Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 50
Filtrar
Mais filtros

Base de dados
País/Região como assunto
Tipo de documento
País de afiliação
Intervalo de ano de publicação
1.
Virol J ; 21(1): 213, 2024 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-39252044

RESUMO

High-risk human papilloma virus (HR-HPV) persistent infection is closely associated with the development of cervical cancer and squamous intraepithelial lesion (SIL).The α-9 HPVs, which is predominantly composed of HR-HPV types, account for 75% of HR-HPV infection in Sichuan. The oncoproteins E6 and E7 of HPV play a crucial role in tumor initiation and progression. Notably, HPV-35 is the only HR-HPV type within the α-9 genus that is not included in the nine-valent HPV prophylactic vaccine. Cervical cell samples obtained from Sichuan were collected for HPV detection and genotyping. Among the 406 HPV-positive samples, 31 HPV-35 were detected, 24 HPV-35 E6 and 26 E7 were successfully amplified and sequenced, five nucleotide mutations in E6 and three in E7 were detected, T232C, T434G of E6 (W78R, I145R) and C67T, G84T of E7 (H23Y, L28F) were non-synonymy mutation. PAML 4.8 server was used to detect positive selection sites of HPV-35 E6, E7, and E6 is W78R. Phyre2 were used to predict and analyze protein structures, W78R made influences on protein structure. IEDB were used to screen epitopes vaccine target for HPV-35 affection therapy, and 5 HPV-35 E6 and 3 HPV-35 E7 most potential epitopes were obtained, the most potential peptides for therapy vaccine design were 79-91YRYSVYGETLEKQ, 45-60FACYDLCIVREGQPY, 124-135RFHNIGGRWTGR of E6; 3-19GEITTLQDYVLDLEPEA, 38-47TIDGPAGQAK, 70-88VQSTHIDIRKLEDLLMGTF of E7 and W78R mainly decreased the epitopes affinity.Conclusions Amino acid substitution in the positive selection sites of HPV-35 E6 and E7 genes have been found to influence protein structure and to decrease the overall affinity of antigen epitopes. This observation aligns with the evolutionary significance of positive selection site, which may confer advantages to the virus by making infected cells more challenging for the immune system to detect, thereby enhancing HPV's adaptability to the host environment. The polymorphism analysis of HPV-35 E6, E7 contributes to the enrichment of α-9 HPV data in Sichuan China, which is instrumental in improving the effectiveness of clinical detection. Furthermore, these findings provide a relevant theoretical foundation for the prevention and treatment of HPV-related diseases.


Assuntos
Proteínas Oncogênicas Virais , Infecções por Papillomavirus , Vacinas contra Papillomavirus , Humanos , Proteínas Oncogênicas Virais/genética , Proteínas Oncogênicas Virais/imunologia , Feminino , China , Infecções por Papillomavirus/virologia , Infecções por Papillomavirus/prevenção & controle , Vacinas contra Papillomavirus/imunologia , Vacinas contra Papillomavirus/genética , Polimorfismo Genético , Proteínas E7 de Papillomavirus/genética , Proteínas E7 de Papillomavirus/imunologia , Genótipo , Adulto , Neoplasias do Colo do Útero/virologia , Neoplasias do Colo do Útero/prevenção & controle , Epitopos/imunologia , Epitopos/genética , Alphapapillomavirus/genética , Alphapapillomavirus/imunologia , Alphapapillomavirus/classificação , Pessoa de Meia-Idade , Mutação , Papillomavirus Humano
2.
Cell Mol Biol (Noisy-le-grand) ; 69(12): 65-71, 2023 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-38063116

RESUMO

The objective of this work was to explore the application value of a new type of fluorescent nucleic acid isothermal amplification (SAT) to detect EV/EV71/CA16-SAT in children with hand-foot-mouth disease (HFMD). For this purpose, from March 2017 to September 2019, Chengdu Children's Specialized Hospital collected throat swabs from children with clinical manifestations of hand, foot and mouth disease, and used SAT technology to screen and detect universal enterovirus (EV) nucleic acid (There were 1860 children with EV-RNA) positive. Patients who are EV-RNA positive at any time: first use the same throat swab specimen to detect EV71/CA16-RNA; secondly, collect venous blood and use the colloidal gold method to detect IgM antibodies in EV71/CA16 serum. The patients with positive EV71/CA16-RNA or EV71/CA16-IgM (or both) were repeated the above two methods 2 weeks and 4 weeks after standard treatment for review and comprehensive analysis. Results showed that 763 cases were enrolled for the first time: 59.76% were male and 40.24% were female; the age ranged from 1 month to 13 years, of which 69.06% were from 1 to 4 years old; CA16-RNA positive 56.23%, EV71-RNA positive 21.89%, CA16/EV71 -RNA were all positive in 1.57%; CA16-IgM was positive in 64.48%, EV71-IgM was positive in 54.26%, and CA16/EV71-IgM were both positive in 18.74%. After 2 weeks, 722 cases were reexamined: 26.73% were positive for CA16-RNA, 7.89% were positive for EV71-RNA, 0.28% were both positive for CA16/EV71-RNA; 66.21% were positive for CA16-IgM, 51.52% were positive for EV71-IgM, and IgM were all positive in 17.73%. Four weeks later, 489 cases were reexamined: among them, CA16-RNA positive 5.73% of which were positive for EV71 color RNA (0.005%), and 12.68% of them were all positive for EV71lym. The strategy of combining SAT technology and colloidal gold method to detect EV/EV71/CA16 nucleic acid (RNA) and serum IgM antibody in children HFMD can improve the early detection rate and accuracy of HFMD; According to the comprehensive analysis of the detection results of children with HFMD at the early stage, 2 weeks and 4 weeks of the present study, it is suggested that EV/EV71/CA16-SAT nucleic acid detection can be used to judge the prognosis, follow-up treatment, set isolation time, return students to school, and community management in children with HFMD. and prevention and control have more clinical application value.


