Molecular characterization of avian influenza virus (H7N8) isolated from poultry in Central China in the mid-1980s

The molecular and pathogenic properties of avian influenza virus (A/duck/Hubei/216/1985/H7N8) isolated from Hubei Province of China in 1985 were characterized.The hemagglutinin gene (HA) of Dk/Hb/216/85/H7N8 had the multiple amino acid se-quences (-PEIPKGRG-) at the connecting peptide between HA1 and HA2, which is considered to be a distinguishing molecular characteristic of low pathogenicity.The key sites of host markers among the genes (M, NP, NS, PA and PB2) of Dk/Hb/ 216/85/H7N8 were similar to those of...     

Molecular patterns of avian influenza A viruses

Avian influenza A viruses could get across the species barrier and be fatal to humans. Highly pathogenic avian influenza H5N1 virus was an example. The mechanism of interspecies transmission is not clear as yet. In this research, the protein sequences of 237 influenza A viruses with different subtypes were transformed into pseudo-signals. The energy features were extracted by the method of wavelet packet decomposition and used for virus classification by the method of hierarchical clustering. The clustering results showed that five patterns existed in avian influenza A viruses, which associated with the phenotype of interspecies transmission, and that avian viruses with patterns C and E could across species barrier and those with patterns A, B and D might not have the abilities. The results could be used to construct an early warning system to predict the transmissibility of avian influenza A viruses to humans.     

Genome evolution of novel influenza A （H1N1 ） viruses in humans

The epidemic situation of A H1N1 flu arose in North America in April 2009, which rapidly expanded to three continents of Europe, Asia and Africa, with the risk ranking up to 5. Until May 13th, the flu virus of A H1N1 had spread into 33 countries and regions, with a laboratory confirmed case number of 5728, including 61 deaths. Based on IRV and EpiFluDB database, 425 parts of A H1N1 flu virus sequence were achieved, followed by sequenced comparison and evolution analysis. The results showed that the current predominant A H1N1 flu virus was a kind of triple reassortment A flu virus： （i） HA, NA, MP, NP and NS originated from swine influenza virus; PB2 and PA originated from bird influenza virus; PB1 originated from human influenza virus. （ii） The origin of swine influenza virus could be subdivided as follows： HA, NP and NS originated from classic swine influenza virus of H1N1 subtype; NA and MP originated from bird origin swine influenza virus of H1N1 subtype. （iii） A H1N1 flu virus experienced no significant mutation during the epidemic spread, accompanied with no reassortment of the virus genome. In the paper, the region of the representative strains for sequence analysis （A/California/04/2009 （H1N1） and A/Mexico/4486/2009 （H1N1）） included USA and Mexico and was relatively wide, which suggested that the analysis results were convincing.     

Molecular characterization of H1N1 influenza A viruses from human cases in North America

Subtypes of H1N1 influenza virus can be found in humans in North America, while they are also associated with the infection of swine. Characterization of the genotypes of viral strains in human populations is important to understand the source and distribution of viral strains. Genomic and protein sequences of 10 isolates of the 2009 outbreak of influenza A （H1N1） virus in North America were obtained from GenBank database. To characterize the genotypes of these viruses, phylogenetic trees of genes PB2, PB1, PA, HA, NP, NA, NS and M were constructed by Phylip3.67 program and N-Linked glycosylation sites of HA, NA, PB2, NS1 and M2 proteins were analyzed online by NetNGlyc1.0 program. Phylogenetic analysis indicated that these isolates are virtually identical but may be recombinant viruses because their genomic fragments come from different viruses. The isolates also contain a characteristic lowly pathogenic amino acid motif at their HA cleavage sites （IPSIQSR↓GL）, and an E residue at position 627 of the PB2 protein which shows its high affinity to humans. The homologous model of M proteins showed that the viruses had obtained the ability of anti-amantadine due to the mutation at the drug-sensitive site, while sequence analysis of NA proteins indicated that the viruses are still susceptible to the neuraminidase inhibitor drug （i.e. oseltamivir and zanamivir） because no mutations have been observed. Our results strongly suggested that the viruses responsible for the 2009 outbreaks of influenza A （H1N1） virus have the ability to cross species barriers to infect human and mammalian animals based on molecular analysis. These findings may further facilitate the therapy and prevention of possible transmission from North America to other countries.     

