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1.
Acta Haematol ; 142(3): 162-170, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31091521

RESUMO

Aplastic anemia (AA) is a hematologic disease characterized by pancytopenia and hypocellular bone marrow, potentially leading to chronic anemia, hemorrhage, and infection. The China Aplastic Anemia Committee and British Committee for Standards in Haematology guidelines recommend hematopoietic stem-cell transplantation (HSCT) or immunosuppressive therapy (IST) comprising antithymocyte globulin (ATG) with cyclosporine (CsA) as initial treatment for AA patients. With limited epidemiological data on the clinical management of AA in Asia, a prospective cohort registry study involving 22 AA treatment centers in China was conducted to describe the disease characteristics of newly diagnosed AA patients and investigate real-world treatment patterns and patient outcomes. Of 340 AA patients, 72.9, 12.6, and 3.5% were receiving IST, traditional Chinese medicine, and HSCT, respectively, at baseline; only 22.2% of IST-treated patients received guideline-recommended ATG with CsA initially. Almost all patients received supportive care (95.6%) as blood transfusion (97.8%), antibiotics (63.7%), and/or hematopoietic growth factors (58.2%). Overall, 64.8% achieved a partial or complete response, and 0.9% experienced relapse. No new safety concerns were identified; serious adverse events were largely unrelated to the treatment regimen. These results demonstrate the need to identify and minimize treatment barriers to standardize and align AA management in China with treatment guideline recommendations and further improve patient outcomes.

2.
Emerg Microbes Infect ; 7(1): 171, 2018 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-30353004

RESUMO

Seasonal H3N2 influenza viruses are recognized as major epidemic viruses, exhibiting complex seasonal patterns in regions with temperate climates. To investigate the influence of viral evolution and mutations on the seasonality of influenza, we performed a genome-wide analysis of samples collected from 62 influenza A/H3N2-infected patients in Shanghai during 2016-2017. Phylogenetic analysis of all eight segments of the influenza A virus revealed that there were two epidemic influenza virus strains circulating in the 2016-2017 winter season (2016-2017win) and 2017 summer season (2017sum). Replication of the two epidemic viral strains at different temperatures (33, 35, 37, and 39 °C) was measured, and the correlation of the mutations in the two epidemic viral strains with temperature sensitivity and viral replication was analyzed. Analysis of the replication kinetics showed that replication of the 2016-2017win strains was significantly restricted at 39 °C compared with that of the 2017sum strains. A polymerase activity assay and mutational analysis demonstrated that the PA I668V mutation of the 2016-2017win viruses suppressed polymerase activity in vitro at high temperatures. Taken together, these data suggest that the I668V mutation in the PA subunit of the 2016-2017win strains may confer temperature sensitivity and attenuate viral replication and polymerase activity; meanwhile, the 2017sum strains maintained virulence at high temperatures. These findings highlight the importance of certain mutations in viral adaptation and persistence in subsequent seasons.

3.
J Med Virol ; 90(4): 721-729, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29247529

RESUMO

Serum Mac-2-binding protein glycosylation isomer (M2BPGi) level was found to be a useful prognostic marker for hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) patients treated with nucleoside/nucleotide analogs (NUCs) therapy, and the aim of our study is to evaluate the clinical implementation of M2BPGi level in the prediction of antiviral responses to pegylated-interferon-α (PEG-IFN-α) treatment in HBeAg-positive CHB patients. Ninety-six CHB patients who received PEG-IFN-α treatment for at least 48 weeks were recruited. The serum M2BPGi, alanine aminotransferase (ALT), hepatitis B surface antigen (HBsAg), HBeAg, and HBV DNA levels at baseline, weeks 4, 12, and 24 after PEG-IFN-α treatment were determined and their associations with antiviral responses were evaluated and the virological response (VR) rate and serological response (SR) rate after 48 weeks of treatment were 65.6% and 35.4%, respectively. Baseline serum M2BPGi level was significantly different between VR and non-VR (P = 0.002) or SR and non-SR groups (P = 0.012). Multivariate analyses suggested that baseline serum M2BPGi level was independently associated with VR and SR of PEG-IFN-α treatment at week 48. The area under the ROC curve (AUC) of baseline M2BPGi was 0.682 in predicting VR, which was superior to HBsAg (AUC = 0.566) or HBV DNA (AUC = 0.567). The AUC of baseline M2BPGi in predicting SR was 0.655, which was also higher than that of HBsAg (AUC = 0.548) or HBV DNA (AUC = 0.583). These results suggested that baseline serum M2BPGi level was a novel predictor of VR and SR for PEG-IFN-α treatment in HBeAg-positive CHB patients.