Assuntos
Enterovirus Humano A , Infecções por Enterovirus , Enterovirus , Doença de Mão, Pé e Boca , Ácidos Nucleicos , Criança , Humanos , Masculino , Feminino , Lactente , Doença de Mão, Pé e Boca/diagnóstico , Enterovirus/genética , Enterovirus Humano A/genética , RNA , Antígenos Virais , Imunoglobulina M , Coloide de Ouro , China
3.
Virol J ; 19(1): 14, 2022 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-35057815

RESUMO

BACKGROUND: The Alphapapillomavirus 9 (α-9 HPV) is a member of the Alphapapillomavirus genus and Papillomaviridae family. These viruses are almost all carcinogenic HPV, which is closely related to 75% of invasive cervical cancer worldwide, and has a high prevalence in Sichuan. The carcinogenic function is mainly realized by its E6 oncoprotein. METHODS: Cell samples were collected by cervical scraped for HPV detecting and typing. HPV-16, HPV-31, HPV-33, HPV-52, HPV-58 5 α-9 genus HPV subtype positive samples were selected, their E6 gene was sequenced and analyzed. The positive selection sites of HPV E6 genes were estimated by PAML 4.8 server. The secondary and tertiary structure of E6 protein were predicted by PSIPred and Swiss-model. The T-cell antigen epitopes of E6 protein were predicted by IEDB. RESULTS: α-9 HPV has a high prevalence in Sichuan, China. From 2012 to 2017, 18,067 cell cervical samples were collected, and 3135 were detected with α-9 HPV infection. Among which, 250 cases HPV-16 E6, 96 cases HPV-31 E6, 216 cases HPV-33 E6, 288 cases HPV-52 E6 and 405 cases HPV-58 E6 were successfully amplified, 17, 6, 6, 13, and 4 non-synonymous nucleotide mutations were respectively detected in HPV-16, 31, 33, 52, and 58 E6, 7 positive selection sites of α-9 HPV E6 were selected out (D32E of HPV-16 E6, K35N, K93N and R145I of HPV-33 E6, K93R of HPV-52 E6, K93N and R145K of HPV-58 E6). The structure and antigen epitopes of E6 protein with amino acid substitution differ from those of wild-type E6 protein, especially for the mutation located in the E6 positive selection site. CONCLUSIONS: HPV E6 nucleotide non-synonymous mutation in the positive selection site influence the protein structure and decrease the antigen epitopes affinity of the E6 protein overall, making it more difficult for the HPV-infected cells to be detected by the immune system, and enhancing the HPV adaptability to the environment. Mutations influence the validity of HPV clinical diagnostic probes, the polymorphism analysis of α-9 HPV E6 enrich the data of HR-risk HPV in Sichuan China, and the detection probes designed with the polymorphism data in mind can improve the efficiency of clinical detection; Mutations influence epitopes affinity, the association of E6 polymorphism and epitope affinity can improve the design of therapeutic vaccine with good immunity and high generality antigen epitope; The above study all provide a good theoretical basis for the prevention and treatment of HPV-related diseases.


Assuntos
Alphapapillomavirus , Proteínas Oncogênicas Virais , Proteínas Repressoras , Alphapapillomavirus/genética , China/epidemiologia , Epitopos de Linfócito T/genética , Feminino , Papillomavirus Humano 16 , Humanos , Proteínas Oncogênicas Virais/química , Proteínas Oncogênicas Virais/genética , Infecções por Papillomavirus , Filogenia , Polimorfismo de Nucleotídeo Único , Proteínas Repressoras/química , Proteínas Repressoras/genética , Neoplasias do Colo do Útero
4.
Virol J ; 18(1): 72, 2021 04 08.
Artigo em Inglês | MEDLINE | ID: mdl-33832494

