The 'normal' range of FMR1 triple CGG repeats may be associated with primary ovarian insufficiency in China.

The aim of this study was to investigate the relationship between normal Fragile X mental retardation gene 1 (FMR1) CGG repeat numbers and primary ovarian insufficiency (POI) occurrence or subsequent resumption of ovarian function. A total of 122 women with POI and 105 controls were followed up and analysed in our centre. The prevalence of premutation and intermediate range of FMR1 CGG repeats in Han Chinese women with POI was only 0.81% (1/122) and 1.64% (2/122), respectively. The risk of POI occurrence for less than 26 CGG repeats and 29 or more CGG repeats in allele1 (smaller allele) was significantly higher than that for 26-28 CGG repeats (odds ratio 13.50, 95% confidence interval: 3.21 to 56.77 and 6.32, 95% confidence interval: 2.49 to 16.09 respectively; both P < 0.001). No significant difference was found in the CGG repeat distribution (<26, 26-28, or ≥29) in FMR1 allele1 between POI cases whose ovarian function resumed and those whose ovarian function did not. It is suggested that the CGG repeat number in allele1, but not that in allele2 (longer allele), was significantly associated with POI occurrence (P < 0.001). Fewer than 26 or more than 28 CGG repeats in FMR1 allele1 were both risk factors of POI occurrence.

Human trophoblast cell during first trimester after IVF-ET differs from natural conceived pregnancy in development and function.

OBJECTIVE: To explore the differences of the trophoblast cell function in first trimester between natural pregnancy and pregnancy after IVF-ET therapy. METHODS: 102 cases with twin to singleton fetal reduction after IVF-ET treatment from July 2010 to August 2013 in Peking University Third Hospital were involved in analysis, and eight specimens were obtained from this group. 10 natural-pregnancy cases undergoing artificial abortion with unwanted pregnancy were recruited as control. Semi-quantitative immunohistochemical method was used to detect the expression of EGFR, Bcl-2, tubulin-α, metallothionein and AFP in villi in both groups. RESULTS: Of the 102 cases, 14 cases (13.73%) were aborted. Preterm birth occurred in seven cases (7.86%). Low birth weight occurred in three patients (3.37%), and extremely low birth weight occurred in four cases (4.49%). The expression of EGFR, tubulin-α, Bcl-2, and metallothionein in the IVF-ET group was significantly lower than that in the control group (P<0.05). However, AFP expression was significantly higher in IVF-ET group than in control group (P<0.05). In IVF-ET group, the miscarriage case had weaker EGFR, tubulin-α, and metallothionein expression than full-term pregnancy; the early preterm labor case had weaker Bcl-2, tubulin-α, and metallothionein expression; and velamentous cord insertion case had weaker tubulin-α expression. CONCLUSIONS: The trophoblast cell function of IVF-ET group in first trimester is different from control group in proliferation, invasion, apoptosis and vascular development, and optimal pregnancy outcome depends on the self-healing balance of trophoblast cells.

[Influence of duration of gonadotropin administration on the clinical outcome of in vitro fertilization embryo transfer].

OBJECTIVE: To investigate the influence of duration of gonadotropin (Gn) administration on the clinical outcome of in vitro fertilization embryo transfer (IVF-ET). METHODS: A total of 3 221 cycles of short protocol or antagonist protocol in our center from January 2012 to December 2012 were included in the retrospective study. According to the different duration of Gn administration, all patients were divided into group A (≤7 days, n=58) and group B (>7 days, n=3 163). The different clinical parameters, such as age, duration of infertility, body mass index (BMI), basis estradiol (E2), follicle-stimulating hormone (FSH), the number of antral follicle, the number of oocytes, endometrium thickness, fertility rate, good quality embryo rate, impatation rate and clinical pregnancy rate were compared between the two groups. RESULTS: There was no significant difference in age, duration of infertility, BMI, basis E2, FSH, the number of antral follicle between the two groups. The number of oocytes in group A was fewer than that in group B [(8.2±5.6)vs.(12.1±8.3), P=0.009]; endometrium thickness on the day of HCG in group A was thinner than that in group B [(9.9±2.1) mm vs.(10.4±1.6) mm,P=0.002]. There was no significant difference in fertility rate, good quality embryo rate, impatation rate and clinical pregnancy rate (36.2% vs. 33.6%, P>0.05). There was no significant difference in clinical pregnancy rate between the two groups in short protocol (33.3% vs. 27.2%, P>0.05). In the same way, there was no significant difference in clinical pregnancy rate between the two groups in antagonist protocol (37.5% vs. 36.6%, P> 0.05). CONCLUSION: Although short duration of gonadotropin administration in short protocol and antagonist protocol has association with fewer number of oocytes, it may not affect the outcome of IVF-ET.