4.
Emerg Microbes Infect ; 6(3): e15, 2017 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-28325923

RESUMO

Coexistence of the hepatitis B surface antigen (HBsAg) and hepatitis B surface antibody (anti-HBs) is an uncommon phenomenon, and the underlying mechanisms remain largely unknown. Amino-acid (aa) substitution from glycine to arginine at aa 145 (G145R), in the major hydrophilic region, has been reported in patients with HBsAg and anti-HBs coexistence. However, there is limited knowledge about the clinical features and viral quasispecies characteristics associated with G145R mutant hepatitis B virus (HBV) infection. We herein describe the dynamic changes in the serological and virological markers in a case of hepatitis B with coexisting HBsAg and anti-HBs, caused by a G145R immune escape mutant (genotype C). Entecavir was administered during the 4th week after admission. Alanine aminotransferase peaked in the 16th week, while both the HBsAg and HBeAg declined rapidly. HBsAg clearance and hepatitis B e antigen (HBeAg)/hepatitis B e antibody (anti-HBe) seroconversion were achieved in the 36th week, and then entecavir was withdrawn. A follow-up of 96 weeks showed that HBV DNA remained undetectable and that anti-HBs was maintained above 100 mIU/mL. The quasispecies characteristics of the G145R mutant HBV were investigated via ultra-deep sequencing. The complexity and genetic distance of the S and RT regions were much higher in the 8th week than at baseline or in the 4th week. Moreover, the frequencies of mutations (L173P, Q181R and A184V) in cytotoxic T lymphocyte epitopes increased before entecavir treatment. These findings extend understanding of the evolution of HBV under host immune pressure and of the clinical outcomes of affected patients.


Assuntos
Antivirais/administração & dosagem , Guanina/análogos & derivados , Anticorpos Anti-Hepatite B/sangue , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/genética , Hepatite B/tratamento farmacológico , Idoso de 80 Anos ou mais , Alanina Transaminase/metabolismo , Substituição de Aminoácidos , Epitopos de Linfócito T/genética , Guanina/administração & dosagem , Hepatite B/metabolismo , Hepatite B/virologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Taxa de Mutação , Análise de Sequência de DNA , Soroconversão
5.
Liver Int ; 37(1): 35-44, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27300763

RESUMO

BACKGROUND & AIMS: Accurate evaluation of liver fibrosis is crucial for predicting progression of chronic hepatitis B virus (HBV) infection. We assessed the utility of a novel fibrosis glycobiomarker Wisteria floribunda agglutinin-positive Mac-2-binding protein (WFA+ -M2BP) for evaluating liver fibrosis and disease progression in patients with chronic HBV infection. METHODS: We enrolled 774 patients with chronic HBV infection, with or without fibrosis, diagnosed by liver biopsy/FibroScan. Patients who underwent liver biopsy (n = 297) were divided into training (n = 221) and validation (n = 76) groups. Serum WFA+ -M2BP values were measured and compared with FIB-4 index, aspartate aminotransferase (AST)-to-platelet ratio (APRI) and AST-to-alanine aminotransferase ratio (AAR) using receiver-operating characteristic (ROC) analysis. RESULTS: Serum WFA+ -M2BP levels increased significantly with fibrosis progression (P < 0.0001). Area under the ROC curve of WFA+ -M2BP for diagnosing significant fibrosis was higher than that of FIB-4 (P = 0.198), APRI (P = 0.017) and AAR (P < 0.001), with sensitivity and specificity in the training set of 60.5% and 79.8% and validation set of 59.5% and 82.1%, respectively. Serum WFA+ -M2BP levels were significantly correlated with FibroScan values (P < 0.0001) and improved the accuracy of FibroScan in assessing significant fibrosis. Changes in WFA+ -M2BP levels were parallel with those in FibroScan values during nucleot(s)ide analogues therapy in patients with chronic HBV infection. CONCLUSIONS: WFA+ -M2BP is an accurate serum indicator for assessing early stages of liver fibrosis and may monitor regression of fibrosis during the treatment of chronic HBV infection. WFA+ -M2BP provides a simple and reliable alternative or complementary method to liver biopsy and FibroScan.


Assuntos
Antígenos de Neoplasias/sangue , Hepatite B Crônica/sangue , Hepatite B Crônica/complicações , Cirrose Hepática/sangue , Glicoproteínas de Membrana/sangue , Adolescente , Adulto , Idoso , Biomarcadores/sangue , China , Progressão da Doença , Técnicas de Imagem por Elasticidade , Feminino , Humanos , Modelos Lineares , Fígado/patologia , Cirrose Hepática/patologia , Masculino , Pessoa de Meia-Idade , Lectinas de Plantas , Curva ROC , Receptores de N-Acetilglucosamina , Estudos Retrospectivos , Adulto Jovem
6.
Springerplus ; 5(1): 1630, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27722049

RESUMO

BACKGROUND: CpG islands in hepatitis B virus (HBV) genome are potential targets for methylation mediated gene silencing, and may be involved in the pathogenesis of HBV infection. To date, their characteristics in HBV quasispecies (QS) remain largely unknown. The purpose of this study was to investigate the characteristics of CpG islands in HBV QS. METHODS: Forty patients diagnosed as acute hepatitis B (AHB, n = 10), immune-tolerant HBV carriers (IT, n = 9), chronic hepatitis B (CHB, n = 11), or acute on chronic liver failure (ACLF, n = 10), were enrolled in this case-control study. A total of 599 clones were isolated, and full-length HBV genomes were sequenced. RESULTS: CpG island II (CGII) in AHB group was shorter in length and its QS heterogeneity was lower than that in the chronic infection group. Among the chronic infection subgroups, CGII and CpG island III (CGIII) in IT group were longer and their heterogeneity was lower compared to CHB and ACLF groups. Length of CGII correlated with HBV DNA levels positively while the complexity and diversity of CGII correlated with HBV DNA levels negatively. Moreover, CGII and CGIII were shorter in genotype B than those in genotype C, while QS complexity and diversity of either CGII or CGIII had no significant difference between genotype B and C. CONCLUSIONS: Overall, our results suggest that the distribution, length and QS heterogeneity of CpG islands in full-length HBV genome differ across clinical phases of infection, of which the mechanism warrants further study.

