The current situation and strategies of snake antivenomimmunoglobulins research and development / 药学学报

Snake antivenomimmunoglobulins are considered to be the most efficient drugs in snake envenomings. Most snake antivenomimmunoglobulins all over the world are still prepared by fragmentation of polyclonal antibodies isolated from hyper-immunized horse serum till now. In this review, we retrospect the history of snake antivenomimmunoglobulins, analysis the present situation and pay the close attention on the key technological links in the process of research and manufacturing, such as properties of IgG and its fragments, selection and preparation of immunogen, optimization of immunization schedule and protein isolation and purification, which can be available for the reference in the research and development of snake antivenom.

Research methods in protein-protein and protein-nucleic acid interactions and application in the study of human enterovirus A71 / 病毒学报

Methods for analyses of protein-protein interactions include: yeast two hybrid (Y2H), phage dis- play (PD), co-immunoprecipitation (Co-IP), glutathione S-transferase pull-down (GST pull-down), cellular co-localization, far-western blotting, virus overlay protein binding assay (VOPBA), surface plasmon resonance (SPR), and fluorescence resonance energy transfer (FRET). Technologies for the detection of protein-nucleic acid interactions include: yeast one hybrid (Y1H), chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA), Southwestern blotting, reporter gene, Co-IP, GST pull-down, and PD. These methods are often used in the study of the human enterovirus A71 (EV-A71) by our research team. Reviews in the Chinese literature in this field are lacking, so we reviewed applications of these methods in the study of EV-A71. This review may impart important knowledge in the research of other viruses with regard to protein-protein and protein-nucleic acid interactions.

Isolation and characterization of rotavirus from bat / 病毒学报

Bats are considered as important animal reservoirs for many pathogenic viruses to humans. The viral metagenomic analysis was performed to study gut and lung tissues of 30 insectivorous bats collected in Yunnan Province and 26 reads were noted to group A rotavirus (RVA). Further RT-PCR screening on bat samples and in vitro viral isolation on cell cultures confirmed the presence of a novel RVA, named as RVA/Bat-tc/MYAS33/2013/G3P[10], in one of 30 Stoliczka's trident bats. The VP7 gene of this strain MYAS33 was closely related to that of an equine RVA strain from Argentina and the nucleotide sequence similarity was 93%, while its VP4 gene was a rare P[10] type and obtained the maximum sequence identity (94.8%) with that of a human strain from Thailand. The present study highlights the potential role of bats as reservoirs for RVAs.

Genetic evolution analysis of matrix protein 2 gene of avian influenza H5N1 viruses from boundary of Yunnan province / 中华预防医学杂志

<p><b>OBJECTIVE</b>To elucidate the variation in characterizations and genetic evolution of the matrix protein 2 or ion channel protein(M2) genes of avian influenza subtype H5N1 viruses in the boundary region of Yunnan province from 2008 to 2012.</p><p><b>METHODS</b>A total of swab samples were collected from foreign poultry such as the junction between Yunnan and Vietnam, Laos,myanmar and wild birds in boundary region of Yunnan province from 2008 to 2012 and screened by H5N1 subtype-specific multiplex RT-PCR. The M genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis of M2 genes were performed with sequences of the known reference strains.</p><p><b>RESULTS</b>A total of 71 positive samples were found out of 1240 samples and the positive rate was 5.72%. A total of 14 different M2 sequences were obtained from 30 positive samples and were divided into 3 distinct clades or sub-clades(1.2.1, 1.2.2 and 2) by phylogenetic analysis, 5, 7 and 2, respectively. The M2 genes and Hemagglutinin(HA) genes of H5N1 viruses from the boundary region of Yunnan province had showed different relationship of genetic evolution. The substitution or mutation of key amino acids sites had been found among the domains of epitope, adamantane-resistance, and poultry or human original viral strains.</p><p><b>CONCLUSION</b>The M2 genes of H5N1 subtype viruses in boundary region of Yunnan province from 2008 to 2012 showed genetic divergence and the virus of clade 1.2.2 had become dominant epidemic strain in this region.</p>

