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OBJECTIVES: To investigate the cumulative incidence of recurrence (CIR) in children with acute lymphoblastic leukemia (ALL) after treatment with the Chinese Children's Cancer Group ALL-2015 (CCCG-ALL-2015) protocol and the risk factors for recurrence. METHODS: A retrospective analysis was conducted on the clinical data of 852 children who were treated with the CCCG-ALL-2015 protocol from January 2015 to December 2019. CIR was calculated, and the risk factors for the recurrence of B-lineage acute lymphoblastic leukemia (B-ALL) were analyzed. RESULTS: Among the 852 children with ALL, 146 (17.1%) experienced recurrence, with an 8-year CIR of 19.8%±1.6%. There was no significant difference in 8-year CIR between the B-ALL group and the acute T lymphocyte leukemia group (P>0.05). For the 146 children with recurrence, recurrence was mainly observed in the very early stage (n=62, 42.5%) and the early stage (n=46, 31.5%), and there were 42 children with bone marrow recurrence alone (28.8%) in the very early stage and 27 children with bone marrow recurrence alone (18.5%) in the early stage. The Cox proportional-hazards regression model analysis showed that positive MLLr fusion gene (HR=4.177, 95%CI: 2.086-8.364, P<0.001) and minimal residual disease≥0.01% on day 46 (HR=2.013, 95%CI: 1.163-3.483, P=0.012) were independent risk factors for recurrence in children with B-ALL after treatment with the CCCG-ALL-2015 protocol. CONCLUSIONS: There is still a relatively high recurrence rate in children with ALL after treatment with the CCCG-ALL-2015 protocol, mainly bone marrow recurrence alone in the very early stage and the early stage, and minimal residual disease≥0.01% on day 46 and positive MLLr fusion gene are closely associated with the recurrence of B-ALL.
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Leucemia-Linfoma Linfoblástico de Células Precursoras , Humanos , Criança , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Feminino , Fatores de Risco , Pré-Escolar , Estudos Retrospectivos , Lactente , Recidiva , Adolescente , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , População do Leste AsiáticoRESUMO
OBJECTIVE: To analyze the efficacy and influencing factors of cyclosporine (CsA) alone in the treatment of children with acquired aplastic anemia (AA). METHODS: The clinical data of children diagnosed with AA and treated with CsA alone from January 1, 2016 to December 31, 2020 in the Children's Hospital of Chongqing Medical University were collected, and the efficacy and influencing factors of CsA treatment were evaluated. RESULTS: Among the 119 patients, there were 62 male and 57 female, with a median age of 7 years and 1 month. There were 45 cases of very severe AA (VSAA), 47 cases of severe AA (SAA), and 27 cases of non-severe AA (NSAA). At 6 months after treatment, the efficacy of VSAA was lower than that of SAA and NSAA, and there was a statistical difference (P < 0.01). 6 cases died early, 16 cases relapsed, 2 cases progressed to AML and ALL. The results of univariate analysis showed that the high proportion of lymphocyte in the bone marrow at 6 months was an adverse factor for the efficacy of CsA, while high PLT count was a protective factor (P =0.008, P =0.002). The ROC curve showed that the cut-off values of PLT count and the proportion of bone marrow lymphocyte at 6 months were 16.5×109 /L, 68.5%, respectively. Multivariate analysis showed that the high proportion of lymphocyte in bone marrow at 6 months was an independent adverse factor for IST (P =0.020, OR =0.062), and high PLT count was a protective factor (P =0.044, OR =1.038). At 3 months of treatment, CsA response and NSAA were the risk factor for recurrence (P =0.001, 0.031). CONCLUSION: The efficacy of NSAA was higher than that of SAA and VSAA after 6 months of treatment with CsA alone. A high PLT count at the initial diagnosis was a good factor for the effectiveness of CsA, and a high proportion of bone marrow lymphocyte was an unfavorable factor. CsA response at 3 months and NSAA were risk factors for recurrence.