7.
J Clin Virol ; 72: 88-94, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26476325

RESUMO

BACKGROUND: The best strategy for chronic hepatitis B patients with poor response to 48 weeks of Peginterferon-based therapy has been controversial and the predictive value of hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and hepatitis B virus (HBV) DNA levels for determining the sustained virological response (SVR) of these patients is uncertain. OBJECTIVES: To optimize management of these patients and evaluate the use of these serobiomarkers to predict SVR. STUDY DESIGN: Eighty-one patients with an unsatisfactory response after 48 weeks of Peginterferon-based therapy were treated with extended Peginterferon therapy with or without nucleo(s) tide analogues (NAs), for a total of 96 weeks of Peginterferon treatment. HBsAg, HBeAg and HBV DNA levels were measured serially during the treatment and follow-up. RESULTS AND CONCLUSIONS: Twenty-six of 81 patients (32.1%) attained SVR during the 72-week follow-up. The SVR rate was not statistically different between groups receiving 1-year prolongation of Peginterferon with or without NAs. The serum HBsAg cut-off of 1800IU/mL at week 48 had area under curve (AUC) of 0.727, and the serum HBsAg cut-off of 1500IU/mL, combined with HBeAg loss at week 72, had AUC of 0.753 to predict SVR during the follow-up. In conclusion, extended treatment with Peginterferon with or without NAs for patients with unsatisfactory response after 48 weeks of Peginterferon-based therapy is a promising strategy to achieve SVR, and quantitative serum HBsAg at week 48 and HBsAg level combined with HBeAg loss at week 72 of therapy can predict SVR to prolongation therapy with Peginterferon.


Assuntos
Antivirais/uso terapêutico , Biomarcadores/sangue , Monitoramento de Medicamentos/métodos , Antígenos de Superfície da Hepatite B/sangue , Antígenos E da Hepatite B/sangue , Hepatite B Crônica/tratamento farmacológico , Interferon-alfa/uso terapêutico , Polietilenoglicóis/uso terapêutico , Adulto , DNA Viral/sangue , Feminino , Humanos , Masculino , Prognóstico , Proteínas Recombinantes/uso terapêutico , Estudos Retrospectivos , Resultado do Tratamento
8.
J Clin Microbiol ; 53(7): 2203-14, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25926495

RESUMO

Hepatitis B virus (HBV) infection results in different clinical presentation due to different levels of immune response. Our study aimed to characterize HBV full-length genome quasispecies (QS) in patients with different phases of infection to better understand its pathogenesis. Forty treatment-naive HBV-infected patients were enrolled, including 10 cases of acute hepatitis B (AHB), 9 cases of immunotolerant (IT) HBV carriers, 11 cases of chronic hepatitis B (CHB), and 10 cases of acute-on-chronic liver failure (ACLF). The present study was conducted by clone-based sequencing. QS heterogeneity within each open reading frame was calculated. The mutation frequency index (MFI) and amino acid variations within the large HBsAg, HBcAg, and HBxAg regions were analyzed based on the different infection phases. In total, 606 HBV full-length sequences were obtained. HBV QS had higher heterogeneity in ACLF and CHB than that in IT among chronically infected individuals. AHB patients had the lower QS heterogeneity at onset than those with chronic infection. ACLF patients had the highest frequency of mutations in the core promoter and precore region. A triple mutation (A1762T/G1764A/G1896A) was observed more frequently in genotype C than in genotype B. The MFI indicated that specific peptides of the studied regions had more frequent mutations in ACLF. Furthermore, several amino acid variations, known as T- and B-cell epitopes, were potentially associated with the immunoactive phase of infection. More HBV genome mutations and deletions were observed in patients with more severe diseases, particularly in specific regions of the core and preS regions, the clinical significance and mechanism of which need to be further investigated.


