肠道病毒71型2A蛋白的研究进展

肠道病毒71(enterovirus 71,EV71)是手足口病(hand,foot and mouth disease,HFMD)主要致病原之一。手足口病临床上常表现为发热,手掌、脚掌及口腔黏膜皮疹或疱疹。然而,EV71感染导致的手足口病易伴随神经系统并发症,甚至死亡。EV71非结构蛋白2A作为蛋白酶和转录激活因子,在EV71生命周期中发挥重要作用。本文对2A的结构与功能研究进展进行综述,揭示2A的双重功能如何促进病毒复制和调控靶细胞,为进一步研究靶向2A的抗病毒疫苗和药物提供理论基础。     

肠道病毒71型北京分离株VP4基因序列及蛋白抗原性分析

为探讨肠道病毒71型(EV71)VP4基因序列与手足口病(HFMD)不同临床类型之间的关系,分析重组蛋白EV71 VP4的抗原性,并初步探讨其与柯萨奇病毒A16(CA16)重组蛋白VP4是否存在交叉反应性,对2007~2009年从北京患HFMD儿童标本分离到的10株EV71的VP4基因进行克隆测序,运用生物学软件对测序结果进行比较分析,并选择其中1株与1株同期分离的CA16的VP4分别进行原核表达;用表达产物对189份正常体检的成人及来首都儿科研究所就医的非HFMD患儿血清中的IgG进行Western Blot检测,并分析14份确诊为EV71感染和12份CA16感染患者急性期血清中的IgM抗体。分析表明这10株EV71 VP4基因核苷酸同源性为94.20%~100.00%,所推导的氨基酸序列则完全相同,从重症与轻症患儿分离的毒株之间VP4的核苷酸序列未见一致性的差异,基于EV71 VP4基因核苷酸序列的进化树分析表明2007~2009年北京地区所流行的毒株均属于C4亚型;本研究中EV71和CA16的VP4核苷酸序列的同源性为69.60%,所推导的氨基酸序列的同源性为78.60%,在运用Western Blot检测189份血清中的VP4特异性IgG时,EV71 VP4的血清阳性率为38.10%,说明其具有良好的抗原性,CA16 VP4的血清阳性率为58.20%,两者差别具有显著统计学意义(2χ=15.30,P<0.01),提示EV71 VP4与CA16 VP4没有交叉反应性;在用表达的VP4检测已确诊为相应病毒的特异性IgM时,两者皆为阴性,提示感染后机体对VP1和VP4产生不同的反应。     

2009年上海市手足口病流行病学和病原学特征

本研究对上海市手足口病(Hand-foot-and-mouth disease,HFMD)的流行病学和病原学特征进行了分析。从国家疾病监测信息报告管理系统获取上海市2009年HFMD的流行病学数据;采用荧光定量RT-PCR方法对来自上海15个区县的799例HFMD进行肠道病毒(Enterovirus,EV)核酸检测;对部分肠道病毒71型(Enterovirus71,EV71)的VP1区和部分其它EV的VP4区序列进行测定和分析。采用Office excel软件进行简单数据统计分析,用BioEdit和MEGA软件进行病毒基因特征分析,通过BLAST服务器的在线比对进行EV型别鉴定。分析显示:上海市18个区县均有病例报告,地区分布无显著特点;小于6岁的婴幼儿期为疾病高发年龄段;4～7月为发病高峰期;EV71和柯萨奇病毒A16(Coxsackie virus A16,CA16)为主要病原,不同地区、不同月份的病原构成各不相同,CA16为轻症病例的主要病原,EV71则为重症病例的主要病原;基于VP1区序列分析,上海EV71毒株与C4a亚型毒株具有最近的亲缘性和最高的同源性;2株其它肠道病毒经鉴定属于CA2和CA10型。结果表明:EV71和CA16为2009年上海HFMD流行的主要病原,EV71属于C4a亚型;除EV71和CA16外,还存在小部分其它EV(如:CA2和CA10)引起的HFMD。     

ANXA2 Facilitates Enterovirus 71 Infection by Interacting with 3D Polymerase and PI4KB to Assist the Assembly of Replication Organelles

Virologica Sinica - Similar to that of other enteroviruses, the replication of enterovirus 71 (EV71) occurs on rearranged membranous structures called replication organelles (ROs)....     