Genetic evolution of non-structural gene among avian influenza H5N1 viruses isolated from the boundary of Yunnan province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To elucidate the characteristics of variation and the genetic evolution of non-structural protein (NS1, NS2) genes related to avian influenza subtype H5N1 viruses isolated from the boundary region of Yunnan province.</p><p><b>METHODS</b>Swab samples were collected from foreign poultry and wild birds in the boundary regions of Yunnan province and screened by H5/N1 subtype-specific multiplex RT-PCR. The NS segment of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis on those available NS1, NS2 genes were performed with sequences of the known reference strains.</p><p><b>RESULTS</b>71 positive samples were identified from 1240 samples, with the positive rate as 5.72%. Fourteen different NS segment sequences were obtained from 30 representative positive samples and could be divided into 3 distinct clades or sub-clades (I-1, I-2 and II), by phylogenetic analysis. The NS1/NS2 genes and Hemagglutinin (HA) genes of H5N1 viruses from the boundary regions of Yunnan province showed different relationships regarding the characteristics on genetic evolution. The substitution or mutation of key amino acids sites had been noticed in the nuclear location signal domains, effect domain, and other pathogenicity markers.</p><p><b>CONCLUSION</b>NS genes of H5N1 subtype viruses in boundary region of Yunnan province showed genetic divergence and the virus of clade I-2 and II had become dominant epidemic strains in this region since 2010.</p>

Genetic diversification of avian influenza H5N1 virus in boundary areas of Yunnan province / 中华流行病学杂志

Objective To elucidate the genetic diversifications of avian influenza subtype H5N1 viruses in the boundary regions of Yunnan province during 2009 to July,2011.Methods Swab samples were collected from foreign poultry and wild birds in boundary regions of Yunnan province during 2009 to July,2011 and tested by H5/N1 subtype-specific multiplex RT-PCR.The HA genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD 18-T vectors for sequencing.Both alignment and phylogenetic analysis were performed with sequences of the known reference strains.Results Fifteen different HA sequences were obtained from 36 representative positive samples and could be divided into 2 distinct Clades (2.3.2 and 2.3.4).Through phylogenetic analysis,Clade 2.3.2 and 2.3.4 could then be further divided into 3 ( Ⅱ -1 to Ⅱ -3 ) and 2 smaller clades ( Ⅰ -1 and Ⅰ -2),respectively.The viruses of Clade 2.3.2 Ⅱ -1 and Ⅱ-2 were new variant strains of H5N 1 virus.The cleavage sites of HA from positive samples all possessed molecular characterization of highly pathogenic avian influenza virus.Mutation of key amino acids had been found among receptor binding sites,potential glycosylation sites,neutralizing epitopes and others.Conclusion It seemed evident that the H5N1 subtype viruses showed genetic diversifications and had undergone the evolution progress of multi-clade (2.3.2,2.3.4) to single caide (2.3.2) in the boundary regions of Yunnan province,during 2009 to July,2011.

Primary clinical result of digital template as navigation to supper cervical pedicle instrumentation / 中华外科杂志

<p><b>OBJECTIVE</b>To observe the primary clinical result of digital template as navigation to the upper cervical pedicle instrumentation.</p><p><b>METHODS</b>CT scan of upper cervical vertebrae was performed. 3-D model of upper cervical vertebrae was reconstructed by software Amira 3.1 and was preserved in STL format. Then 3-D model was run in software UG Imageware 12.0, the best pedicle channel was extracted according to the reverse engineering principle. A virtual navigational template was established according to he lamina anatomic trait, and the best pedicle channel. The virtual vertebrae and navigational template were manufactured using rapid prototyping. The navigational template was sterilized and used intra operative to assist with the placement of pedicle screw. The Accuracy of screw placement was confirmed with postoperative X-ray and CT scanning.</p><p><b>RESULTS</b>The digital navigational template had been established and used in the 3 cases, the good trajectory of cervical pedicle had been showed by the CT scan of post operation. There were not complications of related pedicle screw insertion.</p><p><b>CONCLUSIONS</b>A novel method of upper cervical pedicle location using Reverse Engineering and rapid prototyping has been developed; the navigational template is found to be highly accuracy and has great expectation.</p>

ISOLATION AND IDENTIFICATION OF XJ STRAIN OF CANINE DISTEMPER VIRUS / 微生物学通报

A canine distemper virus strain was isolated from the lung of dog coming from Aksu in Xing Jiang using lung primary M cell during the CDV molecular epidemiological study. It was demonstrated to be a virulent strain of CDV by a series of systematic identification such as morphology , serology neutralization test, canine infection test, and molecular virology test.