ）,对亚太地区１８株汉坦病毒进行了扩增鉴定,并对其中７株汉坦病毒的ＰＣＲ产物进行了测序分析。ＰＣＲ的分型结果表明,Ⅰ型引物只能扩增血清Ⅰ型病毒的ｃＤＮＡ;Ⅱ型引物也只能扩增血清Ⅱ型病毒,其间无交叉反应。采用巢式ＰＣＲ和限制性内切酶验证了ＰＣＲ产物的特异性。序列分析结果表明,Ｒ３６Ｍ片段Ｇ１区的核苷酸序列与血清Ⅰ型病毒代表株７６－１１８的同源性为７８．４％,而与血清Ⅱ型病毒Ｒ２２的同源性为６８．１％;Ｒ３６与汉坦病毒序列同源性的成对比较结果也表明,Ｒ３６与血清Ⅰ型病毒的同源性均高于血清Ⅱ型病毒;Ｌｅａｋｅｙ虽然能被Ⅱ型引物扩增,但其序列与血清Ⅱ型病毒Ｒ２２的同源性仅为４４．９％,故不属于血清Ⅱ型病毒。上述研究结果表明,反转录聚合酶链反应能对多数汉坦病毒准确分型,但最终结果尚有赖于序列分析。     

汉坦病毒肺型综合征

汉坦病毒肺型综合征是由一种新型的汉坦病毒所引起的烈性传染病，该病毒与汉坦病毒有交叉抗，但它们之间有差异。临床表现以发热，肺水肿、呼吸窘迫综合征为特征，病死率高达５０－７０％。     

吉林地区HFRS病原体基因型分析

为调查吉林地区褐家鼠中汉坦病毒病原体基因型及其分布情况,采用免疫荧光法检测吉林市区及吉林市所属各县市收集的鼠肺标本,对汉坦病毒抗原阳性标本以巢式RT-PCR法扩增S基因片段上的核苷酸序列并测序,将扩增片段的核苷酸序列与已知病毒序列进行同源性分析及系统进化树分析.序列分析发现吉林地区褐家鼠携带的汉坦病毒均为汉城病毒,S3...     

我国汉坦病毒基因型和基因亚型的分布研究

为了搞清全国汉坦病毒的基因型和亚型的分布情况,广泛收集了全国各地汉坦病毒毒株、阳性病人血清和阳性鼠肺,并应用RT-PCR的方法,应用汉坦病毒型特异性引物,对这些不同来源的阳性标本中汉坦病毒型特异性M和S片段进行扩增和测序,并与其它已知病毒序列进行比较,以明确其型别和亚型及其在全国的分布情况.结果表明:我国HFRS各疫区仍然为HTNV和SEOV两型病毒,但亚型分布差异较大,其中HTNV可分为9个亚型,SEOV则有4～6个亚型.Q32株的部分M和S片段分属于H9和H2亚型,是一个基因重排病毒,而Nc167株在系统发生上与其它HTNV明显不同,比较核苷酸序列发现,其M片段与其它HTNV的同源性在71.3%～76.7%之间,S片段与其它HTNV的同源性只有52.3%～57.8%,可能是一个新型病毒.     

汉坦病毒陈株S基因编码区的克隆、序列分析及表达

从汉坦病毒陈株感染的Vero-E6细胞裂解液中提取病毒RNA,经逆转录PCR获得病毒S基因编码区约1.3kb cDNA片段,克隆该片段后进行核苷酸序列测定,并与汉坦病毒76-118株进行同源性比较,结果二者核苷酸序列同源性为86%,推导的氨基酸序列同源性为97%.将该基因片段插入原核表达载体pGEX-4T-1,在大肠杆菌中获得高效表达.表达产物为GST-NP融合蛋白.SDS-PAGE检测表达蛋白分子约72kD左右.Western blotting和ELISA试验结果表明,表达产物可与多株抗汉坦病毒核蛋白的McAb发生反应,其抗原表位及McAb反应谱与76-118株相比存在某些差异.     