[Comparison of gonadotropin releasing hormone agonist long protocol and gonadotropin releasing hormone antagonist protocol in infertile women].

OBJECTIVE: To compare the clinical outcomes of GnRH agonist (GnRH-a) long protocol and GnRH antagonist (GnRH-ant)protocol in vitro fertilization (IVF)-embryo transfer (ET) cycles, and to explore the optimized protocol for infertile women. METHODS: From June 2010 to June 2012, 2 444 infertile women underwent their IVF cycles in Peking University Third Hospital, which were divided into 1 706 GnRH agonist long protocol and 738 GnRH antagnist protocol groups. The data of the general demographic, treatment and clinical outcome were compared between the two groups. RESULTS: The age, body mass index(BMI), infertile duration, antral follicle count (AFC) did not reach statistical difference, the level of estradiol on the day of HCG: injection was higher in GnRH agonist group [(10 595±7 368)pmol/L vs. (9 087±7 035) pmol/L], and the mean length of stimulation was longer in GnRH agonist group[(12.5±1.8) d vs.(9.4±1.7) d], The dose of Gn [(3 107±1 377) IU vs. (2 084±903)IU]was higher in GnRH agonist group. The number of ovum was 13.4±6.6 in GnRH agonist group and 11.8±6.4 in GnRH antagonist group. Those clinical parameters all reached statistical difference (P<0.05). The number of the transfer embryos, fertilization rate, and cleavage rate did not reach statistical difference, but the number of the embryos was 5.6±4.5 in GnRH agonist group and 5.1±4.3 in GnRH antagonist group,reached statistical difference (P<0.05). The abortion rate, embryonic death rate, ectopic pregnancy rate, preterm labor rate, postterm pregnancy rate, fatal malformations rate showed no statistical difference, but the GnRH agonist long protocol had higher pregnancy rate (44.0% vs. 38.3%), and higher term pregnancy rate (64.2% vs. 56.9%) compared with GnRH antagonist protocol, thus those parameter reached significant difference (P<0.05). CONCLUSION: Compared with GnRH-antagnist protocol, GnRH agonist long protocol had higher pregnancy rate and better pregnancy outcome.

[Analysis of clinical factors affecting pregnancy rate of intrauterine insemination].

OBJECTIVE: To analyze the factors affecting clinical pregnancy rate of intrauterine insemination in Center of Reproductive Medicine, Peking University Third Hospital, to guide clinical treatment. METHODS: In the study, 5 167 intrauterine insemination cycles were retrospectively analyzed from May 2011 to October 2012 in our reproductive center. The data were collected, the single-factor was analyzed with χ2 test, and the multi-factor was analyzed with Logistic regression with a significant level of 0.05. RESULTS: The cycle clinical pregnancy rate was 12.8%, which decreased with the increase of the female age and infertile duration. The clinical pregnancy rate was low when the sperm density was less than 1×10(6)/mL. In the ovulation group, the clinical pregnancy rate was higher than the natural group. The group with more than 2 dominant follicles had higher clinical pregnancy rate as compared with the single dominant follicle group. The clinical pregnancy rate was the highest in the third cycle but decreased after the fourth cycle. The clinical pregnancy rate was higher in cervical factors, sexual dysfunction, and polycystic ovary than in the group with other reasons. CONCLUSION: The female age, infertile duration, ovarian stimulation and follicle number, cause of infertility were the main factors affecting clinical pregnancy outcome; the sperm density, and cycle numbers have influence too; the insemination timing, and frequency have little effect.

