| Objectives:1. Study the effect of different embryo morphology parameters, provide the basis and support for selecting the good embryo to transfer, enhancing the clinical pregnancy rate and live birth rate.2. Combine use with different embryo morphology parameters, compare the pregnancy outcome, chose the reasonable method to improve the screening criteria of embryo to transfer.3. Study the correlation between other embryo parameters such as fertilization rate,good embryo rate and pregnancy outcome, pick up the parameters which can predict pregnancy outcome of IVF/ICSI as a predictor.Study subjects: The 836 infertility couples who came to the reproductive medical center of First Hospital of Jilin University to receive IVF or ICSI treatment from May2011 to July 2014 were included, 954 cycles were done. Inclusion criteria: female age in year <35, transfer 2 embryo per cycle, number of oocytes collected ≥6. Exclusion criteria: donor semen or oocytes cycle; cancel transfer cycles; female factors that may affect the embryo implantation were exclued, such as polycystic ovary syndrome,endometriosis, adenomyosis. Finally, 509 cycles from 483 couples were included,female average age in year 28.81±3.32, and male average age in year 30.94±4.69.Aetiology of infertility included male factor(n=250), female factor(n=150), multiple factors(n=94) and unexplained(n=15). The protocol for controlled ovarian hyperstimulation included long term protocol(n=442), short term protocol(n=25),minimal ovarian stimulation protocol(n=22), antagonist protocol(n=14), ultra long term protocol(n=6).Methods:1. Collect the patients’ information including age, BMI, duration of infertility and medical history by questionnaire and enquiring. Female endocrine was tested by Roche electrochemical luminescence method. Using IVF or ICSI based on patient’s etiology.2. Embryo assessment method:(1) pronuclear Z score, there was four levels as Z1-Z4;(2) D3 embryo assessed by cell number and grade, there was four levels as I-IV.3. Grouping method(1) Three groups were formed depending on pronuclear Z score, the group in which two embryos with below Z2 pronuclear to transfer(<Z2,<Z2), the group that one embryo with over Z2 pronuclear and one embryo with below Z2 pronuclear to transfer(<Z2,≥Z2), and the group that two embryos with over Z2 pronuclear to transfer(≥Z2,≥Z2). Three groups were formed depending on cell number on D3, the group in which two embryo with below 6 cell number to transfer(<6,<6), the group that one embryo with over 6 cell number and one embryo with below 6 cell number to transfer(<6,≥6), and the group that two embryos with over 6 cell number to transfer(≥6,≥6). Three groups were formed depending on embryo grade on D3, the group in which two embryos with below II grade to transfer(<II,<II), the group that one embryo with over II grade and one embryo with below II grade to transfer(<II,≥II), and the group that two embryos with over II grade to transfer(≥II,≥II).(2) Compare pregnancy outcome when combined using different embryo parameters.Compare was happened in those situation, 1 the same cell number on D3,pregnancy outcome in different embryo grade, 2 the same embryo grade, pregnancy outcome in different cell number on D3, 3 the same cell number on D3,pregnancy outcome in different pronuclear score, 4 the same embryo grade, pregnancy outcome in different pronuclear score. Group method described in(1).(3) According to the group whether 6-10 cell embryo was transferred, to compare the pregnancy outcome.(4) According to pregnancy outcome, cycles were divided into clinical pregnancy group and non-clinical pregnancy, live birth group and non-live birth group,fertilization rate, and good embryo rate etc. were compared in different group.Statistical Method:Using spss17.0 statistical software to analyse. Continuous data are summarized as means±SD, Independent-samplest Test was taken if data fitted normal distribution,and if not, Kruskal-Wallis Test would be taken. The chi-square analysis was taken when count data expressed as a percentage. Correlation analysis using binary logistic regression analysis. And p< 0.05 was considered statistically significant, and p<0.01 was considered significant difference of statistical significance.Results:1. Baseline information of patients in different pregnancy outcome. 509 cycles group into clinical pregnancy(CP) group(n=254) and non-clinical pregnancy(non-CP)group(n=255), and live birth(LB) group(n=213) and non-live birth(non-LB)group(n=296). There was significant difference in the CP group and non-CP group,LB group and non-LB group among female BMI, basal FSH, endometrial thickness on day of embryo transfer(p<0.05). Basal FSH and endometrial thickness on day of embryo transfer were higher in CP group and LB group than non-CP group and LB group(Basal FSH in CP and non-CP group:7.01 ± 1.80IU/L vs 6.64 ± 1.60IU/L;endometrial thickness on day of embryo transfer in CP and non-CP group:11.62±1.91 mm vs 10.96 ± 2.35mm). BMI in CP group and LB group were lower than non-CP group and non-LB group(CP and non-CP group:21.76±3.18kg/m2 vs22.43±3.39kg/m2; LB and non-LB group:21.73±3.10kg/m2 vs 22.37±3.41kg/m2)2. Embryo quality and pregnancy outcome(1) Depended on the pronuclear score, group(<Z2,<Z2) included 342 cycles, group(<Z2,≥Z2) included 136 cycles, group(≥Z2,≥Z2) included 31 cycles. The clinical pregnancy rate was 50.3%, 50.0%, 45.2%, and the live birth rate was 44.2%, 39.0%,29.0%. There were no statistical significance difference in different group.