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Kunkun Zhao,
Min Gu,
Lei Zhong,
Zhiqiang Duan,
Yan Zhang,
Yanmei Zhu, Guo Zhao,
Mingjun Zhao,
Zhaoyang Chen,
Shunlin Hu,
Wenbo Liu,
Xiaowen Liu,
Daxin Peng,
Xiufan Liu
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ABSTRACT: One H5N8 and three H5N5 highly pathogenic avian influenza (HPAI) viruses which derived their HA genes from the Asian H5N1 lineage were isolated from poultry during 2009-2010 in mainland China. Pathogenicity studies showed that these viruses were all highly virulent to chickens, while they varied from moderate to high virulence in mice and from mild to intermediate virulence in mallards. Phylogenetic analyses showed that these viruses were reassortants bearing the H5N1 backbone while acquiring PB1, NP and NA genes from unidentified non-H5N1 viruses, and had developed into three distinct genotypes (B-D). Molecular characterization indicated that all these viruses might resist to antiviral agents. Our findings highlight the emergence and development of HPAI H5 viruses of other NA subtypes in H5N1 endemic areas and their potential threat to poultry industry and public health.
Veterinary Microbiology 01/2013; · 3.33 Impact Factor
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ABSTRACT: Here, we report the complete genomic sequence of a novel reassortant H4N2 influenza virus isolated from domestic ducks in the Jiangsu province of China in 2011. Phylogenetic analysis showed that all the viral genes except for hemagglutinin (HA) were highly homologous to the clade 2.3.4 H5N2 viruses. The data suggest that genetic reassortment occurred between H4 and H5N2 avian influenza viruses, which highlights the role of domestic poultry as a reassortment vessel in China.
Genome announcements. 01/2013; 1(2):e0009113.
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ABSTRACT: The H3 subtype avian influenza virus (AIV) can provide genes for human influenza virus through gene reassortment, which raises great concerns in terms of its potential threat to human health. Here, we report the complete genome sequence of a novel H3N2 AIV isolated from domestic ducks in the Jiangsu province of eastern China in 2004, which is a natural recombinant virus whose genes are derived from H3N8, H5N1, H5N2, H11N2, H4N6, and H1N1 AIVs. This genome will help to understand the epidemiology and molecular characteristics of H3N2 influenza virus in eastern China.
Genome announcements. 01/2013; 1(2).
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ABSTRACT: For the first time we report the complete genomic sequence of an H11N3 influenza virus from domestic ducks in China. Phylogenetic analysis showed that the H11N3 virus was a novel reassortant with its genes from different subtypes of domestic duck-origin avian influenza viruses, which further underlined that domestic ducks play a key role in the genetic reassortment and evolution of influenza viruses in China.
Journal of Virology 11/2012; 86(21):11950-1. · 5.40 Impact Factor
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Guo Zhao,
Xiaobing Gu,
Xinlun Lu,
Jinjin Pan,
Zhiqiang Duan,
Kunkun Zhao,
Min Gu,
Qingtao Liu,
Liang He,
Jian Chen,
Shengqiang Ge,
Yanhong Wang,
Sujuan Chen,
Xiaoquan Wang,
Daxin Peng,
Hongquan Wan,
Xiufan Liu
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ABSTRACT: There has been multiple evidence that domestic poultry may act as a vessel for the generation of novel influenza A viruses. In this study, we have analyzed the evolution and pathogenicity of 4 H5N2 avian influenza viruses isolated from apparently healthy poultry from H5N1 virus endemic areas in China. Phylogenetic analysis revealed that two of these viruses, A/duck/Eastern China/1111/2011 (DK/EC/1111/11) and A/goose/Eastern China/1112/2011 (GS/EC/1112/11) were derived from reassortment events in which clade 2.3.4 highly pathogenic avian influenza (HPAI) H5N1 viruses acquired novel neuraminidase and nonstructural protein genes. Another two isolates, A/chicken/Hebei/1102/2010 (CK/HB/1102/10) and A/duck/Hebei/0908/2009 (DK/HB/0908/09), possess hemagglutinin (HA) gene belong to clade 7 H5 viruses and other genes from endemic H9N2 viruses, or from viruses of various subtypes of the natural gene pool. All of these H5N2 isolates bear characteristic sequences of HPAI virus at the cleavage site of HA, and animal experiments indicated that all of these viruses but DK/HB/0908/09 is highly pathogenic to chickens. In particular, DK/EC/1111/11 and GS/EC/1112/11 are also highly pathogenic to ducks and moderately pathogenic to mice. All of these 4 viruses were able to replicate in domestic ducks and mice without prior adaptation. The emergence of these novel H5N2 viruses adds more evidence for the active evolution of H5 viruses in Asia. The maintenance of the highly pathogenic phenotype of some of these viruses even after reassortment with a new NA subtypes, their ability to replicate and transmit in domestic poultry, and the pathogenicity in the mammalian mouse model, highlight the potential threat posed by these viruses to both veterinary and public health.