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Anemia Aplástica , Ciclosporina , Humanos , Anemia Aplástica/tratamento farmacológico , Ciclosporina/uso terapêutico , Feminino , Masculino , Criança , Resultado do Tratamento , Contagem de Plaquetas , Imunossupressores/uso terapêutico , Pré-Escolar , Adolescente , Medula ÓsseaRESUMO
Background: Severe fever with thrombocytopenia syndrome virus (SFTSV) is a tick-borne bunyavirus that could cause a severe hemorrhagic fever termed SFTS with a high fatality rate of up to 30%. Importantly, SFTSV is frequently transmitted from person-to-person and patients' blood or excreta are considered as the risk factors for transmission of SFTSV. However, the mechanism of person-to-person transmission of SFTSV is still elusive. Methods: In this study, wild-type (WT) C57BL/6 J mice and a lethal SFTSV mouse model IFNAR-/- A129 mice were utilized to evaluate whether SFTSV could be transmitted via oral or ocular routes. C57BL/6 J mice were inoculated with cell-cultured SFTSV via oral and ocular inoculation. IFNAR-/- A129 mice were inoculated with cell-cultured SFTSV or SFTSV infected mouse acute sera via oral and ocular inoculation. Results: We found that SFTSV antibody positive rates in C57BL/6 J mice were 70% (7/10) and 30% (3/10) in the oral inoculation group and ocular inoculation group, respectively on day 21 post SFTSV inoculation. The mortality rates of IFNAR-/- mice with oral and ocular inoculation of cell-cultured SFTSV were 100% and 83.33% (5/6), respectively on day 6 post inoculation. The mortality rates of IFNAR-/- mice with oral and ocular inoculation of SFTSV infected mouse acute serum were 100% and 66.67% (4/6), respectively on day 9 post inoculation. Conclusions: Together, our results show that SFTSV can be transmitted effectively through oral and ocular membrane, suggesting exposure to SFTS positive excreta may be a high-risk factor of nosocomial transmission of SFTSV in hospitals and/or families. Family members and healthcare workers should be protected properly during taking care of SFTS patients to prevent SFTSV nosocomial infection.
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The pathogenicity of the shrew-borne Imjin virus (MJNV) is unknown. The objective of our study was to find serological evidence of MJNV infection in humans. Partial MJNV nucleocapsid protein (NP) was cloned and expressed as an antigen for double-antigen sandwich ELISA, IgM capture ELISA, and dot blot to detect MJNV specific antibodies in hemorrhagic fever with renal syndrome (HFRS) patients' and healthy persons' sera from endemic areas in China. The purified recombinant NP reacted with neither the 90 healthy individuals' sera from non-endemic areas of MJNV nor the 100 antisera to HFRS-causing virus, indicating that the MJNV NP had no cross-reaction with normal human sera and HFRS-causing viral antibodies. As determined by screening ELISA and dot blot analysis, IgG antibodies against MJNV NP were detected in sera from two of 385 healthy individuals from MJNV-endemic areas, suggesting infection with MJNV or MJNV-like thottimvirus. Based on the suggestive evidence, healthcare workers should be alert to febrile diseases occurring among individuals with exposure to shrew-infested habitats.