Assuntos
Variação Genética , Genoma Viral , Genótipo , Vírus da Hepatite B/classificação , Vírus da Hepatite B/genética , Hepatite B/virologia , Epitopos/genética , Hepatite B/patologia , Antígenos da Hepatite B/genética , Vírus da Hepatite B/isolamento & purificação , Humanos , Taxa de Mutação , Mutação de Sentido Incorreto , Deleção de Sequência
9.
Biochem Biophys Res Commun ; 455(1-2): 90-7, 2014 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-25451272

RESUMO

We studied two patients from a nonconsanguineous family with life-long abnormal liver function, hepatomegaly and abnormal fatty acid profiles. Abnormal liver function, hypoglycemia and muscle weakness are observed in various genetic diseases, including medium-chain acyl-CoA dehydrogenase (MCAD) deficiency and glycogen storage diseases. The proband showed increased free fatty acids, mainly C8 and C10, resembling fatty acid oxidation disorder. However, no mutation was found in ACADM and ACADL gene. Sequencing of theamylo-alpha-1, 6-glucosidase, 4-alpha-glucanotransferase (AGL) gene showed that both patients were compound heterozygotes for c.118C > T (p.Gln40X) and c.753_756 del CAGA (p.Asp251Glufsx29), whereas their parents were each heterozygous for one of these mutations. The AGL protein was undetectable in EBV-B cells from the two patients. Transcriptome analysis demonstrated a significant different pattern of gene expression in both of patients' cells, including genes involving in the PPAR signaling pathway, fatty acid biosynthesis, lipid synthesis and visceral fat deposition and metabolic syndrome. This unique gene expression pattern is probably due to the absence of AGL, which potentially accounts for the observed clinical phenotypes of hyperlipidemia and hepatocyte steatosis in glycogen storage disease type IIIa.


Assuntos
Sistema da Enzima Desramificadora do Glicogênio/genética , Doença de Depósito de Glicogênio Tipo III/genética , Mutação , Acil-CoA Desidrogenase/deficiência , Adolescente , Células Cultivadas , Ácidos Graxos/biossíntese , Ácidos Graxos Insaturados/biossíntese , Expressão Gênica , Doença de Depósito de Glicogênio Tipo III/diagnóstico , Doença de Depósito de Glicogênio Tipo III/metabolismo , Humanos , Erros Inatos do Metabolismo Lipídico/diagnóstico , Masculino
10.
Mol Med Rep ; 10(6): 3193-8, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25310131

RESUMO

Forkhead box M1 (FOXM1) is a typical proliferation­associated transcription factor, which is overexpressed in many types of human cancer. We investigated the expression level of FOXM1 in patients with untreated acute leukemia (AL) and explored the correlation between expression levels and AL type. The relationship between the expression of the genes FOXM1 and mammalian target of rapamycin (mTOR) was determined after treatment of ML-2 cells with thiostrepton. The apoptosis, proliferation and cell-cycle progression of ML-2 lines were examined after treatment with metformin. We found that FOXM1 is expressed in the majority of AL patients and that its expression level was associated with the AL type. Thiostrepton is a specific inhibitor of FOXM1, and by inhibiting the FOXM1 expression via thiostrepton, we observed downregulatiion of mTOR; a significant correlation between FOXM1 and mTOR levels was observed. Thus, metformin may be involved in the downregulation of FOXM1. In addition, our study demonstrated that metformin promotes the apoptosis of ML-2 cells, induces cell-cycle arrest at the G0/G1 and G2/M phases, and inhibits proliferation. The potential role of FOXM1 in tumorigenesis renders it an attractive target for anticancer therapy, and metformin may represent a new agent for the treatment of leukemia.


Assuntos
Fatores de Transcrição Forkhead/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Leucemia/genética , Metformina/farmacologia , Apoptose/efeitos dos fármacos , Apoptose/genética , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Transformação Celular Neoplásica/efeitos dos fármacos , Transformação Celular Neoplásica/genética , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Proteína Forkhead Box M1 , Humanos , Serina-Treonina Quinases TOR/genética , Tioestreptona/farmacologia
11.
BMC Med Educ ; 14: 111, 2014 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-24885865

RESUMO

BACKGROUND: Since the global standards for postgraduate medical education (PGME) were published in January 2003, they have gained worldwide attention. The current state of residency training programs in medical-school-affiliated hospitals throughout China was assessed in this study. METHODS: Based on the internationally recognized global standards for PGME, residents undergoing residency training at that time and the relevant residency training instructors and management personnel from 15 medical-school-affiliated hospitals throughout China were recruited and surveyed regarding the current state of residency training programs. A total of 938 questionnaire surveys were distributed between June 30, 2006 and July 30, 2006; of 892 surveys collected, 841 were valid. RESULTS: For six items, the total proportions of "basically meets standards" and "completely meets standards" were <70% for the basic standards. These items were identified in the fields of "training settings and educational resources", "evaluation of training process", and "trainees". In all fields other than "continuous updates", the average scores of the western regions were significantly lower than those of the eastern regions for both the basic and target standards. Specifically, the average scores for the basic standards on as many as 25 of the 38 items in the nine fields were significantly lower in the western regions. There were significant differences in the basic standards scores on 13 of the 38 items among trainees, instructors, and managers. CONCLUSIONS: The residency training programs have achieved satisfactory outcomes in the hospitals affiliated with various medical schools in China. However, overall, the programs remain inadequate in certain areas. For the governments, organizations, and institutions responsible for PGME, such global standards for PGME are a very useful self-assessment tool and can help identify problems, promote reform, and ultimately standardize PGME.


