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1.
Saudi Med J ; 38(4): 391-395, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28397945

RESUMO

OBJECTIVES: To investigate the diagnostic accuracy of ultrasound for evaluation of inflammatory activity in patients with Crohn's disease (CD). Methods: Fifty-six patients with histologically proven CD (39 with active, 17 with inactive disease) and 30 healthy volunteers as a control group were enrolled in the study at WeiFang People's Hospital, Weifang Province, China from October 2012 to December 2014. Bowel wall thickness, and vascularity pattern were measured by Doppler ultrasound. Results: There was a significant difference in flow volume of the superior mesenteric artery (585 ± 235 ml/min) in the patients with active disease, compared with those with inactive disease (401 ± 238 ml/min) and the control group (390 ± 189 ml/min, p less than 0.001). Wall thickness was 5.1 ± 1.5 mm in the active CD group, 3.3 ± 1.6 mm in the inactive disease group (p less than 0.001) and  less than 3 mm in the control group. Resistance index in the thickened bowel wall showed some differences: 0.68 ± 0.05 in the active disease group, 0.78 ± 0.08 in the inactive disease group, and 0.85 ± 0.07 in the control group (p less than 0.05). Conclusion: Doppler ultrasound is a useful diagnostic tool in detecting CD and assessing inflammatory activity.


Assuntos
Doença de Crohn/diagnóstico por imagem , Ultrassonografia Doppler , Adulto , China , Estudos de Coortes , Doença de Crohn/patologia , Feminino , Trato Gastrointestinal/irrigação sanguínea , Trato Gastrointestinal/diagnóstico por imagem , Humanos , Masculino , Artéria Mesentérica Superior/diagnóstico por imagem , Pessoa de Meia-Idade , Estudos Prospectivos , Sensibilidade e Especificidade
2.
J Biol Chem ; 291(28): 14815-25, 2016 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-27226547

RESUMO

Avian metapneumovirus (aMPV) fusion (F) protein mediates virus-cell membrane fusion to initiate viral infection, which requires F protein binding to its receptor(s) on the host cell surface. However, the receptor(s) for aMPV F protein is still not identified. All known subtype B aMPV (aMPV/B) F proteins contain a conserved Arg-Asp-Asp (RDD) motif, suggesting that the aMPV/B F protein may mediate membrane fusion via the binding of RDD to integrin. When blocked with integrin-specific peptides, aMPV/B F protein fusogenicity and viral replication were significantly reduced. Specifically we identified integrin αv and/or ß1-mediated F protein fusogenicity and viral replication using antibody blocking, small interfering RNAs (siRNAs) knockdown, and overexpression. Additionally, overexpression of integrin αv and ß1 in aMPV/B non-permissive cells conferred aMPV/B F protein binding and aMPV/B infection. When RDD was altered to RAE (Arg-Ala-Glu), aMPV/B F protein binding and fusogenic activity were profoundly impaired. These results suggest that integrin αvß1 is a functional receptor for aMPV/B F protein-mediated membrane fusion and virus infection, which will provide new insights on the fusogenic mechanism and pathogenesis of aMPV.


Assuntos
Fusão Celular , Metapneumovirus/fisiologia , Infecções por Paramyxoviridae/fisiopatologia , Receptores de Vitronectina/fisiologia , Proteínas Virais de Fusão/fisiologia , Animais , Linhagem Celular , Infecções por Paramyxoviridae/virologia , Replicação Viral
3.
Virus Genes ; 52(1): 51-60, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26611441

RESUMO

During the course of our continuous surveillance of Gallid herpesvirus 2 (GaHV-2), 44 isolates were obtained from GaHV-2-positive chickens of different flocks in China from 2009 to 2013. The meq gene, considered as a major GaHV-2 oncogene, was sequenced and was found to contain an open reading frame of 1020 nucleotides encoding a 339 amino acid (aa) polypeptide in all isolates. Compared with the GaHV-2 GA strain, the meq genes in 15.9 % (7/44) of the isolates analyzed in this study contained an aa substitution mutation at position 88 (A to T) of which is the first report. The main characteristics of Chinese GaHV-2 isolates meq genes included the substitutions K77E, D80Y, V115A, T139A, P176R, and P217A, and the aa substitution frequency at positions 139 and 176 showed an increase. To test the pathogenicity of the isolates, a pathogenicity study and a vaccination-challenge test were performed on three selected isolates (ZY/1203, WC/1203, and WC/1110) and reference strain GA. The results showed that the three isolates induced gross Marek's disease (MD) lesions in 95.0-100 % cases, which was a higher rate than that obtained for strain GA (82.4 %). Three isolates induced mortality in 10-21.1 % of specific-pathogen-free chickens, which was similar to results with strain GA (23.5 %). The commercially available CVI988 vaccine induced lower protective indices (PIs) against ZY/1203 (82.4) and WC/1110 (83.3) as compared to those against WC/1203 (100) and GA (100). These results showed an evolving trend in the meq genes of the isolates; three isolates exhibited higher morbidity as compared to the reference strain and the vaccine induced lower PIs against two isolates as compared to that against the reference strain.