黄土高原不同降水类型区旱作玉米田土壤干燥化效应与土壤水分承载力模拟研究

在模拟精度验证基础上,应用WinEPIC模型模拟研究了黄土高原不同降水类型区1960～2000年期间不同肥力水平下连作春玉米的产量变化和土壤水分效应.模拟结果表明:(1)洛川、延安、寿阳、榆林连作春玉米产量地区间差异显著,其年平均值分别为4.40、3.75、 2.50、 1.75 t/hm2.施肥增产率和水分利用效率地区间差异均显著,均表现为洛川>延安>寿阳>榆林;同一地区水分利用效率随施肥量的增加而提高,不同地区施肥增产率最大值范围不同.(2)0～7m土层逐月土壤有效含水量地区间差异显著,洛川>延安>寿阳>榆林;同一地区内肥力水平越高,春玉米耗水量越大,逐月土壤有效含水量平均值越低.在春玉米生育期内0～7m土层年度土壤有效水分增减量地区间差异显著,洛川<延安<寿阳<榆林;不同肥力处理间的差异在模拟前期为显著,在模拟后期为不显著.(3)所有地区的春玉米在连作期间0～7m土层土壤湿度剖面分布,在经历了土壤湿度逐年降低、土壤干层逐年加厚的干燥化过程后,均出现了稳定的土壤干层.土壤干燥化速度地区间差异显著,洛川<延安<寿阳<榆林;不同肥力处理间随着施肥水平的提高土壤干燥化速度加快.(4)洛川、延安、寿阳、榆林连作春玉米的适宜产量水平分别为5.25～5.54、4.26～4.58、2.34～2.74、1.37～1.62 t/hm2,相应的施肥水平分别为N240(N 240 kg/hm2,P 120 kg/hm2)～N300(N 300 kg/hm2,P 150kg/hm2)、N180(N 180 kg/hm2,P 90kg/hm2)～N240(N 240 kg/hm2,P 120 kg/hm2)、N120(N 120 kg/hm2,P 60kg/hm2)～N180(N 180 kg/hm2,P 90kg/hm2)、N60(N 60 kg/hm2,P 30kg/hm2)～N120(N 120 kg/hm2,P 60kg/hm2).     

AMMI模型在旱地春小麦稳定性分析中的应用

基因型与环境的互作(GEI)决定了作物在多变环境下性状的稳定性。AMMI模型是一种将方差分析和主成分分析结合于一体,能更有效分析GEI、进而评价基因型稳定性和环境对基因型差异分辨力的有力工具。利用AMMI模型对10个品种(系)、13个试点组成的全国旱地春小麦区域试验产量资料分析表明,试点间平均产量变幅为396.6~4050.2 kg.hm-2,现代品种间的平均产量变幅为1318.6~2315.6 kg.hm-2;基因型间、环境间和GEI引起的产量变异达到极显著水平,三者的变异平方和分别占总处理平方和的6.2%、70.3%、23.5%,表明环境和GEI对产量变化的影响远大于基因型。用前3个代表了90.8%GEI信息的显著主成分计算基因型稳定性参(Di)和试点分辨力(Dj),基因型间Di最大相差达3倍、而试点间Dj最大相差19倍;属于高产、稳产的品种有:定西35、西旱1号、定丰889,而在这两方面均表现最差的品种为蒙麦35号。有些品种对某些试点有特殊适应性,局部推广价值也大。     

广东金山温泉沉积物中原核与真核微生物多样性初步分析

[目的]本研究旨在采用不同的PCR引物对广东省恩平市金山温泉的高温水底沉积物微生物多样性进行初步的分析.[方法]采用改进的玻璃珠法抽提温泉沉积物中环境基因组DNA,通过对用4对引物分别扩增得到的原核微生物16S rRNA基因和真核微生物ITS序列的分析,将所得到的数据与国际基因数据库GenBank进行相似性比较并构建系统发育树.[结果]研究发现原核类群G的 14个优势克隆中7个都属于蛭弧菌属(Bdellovibrio).与它们最相似的序列是从海洋中分离到的两个菌株 Bacteriovorax sp. NE1 (EF092445)和Bdellovibrio sp. JS5 (AF084859),相似性分别为96%和99%.原核类群X的4个序列主要属于蓝细菌类群,其中JS-X2与在美国黄石公园温泉发现的Uncultured Cyanobacterium (L35331)有95%的相似性,并且与已经全基因组测序的嗜热蓝细菌聚球藻Thermosynechococcus elongatus BP-1 (47118315)有89%的相似性.真核类群Z有三个类群,分别是Penicillium sp.,Lodderomyces sp.和Gloeotinia sp..其中大部分序列与青霉属相似性在88%～ 90%之间.[结论]所得到的结果显示金山温泉中的微生物多样性十分丰富.     

