Chinese herbal decoction of Wenshen Yangxue formula improved fertility and pregnancy rate in mice through PI3K/Akt signaling.

OBJECTIVE: Traditional Chinese medicine (TCM) is an effective management to infertility. The association between TCM-mediated fertility and inhibition of phosphatidylinositol-3-kinase (PI3K) would be investigated. METHODS: Institute of Cancer Research mice were treated with three herbal decoctions, named Wenshen Yangxue formula, Wenshen formula, and Yangxue formula, plus with human gonadotropins. PI3K inhibitor wortmannin was administrated to half of mice. Some index such as body weight, fertility ability would be investigated. The expression of P13K/Akt signaling was detected by using Western blot analysis. RESULTS: No difference was observed in body weight among groups. Mice receiving the administration of human gonadotropins and herbal decoctions showed increased follicle numbers, percentage of fertilization, and promoted embryonic development. The treatment of Wenshen Yangxue formula decoction showed the highest efficiency, significant higher than Wenshen and Yangxue formulas. And increased the expression of p-PI3K and p-Akt proteins. CONCLUSION: These results suggested the herbal decoctions promoted the fertilization of mice, which was related to the charge of PI3K/Akt activation.

Myo-inositol supplementation reduces the amount of gonadotropins and length of ovarian stimulation in women undergoing IVF: a systematic review and meta-analysis of randomized controlled trials.

PURPOSE: To evaluate whether oral myo-inositol supplementation (MI) is able to reduce the amount of gonadotropins (GA) and the length of controlled ovarian hyperstimulation (SL) in both Polycystic Ovarian Syndrome (PCOS) and non-PCOS women undergoing in vitro fertilization (IVF). METHODS: We performed a systematic review (PROSPERO ID: CRD42017069439) of randomized controlled trials (RCTs). We searched articles published in English between January 1985 to August 2017, using the combination of the Medical Subject Headings "Inositol" with "Ovulation Induction", "follicle-stimulating hormone, human, with HCG C-terminal peptide", "Reproductive Techniques, Assisted", and "Fertilization in Vitro". We collected data about GA and SL comparing MI to no treatment or D-Chiro-Inositol (DCI) supplementation (controls). A subgroup analysis was performed to evaluate selected outcomes in PCOS and non-PCOS women. RESULTS: We included 8 studies embedding 812 participants. We found a reduction in GA (p < 0.00001) and SL (p = 0.0007) in patients receiving MI with respect to controls. MI was effective in both PCOS (p < 0.00001) and non-PCOS women (p = 0.02) in reducing GA; conversely, MI supplementation decreased the SL only in PCOS women (p < 0.00001). CONCLUSION: During IVF, MI is effective in both PCOS and non-PCOS women in saving gonadotropins, but reduces efficiently SL only in PCOS women.

Gene expression profiling of human blastocysts from in vivo and 'rescue IVM' with or without melatonin treatment.

To evaluate the effect of melatonin supplementation in maturation medium for human 'rescue IVM' and investigate differences in transcriptomic profile of blastocysts developed from oocytes matured in vitro with/without melatonin treatment and in vivo, a total of 314 GV oocytes and 320 MI oocytes were collected from 200 patients younger than 35 years old undergoing ICSI cycle. The oocytes were randomly distributed in the control group (no melatonin) and four other groups of varying melatonin concentrations (10­11, 10­9, 10­7, 10­5 mol/l). Gene profiling was performed on blastocysts developed from in vivo maturation oocytes (in vivo group), and in vitro maturation (IVM) oocytes with an optimal concentration of melatonin treatment (IVM­anti group) or without melatonin (IVM group). The ratio of high quality blastocysts was significantly higher in the groups treated with 10­5 mol/l melatonin compared with others groups. The large­scale analysis of the transcriptome revealed significant differences in mRNA expression levels. In each group, nine blastocysts were selected for gene expression profiling. The differentially expressed genes were involved in cysteine and methionine metabolism, regulation of apoptotic process, mineral absorption, steroid hormone biosynthesis, Wnt signaling, p53 signaling pathway and other functions. The findings indicated that the IVM procedure may potentially affect DNA methylation and the canonical Wnt signaling pathway. Exogenous melatonin positively influenced quality of blastocysts, which may be mediated via upregulation of p53 signaling and correcting DNA methylation changes caused by 'rescue IVM'. However, this study reflected what was generally referred to as 'rescue IVM' and was not a true reflection of clinical IVM techniques. Therefore, melatonin required further investigation as a promising supplement for use in IVM.

[Poor responders: How could we improve our results?] / Mauvaises répondeuses : peut-on améliorer nos résultats ?

