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Hedgehog signaling pathway regulated the target genes for adipogenesis in silkworm Bombyx moil

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Insect Science 2015年第5期22卷 587-596页

作者： Shuang Liang Rui-Ting Chen Deng-Pan Zhang Hu-Hu Xin Yan Lu Mei-Xian Wang Yun-Gen Miao Key laboratory of animal virology of Ministry of Agriculture College of Animal Sciences Zhejiang University Hangzhou 310058 China

Hedgehog （Hh） signals regulate invertebrate and vertebrate development, yet the role of the pathway in adipose development remains poorly understood. In this report, we found that Hh pathway components are expressed in the fat body of silkworm larvae. Functional analysis of these components in a BmN cell line model revealed that activa- tion of the Hh gene stimulated transcription of Hh pathway components, but inhibited the expression of the adipose marker gene AP2. Conversely, specific RNA interference- mediated knockdown of Hh resulted in increased AP2 expression. This further showed the regulation of Hh signal on the adipose marker gene. In silkworm larval models, enhanced adipocyte differentiation and an increase in adipocyte cell size were observed in silkworms that had been treated with a specific Hh signaling pathway antagonist, cyclopamine. The fat-body-specific Hh blockade tests were consistent with Hh signaling inhibiting silkworm adipogenesis. Our results indicate that the role of Hh signaling in inhibiting fat formation is conserved in vertebrates and invertebrates.

关键词： adipocyte differentiation adipogenesis Bombyx mori Hedgehog (Hh) signaling pathway silkworm

Evaluation of Antibody-Dependent Enhancement of SARS-CoV Infection in Rhesus Macaques Immunized with an Inactivated SARS-CoV Vaccine

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Virologica Sinica 2018年第2期33卷 201-204页

作者： Fan Luo Fan-Lu Liao Hui Wang Hong-Bin Tang Zhan-Qiu Yang Wei Hou State Key laboratory of virology/Hubei Province Key laboratory of Allergy and immunology/Institute of Medical virology School of Basic Medical Sciences Wuhan University Department of Clinical laboratory Zhongnan HospitalWuhan University Center for animal Experiment/animal Biosafety Level-Ⅲ laboratory Wuhan University

In 2003,severe acute respiratory syndrome coronavirus（SARS-CoV）emerged in Guangdong Province,China,infected more than 8000 individuals,and resulted in a 10%mortality rate（Rota et al.2003）.Later,in 2012,a novel CoV,Middle East respiratory syndrome coronavirus（MERS-CoV）,was isolated from the sputum of a man in Saudi Arabia（Perl et al.2014）.Notably,MERS-CoV recently reemerged in the Republic of Korea, and killed 36 out of 186 confirmed cases （Korea Centers for Disease Control and Prevention 2015）. Therefore, SARS-CoV still carries the potential for resurgence; efforts are being made to prevent the recurrence of an epidemic.

关键词： SARS-CoV 感染猕猴活性免疫疫苗评估抗体

DNAskew:Statistical Analysis of Base Compositional Asymmetry and Prediction of Replication Boundaries in the Genome Sequences

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Acta Biochimica et Biophysica Sinica 2004年第1期36卷 16-20页

作者： Xiang-Ru MA, Shao-Bo XIAO, Ai-Zhen GUO, Jian-Qiang LU, and Huan-Chun CHEN( laboratory of animal virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China ) laboratory of animal virology College of Veterinary Medicine Huazhong Agricultural University Wuhan 430070 China

Sueoka and Lobry declared respectively that, in the absence of bias between the two DNA strands for mutation and selection, the base composition within each strand should be A=T and C=G (this state is called Parity Rule type 2, PR2). However, the genome sequences of many bacteria, vertebrates and viruses showed asymmetries in base composition and gene direction. To determine the relationship of base composition skews with replication orientation, gene function, codon usage biases and phylogenetic evolution, in this paper a program called DNAskew was developed for the statistical analysis of strand asymmetry and codon composition bias in the DNA sequence. In addition, the program can also be used to predict the replication boundaries of genome sequences. The method builds on the fact that there are compositional asymmetries between the leading and the lagging strand for replication. DNAskew was written in Perl script language and implemented on the LINUX operating system. It works quickly with annotated

