Effect of Progestin-primed Ovarian Stimulation Protocol on Outcomes of Aged Infertile Women Who Failed to Get Pregnant in the First IVF/ ICSI Cycle： A Self-controlled Study

This study aimed to explore the outcomes of progestin-primed ovarian stimulation protocol （PPOS） in aged infertile women who failed to get pregnant in the first IVF/ICSI-ET cycles with GnRH-a long protocol. A self-controlled study was conducted to retrospectively investigate the clinical outcomes of 104 aged infertile patients who didn＇t get pregnant in the first IVF/ICSI-ET treatment by stimulating with GnRH-a long protocol （non-PPOS group）, and underwent PPOS protocol （PPOS group） in the second cycle between January 2016 and December 2016 in the Center for Reproductive Medicine, Renmin Hospital of Wuhan University. The primary outcomes included clinical pregnancy rate of frozen-thawed embryos transfer （FET） in PPOS group, and good-quality embryo rate in both groups. The secondary outcomes were fertilization rate, egg utilization rate and cycle cancellation rate. The results showed that there were no significant differences in basal follicle stimulating hormone （bFSH）, antral follicle count （AFC）, duration and total dosage of gonadotropin （Gn）, number of oocytes retrieved, intracytoplasmic sperm injection （ICSI） rate, fertilization rate, and cycle cancellation rate between the two groups （P〉0.05）. However, the oocyte utilization rate and good-quality embryo rate in PPOS group were significantly higher than those in non-PPOS group （P〈0.05）. By the end of April 2017, 62 FET cycles were conducted in PPOS group. The clinical pregnancy rate and embryo implantation rate were 22.58% and 12.70%, respectively. In conclusion, PPOS protocol may provide better clinical outcomes by improving the oocyte utilization rate and good-quality embryo rate for aged infertile patients who failed to get pregnant in the first IVF/ ICSI-ET cycles.

DS147 Improves Pregnancy in Mice with Embryo Implantation Dysfunction Induced by Controlled Ovarian Stimulation

Summary： The study examined the effect of DS147, the bioactive component of the traditional herbal recipe Bangdeyun, on pregnancy in mice with embryo implantation dysfunction induced by controlled ovarian stimulation （COS）, and the underlying mechanisms. Female mice were superovulated by intrap eritoneal injection of 7.5 IU of pregnant mare serum gonadotropin （PMSG） followed by an additional injection of 7.5 IU hCG 48 h later to establish embryo implantation dysfunction （EID） model. Pregnant mice were randomly divided into normal control group, COS group and DS147-treated groups. The pregnancy rate and the average implantation site were obtained on pregnancy day 8 （PD8）. The side effect of 200 mg/kg of DS147 on naturally pregnant mice was also observed. Further, the uterine and ovarian tissue samples were collected on PD5 for measuring their weights, observing the development of the endometrium and ovary, and detecting the endometrial expression ofMMP-2, TIMP-2, CD34 and angiogenin （ANG）. The female mice treated with DS147 at doses of 100 to 800 mg/kg showed a higher pregnancy rate than those in COS group, and the highest pregnancy rate of 83.3% occurred in the 200 mg/kg DS147-treated group. Moreover, no obvious side effect was found in mice treated with 200 mg/kg DS147 on PD8 and PD16. The ovarian and uterine weights, and the expression levels of MMP-2, ANG and CD34 were significantly increased in DS147-treated groups when compared with COS group. The TIMP-2 expression level was much lower in DS147-treated mice than in COS mice and the ratio of MMP-2/TIMP-2 was much higher in DS147-treated group than in COS group, and even higher than normal control group. In all, these findings suggest that DS147 may improve pregnancy in mice with COS-induced EID by promoting matrix degradation and angiogenesis, and improving the development of corpus luteum and endometrial decidualization around the implantation window.

