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Construction and Immunogenicity of Recombinant Lactococcus lactis Expressing S1 Protein of Porcine Epidemic Diarrhea Virus(PEDV) 被引量:1
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作者 Wang Liping Han Xianjie +3 位作者 Wang Xiaobin Gai Chunyun Li Junwei Shan Hu 《Animal Husbandry and Feed Science》 CAS 2018年第2期115-119,125,共6页
To evaluate the specific immune responses induced by recombinant Lactococcus lactis(L.lactis) which expresses porcine epidemic diarrhea virus(PEDV) S1 protein through oral administration,the spike gene fragment of... To evaluate the specific immune responses induced by recombinant Lactococcus lactis(L.lactis) which expresses porcine epidemic diarrhea virus(PEDV) S1 protein through oral administration,the spike gene fragment of PEDV was amplified from PEDV SDLY strain to construct p MG36 e-S1 recombinant plasmid.The recombinant plasmid was then electro-transferred into competent cells of L.lactis MG1363,to prepare the recombinant L.lactis expressing S1 protein of PEDV.The expression of target protein was identified by SDS-PAGE and Western-blot.New Zealand white rabbits were orally administered with the recombinant strain;the antibody titer in intestinal mucosa and serum was detected by neutralizing test;and the specific Ig G in serum was evaluated by indirect ELISA.The results showed that the recombinant L.lactis could effectively induce high level of Ig G in serum and high level of mucosal immune antibody.The recombinant L.lactis is qualified to be a potential oral vaccine because it could successfully stimulate both humoral and mucosal immune responses against PEDV. 展开更多
关键词 porcine epidemic diarrhea virus pedv Spike protein pMG36e vector Lactococcus lactis MG1363 Immune response
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Screening of Host Proteins Interacting with PorcineEpidemic Diarrhea Virus (PEDV) N Protein by YeastTwo-hybrid System
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作者 Wang Zhongze Qin Cuili +10 位作者 Kong Ning Zuo Yewen Wang Meng Zheng Hao Tong Wu Li Liwei Yu Hai Li Zhili Shan Tongling Tong Guangzhi Li Xue 《Animal Husbandry and Feed Science》 CAS 2018年第4期267-271,共5页
[Objective] The paper was to obtain host proteins interacting with porcine epidemic diarrhea virus (PEDV) N protein. [Method] The re-combinant vector pGBKT7-N of PEDV N gene was constructed and used as the bait plas... [Objective] The paper was to obtain host proteins interacting with porcine epidemic diarrhea virus (PEDV) N protein. [Method] The re-combinant vector pGBKT7-N of PEDV N gene was constructed and used as the bait plasmid to screen the proteins interacting with N protein ofPEDV from the cDNA library of porcine alveolar macrophage (PAM) by yeast two-hybrid method. [Result] There was no toxicity and self activationof bait protein in yeast hybridization system, and six proteins (FTH1, LGALS3, CORO1C, SNRPG, KRTAP5-3, ZNF598) interacting with N proteinwere indentified. It was confirmed that LGALS3 and SNRPG had specific interaction with N protein by return experiment and co-immunoprecipitation(CoIP) test. [Conclusion] The study lays a foundation for further studying the function of PEDV N protein and the pathogenic mechanism of PEDV. 展开更多
关键词 porcine epidemic diarrhea virus pedv Yeast two-hybrid N protein Protein interaction
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Isolation and Identification of Porcine Epidemic Diarrhea Virus(PEDV) HLJ Strain with IPEC-J2 Cells and Phylogenetic Analysis of Its S Gene
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作者 Feng Rui Liu Hai-xin +4 位作者 Zhong Ming Li Xun-liang Huang Xiao-dan Ren Yu-dong Li Guang-xing 《Journal of Northeast Agricultural University(English Edition)》 CAS 2019年第4期63-72,共10页