关键词： strand asymmetry base composition statistics analysis replication origin bioinformatics

Association of swine influenza H1N1 pandemic virus(SIV-H1N1p) with porcine respiratory disease complex in sows from commercial pig farms in Colombia

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Virologica Sinica 2014年第4期29卷 242-249页

作者： Luisa Fernanda Mancipe Jiménez Gloria Ramírez Nieto Victor Vera Alfonso Jairo Jaime Correa laboratory of animal virology Faculty of Veterinary Medicine and Animal Science National University of Colombia

Porcine respiratory disease complex （PRDC） is a serious health problem that mainly affects growing and finishing pigs. PRDC is caused by a combination of viral and bacterial agents, such as porcine reproductive and respiratory syndrome virus （PRRSV）, swine influenza virus （SIV）, Mycoplasma hyopneumoniae （Myh）, Actinobacillus pleuropneumoniae （APP）, Pasteurella multocida and Porcine circovirus 2 （PCV2）. To characterize the specific role of swine influenza virus in PRDC presentation in Colombia, 11 farms from three major production regions in Colombia were examined in this study. Nasal swabs, bronchial lavage and lung tissue samples were obtained from animals displaying symptoms compatible with SIV. Isolation of SIV was performed in 9-day embryonated chicken eggs or Madin-Darby Canine Kidney （MDCK） cells. Positive isolates, identified via the hemagglutination inhibition test, were further analyzed using PCR. Overall, 7 of the 11 farms were positive for SIV. Notably, sequencing of the gene encoding the hemagglutinin （HA） protein led to grouping of strains into circulating viruses identified during the human outbreak of 2009, classified as pandemic H1N1-2009. Serum samples from 198 gilts and multiparous sows between 2008 and 2009 were obtained to determine antibody presence of APP, Myh, PCV2 and PRRSV in both SIV-H1Nlp-negative and -positive farms, but higher levels were recorded for SIV- HI Nlp-positive farms. Odds ratio （OR） and P values revealed statistically significant differences （p〈0.05） in PRDC presentation in gilts and multiparous sows of farms positive for SIV-HINlp. Our findings indicate that positive farms have increased risk of PRDC presentation, in particular, PCV2, APP and Myh.

关键词： SIV-HIN1 swine influenza pandemic porcine respiratory disease complex Colombia

Analysis of Expression Profiles of Long Noncoding RNAs and mRNAs in A549 Cells Infected with H3N2 Swine Influenza Virus by RNA Sequencing

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Virologica Sinica 2020年第2期35卷 171-180页

作者： Yina Zhang Tianqi Yu Yingnan Ding Yahui Li Jing Lei Boli Hu Jiyong Zhou MOA Key laboratory of animal virology and Department of Veterinary Medicine Zhejiang UniversityHangzhou 310058China MOE International Joint Collaborative Research laboratory for animal Health and Food Safety Institute of Immunology and College of Veterinary MedicineNanjing Agricultural UniversityNanjing 210095China

Long noncoding RNAs(lncRNAs)participate in regulating many biological ***,their roles in influenza A virus(IAV)pathogenicity are largely ***,we analyzed the expression profiles of lncRNAs and mRNAs in H3N2-infected cells and mock-infected cells by high-throughput *** results showed that 6129 lncRNAs and 50,031 mRNA transcripts in A549 cells displayed differential expression after H3N2 infection compared with mock *** the differentially expressed lncRNAs,4963 were upregulated,and 1166 were *** annotation and enrichment analysis using gene ontology and Kyoto Encyclopedia of Genes and Genomes databases(KEGG)suggested that target genes of the differentially expressed lncRNAs were enriched in some biological processes,such as cellular metabolism and *** up-or downregulated lncRNAs were selected and further verified by quantitative real-time polymerase chain reaction(RT-qPCR)and reverse transcription PCR(RT-PCR).To the best of our knowledge,this is the first report of a comparative expression analysis of lncRNAs in A549 cells infected with *** results support the need for further analyses of the functions of differentially expressed lncRNAs during H3N2 infection.