Cumulative Live Birth Rate after Three Ovarian Stimulation IVF Cycles for Poor Ovarian Responders According to the Bologna Criteria

Summary： This study explored the cumulative live birth rate after three ovarian stimulation in vitro fer- tilization （IVF） cycles for poor ovarian responders according to the Bologna criteria. In this retrospec- tive cohort study, 479 poor ovarian responders according to the Bologna criteria in the first ovarian stimulation IVF cycle between July 2006 and January 2012 in our IVF centre were included. The cu- mulative live birth rate was calculated by optimistic and pessimistic methods. The cumulative live birth rate after three ovarian stimulation IVF cycles for poor ovarian responders according to the Bologna criteria was 12.7%-20.5%. The three-cycle cumulative live birth rate was 18.5%--24.5%, 13.2%-27.4% and 8.6%-14.9% for poor responders aged ≤35 years, 36-39 years and 〉40 years, re- spectively. In conclusion, poor responders according to the Bologna criteria can receive an acceptable cumulative live birth, rate after three ovarian stimulation IVF cycles, especially poor responders aged 〈40 years.

Timed Intercourse versus Intrauterine Insemination with Mild Ovarian Stimulation for Unexplained Infertility

Background: Infertility is unexplained in about 22% - 28% of infertile couples. Spontaneous pregnancy may occur in them. If did not happen, Expectant management will be via clomiphene citrate (CC) administration, intrauterine insemination (IUI), and IVF & ICSI. Aim: to assess the effectiveness of intrauterine insemination with mild controlled ovarian stimulation compared with expectant management in couples with unexplained infertility more than one year. Methods: 160 couples with unexplained infertility were selected, The couples were randomly divided into two groups: Group (A) “80 couples”: Intrauterine insemination (IUI) with mild controlled ovarian stimulation (combination of CC + hMG) using prepared semen and was performed 36 hours after hCG injection, Group (B) “80 couples”: Couples had no ovulation induction. They encouraged for timed intercourse in the most fertile days of female cycle guided by folliculometry for 6 months. Data were collected quantitatively, coded and analyzed using SPSS. The power of study is 80% and 95% confidence interval. Result: The pregnancy rate was calculated in both groups: In group (A) (IUI/COS): OPR (Ongoing pregnancy rates) was 27.5% and PR (pregnancy rates)/cycle was 8.6%. In group (B) (Expectant management): OPR was 25% and PR/cycle was 5.3%. Conclusion: Mild controlled ovarian stimulation CC + hMG with IUI offers no statistical significance in terms of pregnancy outcomes over expectant management in this study however significance in the response to stimulation.

A Retrospective Study on the Use of High-Dose Letrozole while Undergoing Ovarian Stimulation for Oocyte and Embryo Cryopreservation in Cancer Patients

Objective: To determine the efficacy of letrozole in suppressing estradiol levels during ovarian stimulation in cancer patients. Methods: A retrospective chart review of cancer patients undergoing ovarian stimulation for fertility preservation between 2014-2019 at a private university-affiliated fertility clinic in Canada was conducted. Ovarian stimulation was completed with no letrozole (Group A, n = 10), and adjuvant daily letrozole use at 5.0 (Group B, n = 34) or 7.5 mg (Group C, n = 61). The primary outcomes were peak estradiol levels and oocyte yield. ANOVA with a post hoc two-tailed t-test assuming equal variance was utilized as a statistical method. Result(s): Patient age and AFC count were not different between groups. The yield of mature eggs was not different at each letrozole dose;9.2 ± 6.0, 13.9 ± 6.5 and 12.7 ± 7.2 for Groups A to C respectively (p = 0.18). Mean estradiol levels(pmol/L) were reduced in a dose-dependent manner;7432 ± 4553 for Group A, 2072 ± 1656 for Group B, and 1445 ±1238 for Group C (A vs. C, p vs. C, p Conclusion(s): The use of letrozole during ovarian stimulation for oocyte and embryo cryopreservation in cancer patients can maintain physiologic estradiol levels, while ensuring satisfactory oocyte and embryo yield. Letrozole can, therefore, minimize the theoretical risk of stimulating residual and metastatic diseases, while still optimizing future fertility outcomes.