Porcine epidemic diarrhea(PED)is caused by porcine epidemic diarrhea virus(PEDV),and is characterized by vomiting,diarrhea and dehydration of suckling pigs from 80% to 100% morbidity and 50% to 90% mortality,and resul... Porcine epidemic diarrhea(PED)is caused by porcine epidemic diarrhea virus(PEDV),and is characterized by vomiting,diarrhea and dehydration of suckling pigs from 80% to 100% morbidity and 50% to 90% mortality,and resulted in tremendous economic losses to swine industry.The PEDV mainly infects small intestine of pigs,resulting in vacuolar degeneration and necrosis of mucosal epithelium.The IPEC-J2 is a pig intestine epithelial cell line,which is similar to the intestinal environment of piglets,can be used to isolate and identify the PEDV field isolates.In this study,it appeared the PEDV typical postmortem changes and histopathological lesion of degeneration and destruction of small intestine in infected piglets,and IHC identified that the PEDV distributed in the mucosa and submucosa of small intestine mostly.Furthermore,the PEDV HLJ strain was successfully isolated and characterized in the IPEC-J2 cells,and indicated that the IPEC-J2 cell line was sensitive to isolate and adapt the PEDV field strain,and could be utilized to multiply the PEDV rapidly.The S gene analysis indicated that the PEDV HLJ strain was the prevailed virus,belonged to Group 1 with attenuated virulent DR13,SC1402 and J-S2/2015 strains isolated in South Korea and China from 2014 to 2015.This study had important theoretical and practical significances on analyzing genetic variation of the PEDV,understanding the pathogenic characteristics of the virus and developing new vaccines for the PED. 展开更多
关键词 porcine epidemic diarrhea virus cytopathic effect IPE-J2 cell isolation and identification phylogenetic analysis
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Detection of Porcine Epidemic Diarrhea Virus in Guangxi Province from 2011 to 2014 and Sequence Analysis of Its M Gene 被引量:3
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作者 Lu Bingxia Qin Yibin +12 位作者 He Ying Li Yingying Liang Jiaxing Li Keyu Li Bin Su Qianlian Zhou Yingning Jiang Dongfu Lu Jingzhuan Bi Bingfen Liang Baozhong Duan Qunpeng Zhao Wu 《Animal Husbandry and Feed Science》 CAS 2016年第1期12-17,38,共7页
Detection of pigs epidemic diarrhea virus (PEDV) was conducted on 331 piglets diarrhea fecal samples collected in Nanning, Yulin and other 12 areas of Guangxi Province from January of 2011 to April of 2014 by the me... Detection of pigs epidemic diarrhea virus (PEDV) was conducted on 331 piglets diarrhea fecal samples collected in Nanning, Yulin and other 12 areas of Guangxi Province from January of 2011 to April of 2014 by the method of reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the positive samples of PEDV were 210 and the positive rate was 63.44%. The clone and sequencing of M gene was carried out on 25 positive samples. PEDV reference strains were selected from GeneBank to conduct the sequence homology alignment analysis and the phylogenetic tree of M gene. The M gene homology and amino acid sequence identity between 25 isolated strains and 51 reference strains were 96.0% - 99.6% and 94.3% - 99.6%, respectively. The genetic variation anal- ysis of M gene showed that the genetic relationship of PEDV prevalent strains in Guangxi Province from 2013 to 2014 was close to that of the prevalent strains in Bei- jing, Anhui, Wuhan, Hebei and Guangdong from 2010 to 2013, and which were far from that of the Chinese early isolates CH/S (GenBank number: JN547228 ), vaccine strain CV777 (GenBank number: AF353511 ) and Attenuated DR13 (GenBank number: JQ023162). Indicating that the PEDV strains prevalent in Guan- gxi in recent years showed significant variation with the early isolates. 展开更多
关键词 porcine epidemic diarrhea virus pedv M gene Genetic variation
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Real-time Fluorescence Reverse-transcription Loop-mediated Isothermal Amplification for Detection of Porcine Epidemic Diarrhea Virus 被引量:1
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作者 Yanan LI Jianchang WANG +5 位作者 Bin LI Ruiwen LI Yanhong HOU Lei ZHANG Yun BAI Wanzhe YUAN 《Agricultural Biotechnology》 CAS 2018年第2期137-140,共4页
Porcine epidemic diarrhea,a highly contagious enteric infectious disease caused by the porcine epidemic diarrhea virus(PEDV)with symptoms of vomit,diarrhea,loss of appetite of suckling pig,has led to serious economic ... Porcine epidemic diarrhea,a highly contagious enteric infectious disease caused by the porcine epidemic diarrhea virus(PEDV)with symptoms of vomit,diarrhea,loss of appetite of suckling pig,has led to serious economic loss to the global swine industry.In this study,a real-time fluorescence reverse transcription loop-mediated isothermal amplification(RT-LAMP)assay was developed to detect PEDV RNA.The real-time fluorescence RT-LAMP assay was performed at62℃for 60 min,using a simple and portable device,the ESE-Quant Tube Scanner.The detection limit of RNA was 2.9×10^(6) copies/μl,10 times as sensitive as RT-PCR,and the detection was specific only to PEDV.Application of this method to clinical samples yielded a positivity rate of 93%,which was higher than that of RT-PCR.This technique saves time and is efficient,and is thus expected to be useful for the diagnosis of PEDV infection in the field. 展开更多