RESUMO

BACKGROUND: Human papillomavirus type 39 associated with genital intraepithelial neoplasia and invasive cancers, has a high prevalence in Southwest China. HPV E6, E7 are two main papillomavirus oncoproteins, closely relate to the function of HPV immortalization, cell transformation, and carcinogenesis. L1 is the major capsid protein, can reflect the replication status of the virus in cells and the progression of cervical lesions. The purpose of this study is to reveal the prevalence of HPV 39 and the genetic polymorphisms of HPV39 based on E6, E7 and L1 gene in southwest China. METHODS: Cell samples were collected by cervical scraped for HPV detecting and typing, and HPV39 positive samples were selected out. Important E6, E7 and L1 genes of HPV39 were sequenced and analyzed for the study of HPV39 genetic polymorphisms. Phylogenetic trees were constructed by Maximum-likelihood and Kimura 2-parameters methods in Molecular Evolutionary Genetics Analysis version 6.0. The selection pressures of E6, E7 and L1 genes were estimated by Datamonkey web server. The secondary and three-dimensional structure of HPV39 E6, E7 proteins were created by sopma server and SWISS-MODEL software. RESULTS: 344 HPV39 positive samples were selected from 5718 HPV positive cell samples. Among HPV39 E6-E7 sequences, 20 single nucleotide mutations were detected, including 10 non-synonymous and 10 synonymous mutations; 26 single nucleotide mutations were detected in HPV39 L1 sequences, including 7 non-synonymous and 19 synonymous mutations respectively. 11 novel variants of HPV39 E6-E7 (5 in E6 and 6 in E7) and 14 novel variants of HPV39 L1 were identified in this study. A-branch was the most frequent HPV39 lineage in southwest China during our investigation. Selective pressure analysis showed that codon sites 26, 87, 151 in E6 and 75, 180, 222, 272, 284, 346, 356 in L1 were positively selected sites, as well as codon sites 45, 138, 309, 381 were negative selection sites in L1 gene, E7 has neither positive selection sites nor negative selection sites. A certain degree of secondary and three-dimensional structure dislocation was existed due to the non-synonymous mutations. CONCLUSIONS: Amino acid substitution affected the secondary and three-dimensional structure of HPV39, and resulting in the differences of carcinogenic potential and biological functions as well as the immune response due to the antigen epitopes difference, the antigen epitopes with stronger adaptability in Southwest will be screened out based on the above research results for the later vaccine development. And gene polymorphism of HPV39 in Southwest China may improve the effectiveness of clinical test and vaccine design, specifically for women in Southwest China.


Assuntos
Alphapapillomavirus , Genes Virais , Variação Genética , Infecções por Papillomavirus , Alphapapillomavirus/genética , China , Análise Mutacional de DNA , Epitopos , Feminino , Humanos , Proteínas Oncogênicas Virais/genética , Infecções por Papillomavirus/virologia , Filogenia , Desenvolvimento de Vacinas
5.
Virol J ; 18(1): 94, 2021 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-33941222

RESUMO

BACKGROUND: Variations in human papillomavirus (HPV) E6 and E7 have been shown to be closely related to the persistence of the virus and the occurrence and development of cervical cancer. Long control region (LCR) of HPV has been shown multiple functions on regulating viral transcription. In recent years, there have been reports on E6/E7/LCR of HPV-16 and HPV-58, but there are few studies on HPV-52, especially for LCR. In this study, we focused on gene polymorphism of the HPV-52 E6/E7/LCR sequences, assessed the effects of variations on the immune recognition of viral E6 and E7 antigens, predicted the effect of LCR variations on transcription factor binding sites and provided more basic date for further study of E6/E7/LCR in Chengdu, China. METHODS: LCR/E6/E7 of the HPV-52 were amplified and sequenced to do polymorphic and phylogenetic analysis. Sequences were aligned with the reference sequence by MEGA 7.0 to identify SNP. A neighbor-joining phylogenetic tree was constructed by MEGA 7.0, followed by the secondary structure prediction of the related proteins using PSIPRED 4.0. The selection pressure of E6 and E7 coding regions were estimated by Bayes empirical Bayes analysis of PAML 4.9. The HLA class-I and II binding peptides were predicted by the Immune Epitope Database server. The B cell epitopes were predicted by ABCpred server. Transcription factor binding sites in LCR were predicted by JASPAR database. RESULTS: 50 SNP sites (6 in E6, 10 in E7, 34 in LCR) were found. From the most variable to the least variable, the nucleotide variations were LCR > E7 > E6. Two deletions were found between the nucleotide sites 7387-7391 (TTATG) and 7698-7700 (CTT) in all samples. A deletion was found between the nucleotide sites 7287-7288 (TG) in 97.56% (40/41) of the samples. The combinations of all the SNP sites and deletions resulted in 12 unique sequences. As shown in the neighbor-joining phylogenetic tree, except for one belonging to sub-lineage C2, others sequences clustered into sub-lineage B2. No positive selection was observed in E6 and E7. 8 non-synonymous amino acid substitutions (including E3Q and K93R in the E6, and T37I, S52D, Y59D, H61Y, D64N and L99R in the E7) were potential affecting multiple putative epitopes for both CD4+ and CD8+ T-cells and B-cells. A7168G was the most variable site (100%) and the binding sites for transcription factor VAX1 in LCR. In addition, the prediction results showed that LCR had the high probability binding sites for transcription factors SOX9, FOS, RAX, HOXA5, VAX1 and SRY. CONCLUSION: This study provides basic data for understanding the relation among E6/E7/LCR mutations, lineages and carcinogenesis. Furthermore, it provides an insight into the intrinsic geographical relatedness and biological differences of the HPV-52 variants, and contributes to further research on the HPV-52 therapeutic vaccine development.


Assuntos
Alphapapillomavirus , Proteínas Oncogênicas Virais , Proteínas E7 de Papillomavirus , Infecções por Papillomavirus , Filogenia , Alphapapillomavirus/genética , Teorema de Bayes , China , Epitopos de Linfócito B , Feminino , Humanos , Proteínas Oncogênicas Virais/genética , Proteínas E7 de Papillomavirus/genética , Infecções por Papillomavirus/virologia , Fatores de Transcrição , Neoplasias do Colo do Útero/virologia , Desenvolvimento de Vacinas
6.
Virol J ; 17(1): 106, 2020 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-32677948