Comparison of the Effects and Safety of Mild Ovarian Stimulation and Conventional Ovarian Stimulation with A Long GnRH Agonist Protocol on IVF Outcomes

Objective To conduct a Meta-analysis of studies that compared the efficacies of mild ovarian stimulation and conventional long GnRH agonist protocol in patients under- going IVF or intracytoplasmic sperm injection （ICSI）. Methods Meta-analysis was performed. All studies were published by July 2012 with data related to outcomes following mild ovarian stimulation compared with the conventional protocol. Odds ratios （ORs） and weighted/standardized mean difference （WMD/SMD） from individual study were pooled in fixed and random effect models. Main outcome measure was the efficacy of mild ovarian stimulation. Results Six articles were included in this Meta-analysis. The number of oocytes retrieved was lower, the cycle cancellation rate was higher and the incidence of ovarian hyperstimulation syndrome （OHSS） was lower in the mild stimulation group than in conventional ovarian stimulation group. Clinical pregnancy rates were similar in both mild and conventional stimulation groups. Conclusions The level of evidence supporting the use of mild stimulation protocols in IVF is low, considering the fewer oocytes retrieved and the higher rates of cycle cancellation.

Effect of Progestin-Primed Ovarian Stimulation Protocol in Infertile Women with Basal Follicle-Stimulating Hormone Levels ≥15 IU/L: A Retrospective Analysis

Objective:To evaluate the efficacy of progestin-primed ovarian stimulation(PPOS)protocol in infertile women with high basal follicle-stimulating hormone(FSH)levels≥15 IU/L.Methods:Patients with high basal FSH levels≥15 IU/L with autologous oocytes from September 2016 to March 2019 were reviewed.Either medroxyprogesterone acetate 4 mg/d or clomiphene citrate(CC)50 mg/d was administered daily from day 3 to the trigger day.When serum FSH levels decreased to≤15.0 IU/L,a low dose of human menopausal gonadotropin(hMG)75/150 IU/d was administered to promote late follicular development.Results:Two hundred and twenty women were retrospectively analyzed in this study.Among them,139 patients were administered with PPOS protocol as the study group,and 81 patients were administered with CC protocol as the control group.The numbers of received oocytes and viable embryos were higher in the study group than those in the control group(1.5±1.2 vs.1.2±0.8 and 0.8±0.8 vs.0.5±0.6,respectively,P<0.05).However,hMG duration and dosage were significantly higher in the study group than those in the control group(4.2±2.7 d vs.1.1±2.3 d and 609.1±424.5 IU vs.140.7±231.3 IU,respectively,P<0.01).Incidence of luteinizing hormone surge and cycle cancellation rate were lower in the study group than those in the control group with statistical difference(2.88%vs.16.05%and 36.50%vs.50.63%,respectively,P<0.05).Conclusions:PPOS protocol can effectively downregulate the endogenous FSH levels.Compared with CC protocol,treatment with PPOS protocol in patients with high basal FSH levels≥15 IU/L could receive more oocytes and more viable embryos.