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Infecções por Hantavirus/diagnóstico , Infecções por Hantavirus/virologia , Orthohantavírus , Animais , Anticorpos Antivirais/imunologia , Especificidade de Anticorpos/imunologia , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática , Orthohantavírus/classificação , Orthohantavírus/fisiologia , Infecções por Hantavirus/epidemiologia , Infecções por Hantavirus/imunologia , Febre Hemorrágica com Síndrome Renal/diagnóstico , Febre Hemorrágica com Síndrome Renal/imunologia , Febre Hemorrágica com Síndrome Renal/virologia , Humanos , Camundongos , Proteínas do Nucleocapsídeo/imunologia , Prevalência , Testes SorológicosRESUMO
Autophagy is essential for eukaryotic cell homeostasis and can perform both anti-viral and pro-viral roles depending on the kinds of viruses, cell types and cell environment. Severe fever with thrombocytopenia syndrome phlebovirus (SFTSV) is a newly discovered tick-borne virus in the Phenuiviridae family that causes a severe hemorrhagic fever disease in East Asia. In this study we determined interactions between SFTSV and autophagy. Our results showed that LC3-II (microtubule associated protein 1 light chain 3-II) protein accumulated from 4 h to 24 h after SFTSV infection compared to mock-infected Vero cells, and the use of E64d and pepstatin A did not affect the expression of LC3-II protein, which indicated that the increased LC3-II may be the result of inhibition of autophagic degradation caused by SFTSV infection. However, knockdown of LC3B promotes SFTSV replication, which indicated a negative role of LC3B protein in SFTSV replication. We also detected co-localization of SFTSV non-structure (NSs) protein with LC3B, p62 and Lamp2b respectively in SFTSV infected Vero cells, which indicated the possibility of selective autophagy or chaperone-mediated autophagy involving in SFTSV infection. Our results indicated that SFTSV infection promotes LC3 accumulation and several proteins of the autophagy pathway co-localize with NSs protein during SFTSV infection.
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Proteínas Associadas aos Microtúbulos/metabolismo , Febre por Flebótomos/metabolismo , Phlebovirus/fisiologia , Proteínas não Estruturais Virais/metabolismo , Animais , Autofagia , Chlorocebus aethiops , Interações Hospedeiro-Patógeno , Humanos , Camundongos Endogâmicos BALB C , Proteínas Associadas aos Microtúbulos/análise , Febre por Flebótomos/patologia , Células Vero , Proteínas não Estruturais Virais/análise , Replicação ViralRESUMO
Severe fever with thrombocytopenia syndrome, an emerging hemorrhagic fever, is caused by severe fever with thrombocytopenia syndrome virus (SFTSV), a tick-borne bunyavirus. Information regarding SFTSV animal hosts is very limited. In this study, we showed that 64% (9/14) of hedgehogs in Shandong Province, China were seropositive to SFTSV antibody, suggesting that hedgehog could be a vertebrate parasitifer for SFTSV.
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Infecções por Bunyaviridae/veterinária , Ouriços/virologia , Phlebovirus , Animais , Anticorpos Antivirais/sangue , Infecções por Bunyaviridae/epidemiologia , Infecções por Bunyaviridae/virologia , China/epidemiologia , Reservatórios de Doenças/veterinária , Ouriços/sangue , Estudos SoroepidemiológicosRESUMO
BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) was an emerging hemorrhagic fever that was caused by a tick-borne bunyavirus, SFTSV. Although SFTSV nonstructural protein can inhibit type I interferon (IFN-I) production Ex Vivo and IFN-I played key role in resistance SFTSV infection in animal model, the role of IFN-I in patients is not investigated. METHODS: We have assayed the concentration of IFN-α, a subtype of IFN-I as well as other cytokines in the sera of SFTS patients and the healthy population with CBA (Cytometric bead array) assay. RESULTS: The results showed that IFN-α, tumor necrosis factor (TNF-α), granulocyte colony-stimulating factor (G-CSF), interferon-γ (IFN-γ), macrophage inflammatory protein (MIP-1α), interleukin-6 (IL-6), IL-10, interferon-inducible protein (IP-10), monocyte chemoattractant protein (MCP-1) were significantly higher in SFTS patients than in healthy persons (p < 0.05); the concentrations of IFN-α, IFN-γ, G-CSF, MIP-1α, IL-6, and IP-10 were significant higher in severe SFTS patients than in mild SFTS patients (p < 0.05). CONCLUSION: The concentration of IFN-α as well as other cytokines (IFN-γ, G-CSF, MIP-1α, IL-6, and IP-10) is correlated with the severity of SFTS, suggesting that type I interferon may not be significant in resistance SFTSV infection in humans and it may play an import role in cytokine storm.