RESUMO

BACKGROUND: Long control region (LCR) of human papillomavirus (HPV) has shown multiple functions on regulating viral transcription. The variations of LCR related to different lineages/sub-lineages have been found to affect viral persistence and cervical cancer progression differently. In this study, we focused on gene polymorphism of HPV16/18/58 LCR to assess the effect variations caused on transcription factor binding sites (TFBS) and provided more data for further study of LCR in Southwest China. METHODS: LCR of HPV16/18/58 were amplified and sequenced to do polymorphic and phylogenetic anlysis. Sequences of each type were aligned with the reference sequence by MEGA 6.0 to identify SNPs. Neighbor-joining phylogenetic trees were constructed using MEGA 6.0. Transcription factor binding sites were predicted by JASPAR database. RESULTS: The prevalence of these three HPVs ranked as HPV16 (12.8%) > HPV58 (12.6%) > HPV18 (3.5%) in Chengdu, Southwest China. 59 SNPs were identified in HPV16-LCR, 18 of them were novel mutations. 30 SNP were found in HPV18-LCR, 8 of them were novel. 55 SNPs were detected in HPV58-LCR, 18 of them were novel. Also, an insertion (CTTGTCAGTTTC) was detected in HPV58-LCR between position 7279 and 7280. As shown in the neighbor-joining phylogenetic trees, most isolates of HPV16/18/58 were clustered into lineage A. In addition, one isolate of HPV16 was classified into lineage C and 3 isolates of HPV58 were classified as lineage B. JASPAR results suggested that TFBS were potentially influenced by 7/6 mutations on LCR of HPV16/18. The insertion and 5 mutations were shown effects in LCR of HPV58. CONCLUSION: This study provides more data for understanding the relation among LCR mutations, lineages and carcinogenesis. It also helps performing further study to demonstrate biological function of LCR and find potential marker for diagnosis and therapy.


Assuntos
Papillomavirus Humano 16/genética , Papillomavirus Humano 18/genética , Papillomaviridae/genética , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/virologia , Filogenia , Adulto , Sítios de Ligação , China/epidemiologia , Feminino , Regulação Viral da Expressão Gênica , Humanos , Pessoa de Meia-Idade , Mutação , Papillomaviridae/classificação , Polimorfismo Genético , Prevalência , Lesões Intraepiteliais Escamosas/epidemiologia , Lesões Intraepiteliais Escamosas/virologia , Fatores de Transcrição/genética , Neoplasias do Colo do Útero/epidemiologia , Neoplasias do Colo do Útero/virologia , Adulto Jovem
7.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 51(1): 97-101, 2020 Jan.
Artigo em Zh | MEDLINE | ID: mdl-31950797

RESUMO

OBJECTIVE: To analyse potential genetic cause of a family affected with hereditary elliptocytosis (HE). METHODS: Peripheral blood samples from this HE family were collected. Targeted capture and high-throughput sequencing of 4 813 genetic disease-associated genes was performed in four members of the family. Possible causative genetic variation was obtained and further confirmed by Sanger sequencing. Fifty healthy control subjects were recruited for detection of the candidate variation. RESULTS: High-throughput sequencing detected a nonsense mutation c.1215G>A(p.Trp405Ter)in exon 13 of the EPB41 gene in the proband and his mother presenting with moderate anemia. The pathogenicity of this loss-of-function mutation is very strong, because the G→A transition leads to introduce the premature stop codon instead of tryptophan codon at position 405, which producing a truncating protein with loss of important functional domains. This causative mutation is extremely rare in the population, and it has not yet been reported. The grandmother of the proband was heterozygous for the same mutation. Genotype-phenotype cosegregation was observed in this family. This mutation was not found in the 50 unrelated healthy controls. CONCLUSION: The c.1215G>A mutation of the EPB41 gene probably accounts for the disease in this HE family. This study reports a pathogenic EPB41 mutation in a Chinese HE family for the first time.


Assuntos
Proteínas do Citoesqueleto , Eliptocitose Hereditária , Proteínas de Membrana , Mutação , Proteínas do Citoesqueleto/genética , Eliptocitose Hereditária/genética , Heterozigoto , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Proteínas de Membrana/genética , Linhagem
8.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 51(5): 685-690, 2020 Sep.
Artigo em Zh | MEDLINE | ID: mdl-32975085

RESUMO

OBJECTIVE: To analyze the epidemiological characteristics of hand, foot and mouth disease (HFMD) in Sichuan Province from 2013 to 2019. To study the correlation between meteorological factors and the incidence of HFMD and construct a prediction model. METHODS: The HMFD surveillance data and meteorological data from 2013 to 2019 in Sichuan Province were collected through the Chinese Center for Disease Control and Prevention and the China Meteorological data Network. Spearman correlation was used to analyze the relationship between HFMD incidence and meteorological factors. Multiple regression model and support vector regression (SVR) model were used to construct HFMD incidence prediction models respectively. RESULTS: A total of 615 840 cases of HFMD and 81 deaths were reported from 2013 to 2019. The average annual incidence rate was 107.31/105, and the mortality rate was 0.16/106. Spearman correlation analysis showed that the monthly incidence rate of HFMD was correlated with monthly average relative humidity (r=0.342), monthly average temperature (r=0.284), monthly average water vapor pressure (r=0.304) and monthly average days of precipitation (r=0.259). The prediction effect of the SVR model (R2=0.836) was better than the multiple regression model (R2=0.375). The SVR model provided a good fit to the monthly incidence of HFMD from 2013 to 2018, and can predict the peak incidence of HFMD in 2019. CONCLUSION: Relative humidity has the greatest influence on the incidence of HFMD. The fitting value of SVR model is in good agreement with the actual value, which is valuable in predicting the incidence of HFMD in Sichuan Province.