东北三省6株肾综合征出血热病毒生物学性状的研究

肾综合征出血热(hemorrhagic fever with renal syndrome,HFRS)是我国重点防治的急性传染病,对于流行于不同地区的汉坦病毒(hanatavirus)进行分离和系统地鉴定,可以为HFRS的防治提供科学依据,特别是对流行株血清型及基因型的确定,是研制疫苗和确定防治重点及对策的重要前提,也为研究我国汉坦病毒的遗传和进化特征奠定了基础。研究对6株流行于东北三省的汉坦病毒进行了全面鉴定,其中H8205、H8207株为人源性汉坦病毒,其余均为鼠源性汉坦病毒。研究表明,6株病毒具有病毒型别明确,抗原性较为广谱,免疫原性好,在Vero细胞上适应能力强及病毒滴度高,以及来源及传代历史清楚等特点,适合作为双价HFRS纯化灭活疫苗候选毒株,并且可建立起完善和合格的生产用毒种库,为我国生产以Vero细胞为基质的双价肾综合征出血热纯化灭活疫苗奠定了坚实的基础。     

汉坦病毒感染对Ⅰ型干扰素应答的调节机制

汉坦病毒主要感染人内皮细胞,细胞在病毒感染早期诱导生成的Ⅰ型干扰素(IFN-Ⅰ)可阻断汉坦病毒的复制,但不同的病毒蛋白和不同型别的汉坦病毒在IFN应答的调节机制上可能有所不同。就汉坦病毒感染对IFN应答的调节机制进行了综述。     

流感病毒生命周期研究进展

流感病毒引起急性呼吸道感染,分为季节性流感和大流行流感。季节性流感每年都有发生,感染后可以导致肺炎和急性呼吸道衰竭,往往并发细菌共感染。大流行流感每隔一段时间发生一次,具有高致病性特点。近年来大流行流感频繁暴发,带来了巨大的经济和社会负担。本文从流感病毒生物学、生命周期等方面进行简要综述,有助于为预防和治疗流感病毒感染提供策略。     

TSG101在病毒出芽过程中的作用及其机制

TSG101基因是新发现的抑癌基因候选者,定位于人类 11号染色体 p1511-p1512,其编码产物TSG101蛋白N端区域与泛素结合酶(UBC)同源。近年来研究发现,TSG101基因具有多种重要的功能,与多种病毒出芽密切相关,所以TSG101可作为一个新的抗病毒靶点。本文主要从TSG101在多种病毒（HIV、IAV、MARV、ASV等）出芽过程中扮演的角色,TSG101与多种蛋白(泛素、Nedd4、ARMMs、Tom1、Gag、VP40、NP等)的相互作用进而辅助病毒出芽的机制,以及TSG101抑制剂的研究等方面进行阐述。     

流感病毒在脂筏上组装与出芽

流感病毒造成的季节性流行性疾病给全世界带来沉重的健康负担.近年来,甲型流感病毒的变种H5N1、H7N9给各国带来了很大危害.流感病毒属于正黏附病毒科,它的遗传物质由多个节段的负链RNA组成,其组装和出芽剪切生殖是一个涉及到多种病毒因子,多步骤、复杂的生化过程.流感病毒会使用宿主的细胞膜上的"脂筏"区域作为病毒出芽位点.首先病毒的两种糖蛋白NA蛋白、HA蛋白会在脂筏区域聚集,造成脂筏区膜变形弯曲,并且发动出芽的过程.接着,流感病毒基质蛋白M1的C端与HA、NA结合,其自身在脂筏区域开始多聚化并使膜向外弯曲形成原始病毒体的内部结构,接着招募病毒的核糖核蛋白复合物(VRNP)与M2蛋白,使组装的过程进一步完成.最后,M2蛋白会富集在原始病毒体的底部,完成膜的剪切和病毒体的释放.     

细胞囊泡的研究进展

阐述了细胞囊泡出芽、运输、融合的分子机制,并就最新进展进行了综述．以对这个领域有一初步认识。     

Digestive cell and tentacle number in freshly detached buds of Hydra viridis

Buds were collected from hydras fed four days a week on different schedules. Independent of schedule, parents produced the same number of buds per week, but significant differences appeared in the number of buds detaching on particular days, and in the number of digestive cells present in the buds. Groups of buds collected from parents fed the same number of days (from one to three) during the previous four days contained statistically indistinguishable numbers of digestive cells despite the order or sequence of days on which feedings occurred. The number of digestive cells in all the freshly detached buds collected here can be accounted for by the growth of a bud primordium over a four day period of bud development and growth. Such a primordium would have about 3,600 digestive cells and grow at the rate of 0.33 cells per cell per day of feeding. The numbers of tentacles found on freshly detached buds are correlated with the number of feeding days and digestive cells present in the bud. Tentacles, therefore, may also form from primordia consisting originally of a specific number of cells.     

