Résumé
Objective@#To investigate the distribution of human papillomavirus (HPV) infection characteristics and genotypes in Shenyang area of Liaoning province.@*Methods@#HPV genes were detected in cervical exfoliated cells from 55, 548 patients by amplification and diversion hybridization.@*Results@#A total of 9, 566 patients were positive for HPV infection with a positive rate of 17.22%. Additionally, the positive rate of high risk HPV infection was 14.57% and the positive rate of single genotype HPV infection was 13.63%. Totally, 12, 360 HPV viruses were detected. Among them, 10, 879 HPV viruses were classified into high risk genotypes (10, 879 out of 12, 360, 88.02%). The genotypes in women with ages less than 20 were 16/11/6/51/58/52 genotypes; the susceptible HPV genotypes in other women were 16/58/52/53/39/51/81 genotypes.@*Conclusions@#HPV infections in Shenyang are mainly infections with high risk viruses and single infection. The infection rate and genotype distribution of HPV are different in different age groups. More suitable HPV vaccine prophylaxis can be taken according to the epidemic characteristics of HPV in this area.
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Objective To quantifiably measure the methylation frequency of 18 CpG sites in the 3′region of L1 gene and long control region(LCR) gene of HPVl6 DNA,and study the relationship between HPVl6 DNA methylation and severity of cervical lesions. Methods A total of 10 cases Normal/low?grade squamous intraepithelial lesion(Normal/LSIL),10 cases of high?grade squamous intraepithelial lesion(HSIL),and 10 cases of cervical cancer(CC)were recruited for the study. The relationship between severity of cervical lesions and HPV16 DNA methylation was analyzed by bisultlte?pyrosequencing. Results The methylation rate was highest in Normal/LSIL at position 7 089 located in 3′?L1,followed by CC. The low?est was found in HSIL. The difference in methylation percentage among the three lesions was significant(P=0.006). In 7 134,the proportion meth?ylation was also different among three groups(P=0.01),difference in methylation percentage between Normal/LSIL and CC,as well as Normal/LSIL and HSIL was significant(P=0.038,0.017). Conclusion The methylation status of CpG sites 7 089 and 7 134 in the 3′region of L1gene is asso?ciated with the severity of cervical disease. The quantification of HPV DNA methylation can be used for cervical disease screening in clinical samples.
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Objective To develop and fully validate an UPLC-MS/MS method for simultaneous quantification of six active in?gredients in Rhodiola extract including salidroside,rhodiosin,rhodionin,herbacetin,kaempferol and quercetin. Methods The chro?matographic separation of the analytes was achieved by using Zorbax Eclipse plus C18 column(2.1 mm×100 mm,3.5μm). The mobile phase consisted of 0.2%aqueous formic acid and acetonitrile. Analytes were separated using a linear gradient with a flow rate of 0.4 ml/min. The column temperature was set at 25℃. The mass spectrometric conditions were electrospray negative ionization(ESI)and the scan?ning mode was multi reaction monitoring(MRM). Results The calibration curves of the six active ingredients were linear over 10-20000,10-4000,10-4000,5-2000,10-2000,and 5-2000 ng/ml concentration ranges,respectively. The RSD of the instrument precision was within 3.31%. The current assay was stable and reproducible in quantification of all analytes with RSD within 5.07%and 4.05%,respectively. The average recovery of all analytes was in a range from 99.27%to 108.91%,with an acceptable RSD of no more than 4.92%. Conclusion The UPLC-MS/MS method could be successfully applied to simultaneously quantifying six major active in?gredients in Rhodiola extract with the acceptable specificity,sensitivity and reproducibility.