Assuntos
Doença de Mão, Pé e Boca , China/epidemiologia , Doença de Mão, Pé e Boca/epidemiologia , Humanos , Incidência , Conceitos Meteorológicos , Temperatura
9.
Virol J ; 16(1): 72, 2019 05 28.
Artigo em Inglês | MEDLINE | ID: mdl-31138240

RESUMO

BACKGROUND: Human papillomavirus (HPV) E6 and E7 oncoproteins play a crucial role in HPV-related diseases, such as cervical cancer, and can be used as ideal targets for therapeutic vaccines. Human leukocyte antigen (HLA) participates in the immune response to block HPV infection and invasion by its target/recognition function. HPV-33 and HPV-58 are highly prevalent among Chinese women. Therefore, it is of great significance to study the E6 and E7 region-specific gene polymorphisms of HPV-33 and HPV-58 in Southwest China and to identify ideal epitopes for vaccine design. Both HPV-33 and HPV-58 belong to α-9 genus HPV and are highly homologous, so their correlations are included in our research. METHODS: To study the E6 and E7 variations and polymorphisms of HPV-33 and HPV-58 in Southwest China, we collected samples, extracted and sequenced DNA, and identified variants. Nucleotide sequences were translated into amino acids by Mega 6.0 software. The physical/chemical properties, amino acid-conserved sequences and secondary structure of protein sequences were analysed by the Protparam server, ConSurf server and PSIPRED software. The T and B cell epitopes of the E6/E7 reference and variant sequences in HPV-33 and HPV-58 were predicted by the Immune Epitope Database (IEDB) analysis server and the ABCpred server, respectively. RESULTS: Five and seven optimal HLA-I restricted T cell epitopes were selected from HPV-33 and HPV-58 E6, respectively, and these optimal epitopes are mainly located in 41-58EVYDFAFADLTVVYREGN of HPV-33 E6 and 40-60SEVYDFVFADLRIVYRDGNPF of HPV-58 E6. Six optimal HLA-I-restricted T cell epitopes were selected from HPV-33 and HPV-58 E7, and these epitopes are mainly located in 77-90RTIQQLLMGTVNIV of HPV-33 E7 and 78-91RTLQQLLMGTCTIV of HPV-58 E7. CONCLUSIONS: HPV-33/HPV-58 E6/E7 gene polymorphisms and T/B cell epitopes of their reference and variant sequences were studied, and candidate epitopes were selected by bioinformatics techniques for therapeutic vaccine design for people in Southwest China. This study was the first to investigate the correlation of epitopes between HPV-33 and HPV-58. After experimental validation, these selected epitopes will be employed to induce a wide range of immune responses in heterogeneous HLA populations.


Assuntos
Epitopos/imunologia , Variação Genética , Papillomaviridae/imunologia , Proteínas E7 de Papillomavirus/genética , Proteínas E7 de Papillomavirus/imunologia , Infecções por Papillomavirus/imunologia , Epitopos/genética , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito T/imunologia , Feminino , Humanos , Infecções por Papillomavirus/epidemiologia , Filogenia , Neoplasias do Colo do Útero/virologia
10.
Zhonghua Nan Ke Xue ; 23(12): 1069-1074, 2017 Dec.
Artigo em Zh | MEDLINE | ID: mdl-29738175

RESUMO

OBJECTIVE: To investigate the association of a very common mutation of c.144delC in the aurora kinase C (AURKC) gene with idiopathic teratozoospermia in Chinese infertile men in Sichuan. METHODS: Using polymerase chain reaction (PCR) and next-generation sequencing, we analyzed the correlation between c.144delC polymorphism of the AURKC gene and male infertility in 98 idiopathic teratozoospermia patients in comparison with 162 normal fertile men. RESULTS: Neither c.144delC mutation nor other meaningful mutations were detected in the AURKC gene in the 98 idiopathic teratozoospermia patients or the 162 normal controls. CONCLUSIONS: Teratozoospermia is not correlated with c.144delC mutation in the AURKC gene in the men of the Sichuan area. Therefore, large-scale genotyping of the AURKC gene may not be necessary clinically among Chinese patients with idiopathic teratozoospermia.


Assuntos
Aurora Quinase C/genética , Mutação/genética , Polimorfismo Genético , Teratozoospermia/genética , Humanos , Masculino , Espermatozoides
11.
Virol J ; 13(1): 167, 2016 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-27717385

RESUMO

BACKGROUND: Cervical cancer is associated with infection by certain subtypes of human papillomavirus (HPV). The L1 protein comprising HPV vaccine formulations elicits high-titre neutralizing antibodies and confers protection against specific HPV subtypes. HPV L2 protein is an attractive candidate for cross-protective vaccines. HPV-33 and HPV-58 are very prevalent among Chinese women. METHODS: To study the gene intratypic variations and polymorphisms of HPV-33 and HPV-58 L1/L2 in Sichuan China, HPV-33 and HPV-58 L1 and L2 genes were sequenced and compared with other genes submitted to GenBank. Phylogenetic trees were constructed by maximum-likelihood and the Kimura 2-parameters methods (MEGA 6). The secondary structure was analyzed by PSIPred software, and HPV-33 and HPV-58 L1 homology models were created by SWISS-MODEL software. The selection pressures acting on the L1/L2 genes were estimated by PAML 4.8. RESULTS: Among 124 HPV-33 L1 sequences 20 single nucleotide mutations were observed included 8/20 non-synonymous and 12/20 synonymous mutations. The 101 HPV-33 L2 sequences included 12 single nucleotide mutations comprising 7/12 non-synonymous and 5/12 synonymous mutations. The 223 HPV-58 L1 sequences included 32 single nucleotide mutations comprising 9/32 non-synonymous and 23/32 synonymous mutations. The 201 HPV-58 L2 sequences comprised 26 single nucleotide mutations including 9/26 non-synonymous and 17/26 synonymous mutations. Selective pressure analysis showed that most of the common non-synonymous mutations showed a positive selection. HPV-33 and HPV-58 L2 were more stable than HPV-33 and HPV-58 L1. CONCLUSIONS: HPV-33 and HPV-58 L2 were better candidates as clinical diagnostic targets compared with HPV-33 and HPV-58 L1. Clinical diagnostic probes and second-generation polyvalent vaccines should be designed on the basis of the unique sequence of HPV-33 and 58 L1/L2 variations in Sichuan, to improve the accuracy of clinical detection and the protective efficiency of vaccines.