[Comparison of GnRH antagonist fixed protocol and GnRH agonists long protocol in infertile patients with normal ovarian reserve function in their first in vitro fertilization-embryo transfer cycle].

OBJECTIVE: To compare the clinical outcomes of gonadotropin-releasing hormone (GnRH) antagonist (GnRH-ant) fixed protocol with GnRH agonist (GnRH-a) long protocol in infertile patients with normal ovarian reserve function in their first in vitro fertilization-embryo transfer (IVF-ET) cycle, and to explore the feasibility and advantage of GnRH antagonist protocol performed in normal responders. METHODS: From January 2011 to June 2011, 771 infertile women with normal ovarian reserve function underwent their first IVF or intracytoplasmic sperm injection (ICSI) cycles in Peking University Third Hospital, which were divided into 245 cycles in GnRH-ant fixed protocol group (GnRH-ant group) and 526 cycles in GnRH-a long protocol group (GnRH-a group). The data of general demographic, treatment and clinical outcome were compared between two groups. RESULTS: Age, infertile duration, body mass index (BMI), baseline serum follicle-stimulating hormone (FSH) and estradiol levels between two groups did not reached statistical difference (P > 0.05). The level of estradiol was (12 289 ± 6856) pmol/L in GnRH-ant group and (14 934 ± 8007) pmol/L in GnRH-a group at day of hCG injection. The mean length of stimulation was (10.3 ± 1.2) days in GnRH-ant group and (12.8 ± 1.6) days in GnRH-a group. The dose of gonadotropin was (2013 ± 607) U in GnRH-ant group and (2646 ± 913) U in GnRH-a group. The number of ovum was 15 ± 7 in GnRH-ant group and 17 ± 8 in GnRh-a group. Those clinical parameter all reached statistical difference (P < 0.05). The number of embryo was 7 ± 4 in GnRH-ant group and 8 ± 5 in GnRH-a group, the rate of clinical pregnancy was 40.9% (94/230) in GnRH-ant group and 45.6% (216/474) in GnRH-a group, the rate of implantation was 26.1% (128/490) in GnRH-ant group and 30.9% (307/994) in GnRH-a group, the rate of continuing pregnancy was 38.7% (89/230) in GnRH-ant group and 42.6% (202/474)in GnRH-a group, those parameter did not reach statistical difference (P > 0.05). The rate of moderate or severe ovarian hyperstimulation syndrome was 2.4% (6/245) in GnRH-ant group and 4.2% (22/526) in GnRH-a group, which did not show significant difference (P > 0.05). CONCLUSION: In the first IVF or ICSI cycle of the patients with normal ovarian reserve function, the fixed GnRH-ant protocol could get the same satisfied clinical outcome, and it is more economic, convenient and safer compared with low dose depot GnRH-a long protocol.

Efficacy of local aspiration in the conservative treatment of live interstitial pregnancy coexisting with live intrauterine pregnancy after in vitro fertilization and embryo transfer.

BACKGROUND: Heterotopic pregnancy (HP) is defined as a pregnancy in which one or more embryos is viably implanted in the uterus while the other is implanted elsewhere as an ectopic pregnancy. The occurrence of HP rises dramatically with the increased use of assisted reproductive technology. HP of interstitial pregnancy is one special situation which needs more concern. Here we evaluate the efficacy of local aspiration and instillation of hyperosmolar glucose in the treatment of live interstitial pregnancy complicated with live intrauterine pregnancy after in vitro fertilization and embryo transfer. METHODS: Five female patients were diagnosed with live interstitial pregnancies complicated with intrauterine pregnancies. They were treated with transvaginal ultrasound-guided aspiration of interstitial pregnancy and instillation of hyperosmolar glucose at the Center for Reproductive Medicine of Peking University Third Hospital from January 1st, 2008 to May 30th, 2011. RESULTS: Gemmule embryos in all 5 cases were aspirated successfully and there was no abdominal hemorrhage, threatened abortion or infection in any of the cases. The sac of interstitial pregnancy continued to progress after aspiration and stopped growing between 11 to 20 weeks. By the 30th week of pregnancy, 80% of the interstitial masses had disappeared. Four cases have delivered and one is still in on-going pregnancy. All of the four cases underwent cesarean section and there were nothing special detected in the corner of the uterus. CONCLUSION: Local aspiration and instillation of hyperosmolar glucose may be an effective way to treat live interstitial pregnancy when coexisting with a live intrauterine pregnancy.