Assuntos
Galinhas/virologia , Herpesvirus Galináceo 2/patogenicidade , Animais , China/epidemiologia , Herpesvirus Galináceo 2/classificação , Herpesvirus Galináceo 2/genética , Doença de Marek/epidemiologia , Doença de Marek/virologia , Proteínas Oncogênicas Virais/genética , Filogenia , Virulência
4.
Virus Genes ; 50(3): 418-24, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25850423

RESUMO

Reticuloendotheliosis virus (REV), classified as a gammaretrovirus, has a variety of hosts, including chickens, ducks, geese, turkeys, and wild birds. REV causes a series of pathological syndromes, especially the immunosuppression of the host, which may lead to an increased susceptibility to other pathogens, thus greatly damaging the poultry industry. Mixed infections of REV and Marek's disease virus (MDV) have been reported in many countries, including China. Previous reports revealed that MDV vaccines were not efficacious, and even less-virulent MDV strains would cause some losses due to mixed infections with REV. Additionally, contaminants in the MDV vaccine might be the main source of REV. In this study, two clinical samples were collected from two flocks of chickens that were diagnosed with MDV. Subsequently, two REV isolates were obtained from the clinical samples. The isolates, named CY1111 and SY1209, were further confirmed through an indirect immunofluorescence assay and electron microscopy. Complete genome sequences of the two REV strains were determined to test the relationship between them and other REV strains. Phylogenetic trees showed that the two REV strains were closely related to most REV strains that were isolated from a variety of hosts. Therefore, REVs might spread freely among these hosts under natural conditions. Additionally, most REV strains in China were in the same clade. The present work offers some information regarding REV in China.


Assuntos
Coinfecção/veterinária , Coinfecção/virologia , Genoma Viral , Doenças das Aves Domésticas/virologia , Vírus da Reticuloendoteliose/genética , Vírus da Reticuloendoteliose/isolamento & purificação , Infecções por Retroviridae/veterinária , Animais , Galinhas , China , Análise por Conglomerados , Herpesvirus Galináceo 2/isolamento & purificação , Doença de Marek/complicações , Microscopia , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Infecções por Retroviridae/complicações , Infecções por Retroviridae/virologia , Análise de Sequência de DNA , Homologia de Sequência
5.
Vet Microbiol ; 177(1-2): 62-8, 2015 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-25770895

RESUMO

The virulence of Marek's disease virus (MDV) is continuously evolving, and more virulent MDV pathotypes are emerging, thereby reducing the effectiveness of the existing vaccines. In this study, feather pulps were collected from diseased chickens in commercial chicken flocks in China that presented significant MD visceral tumors in 2011 and were inoculated into a monolayer of duck embryo fibroblasts (DEFs). Three field isolates of MDV were obtained by plaque cloning and identified as MDV via PCR and designated strains LCC, LLY, and LTS. Unvaccinated and CVI988 vaccine-vaccinated specific pathogen-free chickens were challenged at 7 days post vaccination (dpv) with 1000 plaque forming units of each of the respective MDV isolates. These strains induced gross MD lesions in all (100%) of the unvaccinated chickens, and the mortality rates of the unvaccinated chickens were 42.9%, 46.7%, and 23.1% by 60 days post challenge (dpc), respectively. The CVI988 vaccine induced protective indices (PIs) of 85.7, 92.3, and 66.7, respectively. These results showed that the pathogenic characteristics of the Chinese isolates were diverse and that vaccine CVI988 provided different levels of protection against them. These data indicated that the existence of variant MDV strains was a possible reason of immunity failure in China.