OBJECTIVES: Finding an efficient treatment for poor responders still poses a tremendous challenge for assisted reproductive technology. In 2011, an international consensus has been reached in Bologna on how to standardize the definition of poor ovarian response (POR) in a simple and reproducible manner. This article provides an objective assessment of the different treatment options currently available. METHODS: A search of the database PUBMED was carried out for studies published in English between October 2000 and April 2016. RESULTS: There is no ideal protocol to manage poor responders even though the antagonist protocol seems to have an advantage of clinicians. This is thanks to better patient tolerance and reduced total dose of gonadotrophin as well as shorter time of stimulation. It seems that there is no benefit in increasing the gonadotrophin daily doses over 300IU nor using any specific type of gonadotrophin. Today, there is insufficient evidence to recommend any additional treatment for poor responders. Only dehydroepiandrosterone (DHEA) seems to increase embryonic quality and pregnancy rate, however further exploration and complementary prospective studies are necessary. CONCLUSION: New treatment strategies such as "oocyte banking" or double stimulation during the same cycle, could provide new prospects in poor responders management.

The effect of transcutaneous electrical acupoint stimulation on pregnancy rates in women undergoing in vitro fertilization: a study protocol for a randomized controlled trial.

BACKGROUND: The latest meta-analysis demonstrated that acupuncture improves pregnancy rates among women undergoing in vitro fertilization-embryo transfer (IVF-ET), and surface acupoint stimulation, such as transcutaneous electrical acupoint stimulation (TEAS), may have the same or better potential. METHODS/DESIGN: To explore the effect of TEAS on the clinical pregnancy rate (CPR) and live birth rate (LBR) compared with real acupuncture and controls in women undergoing IVF, a multicenter, randomized controlled trial will be conducted. The inclusion criteria are the following: infertile women <40 years of age undergoing a fresh IVF or intracytoplasmic sperm injection cycle, and the study will be restricted to women with the potential for a lower success rate as defined by two or more previous unsuccessful ETs (fresh or frozen). Those who have severe illnesses possibly precluding IVF or pregnancy, have FSH levels greater than 20 IU/L, received donor eggs, had been previously randomized for this study or had undergone acupuncture (in any modality) as infertility treatment will be excluded. The subjects will be randomly assigned to the TEAS group (IVF + TEAS), the electro-acupuncture (EA) group (IVF + EA), or the control group (only IVF). A total sample size of 2,220 women is required to detect differences in CPR among the three groups. TEAS or EA treatments will start once every two or three days from day 3 of menstruation in the ovarian stimulation cycle until the day of ET. The parameters of TEAS or EA will be the following: a frequency of 2/100 Hz, a moderate electrical current of 3 to 5 mA for TEAS and 0.8 to 1.0 mA for EA. The primary outcome is CPR. Secondary outcomes are LBR, the number of oocytes aspirated and the total gonadotropin dose used in the stimulation cycle. DISCUSSION: This study will provide significant evidence for using a new method (TEAS) in IVF. TRIAL REGISTRATION: ClinicalTrials.govID: NCT01608048 (05/24/2012).

Aromatase-null mice expressing enhanced green fluorescent protein in germ cells provide a model system to assess estrogen-dependent ovulatory responses.

Enhanced green fluorescent protein (EGFP) has provided us with valuable approaches for tracking living cells. We established a novel line of transgenic mice, which express EGFP in the testis and ovary. Histological analysis demonstrated that spermatids in the testis and oocytes in ovarian follicles beyond preantral stages were positive for EGFP. By exploiting these features, we evaluated ovulatory responses of aromatase-gene (Cyp19a) knockout mouse expressing the EGFP transgene, which is totally anovulatory due to 17ß-estradiol (E2) deficiency. Ovulation in the knockout mice was induced by sequential injections of E2 on days 1, 4 and 5, pregnant mare serum gonadotropin on day 4 and human chorionic gonadotropin on day 6. Fluorescent oocytes were readily detectable at 15 h after the last gonadotropin injection in the oviduct under a fluorescence stereomicroscope, even when only one oocyte was present. However, when E2 supplementation on day 4 or day 5 in the regimen was omitted, no ovulated oocytes were detected, indicating that exogenous E2 supplementation at the time of gonadotropin stimulation is necessary to induce ovulation in aromatase-gene knockout mice. Our results further demonstrated that the current mouse line can provide an alternative tool to study germ cell biology, including oogenesis, ovulation and senescence.

[Effects of Yupei Qisun Sequential Method of Chinese medicine on correlated indices of repeated implantation failure patients in the fresh cycle].

OBJECTIVE: To study the clinical effects of Yupei Qisun Sequential Method (YQSM, by Shen supplementing and Pi invigorating) of Chinese medicine on correlated indices of repeated implantation failure patients in the fresh cycle. METHODS: Sixty patients with more than three failure cycles of in virto fertilization and embryo transfer (IVF-ET) or intracytoplasmic sperm injection (ICSI) were recruited. They were assigned to the treatment group (treated by IVF/ICSI and Chinese medicine) and the control group (treated by IVF/ICSI alone), 30 in each group. The total dose of gonadotropin (Gn), the days of controlled ovary hyperstimulation (COH), the thickness of endometrium on the day of embryo transplantation, the number of retrieved oocytes, the fertilization number, the embryo number, the high quality embryo number, the pregnancy rate, and the implantation rate were compared. RESULTS: In the treatment group the numbers of embryo and high quality embryo were 7.5 +/- 4.9 and 5.1 +/- 3.2 respectively, which were both higher than those of the control group with significant difference (5.1 +/- 3.2, 3.2 +/- 1.8; P < 0.05). No significant difference existed in aspects of the total numbers of Gn, the days of COH, the thickness of endometrium on the day of embryo transplantation, the numbers of retrieved oocytes, the fertilization number, the pregnancy rate, and the implantation rate between the two groups (P > 0.05). CONCLUSION: YQSM combined with COH could improve the quantity and the quality of embryos, which was promising to increase the accumulative pregnancy rate of RIF patients.

[Effect of bushen tiaojing recipe and xiaoyao pill on expression of cathepsin-L mRNA in gonadotropin-primed mice].

OBJECTIVE: To study the effect of Bushen Tiaojing Recipe (BTR) and Xiaoyao Pill (XP) on cathepsin-L (Cat-L) mRNA in mice. METHODS: Immature mice were randomly divided into the normal group, the control group, the BTR group and the XP group, three in each group. Cat-L mRNA expression in mice was detected using reverse transcription polymerase chain reaction (RT-PCR) at 0, 4, 8 and 12 h after injecting 5 IU (human chorionic gonadotropin, HCG). RESULTS: Cat-L mRNA expression increased gradually after HCG injection, the relative levels in the control group at 0, 4, 8 and 12 h were 0.066 +/- 0.005, 0.383 +/- 0.045, 0.737 +/- 0.024 and 1.036 +/- 0.073 respectively, comparisons between different time-points showed significant difference (P < 0.01). Compared with the control group, the Cat L mRNA expression was higher at 4 h in both BTR and XP groups (P < 0.01), at 8 h in the XP group (P < 0.05), and at 12 h in BTR group after injecting HCG (P < 0.05). Compared with the control group, cat L mRNA expression showed no statistic difference at 8 h in BTR group and at 12 h in XC group. CONCLUSIONS: BTR promoted the ovulation by enhancing the expression of CatL gene, and that of XP by advancing the peak of CatL gene expression.

The role of inositol supplementation in patients with polycystic ovary syndrome, with insulin resistance, undergoing the low-dose gonadotropin ovulation induction regimen.

In an attempt to evaluate the role of inositol supplementation in insulin-resistant patients with polycystic ovary syndrome (PCOS), undergoing gonadotropin ovulation induction using the low-dose step-down regimen, we conducted a prospective longitudinal study comparing the stimulation characteristics of 15 patients treated with inositol, to a cohort, matched by age and body mass index (BMI), without inositol. Inositol nutritional supplementation produced very good clinical results with a significant reduction in cancellation rate (0 vs. 40%) and the consequent improvement in clinical pregnancy rate (PR) (33.3% vs. 13.3%).

Pregnancy and beta-thalassemia: an Italian multicenter experience.

BACKGROUND: Recent advances in the management of thalassemia have significantly improved life expectancy and quality of life of patients with this hemoglobinopathy, with a consequent increase in their reproductive potential and desire to have children. DESIGN AND METHODS: We describe the methods of conception and delivery, as well as the course and outcome of pregnancy including transfusions, iron overload and chelation in 46 women with thalassemia major (58 pregnancies) and in 11 women with thalassemia intermedia (17 pregnancies). Conception was achieved after gonadotrophin-induced ovulation in 33 of the women with thalassemia major and spontaneously in all of those with thalassemia intermedia. RESULTS: Among the women with thalassemia major, 91% of the pregnancies resulted in successful delivery of 45 singleton live-born neonates, five sets of twins and one set of triplets. No secondary complications of iron overload developed or worsened during pregnancy. When considering only the singleton pregnancies, the proportion of babies with intrauterine growth retardation did not differ from that reported in the general Italian population. The high prevalence of pre-term births (32.7%) was mostly related to multiple pregnancies and precautionary reasons. Pregnancy was safe in most women with thalassemia major or intermedia. However, women with thalassemia intermedia who had never previously been transfused or who had received only minimal transfusion therapy were at risk of severe alloimmune anemia if blood transfusions were required during pregnancy. CONCLUSIONS: Provided that a multidisciplinary team is available, pregnancy is possible, safe and usually has a favorable outcome in patients with thalassemia. In women with hypogonadotropic hypogonadism, gonadal function is usually intact and fertility is usually retrievable.

Responses of gonadotropin secretion to short-term dietary supplementation in ovariectomized goats with different body weights.

The responses of luteinizing hormone (LH) and follicle stimulating hormone (FSH) secretion to acute dietary supplementation were studied in goats with different body weights. Ovariectomized Shiba goats (n=11) were used and were maintained with a feed of 100% of their energy requirement. They were implanted subcutaneously with an oestradiol capsule and were divided into light (LBW; 24kg, n=5, mean+/-S.D., 32.0+/-6.3kg) groups on the basis of their body weights at 8 days before the start of treatment. At the start of treatment (Day 1), the level of a feed changed to 250% of their energy requirement and this level was maintained for 7 days in both groups. Blood samples were collected daily from Day -7 to Day 7 for the analysis of FSH, glucose, and insulin profiles in plasma. Frequent blood samples were also collected at 10min intervals for 6h on Day 0, Day 3, and Day 7 for analysis of LH pulses. LH pulse frequency increased significantly on Day 3 as compared with that on Day 0 in both the HBW (7.4+/-0.5pulses/6h vs. 6.2+/-0.8pulses/6h, p<0.05) and LBW (6.5+/-0.8pulses/6h vs. 5.5+/-0.5pulses/6h, p<0.05) groups, whereas it decreased on Day 7 (HBW, 6.4+/-0.9pulses/6h; LBW, 6.3+/-1.6pulses/6h, p>0.05 vs. Day 0). Plasma glucose and insulin concentrations increased temporarily from Day 2 to Day 4 and then decreased to the level before the start of dietary supplementation in both groups. There was no significant difference in the LH pulse frequency or daily concentrations of FSH, glucose, or insulin between the HBW and LBW groups throughout the experimental period. The present study indicated that acute dietary supplementation stimulates pulsatile LH secretion in parallel with a rise of blood glucose and insulin levels. However, the influence of body weight on these responses between light and heavy animals was not observed.

Doxycycline inhibits vascular leakage and prevents ovarian hyperstimulation syndrome in a murine model.

OBJECTIVE: To determine whether doxycycline would inhibit the development of ovarian hyperstimulation syndrome (OHSS) in a murine model. DESIGN: Testing the effect of oral doxycycline treatment on gonadotropin-induced peritoneal capillary leakage in mice. SETTING: Animal research facility. ANIMAL(S): Four-week-old female C57Bl/6 mice. INTERVENTION(S): Ovarian hyperstimulation syndrome was induced in mice by administering gonadotropins for 3 days followed by human chorionic gonadotropin. Peritoneal vascular leakage was quantified using the Miles vascular permeability assay. Mice were treated with daily oral doxycycline or vehicle. MAIN OUTCOME MEASURE(S): The concentration of dye in abdominal fluid extracted by peritoneal lavage was measured spectrophotometrically. Ascites volume was determined using dye dilution. RESULT(S): Doxycycline inhibited peritoneal vascular leakage and ascites accumulation in the hormonally stimulated mice. Doxycycline treatment did not inhibit ovarian stimulation or ovulation when compared with controls, as indicated by similar ovarian morphology, ovarian weights, and corpora lutea counts. Importantly, vessel density within the corpora lutea was similar in the two groups. CONCLUSION(S): Doxycycline prevents OHSS in a murine model without compromising ovarian stimulation. This effect is caused by inhibition of vascular leakage. Doxycycline's potential in preventing human OHSS is promising and warrants further studies.

Ovulation induction with pulsatile gonadotropin-releasing hormone (GnRH) or gonadotropins in a case of hypothalamic amenorrhea and diabetes insipidus.

Hypothalamic amenorrhea is a treatable cause of infertility. Our patient was presented with secondary amenorrhea and diabetes insipidus. Cortisol and prolactin responded normally to a combined insulin tolerance test (ITT) and thyrotropin-releasing hormone (TRH) challenge, while thyroid-stimulating hormone (TSH) response to TRH was diminished, and no response of growth hormone to ITT was detected. Both luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels increased following gonadotropin-releasing hormone (GnRH) challenge. No response of LH to clomiphene citrate challenge was detected. Magnetic resonance imaging findings demonstrated a midline mass occupying the inferior hypothalamus, with posterior lobe not visible and thickened pituitary stalk. Ovulation induction was carried out first with combined human menopausal gonadotropins (hMG/LH/FSH) (150 IU/day) and afterwards with pulsatile GnRH (150 ng/kg/pulse). Ovulation was achieved with both pulsatile GnRH and combine gonadotropin therapy. Slightly better results were achieved with the pulsatile GnRH treatment.