| Objective: The purpose of this study is to retrospectively analyse the correlation between denudation and microinjection time after oocyte retrieval and intracytoplasmic sperm injection(ICSI) and laboratory and pregnancy outcome, discuss the optimal time of denudation and microinjection after human chorionic gonadotropin(HCG) injection and oocyte retrieval and provide bases for improving ICSI process and outcome.Methods: Clinical,laboratory and pregnancy outcome information of cycles undergoing ICSI-ET in the ART Department of the Second Hospital of Hebei Medical University from January 2013 to December 2013 was retrospectively analyzed. The time from HCG injection to ICSI was divided into T1(oocyte retrieval to denudation time),T2(denudation to ICSI time), T3(HCG injection to oocyte retrieval time),T4(HCG injection to denudation time) and T5(HCG injection to ICSI time) according to HCG injection time, oocyte retrieval time, denudation time and ICSI time. Three parts were analysed: ①the correlation between T1-T5 and laboratory information such as MII oocytes rate, fertilization rate, normal fertilization rate, abnormal fertilization rate, damage rate, cleavage rate and good quality embryo rate, ②the correlation between T1-T5 and pregnancy outcome information such as implantation rate, clinical pregnancy rate, abortion rate and live birth rate, ③the correlation between laboratory information and pregnancy outcome information.Results: 1 General condition of ICSI cycles195 ICSI cycles were analysed. The average woman’s age, follicle stimulating hormone(FSH) level, estradiol(E2) level, luteinizing hormone(LH) level, gonadotropin(Gn) days and Gn dose were 28.44 ± 3.76 years old, 7.08 ± 1.59 m IU / ml, 45.09 ± 34.65 pg / ml, 4.30 ± 2.63 m IU / ml,10.12 ± 1.653 days and 2223.32 ± 612.34 iu,respectively. The total number of oocytes was 2480, the average number of oocytes was 12.72 ± 4.84, MII oocytes rate was 82.58%, damage rate was 0.83%, fertilization rate was 90.53%, normal fertilization rate was 84.96%, cleavage rate was 91.21%, good quality embryo rate was 40.85%.The number of implanted embryos were 145 and implantation rate was 35.63%.The clinical pregnancy rate was 51.28% and there were 3 cases of ectopic pregnancy, 8 cases of abortion and 89 cases of childbirth. Childbirth was all live birth. Abortion rate was 8.00% and live birth rate was 45.64%. 2 The correlation between T1-T5 and laboratory outcome 2.1 The correlation between T1(oocyte retrieval to denudation time)and laboratory outcomeThere wasn’t correlation between T1(oocyte retrieval to denudation time) and MII oocytes rate, fertilization rate, normal fertilization rate, abnormal fertilization rate, damage rate, cleavage rate and good quality embryo rate(P>0.05).T1 was divided into 2 groups: 0-1.5h group and >1.5h group. The cycle number is 83 and 112. MII oocytes rate, fertilization rate, normal fertilization rate, cleavage rate and good quality embryo rate of 0-1.5h group were 82%±14%,90%±12%,85%±13%,91%±12% and 35%±22%, respectively. Those of >1.5h group were 83%±13%, 92%±12%, 86%±14%,92%±12% and 41%±25%, respectively. There was no statistical significance on these items but they were higher in >1.5h group. 2.2 The correlation between T2(denudation to ICSI time)and laboratory outcomeThe time between denudation and ICSI(T2) was negatively correlated with normal fertilization rate and good quality embryo rate(Z=-2.293,P=0.022, Z=-2.171,P=0.03)and positively correlated with abnormal fertilization rate and damage rate(Z=2.293,P=0.022, Z=2.019,P=0.043).There wasn’t correlation between T2 and MII oocytes rate, fertilization rate and cleavage rate(P>0.05).T2 was divided into 3 groups: 0-3h, 3-4h and >4h groups. The cycle number is 56,123 and 16. Damage rate of 3 groups were 0.46%±2.1%,1%±7.4% and 2%±4%. Normal fertilization rate were 87%±14%, 85%±13% and 83%±15%. There was no statistical significance but damage rate of 0-3h group was lower than other 2 groups and normal fertilization rate was higher than other 2 groups. 