[Effects of time of recombinant luteinized hormone supplementation on pregnancy outcomes of women with advanced reproductive age during follicular-phase long protocol].

Objective: To investigate the effects of supplementation of recombinant luteinized hormone (rLH) and its timing on pregnancy outcomes of patients at 35 years or older with follicular-phase long protocol. Methods: Clinical data of women undergoing in vitro fertilization or intracytoplasmic sperm injection with follicular-phase long protocol was collected and retrospectively analyzed in the First Affiliated Hospital of Nanjing Medical University from January 2017 to December 2019. There were 558 patients at 35 years or older included in this study, and they were divided into three groups: group A was patients with only recombinant follicle stimulating hormone (rFSH) stimulation (127 cycles), group B was patients with rFSH plus rLH supplementation in the mid-follicular phase (141 cycles), and patients in group C received combined rFSH and rLH from the first day of ovarian stimulation (290 cycles). The basic characteristics of patients of each group were observed and the effects of ovarian simulation and pregnancy outcomes were compared among the three groups. Logistic regression model was performed to explore the association between different groups and pregnancy outcomes. Results: The basic characteristics such as age, duration of infertility, body mass index (BMI) and serum basic follicle stimulating hormone (FSH) were comparable among the three groups (all P>0.05). Anti-Müllerian hormone (AMH), antral follicles count (AFC) and basic luteinized hormone (LH) were significantly lower in group C compared to group A and group B (all P<0.05). There were statistically significant differences in initiation dosage, total dosage and duration of gonadotropin (Gn) among the three groups (all P<0.01), the initiation dosage, total dosage and duration of Gn were higher in group C than the other two groups. The number of oocytes retrieved and available embryos were significantly lower in group B and group C than group A (all P<0.001). In fresh embryo transfer cycles, significantly higher implantation rate (45.3%, 117/258) and clinical pregnancy rate (52.6%, 111/211) were found for group C when compared with group A and group B (P=0.036, P=0.006). The live birth rate in fresh embryo transfer cycles was comparable among the three groups (P=0.098). The implantation rate, clinical pregnancy rate and live birth rate in the subsequent frozen-thawed embryo transfer cycles did not differ significantly among the three groups (all P>0.05). There were no significantly differences in the cumulative pregnancy rate and the cumulative live birth rate among the three groups (all P>0.05). After adjusted for age, BMI, AMH, AFC, basic FSH and LH, total Gn dosage, endometrial thickness at transfer, number of oocytes retrieved, number of embryos transferred and stage of embryo transferred, in fresh embryo transfer cycles, the clinical pregnancy rate (adjusted OR=2.793, 95%CI: 1.512-5.162, P<0.001) and live birth rate (adjusted OR=2.324, 95%CI: 1.241-4.351, P=0.008) were higher in group C, while clinical pregnancy rate and live birth rate were similar between group B and group A in fresh embryo transfer cycles (all P>0.05); there was no significant difference in cumulative live birth rate among the three groups (P>0.05). Conclusions: The supplementation of rLH from the first day of ovarian stimulation improves the pregnancy outcomes of patients at 35 years or older in fresh embryo transfer cycles during follicular-phase long protocol. However, the supplementation of rLH has no benefit on cumulative live birth rate.

[Impact of BMI on fresh cycle embryo transfer outcome of IVF/ICSI in patients with early follicular phase prolonged protocol].