PLoS ONE 01/2012; 7(9):e46183. · 4.09 Impact Factor
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Guo Zhao,
Jinjin Pan,
Xiaobing Gu,
Xinlun Lu,
Qunhui Li,
Jie Zhu,
Chaoyang Chen,
Zhiqiang Duan,
Quangang Xu,
Xiaobo Wang,
Shunlin Hu,
Wenbo Liu,
Daxin Peng,
Xiaowen Liu,
Xiaoquan Wang,
Xiufan Liu
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ABSTRACT: Isolates of the A(H1N1)pdm2009 virus were first identified in asymptomatic swine in Jiangsu province, China in January 2010, indicating that the virus has retro-infected swine after circulating through humans in mainland China. The purpose of this study was to determine whether the avian-origin European H1N1 swine influenza virus (SIV) and the A(H1N1)pdm2009 virus are cocirculating in swine in Jiangsu province of China. From May 2010 to May 2011, 1,030 nasal swab samples were collected from healthy swine in Jiangsu province of China and were tested for influenza A H1N1 using reverse transcription-PCR. Fragments of the complete genomes of viruses from the samples that were positive for influenza A H1N1 were sequenced and analysed. A total of 32 avian-origin European H1N1 SIVs were isolated, and no A(H1N1)pdm2009 viruses were identified; full-length genomes of 18 strains were sequenced. The eight gene segments of some of the isolated H1N1 viruses have 99.1-99.8% sequence identity with the human A/Jiangsu/ALS1/2011(H1N1) isolates in the same region. Our study indicates that the avian-origin European H1N1 SIVs remain endemic in swine and have retro-infected humans after circulating through swine, which may present a risk factor for public health.
Virus Genes 12/2011; 44(2):295-300. · 1.85 Impact Factor
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Qing Sun,
Lixiang Zhao,
Qingqing Song,
Zheng Wang,
Xusheng Qiu,
Wenjun Zhang,
Mingjun Zhao, Guo Zhao,
Wenbo Liu,
Haiyan Liu,
Yunsen Li,
Xiufan Liu
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ABSTRACT: N-linked glycans are composed of three major types: high-mannose (Man), hybrid or complex. The functional role of hybrid- and complex-type N-glycans in Newcastle disease virus (NDV) infection and fusion was examined in N-acetylglucosaminyltransferase I (GnT I)-deficient Lec1 cells, a mutant Chinese hamster ovary (CHO) cell incapable of synthesizing hybrid- and complex-type N-glycans. We used recombinant NDV expressing green fluorescence protein or red fluorescence protein to monitor NDV infection, syncytium formation and viral yield. Flow cytometry showed that CHO-K1 and Lec1 cells had essentially the same degree of NDV infection. In contrast, Lec2 cells were found to be resistant to NDV infection. Compared with CHO-K1 cells, Lec1 cells were shown to more sensitive to fusion induced by NDV. Viral attachment was found to be comparable in both lines. We found that there were no significant differences in the yield of progeny virus produced by both CHO-K1 and Lec1 cells. Quantitative analysis revealed that NDV infection and fusion in Lec1 cells were also inhibited by treatment with sialidase. Pretreatment of Lec1 cells with Galanthus nivalis agglutinin specific for terminal α1-3-linked Man prior to inoculation with NDV rendered Lec1 cells less sensitive to cell-to-cell fusion compared with mock-treated Lec1 cells. Treatment of CHO-K1 and Lec1 cells with tunicamycin, an inhibitor of N-glycosylation, significantly blocked fusion and infection. In conclusion, our results suggest that hybrid- and complex-type N-glycans are not required for NDV infection and fusion. We propose that high-Man-type N-glycans could play an important role in the cell-to-cell fusion induced by NDV.