关键词 porcine epidemic diarrhea virus Real-time fluorescence RT-LAMP DetectionHome
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Porcine NF-κB p65 Subunit:Molecular Characterization,Tissue Expression and Transcriptional Profile in Porcine Epidemic Diarrhea Virus-infected IPEC-J2 Cells
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作者 Liu Hai-xin Wang Hong-wei +8 位作者 Cao Li-yan Dante S Zarlenga Ge Xu-ying Zhang Yue Yin Xue-ting Zhang Rui-li Ren Yu-dong Huang Xiao-dan Li Guang-xing 《Journal of Northeast Agricultural University(English Edition)》 CAS 2020年第2期99-107,共9页
The p65 protein is a functional subunit of NF-κB family and exhibits a crucial role in host immune and inflammatory responses,apoptosis and tumor proliferation if improperly-regulated.Given its ubiquitous association... The p65 protein is a functional subunit of NF-κB family and exhibits a crucial role in host immune and inflammatory responses,apoptosis and tumor proliferation if improperly-regulated.Given its ubiquitous association with nearly all the animal cells and its pleotropic functions,the gene encoding NF-κB p65 subunit was cloned and sequenced from porcine kidney(PK-15)cells.The gene was 1662 bp in length,encoded a 553-amino acid protein and contained the prototypical NF-κB functional domains.Real-time quantitative RT-PCR and Western blot were used to characterize the transcription and expression levels of the p65 in different pig tissues.The results indicated that the p65 gene and protein were both broadly expressed in pig tissues,but most highly expressed in the intestine-associated lymph nodes and the lungs.To localize the recombinant protein in intestinal porcine epithelial cells(IPEC-J2),the gene was subcloned into the vector pEGFP(pEGFP-p65).Using fluorescence microscopy,the protein was found confined to the cytoplasm in normal cells;however,during porcine epidemic diarrhea virus(PEDV)infection,mRNA and protein expression were significantly up-regulated and the protein exhibited an overt tendency for nuclear translocalization consistent with a regulatory role in antiviral innate immunity. 展开更多
关键词 porcine NF-κB p65 tissue expression bioinformatic analysis porcine epidemic diarrhea virus
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Evaluation of purified recombinant spike fragments forassessment of the presence of serum neutralizing antibodiesagainst a variant strain of porcine epidemic diarrhea virus 被引量:5
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作者 Jianwei Hao Yun Zhang +4 位作者 Shengkun Fang Zhifen Wen Xiangbin Zhang Chunyi Xue Yongchang Cao 《Virologica Sinica》 SCIE CAS CSCD 2017年第4期307-316,共10页
Since 2010, variant strains of porcine epidemic diarrhea virus(PEDV) have caused disasters in the pork industry. The spike(S) protein, as the major immunity-eliciting antigen, has previously been used for serological ... Since 2010, variant strains of porcine epidemic diarrhea virus(PEDV) have caused disasters in the pork industry. The spike(S) protein, as the major immunity-eliciting antigen, has previously been used for serological testing and has been found to correlate significantly with the results of the serum neutralization(SN) test. However, further evaluation of this method is needed as new epidemic strains of PEDV emerge. Hence, the main objective of this study was to assess sow sera and determine the correlation between enzyme-linked immunosorbent assay(ELISA) results(involving a newly isolated GDS01 virus-based ELISA and ELISAs based on seven recombinant fragments comprising overlapping S1 and partial S2 sequences) and SN titers. Furthermore, we determined the reliability of the ELISAs based on receiver operating characteristics(ROC) curve analyses. For the most promising ELISA, i.e., the SP4 ELISA, the correlation coefficient(r) and the area under curve(AUC) were determined to be 0.6113 and 0.8538, respectively. In addition, we analyzed the homology of the SP4 sequences obtained from different strains(including vaccine strains) and found that various strains showed a high degree of homology in this region. Thus, we conclude that SP4 is a promising serological testing protein for use in the field. 展开更多