Impact of Progesterone on Inhibins during Controlled Ovarian Stimulation

Background:To explore the impact of progesterone on inhibins during controlled ovarian stimulation in women with normal ovarian reserve and to compare cycle characteristics and pregnancy outcomes in subsequently frozen-thawed embryo transfer(FET)cycles.Methods:A total of 93 patients were randomly divided into two groups,study group(human menopausal gonadotropin[hMG]+medroxyprogesterone acetate[MPA])and control group(short protocol).Serum hormones were detected on day 3 before ovarian and day 10-12,the trigger day,and the day after trigger(approximately 10 h after trigger).Viable embryos were cryopreserved for later transfer in both protocols.Results:In the study group,inhibins signifcantly increased during ovarian stimulation,and the average inhibins level on the trigger day was signifcantly higher than the basal levels.Inhibin A level increased significantly to 2046.7±1280.5 ng/L after trigger 10 h.Serum inhibin B level slightly decreased at the time of trigger 10 h later compared with the trigger time but did not reach a significant difference.The number of oocytes retrieved in study group was similar to that in control(10.5±4.5 vs.9.0±5.2,P<0.05).No statistically significant differences were found in the clinical pregnancy rate(47.4%vs.52.2%,P<0.05),implantation rate(36.5%vs.36%),and live birth rate(43.4%vs.39.1%,P<0.05)between the two groups.Conclusions:The high level of progesterone did not affect the secretion in granulosa cells during the controlled ovarian stimulation.Therefore,sufficient oocytes/embryos can be obtained by hMG and MPA co-treatment in women undergoing in vitro fertilization/intracytoplasmic sperm injection treatments,with optimal pregnancy outcomes in FET cycles.

Effect of Certain Toxicants on Gonadotropin-Induced Ovarian Non-Esterified Cholesterol Depletion and Steroidogenic Enzyme Stimulation of the Common Carp Cyprinus carpio in vitro

Isolated ovarian tissues from the common carp, Cyprinus carpio were incubated in vitro to obtain a discrete effect of four common toxicants of industrial origin, namely phenol, sulfide, mercuric chloride and cadmium chloride, on gonadotropin-induced alteration of nonesterified and esterified cholesterol and steroidogenic enzymes, △5-3β-HSD and 17β-HSD activity. Stage II ovarian tissue containing 30-40% mature oocytes were shown to be most responsive to gonadotropins in depleting only nonesterified cholesterol moiety and stimulating the activity of both. Safe doses of above mentioned toxicants when added separately to stage II ovarian tissue with oLH (1 μg/incubation) gonadotropin-induced depletion of nonesterified cholesterol and gonadotropin-induced stimulation of the activity of both enzymes was significantly inhibited. Esterified cholesterol remained almost unaltered. Findings clearly indicate the impairment of gonadotropin induced fish ovarian steroidogenesis by the four toxicants separately.

Anti-Müllerian hormone and antral follicle count predict ovarian response in women less than 45 years following GnRH antagonist multiple-dose protocol

Objective:To speculate which of the following parameters:antral follicle count(AFC),anti-Müllerian hormone(AMH),follicle-stimulating hormone(FSH)and age can be used as a predictor of ovarian response to gonadotropin-releasing hormone(GnRH)antagonist stimulation multiple-dose protocol in women under 45 years,and to determine the cutoff value of these parameters and their correlations for predicting low and high ovarian response.Methods:This prospective study included 462 women with the mean age of(29.3±6.5)years.All women were subjected to the GnRH antagonist stimulation multiple-dose protocol.On the second day of the menstrual cycle,ultrasonography was conducted to determine AFC in both ovaries.Peripheral blood samples were collected to evaluate the level of estradiol,FSH,luteinizing hormone,prolactin,thyroid-stimulating hormone,and AMH.The women were divided into three groups:low response(AHH<1 ng/mL,n=173),normal response(AMH=1.0-3.5 ng/mL,n=175),and high response(AMH>3.5 ng/mL,n=114).Results:A significant decrease was found in the age and FSH level in the high response group compared to other groups(P<0.001).Conversely,a significant increase was shown in AMH,estradiol on human chorionic gonadotropin(hCG)day,AFC,mature oocytes,fertilized oocytes,and embryos transferred in the high response group compared to the other two groups(P<0.001).The receiver operating characteristic(ROC)curves demonstrated that AFC and AMH had the highest accuracy,followed by basal FSH level and age in the prediction of low ovarian reserves(P<0.001)with cutoff values of≤4.50 and≤0.95 for AFC and AMH,respectively.Moreover,the ROC analysis showed that AFC had the highest accuracy,followed by AMH level and age in the prediction of high ovarian reserves with a cutoff value of≥14.50,≥3.63,and≤27.50 years,respectively(P<0.01).A significant decrease was observed in women's age,estradiol level,and oocyte fertilization rate in pregnant women compared to non-pregnant women(P<0.001).Additionally,significant negative correlations were found between the AFC,the number of mature oocytes,fertilized oocytes,embryos transferred,and the age of pregnant women(P<0.001).Conclusions:AFC and AMH predict low and high ovarian response to GnRH antagonist stimulation multiple-dose protocol in women under 45 years.