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Infecções por Bunyaviridae/imunologia , Infecções por Bunyaviridae/patologia , Citocinas/sangue , Phlebovirus/isolamento & purificação , Índice de Gravidade de Doença , Idoso , Animais , Resistência à Doença , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Severe fever with thrombocytopenia syndrome (SFTS) is an emerging hemorrhagic fever in East Asia, which is caused by a novel bunyavirus-SFTSV. Many studies have reported the clinical characters of SFTS patients, but the reports were not consistent and a systematic summary of clinical manifestations and laboratory parameters are not available. METHOD: A comprehensive literature research of Web of Science, PubMed, Wan Fang Data, and Chinese National Knowledge Infrastructure databases was conducted on articles which have described the clinical characters of SFTS patients. Data from selected studies were pooled by using STATA VERSION 12.0 software. RESULT: Nine articles comprising 844 laboratory-confirmed SFTSV cases were included in this meta-analysis. The pooled case fatality rate was 16% (95% CI: 0.13-0.19). The major clinical characters of patients with SFTSV infection were fever, thrombocytopenia, leucopenia, gastrointestinal symptoms, and central nervous system manifestations. The risk factors for severe disease included bleeding tendency, central nervous system manifestations, elevated serum enzymes, and high viral load. Although there is no specific antiviral therapy for SFTSV infection, symptomatic treatment and supportive therapy including intensive monitoring is the most essential part of case management. CONCLUSION: The major clinical characters of patients with SFTSV infection were fever, thrombocytopenia, leucopenia and gastrointestinal symptoms, and central nervous system manifestations. The risk factors for severity and fatality among SFTS patients included: old age, CNS manifestations, bleeding tendency, elevated serum enzymes, and high vial load.
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Febre por Flebótomos/patologia , Phlebovirus/isolamento & purificação , Fatores Etários , Doenças do Sistema Nervoso Central/patologia , China , Hemorragia , Humanos , Febre por Flebótomos/mortalidade , Fatores de Risco , Análise de Sobrevida , Carga ViralRESUMO
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging hemorrhagic fever in East Asia caused by SFTS virus (SFTSV), a newly discovered phlebovirus. The Haemaphysalis longicornis tick has been suspected to be the vector of SFTSV. To determine whether SFTSV can be transmitted among ticks, from ticks to animals, and from animals to ticks, we conducted transmission studies between developmental stages of H. longicornis ticks and between ticks and mice. Using reverse transcription PCR, we also analyzed the prevalence of SFTSV infection among H. longicornis ticks collected from vegetation in Shandong Province, China. Our results showed a low prevalence of SFTSV among collected ticks (0.2%, 8/3,300 ticks), and we showed that ticks fed on SFTSV-infected mice could acquire the virus and transstadially and transovarially transmit it to other developmental stages of ticks. Furthermore, SFTSV-infected ticks could transmit the virus to mice during feeding. Our findings indicate ticks could serve as a vector and reservoir of SFTSV.
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Vetores Aracnídeos/virologia , Infecções por Bunyaviridae/transmissão , Transmissão de Doença Infecciosa/veterinária , Phlebovirus/genética , Phlebovirus/patogenicidade , Carrapatos/virologia , Viroses/transmissão , Animais , Infecções por Bunyaviridae/virologia , China/epidemiologia , Camundongos , Carrapatos/microbiologia , Viroses/virologiaRESUMO
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging hemorrhagic fever in East Asia with case fatality up to 50%. SFTS is caused by SFTSV, a tick borne bunyavirus. In endemic area in China 1%-3% population was infected with SFTSV, but age is critical risk factor for hospitalization and death of SFTS patients.