云南洱源牛街热泉原核微生物多样性分析

【目的】通过分析富含高分子有机物的云南洱源牛街热泉原核微生物16SrRNA基因克隆文库,丰富对高温热泉原核微生物多样性的认识,为进一步开发和利用该热泉微生物资源奠定基础。【方法】构建洱源牛街高温热泉原核微生物16SrRNA基因克隆文库,通过测序和序列相似性比对以及聚类分析研究该热泉原核微生物的多样性。【结果】该热泉原核微生物以细菌为主,包括变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、梭杆菌门(Fusobacteria)等在内的约10个细菌类群,其中变形菌门中的β-变形菌纲(β-Proteobacteria)为优势菌群,其次为拟杆菌门(Bacteroidetes)、绿菌门(Chlorobi);古菌的生物量和丰度较细菌少,分属广古菌(Euryarchaeota)和泉古菌(Crenarchaeota)两个类群,以广古菌为优势类群。     

半干旱黄土高原地区地膜覆盖和底墒对春小麦生长及产量的影响

在年均降水量为415mm的半干旱地区黄绵土旱地上,以春小麦为供试作物进行大田实验,研究不同底墒(包括低底墒、中底墒和高底墒)下．地膜覆盖(包括不覆膜、播种后覆膜30d、覆膜60d和覆膜120d即全程覆膜)进程对作物生产的影响。结果表明．增加底墒和合理的覆膜进程均会显著增加作物的生长和产量,但底墒不同,其最佳覆膜进程不同：在低底墒时,覆膜处理反而使产量低于不覆膜处理;在中底墒时．覆膜30d产量最高,随着覆膜时间延长．产量呈下降趋势,甚至全程覆膜产量低于不覆膜处理;高底墒以覆膜60d产量最高。综合作物生长和产量,全程覆膜并没有多少实际意义。在同种覆膜处理下,随着底墒的增加,根生物量、地上干物质、叶面积及产量也增加显著,高底墒覆膜60d处理的产量在所有处理中为最高。     

Viruses in recreational water‐borne disease outbreaks: a review

Viruses are believed to be a significant cause of recreationally associated water‐borne disease. However, they have been difficult to document because of the wide variety of illnesses that they cause and the limitations in previous detection methods. Noroviruses are believed to be the single largest cause of outbreaks, which have been documented in the published literature 45% (n = 25), followed by adenovirus (24%), echovirus (18%), hepatitis A virus (7%) and coxsackieviruses (5%). Just under half of the outbreaks occurred in swimming pools (49%), while the second largest outbreak occurred in lakes or ponds (40%). The number of reported outbreaks associated with noroviruses has increased significantly in recent years probably because of better methods for virus detection. Inadequate disinfection was related to 69% (n = 18) of swimming pool outbreaks. A lack of required reporting and nonuniform water quality and chlorination/disinfection standards continues to contribute to water‐borne recreational disease outbreaks.     

The simultaneous detection of both enteroviruses and adenoviruses in environmental water samples including tap water with an integrated cell culture-multiplex-nested PCR procedure

AIMS: To evaluate an integrated cell culture (ICC)-multiplex-nested PCR using the buffalo green monkey kidney (BGMK) cells for the simultaneous detection of both enteroviruses and adenoviruses in surface water and tap water samples and optimize the procedure for more sensitive detection of virus showing no apparent cytopathic effect (CPE). METHODS AND RESULTS: A total of 69 surface water and 50 tap water samples were analysed by the ICC-multiplex-nested PCR. All the PCRs were performed five times with a cell lysate from each flask after at least 2 weeks incubation. Forty-six surface water samples (66.7%) and 23 tap water samples (46.0%) exhibited CPE by the cell culture method. By using an ICC-multiplex-nested PCR, 53 surface water samples (76.8%) and 29 tap water samples (58.0%) were determined as containing infectious enteric viral particles. CONCLUSIONS: An ICC-PCR method with a long incubation time using BGMK cells enables the simultaneous detection of enteroviruses and adenoviruses from environmental water samples, including tap water, even with low numbers of viruses. SIGNIFICANCE AND IMPACT OF THE STUDY: A method capable of detecting small numbers of viral particles is necessary.     