(2) Depended on D3 embryo cell number, group(<6,<6) include 17 cycles, group(<6,≥6) included 52 cycles and group(≥6,≥6) included 440 cycles. The clinical pregnancy rate was 11.8%, 40.4%, 52.5%, there were statistically significant difference in different group on clinical pregnancy rate. and there were statistically significant difference in different group on live birth rate, the live birth rate was11.8%, 28.8%, 44.5%.(3) Depended on D3 embryo score, group(<II,<II) include 13 cycles, group(<II,≥II)include 29 cycles, group(≥II,≥II) include 398 cycles. The clinical pregnancy rate was16.7%, 34.8%, 52.8%, there were statistically significant difference in different group on clinical pregnancy rate. and there were statistically significant difference in different group on live birth rate, the live birth rate was 16.7%, 28.3%, 44.3%.(4) Logistic regression was performed to determine the association between embryo quality and pregnancy outcome suggested that D3 cell number(OR=2.017, p<0.05)and D3 embryo score(OR=2.055, p<0.05) are significantly associated with clinical pregnancy rate after adjusting for other confounding factors. And D3 cell number(OR=1.991, p<0.05) and D3 embryo score(OR=1.742, p<0.05) are also significantly associated with live birth rate.3. Pregnancy outcome of combined different method of embryo assessment(1) In group(≥6,≥6), three subgroups were analysed, subgroup(<II,<II) included 13 cycles, subgroup(<II,≥II) included 29 cycles, subgroup(≥II,≥II) included 398 cycles.There was a statistically significant difference in different group on clinical pregnancy rate, the clinical pregnancy rate was 23.1%, 37.9%, 54.5%.(2) In group(≥II,≥II), three subgroups were analysed, subgroup(<6,<6) included 12 cycles, subgroup(<6,≥6) included 35 cycles, subgroup(≥6,≥6) included 398 cycles.There was a statistically significant difference in different group on clinical pregnancy rate, the clinical pregnancy rate was 16.7%, 45.7%, 54.5%. There was a statistically significant difference in different group on live birth rate, the clinical pregnancy rate was 16.7%, 31.4%, 46.2%.(3) In group(<6,≥6), three subgroups were analysed, subgroup(<Z2,<Z2) included 35 cycles, subgroup(<Z2,≥Z2) included 13 cycles, subgroup(≥Z2,≥Z2) included 4 cycles.There was a statistically significant difference in different group on clinical pregnancy rate, the clinical pregnancy rate was 34.3%, 38.5%, 100%.4. With or without 6-10 cell embryo to transfer and pregnancy outcome. Group with8 cell embryo has 367 cycles, group without 8 cell embryo has 142 cycles. There were statistically significant difference in different group on both clinical pregnancy rate(56.4% vs. 33.1%) and live birth rate(46.6% vs. 29.6%). Clinical pregnancy rate was highest in group with 8 cell embryo when compared with other group that with6,7,9,10 cell embryo. Live birth rate has the similar trend, but there was no significant statistical difference in different group.5. Relationship of fertilization rate, good quality embryo rate and other parameters and pregnancy outcome.(1) Compared different parameters in clinical pregnancy group and non-clinical pregnancy group. MII oocyte rate, utilization embryo rate, good quality embryo rate,the rate of 4 cell embryo on D2, the rate of I grade embryo on D2 and the rate of 8cell embryo on D3 in clinical pregnancy group were significant higher than them of non-clinical pregnancy group(p<0.05). There was no significant statistical difference in oocyte retrieval rate, fertilization rate, 2PN rate, polyspermy rate, 2PN cleavage rate, Z1 zygote rate, Z2 zygote rate, the rate of II grade embryo on D2, the rate of I grade embryo on D3, the rate of II grade embryo on D3 between clinical pregnancy group and non-clinical pregnancy group.(2) Compared different parameters in live birth group and non-live birth group. MII oocyte rate, utilization embryo rate, the rate of 4 cell embryo on D2, the rate of I grade embryo on D2 and the rate of 8 cell embryo on D3 in live birth group were significant higher than them of non-live birth group(p<0.05). There was no significant statistical difference in oocyte retrieval rate, fertilization rate, 2PN rate, polyspermy rate, 2PN cleavage rate, good quality embryo rate, Z1 zygote rate, Z2 zygote rate, the rate of II grade embryo on D2, the rate of I grade embryo on D3, the rate of II grade embryo on D3 between live birth group and non-live birth group.(3) Logistic regression was performed to determine the association between different parameters and pregnancy outcome suggested that MII oocyte rate(OR=5.066,p<0.05), the rate of 4 cell embryo on D2(OR=3.263, p<0.05) are significantly associated with clinical pregnancy rate after adjusting for other confounding factors.And MII oocyte rate(OR=3.830, p<0.05) and the rate of 4 cell embryo on D2(OR=2.993, p<0.05) are also significantly associated with live birth rate.Conclusions:1. Pronuclear score isn’t in correlational relationship with clinical pregnancy and live birth, is not the parameter to predict pregnancy outcome.2. There is an independent positive correlation between embryo cell number on D3 and pregnancy outcome, also embryo grade have the independent positive correlation with pregnancy outcome. Embryo cell number on D3 is much better than embryo grade to select embryo to transfer.3. Embryo with 8 cell number is better than 6 or 7 cell, 6 or 7 cell embryo is better than 9 cell embryo, suggest that 8 cell embryo should be transfered prior.4. MII oocyte rate and the rate of 4 cell embryo on D2 have the independent positive correlation with pregnancy outcome, may be they can used to reflect the whole embryo quality. |
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