Increased substitution rate in H5N1 avian influenza viruses during mass vaccination of poultry

As a means of heated debate,mass vaccination of poultry has been used in some countries to control H5N1 highly pathogenic avian influenza(HPAI),which remains of global economic and public health significance.Theoretically,mass vaccination can act as an evolutionary selective force facilitating the emergence of vaccine-resistant viruses,similar to that widespread use of antibiotics facilitates the emergence of antibiotic-resistant bacteria.To support the hypothesis,the substitution rates in the two subunits,HA1 and HA2,of the viral hemagglutinin gene,were calculated using a Bayesian Markov Chain Monte Carlo(MCMC) approach.It was found that the rate in the HA1 subunit,but not in the HA2 subunit,increased significantly during periods of mass vaccination(2005 2010 in China and 2003 2009 in Indonesia),in contrast to the periods when no mass vaccination programs took place(1996 2004 in China and 2004 2008 in Thailand).Because substitutions in the HA1 subunit rather than in the HA2 subunit can lead to vaccine-resistant viruses,the results support that mass vaccination programs facilitate the emergence of vaccine-resistant viruses,which,in turn,will render mass vaccination programs less effective.Therefore,caution must be taken when adopting mass vaccination as a long-term strategy to control HPAI.     

中国大陆H9N2亚型禽流感病毒HA基因遗传分析及抗原相关性研究

为研究我国大陆H9N2亚型禽流感病毒血凝素(HA)基因的分子进化及抗原相关性, 本研究对来自15个省、市、自治区的34株H9N2亚型禽流感病毒的HA基因进行了测序及系统发育分析, 并采用交叉血凝抑制试验及交叉攻毒保护试验对不同遗传分支下毒株间抗原相关性进行了分析. 结果表明, 所有34个毒株HA基因均符合低致病性禽流感病毒的特征, 但毒株间变异程度增加. 系统发育分析表明, 我国大陆H9N2亚型禽流感病毒主要分为三个系列, 各系列内毒株没有明显的地区及时间特征. 抗原相关性研究表明, 不同遗传系列下的毒株其抗原相关性明显低于同一系列内部毒株间的抗原相关性, 说明我国H9N2亚型禽流感病毒抗原性差异较大. 此外, 本研究同时筛选得到了用于制备多价苗的代表毒株.     

H5N1亚型禽流感毒株A/Duck/HuBei/3/2005的分子特征及致病性研究

采用分子生物学方法,对1株2005年从湖北省某县分离获得的禽流感病毒株(A/Duck/HuBei/3/2005)进行全基因组序列测定.序列分析显示,该分离株为H5N1亚型.HA蛋白在HA1和HA2连接处,含有连续多碱性氨基酸模体(-RRKKR-).根据进化分析结果,分离株A/Duck/HuBei/3/2005的7个基因来源于2004～2005年湖南地区流行株(CK/HN/999/05,DK/HN303/04),但是PA基因片段发生了重排,来源于野禽.动物实验显示DK/HB/3/05对鸡和鸭均具有高致病性;对小鼠有较低致病性.     