关键词： Influenza virus(IAV) Long noncoding RNA(lncRNA) A549 cells High-throughput sequencing

Differential CircRNA Expression Profiles in PK-15 Cells Infected with Pseudorabies Virus Type Ⅱ

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Virologica Sinica 2021年第1期36卷 75-84页

作者： Haimin Li Wen Tang Yulan Jin Weiren Dong Yan Yan Jiyong Zhou MOA Key laboratory of animal virology Center of Veterinary SciencesZhejiang UniversityHangzhou 310058China

Circular RNAs(circ RNAs)belong to a class of non-coding RNAs with diverse biological ***,little is known about their roles in case of pseudorabies virus(Pr V)***,we analyzed the expression profile of host circ RNAs from a virulent Pr V type II strain DX(Pr V-DX)infected and an attenuated g E/TK deficient(g E-TK-Pr V)strain of Pr V infected PK-15 *** RNAs were identified by findcirc and analyzed with DESeq *** with the mock cells,449 differentially expressed(DE)circ RNAs(233 down-regulated and 216 up-regulated)from Pr V-DX infected and578 DE circ RNAs(331 down-regulated and 247 up-regulated)from g E-TK-Pr V infected PK-15 cells were *** addition,459 DE circ RNAs(164 down-regulated and 295 up-regulated)between the Pr V-DX and g E-TK-Pr V infected cells were *** expression patterns of 13 circ RNAs were validated by reverse transcription quantitative real-time PCR(RT-q PCR)and results were similar as of *** putative target mi RNA binding sites of DE circ RNAs were predicted by using mi Randa and ps *** circ RNA-mi RNA-m RNA network was constructed and certain mi RNAs that have possible roles in antiviral immune response,such as mi R-210 and mi R-340,were *** and KEGG pathway analysis demonstrated that DE circ RNAs were enriched in the processes such as cellular metabolism,protein binding,RNA degradation and regulation of actin ***,these findings might provide the useful information for a better understanding of mechanisms underlying the interaction between Pr V-II and host cells.

关键词： Pseudorabies virus infection RNA-seq CircRNA

Construction and immunogenicity of recombinant pseudorabies virus expressing the modified GP5m protein of porcine reproduction and respiratory syndrome virus

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Frontiers in Biology 2007年第1期2卷 85-91页

作者： JIANG Yunbo FANG Liurong XIAO Shaobo ZHANG Hui CHEN Huanchun laboratory of animal virology College of Animal Science and Veterinary MedicineHuazhong Agriculture UniversityWuhan 430070China

Pseudorabies virus(PRV),an alpha-herpesvirus,has been developed as a live viral vector for animal ***,the PRV recombinant virus TK^(-)/gE^(-)/GP5^(+)expressing GP5 of porcine reproductive and respiratory syn-drome virus(PRRSV),based on the PRV genetically depleted vaccine strain TK^(-)/gE^(-)/LacZ^(+),scarcely stimulated the vaccinated animals to produce neutralizing antibodies against *** develop a booster-specific immune response of such PRV recombinants,the ORF5m gene(the modified ORF5 gene having better immune responses)was substituted for the ORF5 gene and introduced into PRV TK^(-)/gE^(-)/LacZ^(+),resulting in a PRV recombinant named TK^(-)/gE^(-)/GP5m^(+),which expressed the modified GP5m *** recombinant virus was confirmed using PCR,Southern blotting and Western ***^(-)/gE^(-)/GP5m^(+)and TK^(-)/gE^(-)/GP5^(+)expressing the authentic GP5 protein were inoculated into Balb/c mice to evaluate their immune *** results indicated that the protecting neutralization antibodies(the 3/6 vaccinated mice obtained 1:16)and cell immune responses induced by TK^(-)/gE^(-)/GP5m^(+)against PRRSV were higher than that induced by TK^(-)/gE^(-)/GP5^(+).Thus,the development of the new PRV recombinant expressing the modified GP5m protein as a candidate vaccine established the basis for the study of bivalent genetic engineering vaccines against PRRSV and PRV.

