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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.
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Objective To establish an effective method for inducing mouse bone marrow mononuclear cells(MNCs) to differentiate into hepatocytes.Methods MNCs were isolated by gradient density centrifugation,and the cells were cultured in Iscove's Modified Dulbecco's medium supplemented with 10 % new bovine serum(NBS),20 ng/ml hepatocyte growth factor(HGF),10 ng/ml fibroblast growth factor-4(FGF-4) and 10 ng/ml oncostatin m(OSM) for 21 days' induction.The medium was changed every 3 days.Results When MNCs were cultured with HGF,FGF-4 and OSM,cuboidal morphology was observed,and cells also expressed marker genes specific for liver cells in a time-dependent manner.?-fetoprotein(AFP) and cytokeratin 19(CK19) were expressed on the day 7,and CK18 and TAT were detected on the day 14-21,in common with the immunofluoresence results.Differentiated cells further demonstrated these cells also acquired functional characteristics of hepatocytes.Conclusion Mouse MNCs can differentiate into hepatocytes when induced by HGF,FGF-4 and OSM.
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Objective To obtain the molecular epidemiological data of Human Papilloma virus(HPV) 16 and HPV 18 and screen new HPV strains. Methods Common type HPV, HPV16 and HPV18 were amplified by PCR and then cloned and sequenced. Results The infection rates of HPV16、 HPV18 accounted for 10.51% of total HPV infection rate in the area. The infection rate of HPV16 was higher than that of HPV18 ( P