ABSTRACT
Objective To analyze the influential factors of clinical pregnancy rate of the frozen-thawed embryo transplantation. Methods The data of 3 192 FET patients in the reproductive medicine center of our hospital up to May 2014 were analyzed retrospectively. According to ages, reasons of infertility, types of infertility, duration of infertility, drug regimen, the number and the time of embryo transplantation, we divided these patients into six groups for comparing the clinical pregnancy rate. Results The FET clinical pregnancy rate of the under 35 years group was higher than the 35~39 years group and the over 39 years group (33.96%vs. 27.58%and 19.35%; P<0.05, respectively). The duration of infertility in the clinical pregnancy group was significantly shorter than the non-pregnancy group (P<0.05). The clinical pregnancy rate in the group with three embryos transplanted was higher than the group with only two embryos transplanted (41.01% vs. 28.75%; P < 0.05). Among the group with the age of over 40 years, those with three embryos transplanted had a higher clinical pregnancy rate than those with only one embryo transplanted (25.49% vs. 0.00%; P < 0.05). The clinical pregnancy rate of the frozen blastocyst transplantation group was higher than that of the cleavage-stage transplantation group (40.00%vs. 26.27%;P<0.05). Conclusion Age, infertility duration, the number and the time of frozen embryo transplantation may affect the clinical pregnancy rate among the FET patients. An individualized transplantation program based on age may improve the patient′s clinical pregnancy rate.
ABSTRACT
Objective To establish the HepG2 cell lines which can stably express and replicate hepatitis 13 virus (HBV).Methods One point two X unit length of HBV genome was cloned intn SalⅠsite of the eukaryotic expression vector pREP10 to construct the recombinant plasmid pREP-HBV. Human hepatoblastoma cell HepG2 was transfected with pREP-HBV by Lipofectamine 2000 and seh,cted by bygromycin at the concentration of 250?g/mL.HBsAg and HBeAg were monitored by enzyme linked immunosorbent assay (ELISA)kits.H13V particles presemed in supernatant were ex- amincd by electronic microscopy.DNA isolated from intracellular HBV core particles was analyzed by Southbern blot using HBV-specific probe.Results The recombinant vector pREP HBV containing 1.2?unit length of HBV DNA was constructed successfully.After transfection of pREP-HBV to HepG2 cells and consistently cultured in hygromycin selective medium.5 drug-resistant cell lines, RHBV1-RHBV5.were established,and all of them could stably express HBsAgand HBeAg.South ern blot analysis revealed that HBV could replicate in all cell lines,as confirmed by the presence of replicateintermediatc DNA in intracellular HBV core particles.Clustered 42 nm Dane particles as well as 22-26 nm spherical H13sAg particles in condensed cuhure supernatant were visualized by elec tronic microsopic analysis.Conclusion HepG2 ceil lines in which HBV can replicate and express specific antigens are successfully established.Up to now,the cells have been passaged every three days for 50 times.