Glycobiology 09/2011; 22(3):369-78. · 3.58 Impact Factor
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Guo Zhao,
Lei Zhong,
Xinlun Lu,
Jiao Hu,
Xiaobing Gu,
Yan Kai,
Qingqing Song,
Qing Sun,
Jinbao Liu,
Daxin Peng,
Xiaoquan Wang,
Xiaowen Liu,
Xiufan Liu
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ABSTRACT: In spring 2009, one strain of H5N1 clade 2.3.2 virus was isolated from wild swans in Shanghai, indicating the importance of the wild swan in the ecology of this highly pathogenic avian influenza virus (HPAIV) in Eastern China. Pathogenicity experiments conducted in this study indicated that the virus was highly pathogenic for chickens but lowly pathogenic for mammalian hosts, as evidenced by reduced infection of mice. The analysis of complete genome sequences and genetic evolution showed that A/Swan/Shanghai/10/09 (SW/SH/09) may be derived from the strain A/silky chicken/Shantou/475/2004 (CK/ST/04), which is homologous to the influenza viruses isolated from chicken, duck, pika, little egret, swan, mandarin duck and bar-headed goose in China Hunan, China Qinghai, Mongolia, Russia, Japan, Korea, Laos and Hong Kong during 2007-2011, indicating that the virus has retro-infected diverse wild birds from chicken, and significant spread of the virus is still ongoing through overlapping migratory flyways. On the basis of the molecular analysis, we also found that there was a deletion of the glycosylation site (NSS) in amino acid 156 of the hemagglutinin (HA) protein when compared with that of the other Clade 2.3.2 viruses isolated between 2007 and 2011. More importantly, the sequence analysis of SW/SH/09 virus displayed the drug-resistant mutations on the matrix protein (M2) and neuraminidase (NA) genes.
Virus Genes 09/2011; 44(1):55-62. · 1.85 Impact Factor
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Wenjun Zhang,
Tao Xue,
Xiaowei Wu,
Pinghu Zhang, Guo Zhao,
Daxing Peng,
Shunlin Hu,
Xiaoquan Wang,
Xiaowen Liu,
Wenbo Liu,
Xiufan Liu
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ABSTRACT: The H5N1 subtype of highly pathogenic avian influenza viruses has spread to over 63 countries in Asia, Europe, and Africa and has become endemic in poultry. Since 2004, vaccination against H5N1 influenza has become common in domestic poultry operations in China. Most influenza vaccines have been produced in embryonated chicken eggs. High yield is the essential feature of a good vaccine candidate virus.
Therefore, the large-scale manufacture of such a vaccine requires that the viral yield of H5N1 reassortant vaccine viruses in eggs and MDCK cells be increased.
We generated two sets of reassortant H5N1 viruses based on backbone viruses A/Chicken/F/98 (H9N2) and A/Puerto Rico/8/34 (H1N1) using reverse genetics. The HAs and NAs of the reassortants were derived from the three epidemic H5N1 strains found in China. We compared the replication properties of these recombinant H5N1 viruses in embryonated chicken eggs and MDCK cells after inserting either 20 or 38 amino acids into their NA stalks.
In this study, we demonstrated that inserting 38 amino acids into the NA stalks can significantly increase the viral yield of H5N1 reassortant viruses in both embryonated chicken eggs and MDCK cells, while inserting only 20 amino acids into the same NA stalks does not. Hemagglutinin inhibition testing and protection assays indicated that recombinant H5N1 viruses with 38 aa inserted into their NA stalks had the same antigenicity as the viruses with wt-NA.
These results suggest that the generation of an H5N1 recombinant vaccine seed by the insertion of 38 aa into the NA stalk may be a suitable and more economical strategy for the increase in viral yield in both eggs and MDCK cells for the purposes of vaccine production.