Assuntos
Herpesvirus Galináceo 2/genética , Vacinas contra Doença de Marek/imunologia , Doença de Marek/virologia , Animais , Galinhas/imunologia , China/epidemiologia , Plumas/virologia , Doença de Marek/epidemiologia , Doença de Marek/prevenção & controle , Reação em Cadeia da Polimerase , Prevalência , Organismos Livres de Patógenos Específicos , Virulência/efeitos dos fármacos
6.
Virus Res ; 178(2): 530-4, 2013 Dec 26.
Artigo em Inglês | MEDLINE | ID: mdl-24076298

RESUMO

SigmaC (σC) protein, which mediates virus attachment to target cells, is the most variable proteins of avian reovirus (ARV). It is responsible for inducing protective antibody immune responses in animals. To understand the antigenic determinants of σC protein, a set of partially overlapping and consecutive peptides spanning σC were expressed and then screened with the monoclonal antibody (mAb) 2B5 directed against σC. The mAb 2B5 recognized peptides with the σC motif (45)ELLHRSISDISTTV(58). Further identification of the displayed B-cell epitope was conducted with a set of truncated peptides expressed as GST fusion proteins. The Western blot and ELISA results indicated that (45)ELLHRSISDI(54) was the minimal determinant of the linear B-cell epitope. Using sequences analysis, we found that this epitope was not a common motif shared among the other members of the ARV and DRV groups. Furthermore, cross reactivity analysis showed that the associated coding motif of other ARV and DRV groups was not recognized by 2B5. These data suggested that (45)ELLHRSISDI(54) was a type-specific linear B-cell epitope of avian reovirus. The results in this study may have potential applications in the development of diagnostic techniques and epitope-based marker vaccines against ARV, which is prevalent in China.


Assuntos
Antígenos Virais/imunologia , Epitopos de Linfócito B/imunologia , Orthoreovirus Aviário/imunologia , Proteínas Estruturais Virais/imunologia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/imunologia , Western Blotting , China , Ensaio de Imunoadsorção Enzimática , Mapeamento de Epitopos , Feminino , Camundongos , Camundongos Endogâmicos BALB C
8.
Sheng Wu Gong Cheng Xue Bao ; 23(4): 719-23, 2007 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-17822051

RESUMO

Infectious bursal disease virus (IBDV), the causative agent of a highly contagious disease in chickens, carries a small nonstructural protein (NS). In this study, vvIBDV Gx-VP5 genes were cloned into plasmid pET30a( + ) and expressed in E. coli with IPTG inducing. BALB/c mice were immunized with the purified recombinant fusion protein. SP2/0 myeloma cells and spleen cells of BALB/c mice were fused by PEG(MW1500), three hybridoma cell lines were examined by indirect ELISA and clone for three times by limited dilution, and were named as 4B4, 6D12, 3E8. The subtype of the monoclonal antibodies were IgG1 with a subtype identified ELISA kit, and light chains were kappa. The ascites titers of monoclonal antibodies were 5 x 10(4), 3.5 x 10(4), 3 x 10(4) by indirect ELISA, respectively. Indirect ELISA and Western blot results showed that the monoclonal antibodies only acted with VP5 protein, IF analysis indicated that three monoclonal antibodies acted with IBDV Gt. There were specific fluorescence in detected Vero E6 cells which transient expressed VP5 protein by IFA. Therefore, monoclonal antibodies specific to IBDV VP5 proteins are specific method for detected VP5 proteins, and base on establish stabilize expressed VP5 protein Vero cell lines to research IBDV VP5 protein function.


Assuntos
Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/biossíntese , Vírus da Doença Infecciosa da Bursa/imunologia , Proteínas não Estruturais Virais/imunologia , Animais , Anticorpos Antivirais/imunologia , Galinhas , Escherichia coli/genética , Escherichia coli/metabolismo , Feminino , Hibridomas/metabolismo , Imunização , Camundongos , Camundongos Endogâmicos BALB C
9.
Avian Dis ; 51(4): 893-9, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18251399

RESUMO

The complete genomic sequence of very virulent infectious bursal disease virus (vvIBDV) Gx strain was determined, including the sequences of segment A, encoding the precursor polyprotein, and segment B, encoding the viral RNA polymerase (VP1) and 5'- and 3'-untranslating regions. Alignment of segment A of Gx with the sequences of 12 other vvIBDV strains showed 97.5% to 99.0% amino acid identity, whereas alignment of segment B of Gx with nine other vvIBDV strains revealed high sequence divergence, ranging from 10.3% to 11%. Phylogenetic analysis of segments A and B showed that they were in different branches, indicating that the reassortment occurred in this strain and that segment A and segment B derived from different pathotype strains. The mutant spectrum analysis of quasispecies virus demonstrated that the mean minimum mutation frequency in VP1 was 8.78-fold higher than in the polyprotein. The most frequent mutations were in the first 1986 nucleotides (nonsynonymous mutations) and the last 660 nucleotides (synonymous mutations), indicating that the 219 amino acid residues in the C-terminal of the VP1 form a functional region.


Assuntos
Vírus da Doença Infecciosa da Bursa/patogenicidade , Vírus Reordenados/genética , Animais , Sequência de Bases , Genoma Viral , Mutação , Filogenia , Virulência
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