Comparison of the major malformation rate of children conceived from cryopreserved embryos and fresh embryos.

BACKGROUND: Cryopreserved embryo transfer has become indispensable in reproductive technology. More and more children are conceived from frozen-thawed embryo transfer (FET). The risk of birth defects associated with frozen-thawed embryo transfer has been evaluated and conflict results are obtained. The aim of this study was to compare the rate of major malformations in children conceived from cryopreserved embryos with that of children from fresh embryos. METHODS: A retrospective analysis was performed on children conceived from frozen-thawed embryos and fresh embryos between January 2005 and December 2008 at the Reproduction Center of the Third Hospital, Peking University. The major malformation rates were compared between two groups for all children, as well as singletons or twins, separately. The frequencies of different subtypes of malformations classified according to different organ system were also compared. RESULTS: Thirty-four of 3125 children from cryopreserved embryos had a major malformation. The malformation rate was 1.09%, which was comparable to that for children after fresh embryos transfer (1.53% (55/3604), OR: 0.71, 95%CI; 0.46-1.09). The malformation rate was also similar when the analysis was limited to children from cryopreserved embryos resulted from in vitro fertilization (IVF) (1.39%) and fresh IVF (1.3%). However, children from cryopreserved embryos resulted from intracytoplasmic sperm injections (ICSI) had much lower malformation rate than from fresh ICSI (0.63% vs.1.83%, OR: 0.34, 95%CI: 0.16-0.75). No difference was found in the incidence of major malformations in singletons from cryo ICSI (0.73%) and fresh ICSI (1.9%), or from cryo IVF (1.49%) and fresh IVF (1.67%). Similar malformation rate was found in multiples from cryo ICSI (0.52%) and fresh ICSI (1.76%), or cryo IVF (1.30%) and fresh IVF (0.90%). The distribution and risk of the subtype of malformations, such as cardiovascular, gastrointestinal, neural tube, urogenital, musculoskeletal and facial abnormalities was not different between the cryo group and fresh group. CONCLUSIONS: The major malformation rate is similar between fetuses/children conceived from cryopreserved embryos and those from fresh embryos. Large prospective and long-term follow-up studies are needed to get exact results concerning the birth defects of the children born after cryopreserved embryos.

[Effect of hyperprolactinemia upon clinical symptoms of patients with polycystic ovary syndrome].

OBJECTIVE: To determine the effects of hyperprolactinemia (hyperPRL) upon the symptoms of patients with polycystic ovary syndrome (PCOS). METHODS: Age, body mass index, levels of hormone, lipid, beta-cell function and family medical history of 224 women with PCOS and 30 women with hyperPRL + PCOS were compared. RESULTS: Patients with hyperPRL + PCOS were younger to develop endocrine disturbances, an increased incidence of acne (64% vs 28% respectively), a high level of androstenedione (20 + or - 7 vs 13 + or - 5) nmol/L respectively and prolactine in serum (1492 + or - 1175 vs 367 + or - 164) mIU/L respectively; The PCOS patients were divided into the groups of hyperandrogenism PCOS and non-hyperandrogenism PCOS depending on the serum level of androgen. A higher level of T and A was found in serum in PRL-PCOS than non-hyperandrogenism patients and similar as hyperandrogenism PCOS patients. They had reduced ApoB (680 + or - 230 nmol/L vs 943 + or - 179 mmol/L respectively) and Lpa level (46 + or - 22 nmol/L vs 162 + or - 194 mmol/L respectively) and high HOMA-IR when compared with non-hyperandrogenism PCOS; Patients' sisters with hyperPRL + PCOS had a significantly greater incidence of acne, higher rates of infertility and PCOS when compared with PCOS patients. Levels of other hormones, metabolic profiles and other family histories did not differ between patients with PCOS and hyper-PRL+PCOS. CONCLUSION: Patients with hyperPRL + PCOS develop the endocrine disturbances at a younger age, a greater incidence rate of acne, level of prolactin and androstenedione, they have reduced ApoB and increased HOMA-IR. Patients' sisters with hyperPRL + PCOS have significantly greater incidence of acne, higher rates of infertility and PCOS as when compared with PCOS patients.