关键词 porcine epidemic diarrhea virus (pedv) SPIKE protein enzyme-linkedimmunosorbent assay SERUM NEUTRALIZATION test
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Isolation and oral immunogenicity assessment of porcine epidemic diarrhea virus NH-TA2020 strain:One of the predominant strains circulating in China from 2017 to 2021 被引量:10
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作者 Xiaowen Li Yang Li +11 位作者 Jiapei Huang Yali Yao Wenying Zhao Yunjing Zhang Jie Qing Jing Ren Zhong Yan Zewei Wang Xiaofang Hu Duli Kang Hongqiang Liu Zhichun Yan 《Virologica Sinica》 SCIE CAS CSCD 2022年第5期646-655,共10页
Porcine epidemic diarrhea(PED)caused by porcine epidemic diarrhea virus(PEDV)is one of the most devastating diseases in the global pig industry due to its high mortality rate in piglets.Maternal vaccines can effective... Porcine epidemic diarrhea(PED)caused by porcine epidemic diarrhea virus(PEDV)is one of the most devastating diseases in the global pig industry due to its high mortality rate in piglets.Maternal vaccines can effectively enhance the gut-mammary gland-secretory IgA axis to boost lactogenic immunity and passive protection of nursing piglets against PEDV challenge.From 2017 to 2021,we collected 882 diarrhea samples from 303 farms in China to investigate the epidemiology of PEDV.The result showed that about 52.15%(158/303)of the farms were positive for PEDV with an overall detection rate of 63.95%(564/882)of the samples.The S1 fragments of S gene from 104 strains were sequenced for the phylogenetic analysis.A total of 71 PEDV strains(68.27%)sequenced in this study were clustered into the predominant G2c subgroup,while the newly-defined G2d strains(9.62%)were identified in three provinces of China.The NH-TA2020 strain of G2c subgroup was isolated and cultured,and its infection to piglets caused watery diarrhea within 24 h,indicating its strong pathogenicity.Oral administration of NH-TA2020 strain to pregnant gilts stimulated high levels of IgA antibody in colostrum.The piglets fed by the gilts above were challenged with NH-TA2020 strain or CH-HeB-RY-2020 strain from G2d subgroup,and the clinical symptoms and virus shedding were significantly reduced compared to the mock group.Our findings suggest that G2c subgroup is the predominant branch circulating in China from 2017 to 2021.Oral administration of NH-TA2020 enhances maternal IgA and lactogenic immune responses,which confer protection against the homologous and emerging G2d PEDV strains challenges in neonates. 展开更多
关键词 porcine epidemic diarrhea virus(pedv) Molecular epidemiological investigation virus isolation Lactogenic immune IGA
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Coexistence of multiple genotypes of porcine epidemic diarrhea virus with novel mutant S genes in the Hubei Province of Chinain 2016 被引量:4
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作者 Zhe Zeng Ting-Ting Li +4 位作者 Xin Jin Fu-Hu Peng Nian-Hua Song Gui-Qing Peng Xing-Yi Ge 《Virologica Sinica》 SCIE CAS CSCD 2017年第4期298-306,共9页
The emergence of highly virulent porcine epidemic diarrhea virus(PEDV) variants in China caused huge economic losses in 2010. Since then, large-scale sporadic outbreaks of PED caused by PEDV variants have occasionally... The emergence of highly virulent porcine epidemic diarrhea virus(PEDV) variants in China caused huge economic losses in 2010. Since then, large-scale sporadic outbreaks of PED caused by PEDV variants have occasionally occurred in China. However, the molecular diversity and epidemiology of PEDV in different provinces has not been completely understood. To determine the molecular diversity of PEDV in the Hubei Province of China, we collected 172 PED samples from 34 farms across the province in 2016 and performed reverse transcription polymerase chain reaction(RTPCR)by targeting the nucleocapsid(N) gene. Seventy-four samples were found to be PEDVpositive.We further characterized the complete spike(S) glycoprotein genes from the positive samples and found 21 different S genes with amino acid mutations. The PEDV isolates here