| Part I Analysis of the related factors of biochemical pregnancy inIVF-ETOBJECTIVETo investigate the relationship of pregnancy outcome with clinical characteristics, ovulation program,serum hormone levels,oocyte and embryo quality of pregnancy patients terminated in biochemical pregnancy(below brief name "biochemical pregnancy") and pregnancy patients which had single implantations at 6 weeks and ongoing singleton pregnancies at term(below brief name "continued pregnancy"). This investigative purpose is to try to look for which factors will affect or reflect to the occurrence of biochemical pregnancy in vitro fertilization and embryo transfer (IVF-ET),explore the reasons for the occurrence of biochemical pregnancy and provide a basis to improve the pregnancy outcome.MATERIALS AND METHODSA retrospective study was performed at Reproductive Medicine Center of Nanfang Hospital during January 2003 and April 2008.The study group included 94 biochemical pregnancy cycles,while the control group consisted of 177 cycles in patients who had single implantations at 6 weeks and ongoing singleton pregnancies at term.Different protocol were applied for superovulation and follicular development was monitored using serial vaginal ultrasound.Oocyte collection scheduled under ultrasound guidance 34～36 hours after HCG injection.IVF,ICSI or IVF+ICSI were performed based on the condition of semen.On the three day of oocyte collection, high-grade embryos were selected to transfer and treatment outcome was followed-up. The clinical characteristics,treatment protocols,oocyte and embryo quality were analyzed in the biochemical pregnancy cycle and control group.SPSS 13.0 statistical package was used for data analysis.Date was expressed by mean±standard deviation or percentage(%),and analyzed statistically using 2 Independent Samples Tests for single factor analysis and using non-conditional Logistic regression analysis for multivariate analysis.The accuracy of predictions of rate was analyzed using aχ2 test.Predictive critical value was determined by the ROC curve and Youden index.All P-values were two-sided,and P＜0.05 was considered statistically significant.RESULTS1.Single factor analysisContrasting to control group,biochemical pregnancy group had higher proportion of composition of women aged≥35 years,frequency of artificial abortion, initial and total dosage of Gn,but had lower antral follicle counting on the day of Gn starting,serum E2 concentration on the day of human chorionic gonadotropin(HCG) injection,the diversities were significant statistically(P＜0.05).The proportion of composition of Gn type,endometrial morphology on the day of HCG injection were also existed significant diversities between the two groups(P＜0.05).Significant diversities were existed that the number of oocyte picked up,Mâ…¡oocyte were decreased and the number of embryo transferred was increased in biochemical pregnancy group than in control group(P＜0.05).There were no significant diversities in the number of high-grade embryo,transferring at least embryos with gradeâ… and 7-8 blastomeres between the two groups(P＞0.05).2.Multivariate analysis and determining the number of Mâ…¡oocyte threshold We did a multi-factor analysis with differences in the factors including woman's age group,frequency of artificial abortion,Gn type,initial and total dosage of Gn, serum E2 concentration on the day of HCG injection,the number of antral follicle counting on the day of Gn starting,oocyte picked up,Mâ…¡oocyte and embryo transferred,endometrial morphology on the day of HCG injection.It showed that Gn type and the number of Mâ…¡oocyte were the independent factors to biochemical pregnancy.Drawing Mâ…¡oocytes for pregnancy diagnosis of ROC curve,ROC area under the curve is 0.630 and 95%confidence interval is 0.561-0.700,the diversities were significant statistically(P＜0.05).Corresponding to cut-off point of the number of Mâ…¡oocytes was 6.CONCLUSIONS1.1.The factors related to biochemical pregnancy in IVF-ET are very complicated and may be related with age,artificial abortion history,the number of antral follicle counting on the day of Gn starting,oocyte picked up,Mâ…¡oocyte and embryo transferred,Gn type,initial and total dosage of Gn,serum E2 concentration on the day of HCG injection,endometrial morphology on the day of HCG injection.Choosing individualized treatment program should be emphasized.It is effective in improving the quality of oocytes and embryos to decrease the dosage of GnRHa or Gn and use pure FSH in controlled ovarian hyperstimulation protocol.It may be beneficial in reducing the incidence of biochemical pregnancy to strengthen perinatal health care,to reduce unintended pregnancies and frequency of artificial abortion,to avoidance injury to endometrium,and to improve endometrial receptivity.2.Gn type and the number of Mâ…¡oocyte were the independent factors associating with biochemical pregnancy.If the number of oocyte retrieved was greater than or equal to 6,the incidence of biochemical pregnancy could be reduced. Partâ…¡The analysis of oocyte and embryo quality of biochemical pregnancy cycle in IVF-ETOBJECTIVETo investigate the relationship of pregnancy outcome with oocyte and embryo quality of pregnancy patients terminated in biochemical pregnancy and pregnancy patients which had single implantations at 6 weeks and ongoing singleton pregnancies at term.This investigative purpose is to try to look for which factors will affect or