Vaccine 08/2011; 29(45):8032-41. · 3.77 Impact Factor
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ABSTRACT: To investigate whether the 2009 pandemic H1N1 influenza A virus was still being transmitted in swine, a total of 1029 nasal swab samples from healthy swine were collected from January to May 2010 in Jiangsu province of China. Eight H1N1 influenza viruses were isolated and identified, and their full length genomes were sequenced. We found that all eight of the H1N1 viruses shared higher than 98.0% sequence identity with the 2009 pandemic virus A/Jiangsu/1/2009 (JS1). In addition, some of these viruses had D225G (3/8) mutations in the receptor binding sites of the hemagglutinin (HA) protein, indicating enhancement of their binding affinity to the sialic α2, 3Gal receptor. In conclusion, the 2009 pandemic H1N1 influenza A virus has retro-infected swine from humans in mainland China, and significant viral evolution is still ongoing in this species.
Research in Veterinary Science 06/2011; 93(1):125-32. · 1.65 Impact Factor
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Min Gu,
Wenbo Liu,
Yongzhong Cao,
Daxin Peng,
Xiaobo Wang,
Hongquan Wan, Guo Zhao,
Quangang Xu,
Wei Zhang,
Qingqing Song,
Yanfang Li,
Xiufan Liu
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ABSTRACT: In China, domestic ducks and wild birds often share the same water, in which influenza viruses replicate preferentially. Isolation of 2 novel reassortant highly pathogenic avian influenza (H5N5) viruses from apparently healthy domestic ducks highlights the role of these ducks as reassortment vessels. Such new subtypes of influenza viruses may pose a pandemic threat.
Emerging Infectious Diseases 06/2011; 17(6):1060-3. · 6.79 Impact Factor
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ABSTRACT: The purpose of this study was to evaluate the soft tissue profile changes of functional Angle Class III malocclusion in mixed dentition, and to evaluate the effectiveness of treatment with Frankel-III appliances.
Fifteen cases of Angle Class III malocclusion in mixed dentition, 8 males and 7 females, were selected. The mean age of the patients was 9.5 years old, ranging from 7 to 12 years old. Lateral cephalometric radiograph was taken before and after the treatments and analyzed. The data was analyzed by paired t test with SPSS17.0 software package.
After the patients were treated with Frankel-III appliances in mixed dentition, Sn-Ns-Si, Ns-Sn-Pos , LiSi-Sn increased, S-Ns-Si, S-Ns-Pos,LL-EP decreased with significant difference (P<0.01). S-Ns-Sn, LsSn-Sn,Sn-Me,Ns-Me,Ul-EP increased with significant difference (P<0.05).
All findings indicate that functional Angle Class III malocclusion can be corrected with Frankel-III appliances. The relationship among nose, upper lip, lower lip and chin become harmonious after treatment. The maxillary and mandibular soft tissue changes are distinct and soft tissue profile changes from Class III to Class I.
Shanghai kou qiang yi xue = Shanghai journal of stomatology 04/2011; 20(2):201-3.
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Guo Zhao,
Xinlun Lu,
Xiaobing Gu,
Kunkun Zhao,
Qingqing Song,
Jinjin Pan,
Quangang Xu,
Zhiqiang Duan,
Daxin Peng,
Shunlin Hu,
Xiaoquan Wang,
Xiufan Liu
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ABSTRACT: Although extensive data demonstrates that the majority of H6 duck isolates belonged to a single H6N2 virus lineage with a single gene constellation in southern China from 2000 to 2005, the prevalence of H6N2 virus in poultry in Eastern China is largely unknown.
Epidemiology revealed that H6N2 viruses were the most frequently detected influenza subtypes in live bird markets from 2002 to 2008 in Eastern China, but from 2009 onwards, they were replaced with novel H6N6 viruses. We phylogenetically and antigenically analyzed 42 H6 viruses isolated mainly in domestic ducks from 2002 to 2010 in Eastern China. Surprisingly, none of these isolates grouped with the previously described H6N2 viruses which belonged to a single H6N2 virus lineage with a single gene constellation in domestic ducks in southern China from 2000 to 2005. Two distinct hemagglutinin lineages were identified and they all underwent frequent reassortment with multiple virus subtypes from the natural gene pool, but few reassortants were persistent or prevalent.
Five subtypes of H6 influenza viruses (H6N1, H6N2, H6N5, H6N6 and H6N8) cocirculated in Eastern China, which form a significant part of the natural influenza virus reservoir in domestic ducks, and significant viral reassortment is still ongoing in this species.
Virology Journal 01/2011; 8:470. · 2.34 Impact Factor