Effects of Bu-Shen-An-Tai Recipe and Its Two Components on Endometrial Morphology during Peri-implantation in Superovulated Mice

The aim of this study was to investigate the effect of Bu-Shen-An-Tai recipe （BSATR） and its two components （Bushen recipe, and Huoxue recipe） on endometrial morphology during peri-implantation in superovulated mice. Mice were randomly divided into five groups, including the normal （N）, model （M）, Bushen （BS）, Huoxue （HX） and Bu-Shen-An-Tai （BH） groups. The uteri were collected on day 4 of pregnancy, and the endometrium thickness, microvessel density （MVD） and number of pinopodes observed. Compared with the M group, the endometrial thickness in the BS, HX and BH groups was significantly increased and there was a significant difference in endometrial thickness between the BS and the BH groups. The mean MVD was significantly lower in the M group than in the N group, and there was a significant increase in MVD in the BS, HX and BH groups as compared with the M group. Compared with the M group, the pinopode scores in the en- dometrium were significantly increased in the HX and BH groups; and the BS group had significantly higher pinipode scores than the HX and BH groups. In conclusion, the results of the present study demonstrated that the recipes （Bushen, Huoxue and BSATR） could improve the endometrial envi- ronment by regulating the endometrial thickness, MVD and the number of pinopodes at the window of implantation. Moreover, the Huoxue recipe and the BSATR were more efficient than the Bushen recipe, with the BSATR tending to have the most beneficial effects.

Pre-hCG variables associated with occurrence of ascites in IVF/ICSI patients at moderate risk of developing OHSS:A pilot investigation

Objective: To identify predictors of ascites collected prior to the hCG administration in patients undergoing IVF/ICSI treatment at moderate risk of developing moderate/severe ovarian hyperstimulation syndrome (OHSS), and, based on these predictors, develop a nomogram for estimation of the probability of presence of ascites. Methods and Materials: Data were derived from 53 patients with 20 - 30 follicles ≥10 mmat end of stimulation. All patients received a single dose of hCG (250 mg) to trigger final follicular maturation when ≥2 follicles of ≥18 mmwere observed. Transvaginal ultrasound to measure ascites (total amount of peritoneal fluid ≥9 cm2inlithotomy position) was performed 2, 5 and 8 days after the hCG administration. Associations between clinical, sonographic and endocrinological variables recorded prior to the hCG administration and presence of ascites were analyzed by univariable and multivariable logistic regression. Results: Thirty-four patients (64%) had ultrasonic evidence of ascites. The multivariable analysis identified the total number of follicles [OR 1.29 (95% CI: 1.02 - 1.69, P = 0.043)], the ovarian volume [OR 1.05 (95% CI: 1.00 - 1.11, P = 0.047)] and BMI [OR 0.76 (95% CI: 0.56 - 0.99, P = 0.053)] as predictors of ascites (AUC = 0.825). A nomogram (PROFET) was designed with these three variables for individual prediction of the probability of development of ascites. Conclusions: This pilot investigation indicates that the risk of peritoneal fluid accumulation in IVF/ICSI patients at moderate risk of developing moderate/severe OHSS is influenced by the number of follicles and the ovarian volume on the day of hCG administration as well as the BMI.