鄱阳湖湿地环境质量分异及风险表征

根据实地采样分析、现场调查及相关资料,获取鄱阳湖相关水质、生态数据,运用统计学、环境风险评价方法对鄱阳湖污染物空间分异及风险表征进行了量化分析,初步得出结论:鄱阳湖各入湖河口,主要污染物为氨氮、石油类、镉、挥发酚等.pH值除乐安河口、信江东支河口较低外,其他入湖河口都在6.0左右浮动.鄱阳湖全湖水质处于中等营养化水平.3月份DO与COD呈正相关,但拟合曲线显著性检验和方差分析不显著,而11月份呈负相关性,显著性检验较好;3月份和11月份DO与BOD,COD与BOD都呈正相关,方差检验不显著.波阳和康山样地重金属含量最高,鄱阳湖样地和东源样地重金属综合含量(东源样地As含量略高)位于最后,湖口和石钟山样地Pb含量普遍高于其他样地.各研究区风险分析综合评价指数大小顺序是:康山>波阳>湖口>九江>南矶山>鄱阳湖样地.     

鄱阳湖微囊藻属形态多样性研究

2011年1月至10月对鄱阳湖的微囊藻 Microcystis 进行采集和分离培养,得到9种常见微囊藻并对其进行了详细的形态学观察和描述,分别是铜绿微囊藻（M. aeruginosa）、水华微囊藻（M. flos-aquae）、鱼害微囊藻（M. ichthyoblabe）、惠氏微囊藻（M. wesenbergii）、假丝微囊藻（M. pseudofilamentosa）、史密斯微囊藻（M. smithii）、放射微囊藻（M. botrys）、绿色微囊藻（M. viridis）、挪氏微囊藻（M. novacekii）,讨论了培养情况下微囊藻的形态学差异,阐述了鄱阳湖水华蓝藻类群的研究现状及微囊藻的潜在危害.     

H5N1亚型禽流感病毒血凝素基因的克隆与表达

 根据已知H5N1亚型禽流感病毒血凝素(HA)基因序列设计、合成克隆引物.自灭活的云南地方H5N1亚型病毒阳性临床组织样品中提取总RNA,反转录后采用高可信度DNA聚合酶(Pyobest^TMDNA Polymerase)扩增HA基因,采用Invitrogen定向表达系统(Champion^TMpET directional TOPO expression system)进行克隆表达,纯化获得N末端携带多聚组氨酸标签的重组HA,分子质量约78ku.采用阳性血清经免疫印迹及ELISA分析重组HA的免疫反应性,结果表明重组HA能与H5N1亚型病毒抗血清发生特异性结合,具有良好的免疫反应性.     

Sequence and epitope analysis of surface proteins of avian influenza H5N1 viruses from Asian patients

Avian influenza virus strain H5N1 is a highly patho- genic type A influenza virus that has caused several outbreaks of severe poultry plagues in the past. H5N1 preferentially binds to receptors with α2-3 linked sialic acids on the surface of avian epithe…     

Antibodies induced by multi?epitope vaccine showed inhibitory activity against heterologous influenza A virus (H3N2)

In this study, recognition of 4 recombinant viral proteins (GST?NHA1) by the antibodies induced by multi?epitope vaccine was testified. Inhibitory activities of these antibodies were also investigated in vitro against four heterologous influenza A viruses (H3N2). Three epitope?specific antibodies purified by affinity chromatography could reduce the plaque formation. Interestingly, the three neutralizing antibodies in combination showed obvious enhancement of inhibitory activity, suggesting that the development of recombinant multi?epitope vaccine might be an effective way against viral mutation.     

Prediction of a common neutralizing epitope of H5N1 avian influenza virus by in silico molecular docking