reflect to the occurrence of biochemical pregnancy in IVF-ET,explore the reasons for the occurrence of biochemical pregnancy and provide a basis to improve the pregnancy outcome.MATERIALS AND METHODSStudy group and control group in this part were required as the same as in the first part of the study.Different protocol were applied for superovulation and follicular development was monitored using serial vaginal ultrasound.Oocyte collection scheduled under ultrasound guidance 34～36 hours after HCG injection.IVF,ICSI or IVF+ICSI were performed based on the condition of semen.4-6 hours after retrieval,granulose cell in the outside layer of oocytes were removed and the oocyte morphous and maturation degree of oocytes were observed.The fertilization rate of oocytes and the status of embryo development were observed and recorded for all the infertile women.On the three day of oocyte collection,high-grade embryos were selected to transfer and treatment outcome was followed-up.The quality of embryos transferred was evaluated by scoring based on the embryo morphology in two-pronuclear stage and on the three day after oocyte collection.Oocyte and embryo quality were analyzed in the biochemical pregnancy cycle and control group.SPSS 13.0 statistical package was used for data analysis.Date was expressed by mean±standard deviation or percentage(%) and analyzed statistically using 2 Independent Samples Tests.The accuracy of predictions of rate was analyzed using aχ2 test.P＜0.05 was considered statistically significant. RESULTSThe comparison of basic characteristics and clinical characteristics in the two groups was the same as in the first part of the study.There were 28 ICSI or IVF + ICSI cycle and a total of 222 oocytes performed ICSI in biochemical pregnancy group.And there were 70 ICSI or IVF + ICSI cycle and a total of 586 oocytes performed ICSI in control group.Normal oocyte percentage were decreased in biochemical pregnancy group than control group,and it was significant in diversity of the percentage of abnormal polar body,perivitelline space,granular cytoplasm and smooth endoplasmic reticulum(SER) increased in biochemical pregnancy group than control group(P＜0.05).But there were no significance differences of the proportion of abnormal zona pellucida,reticulum incidence(RB),vacuolization between the two group(P＞0.05).Significant diversities were existed that the number of oocyte picked up,Mâ…¡oocyte were decreased in biochemical pregnancy group than in control group (P＜0.05).There were no significant diversities in oocyte retrieval rate,but Mâ…¡oocyte percentage was decreased in biochemical pregnancy group than in control group(P＜0.05).There were no significant diversities in the number of high-grade embryo,transferred embryo score,cleavage rate,high-grade embryo rate,transferring at least embryos with gradeâ… and 7-8 blastomeres rate between the two groups (P＞0.05).But the number of embryos transferred was more and frozen embryo rate was lower in biochemical pregnancy group than in control group(P＜0.05).CONCLUSIONS1.Oocyte quality in biochemical pregnancy cycle decreased,manifest mainly in dropped mature degree of cell nuclear and more abnormal morphous of oocytes. Oocyte quality had a direct impact on embryo quality and the differences of embryos quality which derived from Oocyte may be one of the main factors causing biochemical pregnancy.2.The decline of oocyte and embryo quality may be appear to chromosomal abnormalities of oocyte and embryo,particularly the increase in aneuploidy. Partâ…¢Analysis of the related factors impacting on pregnancy out come in the biochemical pregnancy follow-up treatment cycle in IVF-ETOBJECTIVETo investigate the information of biochemical pregnancy follow-up treatment cycle.This investigative purpose is to screen the patients who had experienced a biochemical pregnancy and may be obtain clinical pregnancy in the follow-up treatment cycle,and find out what reasons make her success.It was expected to be helpful for clinical treatment of the women who experienced a biochemical pregnancy.MATERIALS AND METHODSThis study included 29 women who experienced biochemical pregnancy cycle and underwent at least one additional cycle and 32 treatment cycle during January 2003 and April 2008,including 15 clinical pregnancy cycles and 17 non-pregnant cycle.The clinical characteristics,treatment protocols,oocyte and embryo quality were compared in the clinical pregnancy cycle with non-pregnancy cycle and previous biochemical pregnancy cycle.The research methods were required as the same as in the second part of the study.SPSS 13.0 statistical package was used for data analysis.Date was expressed by mean±standard deviation or percentage(%) and analyzed statistically using 2 Independent Samples Tests or 2 Related Samples Test.The accuracy of predictions of rate was analyzed using aχ2 test.P＜0.05 was considered statistically significant.RESULTS1.The comparison of patients who had a clinical pregnancy and patients who had a negative pregnancy test in subsequent treatment cycle.Female age,infertile time and body mass index were decreased and antral follicle counting during the early phase of menstrual cycle were increased in clinical pregnancy group than in non-pregnancy group,the diversities were significant statistically(P＜0.05).Contrasting to non-pregnancy group,clinical pregnancy group had lower initial and total dosage of Gn and had higher antral follicle counting on the day