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Infecções por Bunyaviridae/virologia , Febres Hemorrágicas Virais/virologia , Phlebovirus/genética , Phlebovirus/patogenicidade , Trombocitopenia/virologia , Animais , Vetores Artrópodes , Infecções por Bunyaviridae/complicações , Infecções por Bunyaviridae/epidemiologia , China/epidemiologia , Modelos Animais de Doenças , Febres Hemorrágicas Virais/complicações , Febres Hemorrágicas Virais/epidemiologia , Humanos , Phlebovirus/fisiologia , Síndrome , Trombocitopenia/complicações , Trombocitopenia/epidemiologia , Replicação ViralRESUMO
An Ussuri catfish Pseudobagrus ussuriensis skin (UCS) cell line was developed and subcultured for more than 60 passages. UCS cells consisted of mostly epithelial-like cells and multiplied well in TC199 medium supplemented with 10% fetal bovine serum at 25°C. Chromosome analysis revealed that most UCS cells had a normal diploid karyotype with 2n=52. UCS cells showed differential cytopathic effects (CPEs) after inoculation of spring viremia of carp virus (SVCV, a negative-strand RNA virus), grass carp reovirus (GCRV, a multi-segmented double-stranded RNA virus) and Rana grylio virus (RGV, a large double-stranded DNA virus), and were indicative of high sensitivities to these three aquatic animal viruses by a virus titration study. The CPE caused by SVCV appeared as rounded and granular cells, grape-like clusters and small lytic plaques. Characteristic CPE containing plaque-like syncytia was induced by GCRV. RGV-infected cells produced typical CPE characterized by cells shrinkage and aggregation, formation of clear plaques and cell sheet detachment. Furthermore, significant fluorescent signals were observed after UCS cells were transfected with green fluorescent protein reporter plasmids, and the development of CPE induced by a recombinant RGV, ΔTK-RGV, in UCS cells was illustrated using a combination of light and fluorescence microscopy. The data from this study suggested that UCS cell line can potentially serve as a useful tool for the comparison study of different aquatic animal viruses and the isolation of some newly emerging viruses in Ussuri catfish farming.
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Peixes-Gato , Linhagem Celular , Efeito Citopatogênico Viral , Ranavirus/crescimento & desenvolvimento , Reoviridae/crescimento & desenvolvimento , Rhabdoviridae/crescimento & desenvolvimento , Pele , Animais , Meios de Cultura/química , Células Epiteliais/fisiologia , Células Epiteliais/virologia , Ranavirus/patogenicidade , Reoviridae/patogenicidade , Rhabdoviridae/patogenicidade , Temperatura , Ensaio de Placa ViralRESUMO
OBJECTIVE: To evaluate the diagnostic value of contrast-enhanced ultrasound(CEUS) in calcified thyroid nodules. METHODS: A total of 179 cases with calcified thyroid nodules between February 2009 and September 2012 underwent CEUS by injection of microbubbles via superficial vein was included in the study. Pathological diagnosis as a gold standard, the sensitivity, specificity, positive predictive, negative predictive and diagnostic coincidence rate of CEUS were evaluated. RESULTS: Among 110 cases of benign nodules, 62 cases were nodular goiter (46 cases with macrocalcification and 16 cases with microcalcification); 29 cases were adenoma (28 cases with macrocalcification and 1 case with microcalcification); 17 cases were Hashimoto's thyroid (13 cases with macrocalcification and 4 cases with microcalcification); and 2 cases were granuloma with microcalcification. Among 69 cases of malignant nodules (t = 14.33, P < 0.05), 68 cases were papillary carcinoma (60 cases with microcalcification and 8 cases with macrocalcification), and 1 case of medullary carcinoma with microcalcification. Malignant nodules mainly showed weak inhomogeneous enhancement, with the mean; peak intensity (51.38 ± 14.33)dB that was lower than that (92.37 ± 33.36)dB in benign nodules, and benign nodules showed isoenhancement or hyperenhancement, with significant differences compared to malignant nodules (t = 14.33, P < 0.05) , however, there were significant differences in the enhanced time and the time to peak between benign and malignant nodules. The sensitivity, specificity, positive predictive, negative predictive and diagnostic coincidence rate of CEUS for the diagnoses of thyroid nodules were 92.75%, 90.91%, 86.49%, 95.24% and 91.62%, respectively. CONCLUSION: CEUS has high value in differential diagnosis of benign and malignant calcified thyroid nodules.