The effect of serum luteinizing hormone on trigger day with a GnRH antagonist protocol in IVF/ICSI treatment

Objective:To investigate the effect of serum luteinizing hormone(LH)on trigger day with a Gonadotrophinreleasing hormone(GnRH)antagonist protocol in patients receiving in vitro fertilization/intracytoplasmic sperm injection(IVF/ICSI)treatment for pregnancy outcomes.Methods:We retrospectively reviewed the medical documents of patients receiving IVF/ICSI with fresh embryo transfers from the Reproductive Medicine Center of Peking University People's Hospital between January 2016 and December 2018.730 patients were included and divided into three groups by their serum LH level determined on trigger day.All patients were categorized into Group A,Group B,and Group C based on LH concentrations<1.0 IU/L,1.0–5.0 IU/L,and from 5.0 to 10.0 IU/L on trigger day during the cycle,respectively.Comparisons were made between the three groups.Results:There was a significant difference in implantation rates between Group A and Group C(24.8%versus 40.1%,respectively,P<0.05).The clinical pregnancy rates(39.3%versus 54.3%,respectively,P=0.078)and live birth rate(LBR)(32.1%versus 46.5%,respectively,P=0.116),though the differences were not significant.Multivariate logistic regression analysis showed that the OR of Group C for clinical pregnancy(OR=1.849,P=0.040)and for LBR(OR=1.915,P=0.034)were significant using Group A as the base level.Conclusions:Our study has demonstrated that patients with higher serum LH levels(5.0–10.0 IU/L)on trigger day in the GnRH antagonist protocol may confer better clinical outcomes than those with lower LH levels(<1.0 IU/L).

Clomiphene Citrate or Medroxyprogesterone Acetate with Human Menopausal Gonadotropin in Poor Responders during In vitro Fertilization/Intracytoplasmic Sperm Injection Treatments Combined with Embryo Cryopreservation?

Objective:This study aimed to compare the efficacy of clomiphene citrate(CC)with human menopausal gonadotropin(hMG)and that of medroxyprogesterone acetate(MPA)with hMG in poor responders defined according to the Bologna criteria.Methods:The data of patients with poor ovarian response(POR)according to the Bologna criteria from September 2016 to November 2017 were retrospectively reviewed.All participants received either CC+hMG or the progesterone-primed ovarian stimulation protocol(PPOS)protocol.Results:A total of 340 patients and 563 in vitro fertilization cycles were analyzed in this study.The incidence of spontaneous luteinizing hormone(LH)surge and the mean LH level on trigger day were significantly lower in the PPOS group than in the CC+HMG group(0.04%vs.3.49%and 4.26±3.59 vs.9.38±6.92 mIU/mL,respectively,P<0.05);however,the incidence of premature ovulation was similar between the two groups.The number of viable embryos harvested was not statistically different between the two groups(1.7±1.1 vs.1.5±0.8,P=0.06).The mean dose and duration of hMG were significantly higher in the PPOS group than in the CC+hMG group(908.7±556.6 vs.177.9±214.5 IU and 6.0±3.4 vs.1.2±1.5 days,respectively,P<0.05).However,the number of oocytes retrieved,number of metaphase II oocytes,and fertilization rate were comparable between the two groups.Conclusions:The CC with low-dose gonadotropin strategy was superior to the MPA with hMG protocol for POR.

Source of human oocytes for infertility treatment and fertility preservation

The source of human oocytes is directly associated with the success of infertility treatment and fertility preservation. The number of oocytes obtained is possibly correlated with the success rate in terms of live birth rates. However, oocyte quality must be refined such that the number of oocytes is not positively correlated with the quality of oocytes. Different ovarian stimulation protocols can be used to obtain numerous oocytes. However, oocyte quality should be considered the most important factor affecting infertility treatment and fertility preservation. Infertile women are generally willing to take health-related risks to achieve a live birth, and ovarian stimulation using high-dose hormonal treatments may be harmful for both women and infants. Therefore, successful infertility treatment and fertility preservation should be defined as the birth of a healthy baby at term without compromising the health and safety of the mother and infant. Therefore, the source of high-quality oocytes must be carefully considered prior to infertility treatment and fertility preservation.