Assuntos
Hospitais/estatística & dados numéricos , Internato e Residência/estatística & dados numéricos , Faculdades de Medicina/organização & administração , Adulto , Idoso , China , Coleta de Dados , Feminino , Humanos , Relações Interinstitucionais , Internato e Residência/organização & administração , Masculino , Pessoa de Meia-Idade , Faculdades de Medicina/estatística & dados numéricos , Inquéritos e Questionários , Adulto Jovem
12.
Biochem Biophys Res Commun ; 446(1): 280-5, 2014 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-24582753

RESUMO

Forkhead box M1 (FoxM1) drives cell cycle progression and the prevention of growth arrest and is over-expressed in many human malignancies. However, the characteristics of FoxM1 in acute myeloid leukemia (AML) are not clearly understood. We investigated the expression level of FoxM1 and analyzed the correlation of FoxM1 expression with AML patient characteristics and prognoses. Changes in FoxM1 expression were detected after MV4-11 cells, which have an internal tandem duplication (ITD) of the fms-like tyrosine kinase 3 gene (FLT3-ITD), and control THP1 cells (encoding wild-type FLT3) were treated with the FLT3 receptor tyrosine kinase inhibitor AC220 (quizartinib) or FLT3 ligand (FL). Finally, we determined the apoptosis rates after the addition of the FoxM1 inhibitor thiostrepton (TST) to AML cells with or without FLT3-ITD. The expression of FoxM1 in AML patients was correlated with the presence of FLT3-ITD, genetic groups, and possibly overall survival. Inhibition of FLT3-ITD by AC220 down-regulated FoxM1 expression in MV4-11 cells, and stimulation of FLT3 by FL up-regulated FoxM1 expression in MV4-11 and THP1 cells. TST induced the apoptosis of MV4-11 and THP1 cells in a dose-dependent manner. Thus, FoxM1 is a potential prognostic marker and a promising therapeutic target in AML.


Assuntos
Fatores de Transcrição Forkhead/genética , Leucemia Mieloide Aguda/genética , Tirosina Quinase 3 Semelhante a fms/genética , Apoptose , Benzotiazóis/farmacologia , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Linhagem Celular Tumoral , Feminino , Proteína Forkhead Box M1 , Fatores de Transcrição Forkhead/antagonistas & inibidores , Fatores de Transcrição Forkhead/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Estimativa de Kaplan-Meier , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patologia , Masculino , Compostos de Fenilureia/farmacologia , Prognóstico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Neoplásico/genética , RNA Neoplásico/metabolismo , Sequências de Repetição em Tandem , Tioestreptona/farmacologia , Regulação para Cima , Tirosina Quinase 3 Semelhante a fms/antagonistas & inibidores , Tirosina Quinase 3 Semelhante a fms/metabolismo
13.
J Clin Microbiol ; 52(5): 1556-65, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24574300

RESUMO

Chronic hepatitis B virus (HBV) infection via perinatal transmission is common in the Asia-Pacific region, but related quasispecies (QS) characteristics are not yet defined. To investigate the homologous, full-length HBV QS after perinatal infection and their clinical implications, five pairs of mother-daughter patients with chronic HBV infection (one patient with liver cirrhosis, one with immune tolerance, and eight with chronic hepatitis) were included. Full-length HBV were amplified by PCR from serum samples before antiviral treatment and cloned; an average of 17 clones per sample were sequenced, and the QS complexities, diversities, and evolution patterns were analyzed. Full-length HBV sequence similarities within mother-daughter pairs were 91.3 to 98.3%. The QS complexities of full-length HBV were similar between mothers and daughters (median of 0.9734 compared to 0.9688, P>0.05), as were the diversities (median of 3.396×10(-3) compared to 4.617×10(-3) substitutions/site, P>0.05). However, the distribution patterns of QS complexities and diversities within specific genes were different, and QS genetic distances of the mothers were higher than those of daughters, both more evident in pairs with different antiviral responses and different immune phases or stages. The nucleotide substitution rate of full-length HBV was 14.388×10(-5) substitutions/site/year, whereas the preC/C gene rate was the highest. Mutations and indels were more common in clones from the mothers, which decreased the affinity of epitopes by 6- to 89-fold. The various genes from homologous HBV genomes evolved in different patterns despite numerically comparable full-length QS complexities and diversities. The different patterns may correlate with the immune stages of chronic HBV infection, which warrants further study.


Assuntos
Vírus da Hepatite B/genética , Hepatite B Crônica/transmissão , Transmissão Vertical de Doença Infecciosa , Adulto , Antivirais/uso terapêutico , DNA Viral/genética , Epitopos/genética , Feminino , Genoma Viral/genética , Genótipo , Antígenos do Núcleo do Vírus da Hepatite B/genética , Hepatite B Crônica/tratamento farmacológico , Hepatite B Crônica/virologia , Humanos , Pessoa de Meia-Idade , Mães , Mutação/genética , Núcleo Familiar , Reação em Cadeia da Polimerase/métodos , Estudos Retrospectivos , Análise de Sequência de DNA/métodos , Adulto Jovem
14.
Gene ; 536(2): 362-5, 2014 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-24355556