of Gn starting and serum E2 concentration on the day of HCG injection,the diversities were significant statistically(P＜0.05).Significant diversities were also present that the proportion of composition of Gn type between the two groups (P＜0.05).The number of oocytes picked up,Mâ…¡oocytes,high-grade embryo, embryos transferred and transferring embryos with gradeâ… and 7-8 blastomeres, transferred embryo score,cleavage rate,transferring at least embryos with gradeâ… and 7-8 blastomeres rate were higher in clinical pregnancy group than in non-pregnancy group(P＜0.05).There were no significant diversities in oocyte retrieval rate,Mâ…¡oocyte percentage,normal fertilization rate,high-grade embryo rate and frozen embryo rate between the two groups(P＞0.05).2.The comparison of patients in the biochemical pregnancy cycle previously and in the subsequent clinical pregnancy cycle who experienced a biochemical pregnancy in IVF-ET cycle previously and had a clinical pregnancy in the subsequent cycle.The number of embryos transferred with gradeâ… and 7-8 blastomeres and transferred at least embryos with gradeâ… and 7-8 blastomeres rate were higher in clinical pregnancy cycle than in biochemical pregnancy cycle previously,diversities were significant(P＜0.05).There were no significant diversities in controlled ovarian hyperstimulation protocol,antral follicle counting on the day of Gn starting,the duration,initial and total dosage of Gn,serum E2,P,LH concentration,thickness of endometrium on the day of HCG injection,the proportion of composition of Gn type, endometrial morphology on the day of HCG injection,fertilization methods and luteal support protocol,the number of oocytes picked up,Mâ…¡oocytes,high-grade embryo and embryos transferred,oocyte retrieval rate,Mâ…¡oocyte percentage,normal fertilization rate,Cleavage rate,high-grade embryo rate and frozen embryo rate between the two groups(P＞0.05).CONCLUSIONS1.The patients who are young or have better ovarian reserve function will have a greater likelihood of success in subsequent IVF cycle among the patients who had experienced a history of a biochemical pregnancy.2.It can improve embryo quality and pregnancy rate through the adjustment of ovarian hyperstimulation program and improving endocrine and metabolic environment for the patients who had experienced a history of a biochemical pregnancy and have better ovarian reserve function. Partâ…£Analysis of predictive value of biochemical pregnancy for pregnancy outcome in subsequent IVF cycleOBJECTIVEThe clinical data in the next cycle for women who experienced biochemical pregnancy after IVF-ET were compared with the clinical data for those who had failed to conceive after IVF-ET.The purpose is to try to find out if patients who experienced a biochemical pregnancy after IVF-ET have a greater likelihood of subsequent IVF cycle when compared with patients who fail to conceive.It was expected to be helpful for obtaining clinical information to guide the women who experienced a biochemical pregnancy to make reasonable selection.MATERIALS AND METHODSThis study included 29 women who experienced biochemical pregnancy cycle and underwent at least one additional cycle of IVF and 29 subsequent first treatment cycle during January 2003 and April 2008.Women who had failed to conceive after the first IVF-ET and had a next cycle were recruited to the control group.78 women were recruited to the control group for follow-up biochemical pregnancy cycle.The research methods were required as the same as in the second part of the study.The statistical method was also as the same as in the second part of the study.RESULTSThere were no significant diversities in female age,infertility causes,infertile time,body mass index,frequency of artificial abortion,the number of antral follicle counting,serum E2,FSH,LH concentration and FSH/LH during the early phase of menstrual cycle,initial and total dosage of Gn,thickness of endometrium on the day of HCG injection,the proportion of composition of controlled ovarian hyperstimulation protocol,Gn type,endometrial morphology on the day of HCG injection,fertilization methods,luteal support protocol(P＞0.05).But,frequency of spontaneous abortion were increased in women who experienced a biochemical pregnancy than in women who had a negative pregnancy test in the previous IVF-ET cycle.The diversities were significant(P＜0.05). There were no significant diversities in the number of oocytes picked up, Mâ…¡oocytes,high-grade embryo,embryos transferred,transferring at least embryos with gradeâ… and 7-8 blastomeres,transferred embryo score,Mâ…¡oocyte percentage, normal fertilization rate,cleavage rate,transferring at least embryos with gradeâ… and 7-8 blastomeres rate,frozen embryo rate,clinical pregnancy rate,cumulate pregnancy rate,taken back baby rate and canceled cycle rate(P＞0.05).However,oocyte retrieval rate was lower and high-grade embryo rate were higher in the immediate next cycle when women had a biochemical pregnancy than those women who had a negative pregnancy test.The diversities were significant(P＜0.05).CONCLUSIONSThis study failed to prove that a biochemical pregnancy in IVF is a positive predictor of subsequent IVF success.But,this study also suggested that embryo quality in follow-up treatment cycle in patients with a history of biochemical pregnancy were better than in patients with a history of a negative pregnancy test.
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