Objective: To investigate the impact of body mass index (BMI) on clinical effect and fresh cycle embryo transfer pregnancy outcome of in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) in patients with early follicular phase prolonged protocol. Methods: From January 1st, 2018 to July 1st, 2020, 2 257 cases of early follicular long-term protocol in IVF/ICSI and embryo transfer were collected using the clinical assisted reproductive technologies management system software database of the First Affiliated Hospital of Nanjing Medical University. Patients were divided into three groups according to the recommended Asian BMI cut-off points: low body mass group (BMI<18.5 kg/m2), normal body mass group (18.5≤BMI<24.0 kg/m2), and high body mass group (BMI≥24.0 kg/m2). The ovarian stimulation characteristics among the groups were investigated. Then 1 741 fresh embryo transfer cycles were selected and divided into three groups as above, and then the ovulation induction and clinical outcomes were analyzed among the groups. Results: There were significant differences in the starting dosage of gonadotrophin (Gn), total dosage of Gn and days of Gn used among the low body mass group, normal body mass group, and high body mass group in the 2 257 IVF/ICSI cycles (all P<0.01). The high body mass group needed the most amount of Gn [(2 159±668) U] and longest Gn days [(12.3±2.5) days]. The estradiol and progesterone levels [(7 474±4 852) pmol/L, (3.4±1.9) nmol/L] on hCG trigger day in the high body mass group were lower than those in the low body mass group and normal body mass group (all P<0.01). The oocytes retrieved in high body mass group (8.4±4.1) were significantly lower than normal body mass group (P<0.05). The normal fertilization number, the available embryo number and high quality embryo number were all lower in the high body mass group than other two groups, while no significant difference showed (all P>0.05). In 1 741 cycles of fresh embryo transfer, the average number of transplanted embryos in the low body mass group (1.2±0.4) was decreased compared with the other two groups (P<0.05), while the biochemical pregnancy rate, clinical pregnancy rate and live birth rate in the normal body mass group were higher compared with the other two groups, but the differences showed no statistically significance (all P>0.05). Conclusions: Increased BMI might affect ovulation induction response in early follicular phase prolonged protocol IVF/ICSI patients, leading to the increase of Gn dosage and the extension of Gn induction days. Although there is no significant difference in pregnancy outcome among different BMI groups, considering the increased risk of adverse perinatal outcomes during subsequent pregnancy in overweight or obese patients, certain attention should still be paid to the control of BMI in patients receiving assisted reproduction treatment with early follicular phase prolonged protocol.

[Effects of estradiol on the day of human chorionic gonadotropin administration on adverse perinatal outcomes].

Objective: To investigate the association between estradiol on the day of human chorionic gonadotropin (HCG) administration and birth outcomes among singleton live births following fresh embryo transfers. Methods: Based on the clinical reproduction medicine management system of the First Affiliated Hospital of Nanjing Medical University, this retrospective cohort study collected data of fresh embryo transfer cycles during January 2013 and December 2016, including pregnant women's age, body mass index (BMI), type and cause of infertility, assisted reproductive therapy indicators (fertilization mode, ovulation stimulation protocol, estradiol levels on HCG administration day), adverse birth outcomes[small for gestational age (SGA), premature and low birth weight (LBW)], etc.. A total of 2 060 women with singleton pregnancy (2 061 fresh embryo transfer cycles) were enrolled. Multivariate logistic regression was used to analyze the association between estradiol on HCG administration day and singletons' adverse birth outcomes. Results: The age and BMI of the 2 060 pregnant women were (29.63±3.92) years old and (22.29±2.86) kg/m2. Incidences of SGA, premature and LBW were 9.8% (201/2 061), 6.9% (143/2 061) and 3.5% (73/2 061), respectively. After adjusting for confounders, the risk of LBW in 4 000-4 499 pg/ml group was significantly elevated when compared to estradiol<1 500 pg/ml group [OR (95%CI): 4.42 (1.13-17.24)]. A protective effect of premature was observed in estradiol≥4 500 pg/ml group [OR (95%CI): 0.50 (0.25-0.97)]. Conclusion: The high level of estradiol on HCG administration day might be a risk factor for LBW, but a protective factor for premature.

[Chromosome abnormality rate and related factors of spontaneous abortion in early pregnancy].

Objective: To investigate chromosome abnormality rate and related factors of spontaneous abortion in early pregnancy. Methods: A total of 831 tissue samples of spontaneous abortion in early pregnancy were collected from June 2015 to August 2018 in the First Affiliated Hospital of Nanjing Medical University. Chromosomal copy number was analyzed by next generation sequencing (NGS). The relationships between chromosome abnormality and maternal age, in vitro fertilization-embryo transfer (IVF-ET) pregnancy, number of previous spontaneous abortions, history of live birth were analyzed by statistical methods. Results: Among 831 tissue samples of spontaneous abortion in early pregnancy, 461 (55.5%, 461/831) were found to have chromosome abnormalities. Maternal age (OR=1.107, 95%CI: 1.070- 1.145) and history of live birth (OR=1.909, 95%CI: 1.182-3.083) were the positive correlative factors of chromosome abnormality. Times of previous spontaneous abortion (OR=0.807, 95%CI: 0.702-0.928) and IVF-ET pregnancy (OR=0.554, 95%CI: 0.404-0.760) were the negative correlative factors of chromosome abnormality. Conclusions: Chromosome abnormality is an important cause of spontaneous abortion in early pregnancy. The rate of chromosome abnormality increases with the increase of maternal age and the history of live birth, and decreases with the increase of number of previous spontaneous abortion and IVF-ET pregnancy.

