ABSTRACT
Fertilization is a crucial step for origin of life.During Assisted Reproductive Technologies (ART),total fertilization failure is complex and unpredictable.Total fertilization failure may related to some abnormal cellular mechanistic events,such as:any stage of sperm and cumulus-oocyte-complexes penetration,sperm-zona pellucida binding / penetration,sperm-oocyte membrane binding,oocyte activation,sperm discondensation or pronuclear formation.Most of total fertilization failure could be solved by intracytoplasmic sperm injection.But oocytes of some patient still can't fertilize successfully,even though assisted oocyte activation be used.As for total fertilization failure patients in ART,combining the mature of oocyte,sperm quality and some trail to improve clinical protocol in later cycle may prevent failure to happen again.
ABSTRACT
Objective To observe the influence of patient′s age, and the number of transferred-good-quality-embryos on multiple gestation rates in in vitro fertilization and embryo transfer(IVF-ET) cycles. Methods In this retrospective study, a total of 4395 patients who transferred fresh embryo between Jan 2004 and Nov 2006 was analyzed. According to the age, the patients were divided into 2 groups: aged < 35 (3442 cycles) or aged ≥135(953 cycles). We regularly transferred 2 -3 embryos. If the patients had only one embryo, one was transferred. And those patients who had only 2 embryos, even if they were more than 35 years old or it would be the second time for them to transfer, were transferred 2 embryos. The influence of female age and the number of good quality embryos transferred on the multiple gestation rates in IVF-ET cycle was analyzed. Results (1)The multiple gestation rate of the groups of 1 good quality embryo,2 good quality embryos, or 3 good quality embryos transferred were 21.08% (35/166), 31.41% (413/1315), and 42. 37% (75/177), respectively in women aged < 35, with a significant difference between them. The pregnancy rates of these groups were 29. 64% (166/560) ,51.63% (1315/2547) ,and 52. 84% (177/335), respectively; there were no significant differences between 2 good quality embryos transferred group and 3 good quality embryos transferred group. (2) The multiple gestation rates of the groups of 1 good quality embryo,2 good quality embryos, or 3 good quality embryos transferred were 19. 51% (8/41) ,20. 65% (19/ 92) ,and 40.66% (74/182), respectively, in women aged ≥ 35; there were no significant differences between 1 good quality embryo transferred group and 2 good quality embryos transferred group. The pregnancy rates of these groups were 19. 07% (41/215), 33.70% (92/273), and 39. 14% (182/465), respectively; there were no significant differences between 2 good quality embryos transferred group and 3 good quality embryos transferred group. (3) The pregnancy rate of the patients aged <35 [48. 17% ( 1658/ 3442) ]was significantly higher than in women aged ≥35[33.05% (315/953) ]. Conclusion The transfer of 2 good quality embryos results in similar pregnancy rates and significantly reduced multiple gestation rates when compared to the transfer of 3 good quality embryos in women regardless of their ages.
ABSTRACT
Objective:To explore the influence factors on amplification of single cell duplex-nested PCR.Methods:The mutational loci region CD41-42 and IVS-Ⅱ654 of ?-globin gene were amplified by duplex-nested PCR with different combination of primers concentration, different Taq DNA polymerases, different neutralization buffers and with or without predenaturation at 98 ℃ before the PCR amplification in single lymphocyte or single blastomere, thus, to investigate the influence of these factors on the amplification efficiency of PCR.Results:TaKaRa EX Taq was the most efficient Taq DNA ploymerase among different Taq DNA ploymerases; primer pair R1+F1 at final concertration of 0.25 ?mol/L and R2+F2 at 0.3 ?mol/L were the most efficient ones in amplification among different combinations of primers concentrations; the amplification efficiency in neutralization buffer-1 (200 mmol/L Tricine) was obviously higher than that of neutralization buffer-2 (900 mmol/L Tris-HCl, pH 8.3/300 mmol/L KCl/200 mmol/L HCl)(P0.05). Conclusion:There were remarkable differences of the amplification efficiency of single cell duplex-nested PCR while using different combination of primers concentrations, different Taq DNA polymerases, different neutralization buffers. However, predenaturation at 98 ℃ before the single cell PCR amplification could not improve the PCR amplification efficiency