presented most of the genotypes which were found previously in field isolates in East and SouthEast Asia, North America, and Europe. Besides the typical Genotypes Ⅰ and Ⅱ, the INDEX groups were also found. Importantly, 58 new amino acids mutant sites in the S genes, including 44 sites in S1 and 14 sites in S2, were first described. Our results revealed that the S genes of PEDV showed variation and that diverse genotypes of PEDV coexisted and were responsible for the PED outbreaks in Hubei in 2016. This work highlighted the complexity of the epidemiology of PEDV and emphasized the need for reassessing the efficacy of classic PEDV vaccines against emerging variant strains and developing new vaccines to facilitate the prevention and control of PEDV in fields. 展开更多
关键词 porcine epidemic diarrhea virus (pedv) CORONAvirus spike genetic variation
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Nucleocapsid protein from porcine epidemic diarrhea virus isolates can antagonize interferon-λ production by blocking the nuclear factor-κB nuclear translocation 被引量:10
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作者 Ying SHAN Zi-qi LIU +7 位作者 Guo-wei LI Cong CHEN Hao LUO Ya-jie LIU Xun-hui ZHUO Xing-fen SHI Wei-huan FANG Xiao-liang LI 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2018年第7期570-580,共11页
Porcine epidemic diarrhea virus(PEDV) is a highly infectious pathogen that can cause severe diseases in pigs and result in enormous economic losses in the worldwide swine industry. Previous studies revealed that PED... Porcine epidemic diarrhea virus(PEDV) is a highly infectious pathogen that can cause severe diseases in pigs and result in enormous economic losses in the worldwide swine industry. Previous studies revealed that PEDV exhibits an obvious capacity for modulating interferon(IFN) signaling or expression. The newly discovered type III IFN, which plays a crucial role in antiviral immunity, has strong antiviral activity against PEDV proliferation in IPEC-J2 cells. In this study, we aimed to investigate the effect of PEDV nucleocapsid(N) protein on type III IFN-λ. We found that the N proteins of ten PEDV strains isolated between 2013 and 2017 from different local farms shared high nucleotide identities, while the N protein of the CV777 vaccine strain formed a monophyletic branch in the phylogenetic tree. The N protein of the epidemic strain could antagonize type III IFN, but not type I or type II IFN expression induced by polyinosinic-polycytidylic acid(poly(I:C)) in IPEC-J2 cells. Subsequently, we demonstrated that the inhibition of poly(I:C)-induced IFN-λ3 production by PEDV N protein was dependent on the blocking of nuclear factor-κB(NF-κB) nuclear translocation. These findings might help increase understanding of the pathogenesis of PEDV and its mechanisms for evading the host immune response. 展开更多
关键词 porcine epidemic diarrhea virus Nucleocapsid protein Interferon-λ(IFN-λ) Nuclear factor-κB(NF-κB) Intestinal epithelial cells
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Genome-wide transcriptome analysis of porcine epidemic diarrhea virus virulent or avirulent strain-infected porcine small intestinal epithelial cells 被引量:4
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作者 Ouyang Peng Xiaona Wei +7 位作者 Usama Ashraf Fangyu Hu Yongbo Xia Qiuping Xu Guangli Hu Chunyi Xue Yongchang Cao Hao Zhang 《Virologica Sinica》 SCIE CAS CSCD 2022年第1期70-81,共12页
Porcine epidemic diarrhea virus(PEDV)is the main cause of diarrhea,vomiting,and mortality in pigs,which results in devastating economic loss to the pig industry around the globe.In recent years,the advent of RNAsequen... Porcine epidemic diarrhea virus(PEDV)is the main cause of diarrhea,vomiting,and mortality in pigs,which results in devastating economic loss to the pig industry around the globe.In recent years,the advent of RNAsequencing technologies has led to delineate host responses at late stages of PEDV infection;however,the comparative analysis of host responses to early-stage infection of virulent and avirulent PEDV strains is currently unknown.Here,using the BGI DNBSEQ RNA-sequencing,we performed global gene expression profiles of pig intestinal epithelial cells infected with virulent(GDS01)or avirulent(HX)PEDV strains for 3,6,and 12 h.It was observed that over half of all significantly dysregulated genes in both