Scanning electron microscopic evidence for a budding mode of chloroplast multiplication in higher plants

Scanning electron microscopic observations of leaf sections of certain plants show two distinct types of chloroplast multiplication. In the normal mode, chloroplast division is achieved by binary fission. In Bryophyllum pinnatum Kurz. the chloroplast multiplication occurs by a process similar to the budding of yeast. The advantages of using scanning electron microscopy to follow closely the different stages of chloroplast division are demonstrated.     

The influence of ethylene on proliferation and growth of rose shoot cultures

Ethylene accumulation in four different rose in vitro culture containers was evaluated. Multiplication rate was the highest, and axes most elongated, in the two containers where ethylene accumulation was limited. Pulse treatments of ethylene at various concentrations enhanced proliferation depending on concentration (5 ppm generally was the most favourable) and time of application, while reducing elongation of the shoots. An ethylene trap in the flask atmospheres of the cultures reduced rose shoot proliferation rate but increased elongation of the axes. Inhibitors of ethylene biosynthesis, aminoethoxyvinylglycine (AVG) and cobalt chloride (CoCl₂), increased multiplication rate by providing a higher number of axes of a suitable size for subculture. The ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) had a beneficial effect on multiplication rate, although reducing longitudinal growth of the axes.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglycine - BA benzyladenine - GA₃ gibberellic acid - IBA indolyl-3-butyric acid     

Phospholipase D and membrane traffic: Potential roles in regulated exocytosis,membrane delivery and vesicle budding

It is now well-established that phospholipase D is transiently stimulated upon activation by G-protein-coupled and receptor tyrosine kinase cell surface receptors in mammalian cells. Over the last 5 years, a tremendous effort has gone to identify the major intracellular regulators of mammalian phospholipase D and to the cloning of two mammalian phospholipase D enzymes (phospholipase D1 and D2). In this chapter, we review the physiological function of mammalian phospholipase D1 that is synergistically stimulated by ADP ribosylation factor, Rho and protein kinase Cα. We discuss the function of this enzyme in membrane traffic, emphasising the possible integrated relationships between consumption of vesicles in regulated exocytosis, membrane delivery and constitutive membrane traffic.     

应用正交设计方法优化香蕉外植体直接出芽的条件

利用正交设计方法（L27（3^13)]研究了培养基中的植物生长调节物BAP,NAA和IBA,以及不同外植体对香蕉（Musa acuminata Colla.cv.Williams) 直接出芽频率的影响,外植体的不同是影响香蕉直接出芽频率的最主要因素,NAA和IBA对实验结果的影响相同,均弱于BAP,实验结果的统计分析表明,诱导香蕉外植体直接出芽的优选培养基为不添加BAP,NAA和IBA的MS基本培养基,最佳外植体为芽的顶端分生组织部分。     

MripacC regulates blastosphere budding and influences virulence of the pathogenic fungus Metarhizium rileyi

Fungal dimorphism is the ability of certain fungi to switch between two different cellular forms, yeast and mycelial forms, in response to external environmental factors. The pacC/Pal signal transduction pathway responds to neutral and alkaline environments and is also involved in the fungal dimorphic transition. In this study, we investigated the function of the pacC homolog, MripacC, which regulates the dimorphic transition and modulates virulence of the insect pathogenic fungus Metarhizium rileyi. MripacC expression was upregulated under alkaline condition, with increased number of yeast-like cells compared to the number of hyphae cells. A MripacC deletion mutant (ΔMripacC) was obtained by homologous replacement and exhibited decreased blastospore budding, with direct development of conidia into hyphae without entering the yeast-like stage when cultured on alkaline medium. Observation of host hemolymph morphology and analysis of samples to detect the main immune factors revealed a decreased ability of ΔMripacC to evade the host immune system. The results of insect bioassays showed that ΔMripacC had decreased virulence with extended median lethality time. Together, the results suggested that MripacC not only regulated adaptation to acidic and alkaline environments, but also influenced virulence by budding blastospores. This elucidation of the function of MripacC adds to our understanding of blastospore budding and virulence of this fungal pathogen.