[Analysis on factors related to spontaneous reduction in twin pregnancy following assisted reproductive technology].

Objective: To explorer factors related to spontaneous reduction in twin pregnancy following assisted reproductive technology. Methods: 2 848 twin pregnant women with treatment of vitro fertilization-embryo transfer (IVF-ET) or intra cytoplasmic sperm injection (ICSI) cycles were enrolled at Assisted Reproductive Centre of the First Affiliated Hospital of Nanjing Medical University, Nanjing Maternity Hospital and Shengjing Hospital of China Medical University from January 2013 to December 2016 respectively. Basic features of subjects, relevant clinical indicators, factors of assisted reproductive therapy and pregnancy outcome were collected from clinical assisted reproductive technology management system. According to the pregnancy outcome, the subjects with spontaneous reduction were classified as case group (n=686), and those with normal twin birth were classified as control group (n=2 162). The features of subjects in the two groups were compared. Non-conditional logistics regression model was used to analyze the related factors of the occurrence of spontaneous reduction. Results: The age of case group and control group were (30.6±4.3) and (30.2±4.0) years old respectively. After the adjustment of male sterile factor, compared to the subjects with luteinizing hormone level on the day of human chorionic gonadotropin administration (HCG) <1.43 IU/L, OR (95%CI) of the subjects with value at 2.59-5.10 IU/L was1.62 (1.08-2.42).Compared to the subjects with number of transferred embryo as 1, OR (95%CI) of the subjects with value as 3 was 0.23 (0.07-0.74). Compared to the subjects with stage of transferred embryo as cleavage stage, OR (95%CI) of the subjects with blastula stage was 0.42 (0.27-0.67). Conclusion: Luteinizing hormone level on day of HCG, number and stage of embryo transfer are related factors to spontaneous reduction in twin pregnancy following assisted reproductive technology.

[Cumulative live birth rates per oocytes retrieved cycle: evaluation of clinical outcomes of IVF/ICSI].

Objective: Using of cumulative live birth rate (CLBR) per oocytes retrieved cycle, to assess the clinical outcomes of in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI) , and to explore impact factors on CLBR following utilization of all fresh and frozen embryos in one complete IVF/ICSI cycle using gonadotropin-releasing hormone (GnRH) agonist, GnRH-antagonist and clomiphene mild stimulation protocols. Methods: Of the patients who underwent IVF/ICSI from January 1st, 2014 to December 31st, 2015 in the First Affiliated Hospital, Nanjing Medical University, a total of 6 142 oocytes retrieved cycles were included. The clinical and laboratory parameters of different ovarian stimulation protocols, and the effects of the age, number of oocytes retrieved and number of embryos available on the CLBR of each oocytes retrieved cycle were analyzed. Results: The CLBR was 69.0% (2 004/2 906) in the GnRH-agonist protocol versus 67.4% (644/955) in the GnRH-antagonist protocol (P>0.05); the CLBR of clomiphene mild stimulation protocol was 53.2% (1 215/2 281) , significantly lower than those of the other two protocols (all P<0.05). The CLBR significantly decreased with age increased. When divided into four groups according to the patients' age, we found that CLBR were not statistically significant using three different protocols in the 20-25 years old group (all P>0.05). There was a strong association between the number of oocytes retrieved and embryos available on CLBR. CLBR rose significantly with an increasing number of oocytes up to 6, then the rising trend slowed down. Patients were categorized into four groups according to the number of oocytes retrieved, CLBR was significantly higher using GnRH-antagonist protocol (50.0%) than mild stimulation protocol (37.0%) in low ovarian responder (0-4 oocytes) group (P<0.05) . The CLBR were no significant difference among three protocols in normal (10-15 oocytes) and high responders (≥15 oocytes) group (all P>0.05) . The incidence rate of ovarian hyperstimulation syndrome in GnRH-agonist protocols (5.2%, 152/2 906) were significantly higher than those of GnRH-antagonist (4.4%, 42/955) and clomiphene mild stimulation protocols (1.5%, 34/2 281; all P<0.05) . Conclusions: CLBR is an important index to assess the clinical outcomes of IVF/ICSI. Age, number of oocytes retrieved and embryos available could affect CLBR obviously. According to the different age and ovarian response of patients, we should design ovarian stimulation protocols based on target oocytes number in order to get higher CLBR and reduce complications.