1.Analysis on the non-structural protein genes of avian influenza viruses subtype H5N1 isolated from chicken in China
Yanyan HUANG ; Sanjie CAO ; Beixia HU ; Qijing DU ; Xiumei ZHANG ; Xintian WEN
Chinese Journal of Zoonoses 2010;(2):154-157
Fourteen H9N2 avian influenza viruses (AIV) were isolated from sick chickens in China from 1998 to 2008. The sequences of the Non-structural(NS) gene of these isolates were determined by RT-PCR and sequencing, and the entire ORF sequences of NS1 and NS2 protein were obtained.-The homology of these nucleotide sequences and the putative amino acid sequences were compared with several classic reference viruses of H9N2. These isolates were proved to be highly homologous in NS gene (92.9%-99.9% identity) and all belonged to A/Chicken/Beijing/1/1994-like group in the Asia bird-swine branch of allele A of HS gene phylogenetic tree.-According to this study and previous reports of other researchers, NS gene of H9N2 subtype AIV in chickens of China is genetically stable and there is no enough evidence to support the establishment of other sub-lineages in chickens.
2.Cloning and expression of the σC gene of avian reovirus
Xiaobo HUANG ; Shanshan LING ; Sanjie CAO ; Xintian WEN ; Cunwei WANG ; Yuankun CHEN
Chinese Journal of Veterinary Science 2009;29(7):836-840
The σC gene of ARV S1133 was designed to amplify by reverse transcription chain reaction(RTPCR).The σC gene was inserted into the vector pMD19-T,identified by PCR method and restriction enzyme,and sequenced.It showed that the insert cloned gene fragment was the σC gene of ARV.Then the gene was inserted intc the pET32a(+) and indicated that fusion expression vector pET32a-σC was constructed.The recombinant fusion protein was highly expressed in E.coli BL21 induced by 1.0 mmol/L IPTG for 5 hours in the form of inclusion bodies.The weight of recombinant fusion protein molecular is 54 000.Western-blot with ARV antibodies against the fusion protein showed the recombinant protein has a favourable reactivity.
3.Effect of oxycodone on migration of human colon cancer cells : the role of μ and κ receptors
Xueqin ZHENG ; Meijuan LIAO ; Chengxiang YANG ; Xintian LIANG ; Yun LI ; Xianjie WEN ; Yueyuan CHEN
Chinese Journal of Anesthesiology 2017;37(4):439-442
Objective To evaluate the effect of oxycodone on migration of human colon cancer cells and the role of μ and κ receptors.Methods The human colon cancer HCT116 cells at the logarithmic growth phase were seeded in 24-well or in 6-well plates at a density of 1 × 106 cells/mnl (0.5 ml/well or 2 ml/well,144 wells in total).The cells were divided into 6 groups (n=24 each) using a random number table:control group (group C),1,5 and 10 μmol/L oxycodone groups (group O1,group O2 and group O3),oxycodone plus μ receptor antagonist CTOP group (group O2+CTOP) and oxycodone plus κ receptor antagonist nor-binaltorphimine group (group O2+BNI).The cells were incubated for 24 h with oxycodone 1,5 and 10 μmol/L in O1,O2 and O3 groups,respectively.The cells were incubated for 24 h with 5 μmol/L oxycodone plus 20 μmol/L CTOP and 5 μmol/L oxycodone plus nor-binahorphimin 20 μmol/L in O2+CTOP and O2+BNI groups,respectively.The invaded and migrated cells were counted,and the levels of Ras homolog gene family member A (RhoA),Rho-associated protein kinase 1 (ROCK1),matrix metalloproteinase-2 (MMP-2) and MMP-9 were detected.Results Compared with group C,the number of invaded and migrated cells was gradually decreased,and the levels of RhoA,ROCK1,MMP-2 and MMP9 were gradually decreased in O1,O2 and O3 groups (P<0.05),and no significant change was found in the parameters mentioned above in group O2+BNI (P>0.05).Compared with group O2,the number of invaded and migrated cells was significantly increased,and the levels of RhoA,ROCK1,MMP-2 and MMP9 were increased in group O2 + BNI (P<0.05),and no significant change was found in the parameters mentioned above in group O2+CTOP (P>0.05).Conclusion Oxyc odone can inhibit the migration of human colon cancer cells,and the mechanism is totally related to inhibition of RhoA/ROCKl signaling pathway activation after activating κ receptors,but not related to μ receptors.
4.Serological and molecular epidemiology of Japanese encephalitis virus infections in swine herds in China, 2006–2012
Chunxia CHAI ; Qiao WANG ; Sanjie CAO ; Qin ZHAO ; Yiping WEN ; Xiaobo HUANG ; Xintian WEN ; Qiguai YAN ; Xiaoping MA ; Rui WU
Journal of Veterinary Science 2018;19(1):151-155
Japanese encephalitis virus (JEV) is a mosquito-borne, zoonotic flavivirus causing viral encephalitis in humans and reproductive disorder in swine. JEV is prevalent throughout China in human; however, spatiotemporal analysis of JEV in Chinese swine herds has not been reported previously. Herein, we present serological and molecular epidemiological results and estimates of prevalence of JEV infections among swine herds in various regions of China. The results suggest that JEV infections are widespread and genotype I and III strains co-exist in the same regions. Therefore, there is an urgent need to monitor JEV infection status among swine herds in China.
Asian Continental Ancestry Group
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China
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Encephalitis Virus, Japanese
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Encephalitis, Japanese
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Encephalitis, Viral
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Flavivirus
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Genotype
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Humans
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Molecular Epidemiology
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Prevalence
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Spatio-Temporal Analysis
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Swine