Sulfur-metabolizing bacterial populations in microbial mats of the Nakabusa hot spring, Japan

At the Nakabusa hot spring, Japan, dense olive-green microbial mats develop in regions where the slightly alkaline, sulfidic effluent has cooled to 65 °C. The microbial community of such mats was analyzed by focusing on the diversity, as well as the in situ distribution and function of bacteria involved in sulfur cycling. Analyses of 16S rRNA and functional genes (aprA, pufM) suggested the importance of three thermophilic bacterial groups: aerobic chemolithotrophic sulfide-oxidizing species of the genus Sulfurihydrogenibium (Aquificae), anaerobic sulfate-reducing species of the genera Thermodesulfobacterium/Thermodesulfatator, and filamentous anoxygenic photosynthetic species of the genus Chloroflexus. A new oligonucleotide probe specific for Sulfurihydrogenibium was designed and optimized for catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH). In situ hybridizations of thin mat sections showed a heterogeneous vertical distribution of Sulfurihydrogenibium and Chloroflexus. Sulfurihydrogenibium dominated near the mat surface (50% of the total mat biovolume), while Chloroflexus dominated in deeper layers (up to 64% of the total mat biovolume). Physiological experiments monitoring in vitro changes of sulfide concentration indicated slight sulfide production by sulfate-reducing bacteria under anoxic-dark conditions, sulfide consumption by photosynthetic bacteria under anoxic-light conditions and strong sulfide oxidation by chemolithotrophic members of Aquificae under oxic-dark condition. We therefore propose that Sulfurihydrogenibium spp. act as highly efficient scavengers of oxygen from the spring water, thus creating a favorable, anoxic environment for Chloroflexus and Thermodesulfobacterium/Thermodesulfatator in deeper layers.     

半干旱区春小麦生长系统的人工神经网络模型与产量预测

以半干旱区春小麦生长系统为研究对象。探讨了作物生长系统中水分、土壤养分等生态因子的时空变化特征及春小麦产量形成机制,应用人工神经网络方法建立了半干旱区春小麦生长系统的产量随环境因子变化的神经网络模型,并与传统的CTM模型进行了比较。模拟结果表明,人工神经网络模型可适用于半干旱区春小麦生长系统产量随环境因子变化规律描述,且优于传统模型,从而为春小麦产量预测提供了新的途径,也为作物生态系统的人工调控提供了新的模式与定量依据。     

Biodiversity and distribution of water mites (Acari, Hydrachnidia) in spring habitats

1. The distribution, species richness and ecology of spring‐dwelling water mites in Italy were investigated with the aim to better elucidate the role of spring habitats to sustain high levels of biodiversity and their contribution to freshwater biodiversity at a local and regional scale. 2. More than 300 springs in different geographic areas (Alps, Central and Southern Apennines, Sicily and Sardinia), were examined with a total of 163 water mite species recorded. 3. Species richness in each area ranged from 33 species on Sardinia to 77 on Sicily. The highest diversity was found in the Gran Sasso (Central Apennines). The proportion of crenobionts (species strictly bound to this type of habitat) exceeded 50% in almost all the areas investigated. 4. A diverse (up to 20 species per spring) and highly specialised mite fauna was observed in undisturbed rheocrenes and in natural springs of intermediate typology (rheohelocrenes and rheopsammocrenes). In springs subjected to human impacts (pasture, deforestation, alteration and transformation of spring sources) species richness declined and crenobionts were replaced by unspecialised crenoxenes. 5. The zoogeographic importance of spring habitats is confirmed by the presence of 18 endemic species and by members of genera with an interesting disjunct and relict distribution. 6. A comparison with other geographic areas suggests that springs contain a significant fraction of the total number of species found in freshwater habitats and may contribute almost one third of regional freshwater biodiversity. The presence of endemic crenobionts and rare taxa highlights the importance of these habitats in maintaining high levels of biodiversity as well as their contribution to a better understanding of biodiversity patterns in freshwaters.