RESUMO

Glycogen storage disease type Ia (GSD-Ia) is an autosomal recessive genetic disorder resulting in hypoglycemia, hepatomegaly and growth retardation. It is caused by mutations in the G6PC gene encoding Glucose-6-phosphatase. To date, over 80 mutations have been identified in the G6PC gene. Here we reported a novel mutation found in a Chinese patient with abnormal transaminases, hypoglycemia, hepatomegaly and short stature. Direct sequencing of the coding region and splicing-sites in the G6PC gene revealed a novel no-stop mutation, p.*358Yext*43, leading to a 43 amino-acid extension of G6Pase. The expression level of mutant G6Pase transcripts was only 7.8% relative to wild-type transcripts. This mutation was not found in 120 chromosomes from 60 unrelated healthy control subjects using direct sequencing, and was further confirmed by digestion with Rsa I restriction endonuclease. In conclusion, we revealed a novel no-stop mutation in this study which expands the spectrum of mutations in the G6PC gene. The molecular genetic analysis was indispensable to the diagnosis of GSD-Ia for the patient.


Assuntos
Grupo com Ancestrais do Continente Asiático/genética , Glucose-6-Fosfatase/genética , Doença de Depósito de Glicogênio Tipo I/genética , Mutação/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Células COS , Linhagem Celular , Cercopithecus aethiops/genética , Criança , Feminino , Homozigoto , Humanos , Dados de Sequência Molecular , Processamento de RNA
15.
J Hepatol ; 60(3): 515-22, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24239777

RESUMO

BACKGROUND & AIMS: HBV immune escape represents a challenge to prevention, diagnosis, and treatment of hepatitis B. Here, we analyzed the molecular and clinical characteristics of HBV immune escape mutants in a Chinese cohort of chronically infected patients. METHODS: Two hundred sixteen patients with HBsAg and anti-HBs were studied, with one hundred eighty-two HBV carriers without anti-HBs as a control group. Recombinant HBsAg bearing the most frequent N-glycosylation mutations were expressed in CHO and HuH7 cells. After confirming N-glycosylation at the most frequent sites (129 and 131), together with inserted mutations, functional analysis were performed to study antigenicity and secretion capacity. RESULTS: One hundred twenty-three patients had the wild-type HBs gene sequence, 93 patients (43%) had mutants in the major hydrophilic region (MHR), and 47 of the 93 patients had additional N-glycosylation mutations, which were transmitted horizontally to at least 2 patients, one of whom was efficiently vaccinated. The frequency of N-glycosylation mutation in the case group was much higher than that of the control group (47/216 vs. 1/182). Compared with wild-type HBsAg, HBsAg mutants reacted weakly with anti-HBs using a chemiluminescent microparticle enzyme immunoassay. Native gel analysis of secreted virion in supernatants of transfected HuH7 cells indicated that mutants had better virion enveloping and secretion capacity than wild-type HBV. CONCLUSIONS: Our results suggest that specific HBsAg MHR N-glycosylation mutations are implicated in HBV immune escape in a high endemic area.


Assuntos
Antígenos de Superfície da Hepatite B/genética , Evasão da Resposta Imune , Mutação , Adolescente , Adulto , Idoso , Feminino , Glicosilação , Antígenos de Superfície da Hepatite B/imunologia , Humanos , Interações Hidrofóbicas e Hidrofílicas , Masculino , Pessoa de Meia-Idade
16.
Zhonghua Gan Zang Bing Za Zhi ; 21(7): 510-3, 2013 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-24074709

RESUMO

OBJECTIVE: To evaluate the influence of insertion mutations occurring in the hydrophobic region, between amino acids 114 and 115, of the hepatitis B surface antigen (HBsAg) on viral antigenicity and replication. METHODS: Hepatitis B virus (HBV) DNA was obtained from patients with HBsAg-positive chronic hepatitis B (CHB) infection and subjected to sequence analysis and comparison to GenBank reference sequences for HBV genotype B (AB073826) and genotype C (AF286594). Insertion mutations detected in the HBsAg region were used to make recombinant expression plasmids via site-directed mutagenesis. After transfecting the recombinant HBsAg into Huh7 cells, the mutants' effects on viral antigenicity and replication were evaluated by chemiluminescence microparticle immunoassay (CMIA) and Southern blot hybridization, respectively. The viral antigenicity of each mutant was predicted by bioinformatic analysis, using the Jameson-Wolf method to predict the antigenic index, the Hopp-Woods method to predict hydrophilicity, the Emini method to predict the probability of a region lying of the protein's surface, and the Karplus-Schulz method to predict the flexibility of the protein backbone. RESULTS: Two CHB patients harbored HBV with insertion mutations in HBsAg: one with two (NT) and one with three (NTT) inserted amino acids between 114 and 115. The NTT recombinant HBsAg mutant showed no impact on viral replication and reacted weakly with anti-HBs in CMIA (P = 0.02). The antigen indices for the insertion of NTT were 1.00, -0.16, and 0.18, and insertion of the three amino acids affected the index values of five proximal amino acid sites (with an average increase of 0.13). The hydrophilic indices for the insertion of NTT were 0.2, -0.4, and -0.4, with no significant effect on the proximal amino acids. The insertion of the three amino acids changed both the surface probability (range: -0.55 to 2.97; affecting eight proximal amino acids) and the flexibility (range: -0.01 to 1.1; affecting five proximal amino acids). CONCLUSION: The NTT three amino acids insertion in the HBsAg open reading frame, between 114 and 115 of the wild-type sequence, detected in a CHB patient may play a role in HBV immune escape without influencing the viral replicative capacity.