The H5N1 avian influenza virus （AIV） has widely spread in Asia, Europe and Africa, making a large amount of economic loss. Recently, our research group has screened a common neutralizing mono-clonal antibody named 8H5, which can neutralize almost all H5 subtype AIV ever isolated so far. Obviously, this monoclonal antibody would benefit for research and development of the universal AIV vac-cine and design of the drug against H5N1 AIV in high mutation rate. In this study, the homology modeling was applied to generate the 3D structure of 8H5 Fab fragment, and ＂canonical structure＂ method was used to define the specified loop conformation of CDR regions. The model was subjected to energy minimization in cvff force field with Discovery module in Insight II program. The resulting model has correct stereochemistry as gauged from the Ramachandran plot calculation and good 3D-structure compatibility as assessed by interaction energy analysis, solvent accessible surface （SAS） analysis, and Profiles-3D approach. Furthermore, the 8H5 Fab model was subjected to docking with three H5 subtype hemagglutinin （HA） structures deposited in PDB （ID No： ljsm, 2ibx and 2fk0） respectively. The result indicates that the three docked complexes share a common binding interface, but differ in binding angle related with HA structure similarity between viral subtypes. In the light of the three HA inter-faces with structural homology analysis, the common neutralizing epitope on HA recognized by 8H5 consists of 9 incontinuous amino acid residues： Asp^58, Asn^72, Glu^112, Lys^113, lie^114, Pro^118, Ser^120, Tyr^137, Tyr^252 （numbered as for ljsm sequence）. The primary purpose of the present work is to provide some insight into structure and binding details of a common neutralizing epitope of H5N1 AIV, thereby aiding in the structure-based design of universal AIV vaccines and anti-virus therapeutic drugs.     

Prediction of a common neutralizing epitope of H5N1 avian influenza virus by in silico molecular docking

The H5N1 avian influenza virus (AIV) has widely spread in Asia, Europe and Africa, making a large amount of economic loss. Recently, our research group has screened a common neutralizing mono- clonal antibody named 8H5, which can neutralize almost all H5 subtype AIV ever isolated so far. Obvi- ously, this monoclonal antibody would benefit for research and development of the universal AIV vac- cine and design of the drug against H5N1 AIV in high mutation rate. In this study, the homology mod- eling was applied to generate the 3D structure of 8H5 Fab fragment, and "canonical structure" method was used to define the specified loop conformation of CDR regions. The model was subjected to en- ergy minimization in cvff force field with Discovery module in Insight II program. The resulting model has correct stereochemistry as gauged from the Ramachandran plot calculation and good 3D-structure compatibility as assessed by interaction energy analysis, solvent accessible surface (SAS) analysis, and Profiles-3D approach. Furthermore, the 8H5 Fab model was subjected to docking with three H5 subtype hemagglutinin (HA) structures deposited in PDB (ID No: 1jsm, 2ibx and 2fk0) respectively. The result indicates that the three docked complexes share a common binding interface, but differ in bind- ing angle related with HA structure similarity between viral subtypes. In the light of the three HA inter- faces with structural homology analysis, the common neutralizing epitope on HA recognized by 8H5 consists of 9 incontinuous amino acid residues: Asp68, Asn72, Glu112, Lys113, Ile114, Pro118, Ser120, Tyr137, Tyr252 (numbered as for 1jsm sequence). The primary purpose of the present work is to provide some insight into structure and binding details of a common neutralizing epitope of H5N1 AIV, thereby aiding in the structure-based design of universal AIV vaccines and anti-virus therapeutic drugs.     

姜黄素体外抗流感病毒H1N1、H3N2实验研究

目的：观察姜黄素提取物体外对流感病毒H1N1、H3N2的抑制作用。方法：用狗肾细胞（MDCK）观察姜黄素体外对A型流感病毒H1N1亚型、A型流感病毒H3N2亚型病毒的直接杀灭作用。结果：姜黄素最大无毒浓度为12．5g／L,对H1N1有效抑制浓度为6．25g／L,对H3N2有效抑制浓度为1．56g／L。结论：姜黄素提取物确有明显的抗H1N1、H3N2复制作用。     

Orisin and future distribution of the new A （H 1N1 ） influenza virus emerging in North America in 2009

The origin of the new A （H1N1） influenza virus recently emerging in North America is a hot controversial topic of significance in disease control and risk assessment. Some experts claimed that it was an unusually mongrelized mix of human, avian and swine influenza viruses, while some others concluded that it was totally a simple re-assortment hybrid of two lineages of swine influenza viruses. Here the phylogenetic diversity of the viral PB1, PA and PB2 gene sequences using online web servers, and the results suggest that all the 8 genetic segments of the new virus were possibly from two lineages of swine influenza viruses, and one of the lineage was a mongrelized mix of human, avian and swine influenza viruses emerging in the world approximately 10 years ago. Considering the recent epidemiological trends of the new virus, we believe it will spread more widely in the world and persist long in human populations. It also could spread among swine populations. The future wide spreading of the new virus may coincide the disappearance of a subtype of previous human influenza A virus.     