[Comparison of multiple pregnancy rate following transfer of frozen-thawed or fresh embryos and analysis of related factors].

OBJECTIVE: To compare the multiple pregnancy rates of frozen-thawed embryos transfer (FET) or fresh embryos transfer, analyze the factors related to multiple pregnancies after IVF and study pregnancy rates with different number of embryos transferred in FET cycle. METHODS: A retrospective analysis was performed upon multiple pregnancies from 1235 and 1561 clinical pregnancies conceived by FET or fresh embryo transfers. RESULTS: No correlation was found between fresh or cryopreserved embryos transfer and multiple pregnancy rates. There were significant effects of woman's age, number of embryos transferred and stage of embryos upon multiple pregnancy rates. When the same number of cleavage-stage embryos was transferred to women with the same age, twin pregnancy rate or triplet pregnancy rate was the same between FET and fresh cycles. Triplet pregnancy rate with three embryos transferred was significantly higher than that of two embryos transferred. In women under 35 years old, the pregnancy rate with two embryos transferred reached 36.1%. CONCLUSION: Frozen-thawed embryos or fresh embryos transfer has no effect upon the multiple pregnancy rate. Women under 35 years old can achieve acceptable pregnancy rates when two cryopreserved embryos are transferred. It is helpful to reduce the triplet pregnancy rate.

Normal birth from cryopreserved embryos after intracytoplasmic sperm injection of frozen semen into vitrified human oocytes.

OBJECTIVE: To describe the birth achieved from frozen embryos after intracytoplasmic sperm injection (ICSI) of donor sperm into vitrified oocytes. PATIENT: A 25-year-old woman whose husband was azoospermic undergoing IVF therapy. METHODS: Oocytes collected after ovarian stimulation were vitrified, thawed, and fertilized by frozen donor sperm. RESULTS: Nineteen oocytes were vitrified and all survived after thawing. Thirteen of the 19 oocytes that underwent ICSI with donors sperm were successfully fertilized. Twelve embryos were cryopreserved again by conventional slow-freezing protocol because of uterine bleeding on the day of transfer. Three thawed embryos were transferred, and a normal male with an infant karyotype of 46,XY was delivered. CONCLUSION: This case report demonstrates effective oocyte cryopreservation by vitrification.

[Follicular stimulating hormone receptor gene C566T mutation in premature ovarian failure].

OBJECTIVE: To investigate the incidence of follicular stimulating hormone receptor (FSHR) gene C566T mutation in Chinese women with premature ovarian failure (POF) and to explore the etiologies of POF. METHODS: This case-control study was carried out between 73 Chinese women with idiopathic POF (POF group) and 35 controls (control group), including 25 normal females with a regular menstrual history and 10 normal post-menopause women. DNA was extracted from the peripheral blood of patients and controls. The exon 7 of FSHR gene was amplified by PCR. PCR products were subsequently digested by the enzyme BsmI and then subjected to electrophoresis on agarose gels and stained with ethidium bromide to determine the C566T mutation. DNA samples of random sampling were further analysed by sequencing the PCR products to confirm the mutation. RESULTS: BsmI digestion resulting in two fragments of 51 and 27 base pairs was noted for all 73 POF patients and 35 controls. PCR sequencing confirmed that the 566 allele of FSHR gene is C, demonstrating normal FSHR allele. CONCLUSIONS: No FSHR gene C566T mutation is present in POF patients and controls. FSHR C566T mutation may be rare in Chinese women with POF.