Assuntos
Antígenos de Superfície da Hepatite B/genética , Vírus da Hepatite B/genética , Vírus da Hepatite B/imunologia , Mutagênese Insercional , Adulto , DNA Viral/genética , Genótipo , Vírus da Hepatite B/fisiologia , Humanos , Masculino , Pessoa de Meia-Idade , Replicação Viral
17.
Zhonghua Yi Xue Za Zhi ; 93(19): 1458-62, 2013 May 21.
Artigo em Chinês | MEDLINE | ID: mdl-24029567

RESUMO

OBJECTIVE: To investigate the characteristics of high density lipoprotein cholesterol and the relationship between high density lipoprotein cholesterol and the severity of coronary artery lesions in young men with acute myocardial infarction (AMI). METHODS: We retrospectively studied 278 young men with acute myocardial infarction and compared with 208 non-CHD young men, 137 old men with AMI. All patients were admitted to hospital from Jan 2009 to Dec 2011 and undergone coronary angiography, and the clinic and coronary angiographic features were assessed.According to the result of coronary angiography, the patients were divided into three groups:the single, double and triple vessel lesions. The relation between systolic body mass index (BMI), hemoglobin (Hb), serum uric acid (UA), total cholesterol(TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), smoking history, essential hypertension, type 2 diabetes mellitus, familial history of early coronary artery disease with acute myocardial infarction and severity of coronary artery disease are observed.And observe the characteristics of HDL-C in the obesity group and the smoking group in young men based on body mass index and smoking history. RESULTS: (1) In young men with AMI group, the HDL-C levels was significantly lower than those in non-CHD young men group((1.00 ± 0.28) mmol/L vs (1.05 ± 0.23) mmol/L, P < 0.05).In young men with AMI group, the HDL-C levels was significantly lower than those in old men group with AMI ((1.00 ± 0.28) mmol/L vs (1.07 ± 0.30) mmol/L, P < 0.05);the HDL-C level in young AMI men with smoking history was significantly lower than those in young AMI men without smoking history ((0.98 ± 0.25) mmol/L vs (1.09 ± 0.40) mmol/L, P < 0.05);the HDL-C level in normal weight group is significantly higher than those in overweight and obesity groups in young AMI men ((1.30 ± 0.55) mmol/L vs (0.99 ± 0.22) mmol/L, (0.98 ± 0.29) mmol/L, P < 0.05). (2)The HDL-C level in the single lesions group was significantly lower than those in the double and triple vessel lesions groups ((1.06 ± 0.29) mmol/L vs (0.92 ± 0.20) mmol/L, (0.91 ± 0.26) mmol/L, P < 0.05). (3) Applying Logistic regression analysis, type 2 diabetes mellitus (OR = 35.784), essential hypertension (OR = 7.782), familial history of early coronary artery disease (OR = 4.613), low density lipoprotein cholesterol (OR = 2.496), smoking history (OR = 2.241), hemoglobin (OR = 1.042) and serum uric acid (OR = 1.005) are independent risk factors (P < 0.05) for young men with AMI, while high density lipoprotein cholesterol (OR = 0.147, P < 0.05) is a protective factor; low density lipoprotein cholesterol (OR = 2.095) and essential hypertension (OR = 1.042) are independent risk factors (P < 0.05) for young men with multiple vessel lesions in AMI, while high density lipoprotein cholesterol (OR = 0.071, P < 0.05) is a protective factor. CONCLUSION: High density lipoprotein cholesterol are protective factors for young men with AMI and multiple vessel lesions in young men with AMI.


Assuntos
HDL-Colesterol/sangue , Doença da Artéria Coronariana/etiologia , Infarto do Miocárdio/sangue , Infarto do Miocárdio/patologia , Adolescente , Adulto , Estudos de Casos e Controles , Angiografia Coronária , Doença da Artéria Coronariana/diagnóstico por imagem , Humanos , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/diagnóstico por imagem , Estudos Retrospectivos , Adulto Jovem
18.
J Huazhong Univ Sci Technolog Med Sci ; 33(1): 37-42, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23392705

RESUMO

The molecular pathogenesis of leukemia is poorly understood. Earlier studies have shown both Wilms' tumor 1 suppressor gene (WT1) and CML28 abnormally expressed in malignant diseases of the hematopoietic system and WT1 played an important role in leukemogenesis. However, the relationship between molecular CML28 and WT1 has not been reported. Here we described the use of small interfering RNA (siRNA) against WT1 and CML28 in leukemic cell line K562 to examine the interaction between CML28 and WT1. WT1 and CML28 gene expression in transfected K562 cells was detected by using RQ-PCR and Western blotting. K562 cells transfected with WT1-siRNA could greatly decrease both mRNA and protein expression levels of WT1 and CML28. In contrast, CML28-siRNA did not exert effect on WT1. Further, subcellular co-localization assay showed that the two proteins could co-localize in the cytoplasm of K562 cells, but WT1/CML28 complexes were not detected by using immunoprecipitation. It was suggested that there exists the relationship between CML28 and WT1. CML28 may be a downstream target molecule of WT1 and regulated by WT1, which will provide important clues for further study on the role of CML28 and WT1 in leukemic cells.