关键词： porcine reproductive and respiratory syndrome virus GP5m the recombinant pseudorabies virus

Identification of one peptide which inhibited infectivity of avian infectious bronchitis virus in vitro

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Science China(Life Sciences) 2006年第2期49卷 158-163页

作者： PENG Bo1, CHEN Hanyang1, TAN Yadi1,2, JIN Meilin1,2, CHEN Huanchun1,2 & GUO Aizhen1,2 1. laboratory of animal virology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China 2. National Key laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China laboratory of animal virology College of Veterinary Medicine Huazhong Agricultural University Wuhan 430070 China.

Purified avian infectious bronchitis virus (IBV) was used to screen a random phage dis-play peptide library. After the fourth panning, 10 positive phages were sequenced and characterized. The phages specifically inhibited IBV infectivity in HeLa cells and blocked IBV haemagglutination. One linear peptide “GSH HRH VHS PFV” from the positive phages with the highest neutralization titer was synthesized and this peptide inhibited IBV infection in HeLa as well. The results may contribute to development of antiviral therapeutics for IBV and studying the determinants for viral and cell interac-tion.

关键词： avian infectious bronchitis virus, phage display, peptides.

The Effect of Bovine IFN-α on the Immune Response in Guinea Pigs Vaccinated with DNA Vaccine of Foot-and-Mouth Disease Virus

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 701-706页

作者： Hui-Chen GUO, Zai-Xin LIU*, Shi-Qi SUN, Qing-Wen LENG1, Dong LI, Xiang-Tao LIU, and Qing-Ge XIE* Key laboratory of animal virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China College of animal Science and Technology, Shihezi University, Shihezi 832003, China 1 Key laboratory of animal virology of Ministry of Agriculture Lanzhou Veterinary Research Institute Chinese Academy of Agricultural SciencesLanzhou 730046 China College of animal Science and Technology Shihezi UniversityShihezi 832003 China

In this study, we constructed recombinant plasmid pcDNA3.1/P12X3C3D including P1, 2A, 3C, 3D and part of 2B gene of FMDV and pcDNA3.1/IFN containing the gene encoding bovine IFN-α. We inoculated the DNA vaccine pcDNA3.1/P12X3C3D with or without pcDNA3.1/IFN to evaluate the effi- ciency of this DNA vaccine and the immunogenicity of DNA vaccine enhanced by the co-delivery with pcDNA3.1/IFN. After two times of vaccination with DNA vaccine, all of guinea pigs were challenged with 103 ID50 FMDV type O. Anti-FMDV antibody levels were detected by ELISA and T lymphocyte proliferation response was tested by MTT assay. The result shows that guinea pigs inoculated by pcDNA3.1/P12X3C3D alone or with pcDNA3.1/IFN generated specific antibodies and induced an FMDV-specific T lymphocyte proliferation response. FMDV challenge tests showed that one in four guinea pigs immunized by pcDNA3.1/ P12X3C3D with pcDNA3.1/IFN was protected from the FMDV serotype O infection. This result indicated that the efficiency of the DNA vaccine was enhanced by co-delivery with pcDNA3.1/IFN. However, the protection rate was considerably lower than that immunized with conventional FMD vaccine.

关键词： foot-and-mouth disease virus (FMDV) DNA vaccine IFN-α gene vaccination