infection groups exhibited a down-regulated expression pattern.Functional enrichment analyses indicated that the differentially expressed genes(DEGs)in the GDS01 group were predominantly related to autophagy and apoptosis,whereas the genes showing the differential expression in the HX group were strongly enriched in immune responses/inflammation.Among the DEGs,the functional association of TLR3 and IFIT2 genes with the HX and GDS01 strains replication was experimentally validated by TLR3 inhibition and IFIT2 overexpression systems in cultured cells.TLR3 expression was found to inhibit HX strain,but not GDS01 strain,replication by enhancing the IFIT2 expression in infected cells.In conclusion,our study highlights similarities and differences in gene expression patterns and cellular processes/pathways altered at the early-stage infection of PEDV virulent and avirulent strains.These findings may provide a foundation for establishing novel therapies to control PEDV infection. 展开更多
关键词 CORONAvirus porcine epidemic diarrhea virus(pedv) viruLENCE TRANSCRIPTOME Apoptosis Autophagy Immunity
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Significant Inhibition of Porcine Epidemic Diarrhea Virus In Vitro by Remdesivir,Its Parent Nucleoside and β-D-N^(4)-hydroxycytidine 被引量:3
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作者 Yuanchao Xie Xiaozhen Guo +5 位作者 Tianwen Hu Daibao Wei Xiuli Ma Jiaqiang Wu Bing Huang Jingshan Shen 《Virologica Sinica》 SCIE CAS CSCD 2021年第5期997-1005,共9页
Porcine epidemic diarrhea(PED)caused by porcine epidemic diarrhea virus(PEDV)is widespread in the world.In recent years,the increased virulence of the virus due to viral variations,has caused great economic losses to ... Porcine epidemic diarrhea(PED)caused by porcine epidemic diarrhea virus(PEDV)is widespread in the world.In recent years,the increased virulence of the virus due to viral variations,has caused great economic losses to the pig industry in many countries.It is always worthy to find effective therapeutic methods for PED.As an important class of antivirals,nucleoside drugs which target viral polymerases have been applied in treating human viral infections for half a century.Herein,we evaluated the anti-PEDV potential of three broad-spectrum antiviral nucleoside analogs,remdesivir(RDV),its parent nucleoside(RDV-N)andβ-D-N^(4)-hydroxycytidine(NHC).Among them,RDV-N was the most active agent in Vero E6 cells with EC_(50) of 0.31μmol/L,and more potent than RDV(EC_(50)=0.74μmol/L)and NHC(EC_(50)=1.17μmol/L).The activity of RDV-N was further confirmed using an indirect immuno-fluorescence assay.Moreover,RDV-N exhibited a good safety profile in cells and in mice.The high sequence similarity of the polymerase functional domains of PEDV with other five porcine coronaviruses indicated a broader antiviral spectrum for the three compounds.Generally,RDV-N is a promising broad-spectrum antiviral nucleoside,and it would be worthy to make some structural modifications to increse its oral bioavailability. 展开更多
关键词 porcine epidemic diarrhea virus(pedv) Nucleoside analog RNA dependent RNA polymerase(RdRp) Antiviral activity
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Identification of niclosamide as a novel antiviral agent against porcine epidemic diarrhea virus infection by targeting viral internalization 被引量:1
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作者 Yue Wang Huimin Huang +8 位作者 Dongliang Li Chenxu Zhao Shuai Li Panpan Qin Yaqin Li Xia Yang Wenjuan Du Wentao Li Yongtao Li 《Virologica Sinica》 SCIE CAS CSCD 2023年第2期296-308,共13页
Porcine epidemic diarrhea virus(PEDV),an enteropathogenic coronavirus,has catastrophic impacts on the global pig industry.However,there remain no effective drugs against PEDV infection.In this study,we utilized a reco... Porcine epidemic diarrhea virus(PEDV),an enteropathogenic coronavirus,has catastrophic impacts on the global pig industry.However,there remain no effective drugs against PEDV infection.In this study,we utilized a recombinant PEDV expressing renilla luciferase(PEDV-Rluc)to screen potential anti-PEDV agents from an FDAapproved drug library in Vero cells.Four compounds were identified that significantly decreased luciferase activity of PEDV-Rluc.Among them,niclosamide was further characterized because it exhibited the most potent antiviral activity with the