Assuntos
Antígenos de Neoplasias/metabolismo , Complexo Multienzimático de Ribonucleases do Exossomo/metabolismo , Leucemia Eritroblástica Aguda/metabolismo , Proteínas de Neoplasias/metabolismo , Proteínas de Ligação a RNA/metabolismo , Frações Subcelulares/metabolismo , Proteínas WT1/metabolismo , Linhagem Celular Tumoral , Humanos , Células K562 , Mapeamento de Interação de Proteínas
19.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 20(6): 1302-6, 2012 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-23257421

RESUMO

This study was aimed to investigate the expression of GST-CML28 in Escherichia Coli and to prepare its antibody. The constructed recombinant expression vectors CML28-pGEX-3X were transformed into Escherichia Coli BL21 under IPTG induction. The protein was abstracted from the transformers, and purified by a GSTrap FF column. The rabbits were immunized by the purified fusion protein to produce serum with anti-CML28 antibody. The serum was purified by chromatographic column stuffed with glutathione Sephamse 4B to get the antibody. The specific antibody against CML28 was further identified by ELISA, Western blot, immunohistochemistry and quantum dot luminescence. The results indicated that GST-CML28 fusion protein was expressed in Escherichia coli and its specific polyclonal antibody was obtained. It is concluded that the anti-CML28 polyclonal antibodies with high titer and specificity are successfully prepared. These antibodies provide an useful experimental tool to profoundly research the physiological significance and biological function of the CML28 gene.


Assuntos
Anticorpos/metabolismo , Antígenos de Neoplasias/biossíntese , Complexo Multienzimático de Ribonucleases do Exossomo/biossíntese , Glutationa Transferase/biossíntese , Proteínas de Ligação a RNA/biossíntese , Proteínas Recombinantes de Fusão/biossíntese , Animais , Anticorpos/imunologia , Anticorpos/isolamento & purificação , Antígenos de Neoplasias/imunologia , Antígenos de Neoplasias/isolamento & purificação , Células Cultivadas , Escherichia coli/metabolismo , Complexo Multienzimático de Ribonucleases do Exossomo/imunologia , Complexo Multienzimático de Ribonucleases do Exossomo/isolamento & purificação , Vetores Genéticos , Glutationa Transferase/isolamento & purificação , Células Endoteliais da Veia Umbilical Humana/citologia , Humanos , Proteínas de Ligação a RNA/imunologia , Proteínas de Ligação a RNA/isolamento & purificação , Coelhos , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/isolamento & purificação
20.
Zhonghua Xue Ye Xue Za Zhi ; 33(8): 632-6, 2012 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-23134857

RESUMO

OBJECTIVE: To analyze the correlation between early lymphocyte count (lymphocyte count on day 30, LC30) post-allogeneic hematopoietic stem cell transplantation (allo-HSCT) and transplant prognosis in leukemia patients. METHODS: The data from 124 consecutive patients undergoing allo-HSCT for leukemia from January 2003 to April 2011 was analyzed retrospectively. LC30 post-allo-HSCT correlated with 5-year overall survival (OS), 5-year relapse rate (RR), 5-year nonrelapse mortality (NRM), accumulative rate of acute graft versus host disease (aGVHD) and chronic graft versus host disease (cGVHD) was studied. RESULTS: Univariate analysis indicated that patients with LC30 ≥ 0.40×10(9)/L had higher 5-year OS than those with LC30 < 0.40×10(9)/L \[(62.2 ± 5.8)% vs (37.0 ± 8.6)%, P = 0.003\], lower 5-year RR\[(13.9 ± 4.7)% vs (32.0 ± 8.4)%, P = 0.027\], lower 5-year NRM \[(31.3 ± 5.8)% vs (45.0 ± 9.3)%, P = 0.048)\], and higher cGVHD cumulative incidence \[(82.9 ± 4.6)% vs (62.7 ± 11.1)%, P = 0.042)\]. Multivariate analysis also suggested that LC30 was associated with 5-year OS, 5-year RR, 5-year NRM, and cGVHD cumulative incidence. At the same time disease risk stratification was associated with prognosis. CONCLUSIONS: Early lymphocyte count (LC30) post-allogeneic hematopoietic stem cell transplantation in leukemia is highly associated with prognosis, which can be the independent prognosis index after allo-HSCT in leukemia and can identify a group of patients who might be suitable candidates for early interventions treatment.


Assuntos
Transplante de Células-Tronco Hematopoéticas , Leucemia/diagnóstico , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Leucemia/cirurgia , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Adulto Jovem
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