2.3 The correlation between T3(HCG to oocyte retrieval time)and laboratory outcomeThe time between HCG and oocyte retrieval(T3) was negatively correlated with fertilization rate(Z=-2.445,P=0.015)and had no correlation with MII oocytes rate, normal fertilization rate, abnormal fertilization rate, damage rate, cleavage rate and good quality embryo rate(P>0.05). 2.4 The correlation between T4(HCG to denudation time)and laboratory outcomeThe time between HCG and denudation(T4) was negatively correlated with fertilization rate and cleavage rate(Z=-4.514,P<0.001, Z=-3.379, P=0.001)and had no correlation with MII oocytes rate, normal fertilization rate, abnormal fertilization rate, damage rate and good quality embryo rate(P>0.05).T4 was divided into 2 groups: 38-38.5h group and >38.5h group. The cycle number is 61 and 134.Fertilization rate of the 2 groups were 94%±8.7 %and 89%±13%. Cleavage rate were 94%±8.8% and 90%±13%.There was statistical significance on fertilization rate and cleavage rate(u=-2.769,P=0.006, u=-2.136,P=0.033). 2.5 The correlation between T5(HCG to ICSI time)and laboratory outcomeThe time between HCG and ICSI(T5) was negatively correlated with fertilization rate, normal fertilization rate and cleavage rate(Z=-4.043,P<0.001, Z=-2.515,P=0.012,Z=-3.359,P=0.001)and positively correlated with abnormal fertilization rate(Z=2.515,P=0.012). It had no correlation with MII oocytes rate, damage rate and good quality embryo rate(P>0.05).T5 was divided into 2 groups: 40-42.5h group and >42.5h group. The cycle number is 161 and 34.Fertilization rate of the 2 groups were 92%±10% and 84%±17%. Normal fertilization rate were 97%±13% and 79%±16%.Cleavage rate were 93%±10% and 85%±17%.There was statistical significance on fertilization rate, normal fertilization rate and cleavage rate(u=-2.99,P=0.003, u=-3.13, P=0.002, u=-2.91,P=0.004). 3 The correlation between T1-T5 and pregnancy outcomeThere wasn’t correlation between T1(oocyte retrieval to denudation time) and implantation rate, clinical pregnancy, abortion rate and live birth rate(P>0.05).T1 was divided into 2 groups: 0-1.5h group and >1.5h group. Clinical pregnancy rate, abortion rate and live birth rate of 0-1.5h group were 45.78%, 7.89% and 40.96%.Those of >1.5h group were 55.36%, 8.06% and 49.11%. Clinical pregnancy rate, abortion rate and live birth rate of >1.5h group were higher than 0-1.5h group and there was statistical significance(u=-12.36,P<0.001, u=-8.107,P<0.001, u=-12.36, P<0.001). Implantation rate were 33%±40% and 38%±40%.There was no statistical significance(P>0.05), but >1.5h group had higher implantation rate.There wasn’t correlation between T2-T5 and implantation rate, clinical pregnancy, abortion rate and live birth rate(P>0.05). 4 the correlation between laboratory outcome and pregnancy outcomeClinical pregnancy rate was positively correlated with good quality embryo rate(χ2=4.16,P=0.041,B=1.293,OR=3.64,95%CI[1.05,12.62])and had no correlation with MII oocytes rate, fertilization rate, normal fertilization rate, abnormal fertilization rate, damage rate, cleavage rate(P>0.05).Implantation rate, abortion rate and live birth rate had no correlation with MII oocytes rate, fertilization rate, normal fertilization rate, abnormal fertilization rate, damage rate, cleavage rate and good quality embryo rate(P>0.05).Conclusion: The time of denudation and sperm injection may be closely related with fertilization and pregnancy outcome. Denudation over 1.5h after oocyte retrieval may improve the potential of oocyte maturity, fertilization and embryo development and affect pregnancy outcome. ICSI within 3h after denudation may be beneficial to the normal fertilization and embryo development. Delaying the time of denudation and ICSI after HCG injection too much may influence fertilization rate, cleavage rate and ICSI outcome. |
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