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Objective To analyze hepatitis B virus( HBV)genotype and the correlation between variation of P gene region and genotyping of Lamivudine-resistant HBV,in order to provide molecular virology basis for anti-virus individualized treatment to different HBV genotypes. Methods Serum markers and HBV DNA of 187 Lamivudine -resistant chronic HBV patients were detected by ELISA and fluorescent quantitative polymerase chain reaction. Moreover,DNA Star and viral genotyping tool were used. Results Of the 187 cases infected with HBV genotype B and C,51 cases were B gene (27. 27%),134 cases were C gene(71. 66%),and 2 cases were mixed infections(1. 07%). HBeAg levels were significantly different between genotype B and C(P<0. 05). P gene mutation results showed that genotype B dominated by YMDD mutation alone and genotype C dominated by YMDD and rtL180M mutation. Conclusion The main HBV genotype of this study is genotype C,and different genotypes may determine the variation patterns of P region which associated with resistance.
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Objective To develop a method detecting hMAM gene expression of breast cancer cell lines in peripheral blood.Methods The hMAM gene was cloned and confirmed by agarose gel electrophoresis and sequence analysise,the standard substance of real-time quantitative polymerase chain reaction (FQ-PCR )was prepared. The breast cancer cell lines MDA-MB453、MCF7 were cultured and the blood sample model was made.The hMAM expression of breast cancer cell lines blood model and 10 cases of breast cancer patient peripheral blood were detected by RT-PCR and FQ-PCR,GAPDH gene expression as contrast.Results The production of PCR was cloned and testified by sequence.The correlation coefficient(r) of standard curve line of FQ-PCR was -1.0. The x?s and CV were 718.9?120.5 and 16.7% respectively.The hMAM expression of one breast cancer cell lines MDA-MB453 in 104 blood cells could be detected, but not detected in the breast cancer cell lines MCF7. The hMAM expression was detected in 3 of 5 patients with sentinel nodes and in 1 of 5 patients with non-sentinel nodes.Conclusions It might be a good method for detecting breast cancer cell micrometastased in peripheral blood. The diffrernce of hMAM gene expression correlates with the type of breast cancer cell lines.
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OBJECTIVE To learn the molecular epidemiologic character of 21 genotypes of human papillomavirus(HPV).METHODS Totally 500 samples from women genital tract secretion were monitored by flow-through hybridization and gene chip,then studying the molecular epidemiologic character of HPV included HPV6,11,16,18,31,33,35,39,42,43,44,45,51,52,53,56,58,59,66,68,and CP8304.RESULTS The total positive rate of 21 genotypes of HPV was 53% in women cervicitis patients,high-risk HPV genotypes were 60% in all positive samples.Among 21 genotypes of HPV,the positive rate of HPV11 was the first(12.4%),but HPV16 positive rate was the first among high-risk genotypes(11.2%).Among the positive rates in different stage of years,the 21-30 years old group was the highest,the 31-40 years old group was the second,total of two groups was 76.6%.CONCLUSIONS The molecular epidemiologic results can be used in development of HPV vaccine,and in prevention and cure of HPV infection.