鄱阳湖松门山岛砂质沉积颗粒表面特征及环境指示意义

沉积物沉积环境的判别对于沉积物物源的分析、古地理重建、古环境和古气候的恢复均具有重要的意义。通过研究发现鄱阳湖松门山岛沉积颗粒表面具有丰富的特征,主要是:贝壳状断口、V形撞击坑、撞击沟、碟形撞击坑与新月形撞击坑等。利用扫描电镜对鄱阳湖松门山岛的样品进行了研究,发现鄱阳湖松门山岛的水下砂体样品普遍可见贝壳状断口、V形撞击坑与撞击沟等,说明该类样品为水成环境;沙丘样品普遍可见标准的碟形撞击坑与新月形撞击坑等,说明为风成环境;处于水下砂体和沙丘过渡带之间的样品上述现象几乎均存在,说明这些沉积物经过风成和水成两种环境的共同改造。分析水成和风成环境下的石英颗粒的不同特征,对这些特征利用线测法进行统计分析,发现不同环境下石英颗粒的特征不仅可以定性判别,还可以半定量区分:在水成环境下的样品撞击沟所占的比值高,通常大于30%,而在风成环境中的样品碟形坑所占的比值高,通常大于40%。这一发现对该地区的沉积环境分析有指导作用,并且对于该地区沉积微相划分具有一定意义。     

鄱阳湖水面蒸发模型的研究与应用

本文根据都昌蒸发实验站近8年实验资料,建立了鄱阳湖3个单站水面蒸发模型,并应用它们计算了鄱阳湖大水面1962～1990年水面蒸发量,所得模型不仅可用于鄱阳湖地区,还可供江西省其它地区引用。     

鄱阳湖湿地洲滩前缘浅层土壤碳-氮-磷的时空特征

湿地洲滩土壤碳、氮、磷是重要的营养元素,其分布特征直接影响湿地生态系统的生产力和生态系统服务功能.通过2014-2017年对鄱阳湖湿地洲滩前缘浅层土壤(0～20 cm)有机碳、全氮、全磷观察实验分析,结果表明:鄱阳湖湿地洲滩前缘浅层土壤有机碳、全氮、全磷的年际变化特征不同,有机碳变化不显著,全氮、全磷变化显著; 浅层土壤有机碳、全磷、全氮的高程梯度变化极显著; 浅层土壤碳氮比、碳磷比年际变化极显著,氮磷比不显著; 浅层土壤碳氮比高程梯度变化不显著,碳磷比、氮磷比的高程梯度变化极显著.浅层土壤氮磷含量较其他     

多介质逸度模型研究鄱阳湖流域p,p'-DDT

以鄱阳湖流域为研究对象,利用TaPL3模型(Version 3.00)计算了该区域p,p’-DDT通过大气和水体的长距离传输潜力(LRTP),同时得出p,p’-DDT在水和大气环境中的持续时间以及在各相间的迁移通量和最终的相间分配结果。结果表明,鄱阳湖中p,p’-DDT通过大气的LRTP为385.6 km~678.7 km,均值为503.3 km,停留时间为1 761 d;通过水体的LRTP为24 395.7 km~184 154.3 km,均值为37 881 km,停留时间为1 173 d。土壤和沉积物相是鄱阳湖流域p,p’-DDT最大的汇,该区域最主要的界面迁移过程是气-植被界面、气-土壤界面、水-沉积物界面的扩散和沉降,植被的吸收、土壤和沉积物中的降解是鄱阳湖流域p,p’-DDT消失的最主要途径。