microRNA let-7调控动物个体发育的研究进展

microRNA(miRNA)是一类广泛存在于多细胞动物中的进化保守的大小为18~25nt的非编码小分子RNA,可以通过与靶基因mRNA的非编码区(3′UTR)结合导致mRNA降解,或阻断mRNA翻译而调节基因表达。let-7是在线虫中发现的具有转录后调节功能的小分子RNA,具有高度的保守性,研究发现,let-7参与动物个体多个器官组织的发育过程。作者综述了近年来let-7参与调控脑、神经系统、心肺系统和肌肉发育等组织器官的研究成果,初步阐述了let-7调控组织器官发育的作用机制,以期为进一步探索let-7在动物体内的功能奠定基础。     

Isolation and Identification of one Strain of H9 Subtype Avian Influenza Virus and Study on its Biological Characteristics

In 2013,one case of suspected H9 subtype avian influenza occurred in a chicken farm of Jilin povince.Clinical samples were collected from the diseased farm,inoculated into the allantoic cavity of 9-day-old SPF chicken embryo,and then one strain of virus was isolated.The results of HA test,HI test and molecular biology test all showed that the isolate belonged to H9 subtype avian influenza virus (AIV).The HA cleavage site of the isolate was RSSR↓GLF,which was consisted with the molecular characteristic of low pathogenic AIV.The HA peptide chain had 9 potential glycosylation sites which were same as other isolates of recent years.The isolate had 8 receptor binding sites,including 234 receptor binding site by glutamine (Q) mutating into threonine (T).The phylogenetic tree revealed the isolate belonged to Eurasian lineages and it had far genetic relationship with the earliest domestic isolate (A/Chicken/Beijing/1/94(H9N2)),but had close genetic relationship with the representative strain (A/Chicken/Guangxi/55/2005(H9N2)) of major epidemic branch since 2007.We prepared an inactivated oil-emulsion vaccine of the isolate,and then vaccinated SPF chicken.21 days after vaccination,the HI titer of chicken serum antibody reached up to 10log2.The result suggested the isolate had good immunogenicity.     

Cloning and Sequence Analysis of N and M Genes of Avian Infectious Bronchitis Virus Guangxi Strain

To investigate genetic variation of avian infectious bronchitis virus (IBV) in Guangxi province,one strain of IBV was isolated from chicken.Two pairs of primers for amplifying the N and M genes of IBV were designed according to the sequences in GenBank.The N and M genes of the strain were amplified by RT-PCR,and they were proved to be the N and M genes of IBV by cloning,sequencing and compared with reference IBV strains published in GenBank.The results showed that the N gene from the IBV isolate consisted of 1 230 bp,coding 409 amino acids.The M gene from the IBV isolate consisted of 678 bp,coding 225 amino acids.The sequence analysis of N gene showed that it shared 87.2% to 93.3% nucleotide homologies and 90.0% to 94.4% deduced amino acid sequence homologies with IBV strains from GenBank.The M gene sequence analysis showed that it shared 83.6% to 91.0% nucleotide homologies and 82.7% to 92.9% deduced amino acid sequence homologies.The phylogenetic tree analysis showed that it was closely related to BJ and LX4 strains,and were clustered into one group;But with the distant relatives from other strains of IBV.These results suggested that the isolate was a new variant of IBV.     

Construction and Identification of PRRSV ORF5 Gene Prokaryotic Expression Vector

The present study was designed to construct recombinant plasmids,which could express porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 gene.RNA was extracted from spleen and lung samples of the suspected pigs which were infected with PRRSV.According to PRRSV ORF5 gene,a pair of primers was designed for RT-PCR amplification.The ORF5 target gene was cloned into pMD19-T vector and then the recombinant pMD19-ORF5 was achieved.According to the sequencing results and the characteristics of expression vectors,a pair of primers with NcoⅠand XbaⅠenzyme cleavage sites was designed.Target fragment dORF5 was amplified and then connected to pProEXHTb and pNZ8149 vectors,respectively.And recombinant HTb-dORF5/DE3 and pNZ8149-dORF5/NZ3900 was induced by IPTG and Nisin,respectively,and analyzed by SDS-PAGE and Western blotting.Recombinant HTb-dORF5/DE3 induced by 1.5 mmol/L IPTG was expressed in the highest quantity.There were specific band at about 22 ku with reactionogenicity when it was tested by SDS-PAGE and Western blotting.Recombinant pNZ8149-dORF5/NZ3900 induced by 20 ng/mL Nisin was expressed in the highest quantity.There were specific band at about 19 ku with reactionogenicity when it was tested by SDS-PAGE and Western blotting.The IFA result showed specific green fluorescence.This study successfully constructed recombinant plasmids HTb-dORF5 and pNZ8149-dORF5 and expressed,the result laid a solid foundation for further development of PRRS vaccines.     