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Meios de Contraste , Nódulo da Glândula Tireoide , Bócio Nodular , Humanos , Microbolhas , Nódulo da Glândula Tireoide/diagnóstico por imagem , UltrassonografiaRESUMO
OBJECTIVE: To evaluate the contrast-enhanced ultrasound(CEUS) and ultrasonic elastography (UE) in diagnosis of thyroid nodules by a binary Logistic regression model. METHODS: A total 149 cases with thyroid nodules were examed by CEUS and UE and were confirmed by surgical pathology. A Logistic model was obtained on the basis of ultrasonographic features. Receiver operator characteris(ROC) curve was constructed to assess the performance of the Logistic model. RESULTS: Four ultrasonographic features including calcification, enhancement degree, biggest perfusion strength and elastic score were finally entered into the Logistic model. The percentage of correct prediction was 91.90%. CONCLUSION: Both CEUS and UE have high value in differential diagnosis of thyroid nodules and the Logistic regression model has high diagnostic rate.
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Técnicas de Imagem por Elasticidade , Nódulo da Glândula Tireoide/diagnóstico por imagem , Calcinose , Meios de Contraste , Diagnóstico Diferencial , Humanos , Modelos Logísticos , Curva ROC , Nódulo da Glândula Tireoide/diagnóstico , UltrassomRESUMO
Interferon-inducible transmembrane (IFITM) protein family is novel viral restriction factors with representative transmembrane structure. These proteins also exist in fish, however, their roles in the innate immune response remain unknown. Here, we report a characterization of teleost IFITM1 from flounder Paralichthys olivaceus (PoIFITM1), which exhibits conserved structure characteristic of the IFITM family but comprises a relatively longer N-terminal region. The expression and promoter activity of PoIFITM1 are markedly induced by aquatic animal viruses: Rana grylio virus (RGV) and Scophthalmus maximus rhabdovirus (SMRV). Overexpression and siRNA-mediated knockdown demonstrate that PoIFITM1 exhibits strong antiviral effects against both DNA virus (RGV) and RNA virus (SMRV), expanding the spectrum of viruses inhibited by IFITM proteins. Further analysis shows that PoIFITM1 suppresses viral entry into host cells, confirming that the IFITM-mediated restriction is conserved from lower vertebrates to mammals. Deletion mutagenesis reveals that PoIFITM1 exerts antiviral activity by targeting to Golgi complex and the N-terminal region is required for its subcellular localization, which is not observed in other known IFITM family members. Our current data provide the first evidence that IFITM1 functions as a key effector of the innate immune to restrict virus replication in lower vertebrates, through the action of impeding viral entry.
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Antígenos de Diferenciação/metabolismo , Linguado/imunologia , Linguado/metabolismo , Iridoviridae/fisiologia , Rhabdoviridae/fisiologia , Internalização do Vírus , Sequência de Aminoácidos , Animais , Antígenos de Diferenciação/genética , Antivirais , Sequência de Bases , Células Cultivadas , Linguado/embriologia , Linguado/genética , Regulação da Expressão Gênica/imunologia , Imunidade Inata , Dados de Sequência Molecular , Filogenia , Regiões Promotoras Genéticas , Replicação Viral/fisiologiaRESUMO
BACKGROUND: The complete genome of Rana grylio virus (RGV) was sequenced and analyzed recently, which revealed that RGV 50 L had homologues in many iridoviruses with different identities; however, the characteristics and functions of 50 L have not been studied yet. METHODOLOGY/PRINCIPAL FINDINGS: We cloned and characterized RGV50L, and revealed 50 L functions in virus assembly and gene regulation. 50 L encoded a 499-amino acid structural protein of about 85 kDa in molecular weight and contained a nuclear localization signal (NLS) and a helix- extension-helix motif. Drug inhibition assay demonstrated that 50 L was an immediate-early (IE) gene. Immuno-fluorescence assay revealed that 50 L appeared early and persisted in RGV-infected cells following two distribution patterns. One pattern was that 50 L exhibited a cytoplasm-nucleus- viromatrix distribution pattern, and mutagenesis of the NLS motif revealed that localization of 50 L in the nucleus was NLS-dependent; the other was that 50 L co-localized with viral matrix which plays important roles in virus assembly and the life circle of viruses. CONCLUSIONS/SIGNIFICANCE: RGV 50L is a novel iridovirus IE gene encoded structural protein which plays important roles in virus assembly.