Assuntos
Proteínas do Capsídeo/genética , Papillomaviridae/genética , Infecções por Papillomavirus/virologia , Vacinas contra Papillomavirus/genética , Polimorfismo Genético , Adolescente , China/epidemiologia , Análise por Conglomerados , DNA Viral/química , DNA Viral/genética , Feminino , Humanos , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/epidemiologia , Filogenia , Estrutura Secundária de Proteína , Estrutura Terciária de Proteína , Seleção Genética , Análise de Sequência de DNA , Adulto Jovem
12.
J Assist Reprod Genet ; 33(2): 181-7, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26584823

RESUMO

PURPOSE: The purpose of this research was to study the association between the single nucleotide polymorphisms (SNPs) of the tektin-t gene and idiopathic asthenozoospermia. METHODS: We conducted sequence analyses of the tektin-t gene in 104 idiopathic asthenozoospermia and 102 fertile men with normospermic parameters in Sichuan, China. RESULTS: In this study, we found that allele 136 T (odds ratio [OR] 1.745, 95 % confidence interval [CI] 1.146-2.655, P = 0.009) was significantly increased in idiopathic asthenozoospermic patients compared with fertile men. This mutation substitutes a highly conserved arginine at position 46 to cysteine. Moreover, PolyPhen-2 analysis predicted that this variant was "probably damaging". In addition, a novel heterozygous mutation, R207H (c.620G >A), was detected in five asthenozoospermic patients, while there was no detection of this genotype among the fertile candidates, indicating that the mutation was located within a conserved domain predicted by PolyPhen-2 analysis as "probably damaging" to the protein. CONCLUSIONS: These results suggested that tektin-t variants (Arg/Cys + Cys/Cys) were probably one of the high risk genetic factors for idiopathic asthenozoospermia among males in Sichuan, China, while the R207H polymorphism may be associated with idiopathic asthenozoospermia risk.


Assuntos
Astenozoospermia/genética , Estudos de Associação Genética , Proteínas dos Microtúbulos/genética , Motilidade dos Espermatozoides/genética , Adulto , Alelos , Astenozoospermia/patologia , China , Genótipo , Humanos , Masculino , Mutação , Polimorfismo de Nucleotídeo Único , Fatores de Risco , Espermatozoides/patologia
13.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 32(2): 269-73, 2015 Apr.
Artigo em Zh | MEDLINE | ID: mdl-25863102

RESUMO

OBJECTIVE: To assess the association of single nucleotide polymorphisms (SNPs) of ubiquitin-specific protease 8 gene (USP8) with male infertility among ethnic Han Chinese from Sichuan. METHODS: A total of 316 infertile males were recruited (case group), which included 72 severe oligozoospermic (SO) cases and 244 non-obstructive azoospermic (NOA) cases. The control group consisted of 149 fertile males. The genotypes of 4 SNPs (rs2241769, rs11857513, rs7174015 and rs3743044) were determined with a Sequenom MassArray technique. The frequencies of genotype, allele and haploptye were analyzed. RESULTS: No significant difference was detected in the allelic or genotypic frequencies of the 4 SNPs between the two groups (P>0.05). Based on linkage disequilibrium analysis and haplotype construction, the frequency distribution of haplotype CAAG showed a significant difference between non-obstructive azoospermic patients and the controls (P=0.021). CONCLUSION: The 4 SNPs (rs2241769, rs11857513, rs7174015 and rs3743044) of USP8 gene may not be associated with male infertility in ethnic Hans from Sichuan. While the haplotype CAAG may be a down-regulating factor for the risk of NOA.


Assuntos
Povo Asiático/genética , Endopeptidases/genética , Complexos Endossomais de Distribuição Requeridos para Transporte/genética , Infertilidade Masculina/genética , Polimorfismo de Nucleotídeo Único , Ubiquitina Tiolesterase/genética , Adulto , Povo Asiático/etnologia , Azoospermia/genética , Sequência de Bases , Estudos de Casos e Controles , China/etnologia , Predisposição Genética para Doença/etnologia , Genótipo , Humanos , Infertilidade Masculina/etnologia , Masculino , Dados de Sequência Molecular
14.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 31(1): 69-73, 2014 Feb.
Artigo em Zh | MEDLINE | ID: mdl-24510567

RESUMO

OBJECTIVE: To assess the association of single nucleotide polymorphisms (SNPs) in exons of H2BFWT gene with male infertility in southwest China. METHODS: Three hundred and twelve infertile men and 211 fertile men were recruited. PCR was employed to amplify the target fragments of H2BFWT, and PCR products were sequenced. Prevalence of SNPs in the two groups was analyzed by statistical method. RESULTS: The detected SNPs have mainly distributed in the first exon of the H2BFWT gene. The ratios of 368G/A (rs553509) and -9C/T (rs7885967) were significantly higher in infertile group than fertile group. Additionally, a context-dependent effect was observed between 368G/A and -9C/T which the allele 368G combined with allele -9T will considerably increase the risk of male infertility. CONCLUSION: The present study has revealed that the SNPs in H2BFWT are associated with male infertility, and may increase the susceptibility of male infertility in southwest China.