Geographic Information System‐Based Analysis of the Spatial and Spatio‐Temporal Distribution of Zoonotic Cutaneous Leishmaniasis in Golestan Province,North‐East of Iran

Zoonotic cutaneous leishmaniasis (ZCL), a vector‐borne disease, poses serious psychological as well as social and economic burden to many rural areas of Iran. The main objectives of this study were to analyse yearly spatial distribution and the possible spatial and spatio‐temporal clusters of the disease to better understand spatio‐temporal epidemiological aspects of ZCL in rural areas of an endemic province, located in north‐east of Iran. Cross‐sectional survey was performed on 2983 recorded cases during the period of 2010–2012 at village level throughout the study area. Global clustering methods including the average nearest‐neighbour distance, Moran's I, general G indices and Ripley's K‐function were applied to investigate the annual spatial distribution of the existing point patterns. Presence of spatial and spatio‐temporal clusters was investigated using the spatial and space–time scan statistics. For each year, semivariogram analysis and all global clustering methods indicated meaningful persistent spatial autocorrelation and highly clustered distribution of ZCL, respectively. Eight significant spatial clusters, mainly located in north and northeast of the province, and one space–time cluster, observed in northern part of the province and during the period of September 2010–November 2010, were detected. Comparison of the location of ZCL clusters with environmental conditions of the study area showed that 97.8% of cases in clusters were located at low altitudes below 725 m above sea level with predominantly arid and semi‐arid climates and poor socio‐economic conditions. The identified clusters highlight high‐risk areas requiring special plans and resources for more close monitoring and control of the disease.     

The potential drivers in forming avian biodiversity hotspots in the East Himalaya Mountains of Southwest China

Little has been published to describe or interpret Asian biodiversity hotspots, including those in the East Himalayan Mountains of Southwest China (HMSC), thus making necessary a review of the current knowledge. The Pliocene and Pleistocene geological and glacial histories of the Asian continent differ from those of Europe and North America, suggesting different mechanisms of speciation and extinction, and, thus, different responses to climate changes during the Quaternary glaciations. This short review summarizes potential drivers in shaping and maintaining high species richness and endemism of birds in the HMSC. The geographical location at the junction of different biogeographical realms, the wide range of habitats and climates along the extensive elevational range, the complex topography and the distinct geological history of this region have probably contributed to the evolution of an exceptionally species‐rich and endemic‐rich, specialized montane avian fauna. The Mountain systems in the HMSC may have provided refugia where species survived during the glacial periods and barriers for preventing species dispersal after the glacial periods. More studies are required to further test this refugia hypothesis by comparing more cold‐tolerent and warm‐tolerent species.     

The face of conservation responding to a dynamically changing world

In its 40‐year history, the science of conservation has faced unprecedented challenges in terms of environmental damage and rapid global change, and environmental problems are only increasing as greater demands are placed on limited natural resources. Conservation science has been adapting to keep pace with these changes. Here, we highlight contemporary and emerging trends and innovations in conservation science that we believe represent the most effective responses to biodiversity threats. We focus on specific areas where conservation science has had to adjust its approach to address emerging threats to biodiversity, including habitat destruction and degradation, climate change, declining populations and invasive species. We also document changes in attitudes, norms and practices among conservation scientists. A key component to success is engaging and maintaining public support for conservation, which can be facilitated through the use of technology. These recent trends in conservation and management are innovative and will assist in optimizing conservation strategies, increasing our leverage with the general public and tackling our current environmental challenges.     

仙游县猪口蹄疫免疫效果检测与分析

通过对仙游县2014年秋季猪O型口蹄疫免疫抗体检测,了解和掌握仙游县猪口蹄疫免疫效果,以更好地指导今后动物疫病防控工作。     

貉源犬瘟热病毒CDV-ZC株分离鉴定、全基因测序与分析

应用非洲绿猴肾细胞(Vero)从山东省诸城市疑似犬瘟热(canine distemper,CD)感染貉的肝脏、脾脏、肺脏等组织病料的研磨上清液中分离出1株病毒。分离株经Vero细胞传至第4代出现典型的细胞病变效应(CPE),经毒力测定、血清学、RT-PCR检测及测序、回归动物试验证明为犬瘟热病毒(Canine distemper virus,CDV),命名为CDV诸城分离株(CDV-ZC)。采用RT-PCR方法分段克隆分离株的全基因序列并测序,测序成功的各序列依次拼接得到全基因序列并进行序列比对。结果表明,CDV-ZC株全基因(KJ994343)与标准美洲型犬瘟热强毒株A75/17核苷酸同源性高达96.5%,而与疫苗株Onderstepoort、CDV3等亲缘关系较远,同源性为91.6%~92.0%。血凝素(H)基因序列分析表明,CDV-ZC株与国内野毒株LN(13)2、GP株等以及日本野毒株UENO、HAMA等共同归属于Asia-1型,在H蛋白信号淋巴细胞激活因子(SLAM)受体结合区即542~544位增加了1个潜在N-连接糖基化位点(N-X-S/T)。致病性强毒株CDV-ZC的成功分离及全基因序列测定加深了我们对当前中国CDV流行株遗传变异情况的了解,为CD的有效预防、诊断及控制提供理论依据。     

Pathogenicity of emerging Japanese type 1 porcine reproductive and respiratory syndrome virus in experimentally infected pigs

To clarify the pathogenicity of Japanese type 1 porcine reproductive and respiratory syndrome virus (PRRSV) isolate in experimentally infected pigs, we evaluated clinical signs and monitored viremia for 21 days post-inoculation (dpi). Lungs were mottled, tanned and reddish in appearance; had lesions predominantly in the cranial, middle and accessory lobes; and failed to collapse at 10 dpi. Although microscopic lesions of lungs were reproduced using the Japanese emerging type 1 PRRSV isolate under experimental conditions, no significant differences were noted between the challenge and control groups regarding mean rectal temperature and daily weight gain. These results provide useful insights into the limited pathogenicity of single infection with the Japanese type 1 PRRSV isolate in piglets, which differ from findings in reported field cases.