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Regulação Viral da Expressão Gênica/genética , Genes Precoces/genética , Sinais de Localização Nuclear/metabolismo , Ranavirus/genética , Proteínas Virais/metabolismo , Montagem de Vírus/genética , Sequência de Bases , Western Blotting , Clonagem Molecular , Imunofluorescência , Perfilação da Expressão Gênica , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Análise de Sequência de DNA , Proteínas Virais/genéticaRESUMO
Infection with Rana grylio virus (RGV), an iridovirus isolated in China in 1995, resulted in a high mortality rate in frogs. The complete genome sequence of RGV was determined and analyzed. The genomic DNA was 105,791 bp long, with 106 open reading frames (ORFs). Dot plot analysis showed that the gene order of RGV shared colinearity with three completely sequenced ranaviruses. A phylogenetic tree was constructed based on concatenated sequences of iridovirus 26 core-gene-encoded proteins, and the result showed high bootstrap support for RGV being a member of the genus Ranavirus and that iridoviruses of other genera also clustered closely. A microRNA (miRNA) prediction revealed that RGV could encode 18 mature miRNAs, many of which were located near genes associated with virus replication. Thirty-three repeated sequences were found in the RGV genome. These results provide insight into the genetic nature of RGV and are useful for laboratory diagnosis for vertebrate iridoviruses.
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Genoma Viral , Ranavirus/classificação , Ranavirus/genética , Ranidae/virologia , Proteínas Virais/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Filogenia , Alinhamento de Sequência , Análise de Sequência de DNA , Proteínas Virais/químicaRESUMO
Plasmid DNAs containing Siniperca chuatsi rhabdovirus (SCRV) glycoprotein gene (pcDNA-G) and nucleoprotein gene (pcDNA-N) were constructed, and used to determine the antiviral immune response elicited by DNA vaccination in mandarin fish. In vitro and in vivo expression of the plasmid constructs was confirmed in transfected cells and muscle tissues of vaccinated fish by Western blot, indirect immunofluorescence or RT-PCR analysis. Fish injected with pcDNA-G exhibited protective effect against SCRV challenge with a relative percent survival (RPS) of 77.5%, but no significant protection (RPS of 2.5%) was observed in fish vaccinated with pcDNA-N. Immunohistochemical analysis showed that vaccination with pcDNA-G decreased histological lesions and suppressed the virus replication in fish target organs, e.g. kidney, liver, spleen, gill and heart. Transcriptional analysis further revealed that the expression levels of type I IFN system genes including interferon regulation factor-7 (IRF-7) gene, myxovirus resistance (Mx) gene and virus inhibitory protein (Viperin) gene were strongly up-regulated after injection with pcDNA-G, whereas the level of transcription of immunoglobulin M (IgM) gene did not show a statistically significant change. These results reveal that type I IFN antiviral immune response is rapidly triggered by the plasmid DNA containing rhabdovirus glycoprotein gene in fish, which offers an explanation of molecular mechanisms for DNA vaccination inducing mandarin fish resist to SCRV disease.
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Doenças dos Peixes/prevenção & controle , Perciformes/imunologia , Infecções por Rhabdoviridae/veterinária , Rhabdoviridae/patogenicidade , Vacinas de DNA/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Doenças dos Peixes/imunologia , Proteínas de Ligação ao GTP/imunologia , Glicoproteínas/genética , Glicoproteínas/imunologia , Imunoglobulina M/imunologia , Fator Regulador 7 de Interferon/imunologia , Interferon Tipo I/imunologia , Proteínas de Resistência a Myxovirus , Perciformes/virologia , Plasmídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Infecções por Rhabdoviridae/imunologia , Infecções por Rhabdoviridae/prevenção & controle , Vacinação/veterinária , Vacinas de DNA/genética , Proteínas Virais/genética , Proteínas Virais/imunologia , Vacinas Virais/genéticaRESUMO
A moribund juvenile Yangtze finless porpoise (Neophocaena phocaenoides) with skin lesions and ulceration was found in Dongting Lake, China. Pathologic examination, electron microscopy, and polymerase chain reaction of liver tissue revealed widely distributed necrotic lesions, sinusoidal dilatation, congestion and herpes-like virus particles.