Assuntos
Infertilidade Masculina/genética , Adulto , Alelos , Sequência de Bases , China , Humanos , Masculino , Polimorfismo de Nucleotídeo Único , Adulto Jovem
15.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 30(1): 102-5, 2013 Feb.
Artigo em Zh | MEDLINE | ID: mdl-23450492

RESUMO

OBJECTIVE: To assess the association between glutathione-S-transferase gene polymorphisms GSTT1, GSTM1 and GSTP1 and onset of azoospermia. METHODS: Multi-PCR was used to detect GSTM1 and GSTT1 gene deletions. Polymorphisms of GSTP1 were determined with restriction fragment length polymorphism (RFLP) method in 236 azoospermia patients and 142 healthy fertile male controls. RESULTS: The frequency of M1 (-/-) and P1 (Ile/Val or Val/Val) genotype was 24.65% in the control group, which was significantly higher than that of the patient group (15.68%, P=0.031). Frequency of M1 (-/-), T1 (+/+) and P1 (Ile/Val or Val/Val) genotype was 12.68% in the control group, which was significantly higher than that of the patient group (5.51%, P=0.014). CONCLUSION: The M1(-/-) and P1(Ile/Val or Val/Val) genotype and the M1(-/-), T1(+/+) and P1 (Ile/Val or Val/Val) genotype are associated with reduced risk of azoospermia in ethnic Chinese Han population.


Assuntos
Azoospermia/genética , Glutationa S-Transferase pi/genética , Glutationa Transferase/genética , Polimorfismo Genético , Adulto , Povo Asiático , Estudos de Casos e Controles , China , Genótipo , Humanos , Masculino , Fenótipo
16.
Yi Chuan ; 35(1): 73-8, 2013 Jan.
Artigo em Zh | MEDLINE | ID: mdl-23357267

RESUMO

Idiopathic azoospermia and oligospermia are one of the most important reasons for male infertility. Abnormal karyotype and azoospermia factor (AZF) microdeletion are two widely acknowledged reasons, but the most causes remain unclear. Y chromosome, as the male-specific chromosome, is closely related to the development of male reproductive system. To understand better the etiology of idiopathic azoospermia and oligospermia, we investigated the possible association between Y-haplogroup distributions and susceptibility to idiopathic azoospermia and severe oligospermia. Peripheral blood was collected from 193 men with normal reproductive history, 193 men with idiopathic azoospermia, and 72 men with idiopathic severe oligospermia. All the subjects underwent karyotyping and AZF deletion analysis to screen out those with AZF deletion and abnormal karyotype. The comparison of Y-haplogroup distribution between experimental group and control group was performed with SPSS V.18.0 software. Significant difference of Y-haplogroup distribution was observed in D1*, F*, K*, N1* and O3*(P=0.032, 0.022, 0.009, 0.009, 0.017, <0.05). The results suggest that Y chromosome haplogroup plays a important role in spermatogenic impairment.


Assuntos
Azoospermia/genética , Cromossomos Humanos Y/genética , Oligospermia/genética , Espermatogênese , Adulto , Povo Asiático/etnologia , Povo Asiático/genética , Azoospermia/etnologia , Azoospermia/fisiopatologia , China/etnologia , Predisposição Genética para Doença/etnologia , Humanos , Cariotipagem , Masculino , Pessoa de Meia-Idade , Oligospermia/etnologia , Oligospermia/fisiopatologia , Adulto Jovem
17.
J Mol Model ; 29(5): 138, 2023 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-37055578

RESUMO

CONTEXT: In the replication of SARS-CoV-2, the main protease (Mpro/3CLpro) is significant. It is conserved in a number of novel coronavirus variations, and no known human proteases share its cleavage sites. Therefore, 3CLpro is an ideal target. In the report, we screened five potential inhibitors (1543, 2308, 3717, 5606, and 9000) of SARS-CoV-2 Mpro through a workflow. The calculation of MM-GBSA binding free energy showed that three of the five potential inhibitors (1543, 2308, 5606) had similar inhibitor effects to X77 against Mpro of SARS-CoV-2. In conclusion, the manuscript lays the groundwork for the design of Mpro inhibitors. METHODS: In the virtual screening phase, we used structure-based virtual screening (Qvina2.1) and ligand-based virtual screening (AncPhore). In the molecular dynamic simulation part, we used the Amber14SB + GAFF force field to perform molecular dynamic simulation of the complex for 100 ns (Gromacs2021.5) and performed MM-GBSA binding free energy calculation according to the simulation trajectory.