highest selectivity index.It can efficiently inhibit viral RNA synthesis,protein expression and viral progeny production of classical and variant PEDV strains in a dose-dependent manner.Time of addition assay showed that niclosamide exhibited potent anti-PEDV activity when added simultaneously with or after virus infection.Furthermore,niclosamide significantly inhibited the entry stage of PEDV infection by affecting viral internalization rather than viral attachment to cells.In addition,a combination with other small molecule inhibitors of endosomal acidification enhanced the anti-PEDV effect of niclosamide in vitro.Taken together,these findings suggested that niclosamide is a novel antiviral agent that might provide a basis for the development of novel drug therapies against PEDV and other related pathogenic coronavirus infections. 展开更多
关键词 CORONAvirus porcine epidemic diarrhea virus(pedv) Niclosamide(NIC) Antiviral virus entry ENDOCYTOSIS Host-targeted antivirals
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PEDV乳酸菌工程菌株对小鼠的免疫效果研究
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作者 安琦 牛彦波 +3 位作者 吴皓琼 樊川 曹亚彬 原韬 《农学学报》 2024年第6期67-71,共5页
本研究以引起猪流行性腹泻的猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)为研究目标,利用含有PEDV S基因的植物乳杆菌活载体菌株LP1522-PEDS口服免疫小鼠,应用ELISA试验检测免疫小鼠粪便中SIgA含量、血清中IgG、IL-4、IL-2... 本研究以引起猪流行性腹泻的猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)为研究目标,利用含有PEDV S基因的植物乳杆菌活载体菌株LP1522-PEDS口服免疫小鼠,应用ELISA试验检测免疫小鼠粪便中SIgA含量、血清中IgG、IL-4、IL-2和IFN-γ含量,评价乳酸菌重组菌株LP1522-PEDS对小鼠的免疫效果。结果表明,首次免疫后,商品灭活疫苗组和工程菌株免疫组粪便中的SIgA、血清中的IgG、IL-4、IL-2和IFN-γ含量较对照组显著升高(P<0.05)。在免疫42~56 d,灭活疫苗组和重组菌株组抗体水平均达到最大值,抗体和免疫因子水平依次为:商品灭活疫苗组>LP1522-PEDS组>空载组≈对照组。随后商品灭活疫苗组、LP1522-PEDS组抗体和免疫因子水平逐渐下降。免疫70 d,商品灭活疫苗组抗体水平降幅度相对较小,工程菌株免疫组降幅较大。结果表明,小鼠口服携带PEDV S基因的基因工程植物乳杆菌LP1522-PEDS后能够诱导机体产生PEDV免疫应答。 展开更多
关键词 pedv 乳酸菌 活载体疫苗 小鼠免疫
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猪lncRNA LOC102157897表达、定位及其对PEDV复制的调控作用
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作者 黄东璋 包文斌 吴正常 《扬州大学学报(农业与生命科学版)》 CAS 北大核心 2024年第5期82-89,共8页
猪流行性腹泻(porcine epidemic diarrhea,PED)是由猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)引起的高度传染性疾病,前期基于组学测序筛选出1个关键lncRNA(LOC102157897),有关其表达定位以及对PEDV复制的调控作用目前尚... 猪流行性腹泻(porcine epidemic diarrhea,PED)是由猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)引起的高度传染性疾病,前期基于组学测序筛选出1个关键lncRNA(LOC102157897),有关其表达定位以及对PEDV复制的调控作用目前尚不清楚。首先在组织和细胞水平上测定PEDV感染前后LOC102157897的表达水平;其次利用lncLocator软件预测和核质分离试验分析其细胞定位情况,并验证其表达水平与PEDV复制的关系;最后利用转录组测序筛选其下游靶基因及信号通路。结果表明:PEDV感染可极显著提高猪空肠组织和IPEC-J2细胞中LOC102157897的表达水平;LOC102157897主要定位于细胞核中,这可能与其生物学功能有关;成功构建了LOC102157897干扰细胞系,且干扰后IPEC-J2细胞中PEDV复制水平呈极显著下降。表达水平下调有利于提高宿主细胞对PEDV感染的抵抗能力。转录组测序分析显示,LOC102157897干扰前后存在151个差异表达基因,主要参与病毒蛋白互作、趋化因子信号、白细胞受体互作、白细胞介素17信号等免疫通路,并根据功能注释和差异程度,筛选出2个组蛋白基因H2AC18和H3C14。综上,这一研究确定了1个PEDV抗性相关lncRNA分子LOC102157897,并初步探究了其功能和下游调控机制,为揭示lncRNA在PEDV感染宿主过程中的重要作用以及为今后制定PEDV的抗病育种工作策略和筛选分子标记物奠定基础。 展开更多
关键词 流行性腹泻病毒 长链非编码RNA 表达调控
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碱性清洁剂安全性及对PEDV杀灭效果的评估
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作者 欧阳康 黄夏玲 +8 位作者 黄家宝 秦秋英 曾令佑 冯晋平 牛宇峰 覃一峰 陈樱 韦祖樟 黄伟坚 《广西农学报》 2024年第3期34-40,共7页
近年来,猪病毒性腹泻在我国流行日趋严重,引起仔猪大量腹泻和死亡,给养猪业带来巨大的经济损失。研究评估了碱性清洁剂对猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)的杀灭效果,为猪场选用合适的清洁消毒剂提供参考。通过p... 近年来,猪病毒性腹泻在我国流行日趋严重,引起仔猪大量腹泻和死亡,给养猪业带来巨大的经济损失。研究评估了碱性清洁剂对猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)的杀灭效果,为猪场选用合适的清洁消毒剂提供参考。通过pH值测定、细胞增殖和细胞毒性实验以及细胞病变观察测定不同浓度碱性清洁剂A、B和NaOH对Vero细胞增殖活性的影响以及在有机物干扰条件下碱性清洁剂对PEDV的杀灭作用。NaOH对Vero细胞增殖活性无影响的最高稀释浓度为0.03125%,而碱性清洁剂A和B分别为2%和0.5%,在安全性上优于NaOH;在含有不同浓度血清的环境下,碱性清洁剂A、B浓度达到0.5%时均能杀灭病毒。研究证明,碱性清洁剂在体外对PEDV有一定的杀灭效果,可用于猪场的清洗消杀,帮助养殖场做好疫病防控。 展开更多
关键词 碱性清洁剂 猪流行性腹泻病毒 PH值 病毒杀灭 猪场应用
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单月桂酸甘油酯对PEDV感染3D4/21巨噬细胞基因表达的影响 被引量:1
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作者 徐欢欢 王倩 +5 位作者 王宇杰 刘佳乐 王蕾 赵迪 张倩 侯永清 《饲料工业》 CAS 北大核心 2024年第15期120-127,共8页
试验旨在研究单月桂酸甘油酯(ML)对猪流行性腹泻病毒(PEDV)感染3D4/21巨噬细胞基因表达的影响。通过ML对3D4/21细胞活力试验和PEDV在3D4/21细胞中的生长曲线确定最适浓度和最佳时间后,将3D4/21细胞随机分为3组(control组、PEDV组、PEDV... 试验旨在研究单月桂酸甘油酯(ML)对猪流行性腹泻病毒(PEDV)感染3D4/21巨噬细胞基因表达的影响。通过ML对3D4/21细胞活力试验和PEDV在3D4/21细胞中的生长曲线确定最适浓度和最佳时间后,将3D4/21细胞随机分为3组(control组、PEDV组、PEDV+ML组),各组分别以全时添加10µmol/L ML和PEDV感染细胞后添加10µmol/L ML两种方式进行处理,并检测相关基因相对表达量。结果表明:添加10µmol/L的ML对3D4/21细胞生长具有显著的促进作用(P<0.05),病毒感染3D4/21细胞的48 h内PEDV-S、M、N基因相对表达量呈现先增加再降低的趋势,12 h时处于最高;与PEDV组相比,PEDV+ML组全时添加ML处理12 h使PEDV-S、M、N、IL-8、IFN-β、IFITM3基因相对表达量显著下调(P<0.05),处理24 h使PEDV-S、M、N、IL-8、IFN-β、MX1、ISG15、IFIT1和IFITM1基因相对表达量显著下调(P<0.05),KCNJ13基因相对表达量显著上调(P<0.05)。与PEDV组相比,PEDV+ML组在感染后添加ML处理12 h使MMP13基因相对表达量显著下调(P<0.05),PEDV-M基因相对表达量显著上调(P<0.05),处理24 h使IL-6、IL-8、IFN-β、MX1、ISG15、IFIT1、IFITM1、IFITM3基因相对表达量显著下调(P<0.05),PEDV-S、M、N、KCNJ13基因相对表达量显著上调(P<0.05)。综上所述,PEDV感染前ML预处理具有一定的抗病毒效果;PEDV感染使细胞处于免疫应激状态,ML有一定的缓解作用。 展开更多
关键词 单月桂酸甘油酯(ML) 猪流行性腹泻病毒(pedv) 3D4/21细胞 生长曲线 免疫功能
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葛根素与茯苓多糖复合物对PEDV感染幼龄仔猪结肠的保护作用 被引量:1
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作者 谌梦 胡锦超 +5 位作者 田晓榕 赵迪 王蕾 吴涛 张焱焱 侯永清 《饲料工业》 CAS 北大核心 2024年第16期47-54,共8页
试验旨在探究由葛根素(Puerarin,PR)和茯苓多糖(Poria cocos polysaccharide,PCP)组成的复合物(PR+PCP,PP)对猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)感染仔猪结肠的保护作用。采用单因素试验设计,通过使用PEDV感染仔... 试验旨在探究由葛根素(Puerarin,PR)和茯苓多糖(Poria cocos polysaccharide,PCP)组成的复合物(PR+PCP,PP)对猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)感染仔猪结肠的保护作用。采用单因素试验设计,通过使用PEDV感染仔猪肠道损伤模型进行试验,选用30头体重相近的7日龄健康仔猪,随机分成三个处理组:对照组、PEDV组和PP+PEDV组,每组10个重复,每个重复1头猪。试验期为14 d,其中预试期为4 d,正试期为10 d,试验期间各组饲喂相同的日粮,正试期内每晚给PP+PEDV组仔猪口腔灌服PP,将PR(0.5 mg/kg BW),PCP(20 mg/kg BW)混合溶于人工奶中;其余两组灌服相同体积人工奶。第10天向PEDV组和PP+PEDV组的仔猪口腔灌服PEDV(毒价为105.5 TCID50,用PBS溶液稀释),对照组仔猪灌服相同体积的PBS溶液。结果表明:①与对照组相比,PEDV组仔猪血清中总胆固醇(TC)、高密度脂蛋白(HDL)和低密度脂蛋白(LDL)含量显著降低(P<0.05);三酯甘油(TG)、总胆红素(TB)、直接胆红素(DB)和尿素氮(BUN)的含量显著升高(P<0.05);结肠隐窝深度(Crypt depth,CD)显著升高(P<0.05);结肠中总超氧化物歧化酶(T-SOD),髓过氧化物酶(MPO)活性显著升高(P<0.05);结肠中干扰素刺激基因15(ISG15)相对表达量显著升高(P<0.05);结肠白细胞介素(IL)-1β、白细胞介素-6(IL-6)、C-X-C基序趋化因子配体2(CXCL2)和再生胰岛衍生蛋白3γ(REG3G)的相对表达量显著升高(P<0.05)。②与PEDV组相比,PP+PEDV组仔猪血清中TG、谷草转氨酶(AST)和BUN的水平显著降低(P<0.05);结肠CD显著降低(P<0.05);结肠中ISG15基因相对表达量显著降低(P<0.05);结肠中IL-1β、IL-6、CXCL2和REG3G基因相对表达量显著降低(P<0.05)。综上所述,在试验条件下灌服PP能够缓解PEDV感染仔猪血清生化指标异常情况,并通过改善PEDV感染仔猪结肠的隐窝深度,缓解结肠炎性反应及影响机体免疫反应来缓解PEDV感染造成的仔猪结肠肠道损伤。 展开更多
关键词 仔猪 结肠 葛根素 茯苓多糖 猪流行性腹泻病毒 免疫反应
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PEDV、PoRVA和PDCoV TaqMan三重RT-qPCR检测方法的建立与初步应用 被引量:1
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作者 胡泽奇 李润成 +5 位作者 谭祖明 谢秀艳 王江平 秦乐娟 李荣 葛猛 《畜牧兽医学报》 CAS CSCD 北大核心 2024年第5期2267-2272,共6页
旨在建立一种可快速同时检测猪流行性腹泻病毒(porcine epidemic diarrhea virus, PEDV)、猪A群轮状病毒(porcine rotavirus type A, PoRVA)和猪丁型冠状病毒(porcine deltacoronavirus, PDCoV)三重荧光定量PCR方法。针对PEDV-N、PoRVA-... 旨在建立一种可快速同时检测猪流行性腹泻病毒(porcine epidemic diarrhea virus, PEDV)、猪A群轮状病毒(porcine rotavirus type A, PoRVA)和猪丁型冠状病毒(porcine deltacoronavirus, PDCoV)三重荧光定量PCR方法。针对PEDV-N、PoRVA-VP6和PDCoV-M基因设计了引物和探针,条件优化后进行性能评估,并与商品化试剂盒检测对比。结果显示:本研究建立的三重RT-qPCR方法具有良好特异性,对PRRSV、PCV_2和PRV等阳性核酸不发生扩增;具有较高的敏感性,PEDV、PoRVA和PDCoV的最低检测限均达1 copies·μL^(-1);重复性良好,组内和组间变异系数均小于1%;样本适应性广,检测不同样本类型时,变异系数均小于1%。与商品化试剂盒对比,本研究建立的方法PEDV和PoRVA的检测符合率为92.5%和97.5%,并对PDCoV的检测范围更广,对其变异毒株仍有较好的检测效果。本研究建立的检测方法具有特异性好、敏感性高、稳定性强和样本适应性广等优势,为临床腹泻样本提供了一种好的检测方法。 展开更多
关键词 猪流行性腹泻病毒 猪A群轮状病毒 猪丁型冠状病毒 三重RT-qPCR
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基于网络药理学和分子对接探究白杨素和柚皮素抗PEDV的作用机制及试验验证 被引量:1
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作者 植玉鹏 刘雨桐 +3 位作者 陈弟诗 宫萌菲 夏学妹 任玉鹏 《中国畜牧兽医》 CAS CSCD 北大核心 2024年第4期1757-1772,共16页
【目的】探讨白杨素和柚皮素抗猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)的作用靶点和机制,为进一步以白杨素和柚皮素开发新型抗PEDV的治疗药物提供理论依据。【方法】利用PharmMapper、TCMSP、SEA Search Server和STITC... 【目的】探讨白杨素和柚皮素抗猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)的作用靶点和机制,为进一步以白杨素和柚皮素开发新型抗PEDV的治疗药物提供理论依据。【方法】利用PharmMapper、TCMSP、SEA Search Server和STITCH在线数据库获得白杨素和柚皮素的潜在作用靶点,同时检索GeneCards数据库获得PEDV对宿主的作用靶点,利用在线程序Draw Venny Diagram获取白杨素、柚皮素与疾病的交集靶点。通过STRING数据库和Cytoscape 3.9.1软件构建靶蛋白互作(PPI)网络并筛选关键靶点,对靶点进行GO功能和KEGG通路富集分析。通过AutoDock Vina v 1.2.0软件对核心靶点及小分子进行分子对接,并分析白杨素和柚皮素与靶蛋白的结合能和结合模式,利用PyMOL v 2.5实现对接结果的可视化。采用实时荧光定量PCR和Western blotting检测白杨素和柚皮素对核心靶蛋白表达的影响。【结果】白杨素潜在抗PEDV的靶点有12个,其中白蛋白(ALB)、雌激素受体1(ESR1)、转化生长因子β-1蛋白(TGF-β1)可能是白杨素抗PEDV的核心靶点;柚皮素潜在抗PEDV的靶点有18个,其中ALB、胱天蛋白酶3(Caspase-3,CASP3)、过氧化物酶体增殖物激活受体γ(PPARG)可能是柚皮素抗PEDV的核心靶点。白杨素抗PEDV靶点涉及21种生物过程、5种细胞组分和6种分子功能,共获得11条信号通路;柚皮素抗PEDV靶点涉及31种生物过程、8种细胞组分和19种分子功能,共获得13条信号通路。核心靶点与两种天然化合物之间以氢键和疏水作用力为主,有较强的相互作用。白杨素和柚皮素能极显著降低Caspase-3表达量(P<0.01),可能通过影响Caspase-3的表达和活化来颉颃PEDV诱导的细胞凋亡;极显著升高TGF-β1蛋白表达量(P<0.01),可能通过影响TGF-β1的表达抗PEDV诱导的炎症反应而治疗PEDV的感染。【结论】本研究揭示了白杨素和柚皮素分别通过潜在核心靶点ALB、ESR1、TGF-β1和ALB、Caspase-3、PPARG,以及IL17、PI3K-Akt和AMPK信号通路发挥抗PEDV作用的机制,为新型抗PEDV药物的研发提供新的思路。 展开更多
关键词 猪流行性腹泻病毒(pedv) 网络药理学 分子对接 白杨素 柚皮素
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