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Infecções por Herpesviridae/veterinária , Herpesviridae , Toninhas/virologia , Animais , China , Evolução Fatal , Herpesviridae/genética , Herpesviridae/isolamento & purificação , Herpesviridae/ultraestrutura , Infecções por Herpesviridae/patologia , Infecções por Herpesviridae/virologiaRESUMO
A red-spotted grouper Epinephelus akaara skin (RGS) cell line was established and characterized. RGS cells had a normal diploid chromosome number of 2n = 48, the morphology of which was fibroblastic-like in 3 days and epithelial-like over 5 after 16 passages. The cells multiplied well in Dulbecco's modified Eagle's medium supplemented with 10% of fetal bovine serum at 25°C. Susceptibilities of RGS and grass carp ovary (GCO) cells to two viruses were tested, and the results showed that the titer of an iridovirus Rana grylio virus (RGV) in RGS cells was 10³·5 TCID50 ml⻹, which was much higher than a rhabdovirus spring viremia of carp virus (SVCV) in the cells (10°·5 TCID50 ml⻹). The titers of RGV and SVCV in GCO were 106·° TCID50 ml⻹ and 108·° TCID50 ml⻹, respectively, which were higher than those in RGS cells. The data may imply that RGS cells could be selectively resistible to some viruses during infection. RT-PCR analysis of RGV-infected RGS cells showed that RGV could replicate in RGS cells. Further study of virus replications in RGS cells was conducted by electron microscopy and immunofluorescence microscopy has shown that virus particles scattered in the cytoplasm and virus protein appeared in both the cytoplasm and nucleus. The results suggested that RGS cells could be used as a potential in vitro model to study the cutaneous barrier function against virus infection.
Assuntos
Linhagem Celular , Perciformes/imunologia , Pele/citologia , Animais , Técnicas de Cultura de Células , Linhagem Celular/imunologia , Linhagem Celular/ultraestrutura , Linhagem Celular/virologia , Proliferação de Células , Infecções por Vírus de DNA/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/virologia , Iridovirus/imunologia , Cariótipo , Microscopia de Fluorescência , Perciformes/virologia , Rhabdoviridae/imunologia , Pele/imunologia , Pele/virologiaRESUMO
Genomic sequence of Scophthalmus maximus rhabdovirus (SMRV) isolated from diseased turbot has been characterized. The complete genome of SMRV comprises 11,492 nucleotides and encodes five typical rhabdovirus genes N, P, M, G and L. In addition, two open reading frames (ORF) are predicted overlapping with P gene, one upstream of P and smaller than P (temporarily called Ps), and another in P gene which may encodes a protein similar to the vesicular stomatitis virus C protein. The C ORF is contained within the P ORF. The five typical proteins share the highest sequence identities (48.9%) with the corresponding proteins of rhabdoviruses in genus Vesiculovirus. Phylogenetic analysis of partial L protein sequence indicates that SMRV is close to genus Vesiculovirus. The first 13 nucleotides at the ends of the SMRV genome are absolutely inverse complementarity. The gene junctions between the five genes show conserved polyadenylation signal (CATGA(7)) and intergenic dinucleotide (CT) followed by putative transcription initiation sequence A(A/G)(C/G)A(A/G/T), which are different from known rhabdoviruses. The entire Ps ORF was cloned and expressed, and used to generate polyclonal antibody in mice. One obvious band could be detected in SMRV-infected carp leucocyte cells (CLCs) by anti-Ps/C serum via Western blot, and the subcellular localization of Ps-GFP fusion protein exhibited cytoplasm distribution as multiple punctuate or doughnut shaped foci of uneven size.