Assuntos
Proteases 3C de Coronavírus , Inibidores de Protease de Coronavírus , Simulação de Dinâmica Molecular , SARS-CoV-2 , Humanos , Endopeptidases , Simulação de Acoplamento Molecular , Farmacóforo , SARS-CoV-2/enzimologia , Proteases 3C de Coronavírus/antagonistas & inibidores , Inibidores de Protease de Coronavírus/química , Inibidores de Protease de Coronavírus/farmacologia
18.
J Assist Reprod Genet ; 29(6): 521-7, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22415247

RESUMO

PURPOSES: To investigate the frequency and type of both chromosomal abnormalities and Y chromosome microdeletions and analyze their association with defective spermatogenesis in Chinese infertile men. METHODS: This is a single center study. Karyotyping using G-banding and screening for Y chromosome microdeletion by multiplex polymerase chain reaction(PCR)were performed in 200 controls and 1,333 infertile men, including 945 patients with non-obstructive azoospermia and 388 patients with severe oligozoospermia. RESULTS: Out of 1,333 infertile patients, 154(11.55%) presented chromosomal abnormalities. Of these, 139 of 945 (14.71%) were from the azoospermic and 15 of 388 (3.87%) from the severe oligozoospermic patient groups. The incidence of sex chromosomal abnormalities in men with azoospermia was 11.53% compared with 1.03% in men with severe oligozoospermia (P < 0.01). Also 144 of 1,333(10.80%) patients presented Y chromosome microdeletions. The incidence of azoospermia factor(AZF) microdeletion was 11.75% and 8.51% in patients with azoospermia and severe oligozoospermia respectively. Deletion of AZFc was the most common and deletions in AZFa or AZFab or AZFabc were found in azoospermic men. In addition, 34 patients had chromosomal abnormalities among the 144 patients with Y chromosome microdeletions. No chromosomal abnormality and microdeletion in AZF region were detected in controls. CONCLUSIONS: There was a high incidence (19.80%) of chromosomal abnormalities and Y chromosomal microdeletions in Chinese infertile males with azoospermia or severe oligozoospermia. These findings strongly suggest that genetic screening should be advised to infertile men before starting assisted reproductive treatments.


Assuntos
Aberrações Cromossômicas , Cromossomos Humanos Y , Infertilidade Masculina/genética , Povo Asiático/genética , Azoospermia/genética , Estudos de Casos e Controles , Deleção Cromossômica , Testes Genéticos , Humanos , Cariótipo , Masculino , Reação em Cadeia da Polimerase Multiplex , Oligospermia/genética , Aberrações dos Cromossomos Sexuais , Transtornos do Cromossomo Sexual no Desenvolvimento Sexual , Espermatogênese/genética
19.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 29(3): 289-92, 2012 Jun.
Artigo em Zh | MEDLINE | ID: mdl-22678791

RESUMO

OBJECTIVE: To rapidly detect SOX2 gene using primed in situ labeling (PRINS). METHODS: Human peripheral blood samples were cultured using an optimized method. Sequence of the SOX2 gene was amplified in situ with biotin-labeled specific primers and processed with a tyramide signal amplification (TSA) biotin system. Subsequently, fluorescence-stained signal was detected by streptavidin-Texas red. For the control group, MCF-10F cells were transfected with Lentivirus hSox2. RESULTS: By VideoTesT-FISH software analysis, the long arm of chromosome 3 in the experimental group showed a specific red fluorescence signal, whilst the control samples showed no specific signals for SOX2. Transfected MCF-10F cells showed various efficiency of SOX2 gene integration. CONCLUSION: PRINS utilizes a highly sensitive in situ PCR technique combined with fluorescence labeled oligodeoxynucleotides can synthesize probes in situ, thus greatly reducing the cost of probe and time for detection. It can facilitate identification and classification of induced pluripotent stem cells, and has many potential applications in this prospect.


Assuntos
Marcação in Situ com Primers/métodos , Fatores de Transcrição SOXB1/química , Humanos , Hibridização in Situ Fluorescente/métodos , Masculino
20.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 29(2): 184-7, 2012 Apr.
Artigo em Zh | MEDLINE | ID: mdl-22487830

RESUMO

OBJECTIVE: To investigate the prevalence and subtypes of microdeletions in azoospermia factor (AZF) region in infertile men from Sichuan in order to correlate genotypes with phenotypes. METHODS: Multiplex-PCR was used to detect sequence tagged sites (STS) of AZF microdeletions in 1011 infertile men including 713 cases of non-obstructive azoospermia and 298 cases of severe oligospermia. RESULTS: The overall prevalence of microdeletions was 10.48% (106/1011), and the deletion rates were 11.08% (79/713) in non-obstructive azoospermia and 9.06% (27/298) in severe oligospermia. Complete AZFa or AZFb deletions were associated with azoospermia, whereas AZFc deletion (60.38%) was the most frequent deletion. The deletions were associated with variable spermatogenic phenotypes, and 37.50% of the patients with a deletion had sperms in the ejaculate. A mild decline in sperm concentration was found in two cases with partial AZFb deletion and one case with partial AZFb-c deletion. CONCLUSION: Deletions of the AZFc region were most commonly found in our patients. All cases with complete AZFa or AZFb deletions and a proportion of cases with AZFc deletion were associated with azoospermia. Our study has provided more insight into the genotype-phenotype correlation, and confirmed that Yq microdeletion screening has a significant value for the diagnosis for male infertility.


Assuntos
Azoospermia/genética , Deleção Cromossômica , Cromossomos Humanos Y , Infertilidade Masculina/genética , Adulto , Estudos de Associação Genética/métodos , Humanos , Masculino , Fenótipo , Adulto Jovem
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA