Outcomes of follicle-stimulating hormone priming and nonpriming in in vitro maturation of oocytes in infertile women with polycystic ovarian syndrome: a single-blinded randomized study.

BACKGROUND/AIMS: Follicle-stimulating hormone (FSH) priming has been studied in in vitro matured oocytes for oocyte maturation rate, embryo quality, and pregnancy rate with discouraging results. This study aimed to initiate FSH stimulation later, i.e. on day 6, to prolong natural endometrial priming and promote oocyte maturation. METHODS: Forty polycystic ovarian syndrome (PCOS) patients were enrolled into a single, blinded (investigator), randomized, controlled study, and randomly allocated to group 1 (no FSH priming) or group 2 (day 6 recombinant FSH priming). Oocytes were retrieved after human chorionic gonadotropin injection on day 10. After 27 or 51 h of incubation, only mature oocytes were denuded and fertilized by intracytoplasmic sperm injection. Two day 3 embryos were transferred in most patients. Rates of oocyte maturation, cleavage, and pregnancy were compared. RESULTS: The oocyte maturation rates within 51 h were 62.6 and 72.7% in groups 1 and 2, respectively (p < 0.01). The embryo cleavage rate was significantly higher in group 2 than in group 1 (77.3 vs. 63.6%, p < 0.05). The pregnancy rate was higher in group 1 than in group 2 (50 vs. 30%, p > 0.05). CONCLUSION: FSH priming is beneficial for promotion of the maturation and quality of oocytes, leading to a higher embryo cleavage rate and lower rate of pregnancy loss.

Ovarian reserve evaluation by anti-mullerian hormone in women undergoing laparoscopic cystectomy of endometrioma.

OBJECTIVE: To study the serial changes of serum AMH to determine ovarian reserve recovery after laparoscopic cystectomy of endometrioma. MATERIAL AND METHOD: Forty-three endometrioma patients who underwent laparoscopic cystectomy of endometrioma were tested for levels of serum AMH at preoperation, 1 week, and 3 months postoperation. RESULTS: Median serum AMH was 2.11 ng/mL (range = 0.22 to 9.24 ng/mL) before surgery. This level also reduced at first week postoperation (p < 0.01) but did not reach a significant difference between the first week and the third month (1.02 ng/mL and 1.06 ng/mL, respectively). The recovery rate of AMH level in unilateral endometrioma was higher than bilateral endometrioma (32.4% vs. -3.6%, p = 0.02). CONCLUSION: Ovarian reserve was decreased after laparoscopic cystectomy of endometrioma and did not significantly restore after three months of postoperation. The recovery of ovarian reserve after unilateral endometriotic cystectomy was faster than that after bilateral endometriotic cystectomy.

Outcome of sperm preparation using double-gradients technique study in Siriraj Hospital.

OBJECTIVE: To determine the succession of sperm preparation using the double-gradients technique on sperm quality: sperm recovery rate, sperm concentration, sperm motility and percentage of post wash total motile sperm count. STUDY DESIGN: Retrospective descriptive study. SETTINGS: Infertility clinic, Faculty of Medicine, Siriraj hospital. MATERIAL AND METHOD: During the period of January 1, 2002 through December 31, 2007, data including semen analysis before and after IUI procedure were reviewed in all male patients who were referred to the andrology laboratory for sperm washing and IUI. Comparison of semen parameters such as total sperm concentration, total motile sperm count before and after sperm preparation as well as total sperm recovery rate and total motile sperm recovery rate was evaluated. RESULTS: After sperm preparation, both sperm concentration and progressive sperm motility significantly increased, while total motile sperm count significantly decreased. Moreover, the percentage of motile sperm recovery rate and total sperm recovery rate was higher after sperm preparation at around 59.88 +/- 19.26% and 34.03 +/- 14.58% respectively. When categorizing semen parameters to 4 groups: normozoospermia, oligozoospermia, astenozo-ospermia and oligo-astenozoospermia, sperm motility in each group, comparing with sperm motility prior preparation, significantly improved after sperm preparation. Furthermore, motile sperm recovery rate in each group significantly increased except for astenozoospermia. Total sperm recovery rate in oligozoospermia was significantly higher than normozoospermia, yet the others were significantly lower. CONCLUSION: Sperm preparation using double gradient percoll provided a high percentage of motile sperm recovery rate and total sperm recovery rate. It also dramatically improved progressive sperm motility in normozoospermia, oligozoospermia, astenozoospermia and oligo-astenozoospermia.

Epigenetic analysis and suitability of amniotic fluid stem cells for research and therapeutic purposes.

Amniotic fluid stem cells (AFSs) are interesting mesenchymal stem cells (MSCs) that are characterized by their great potential for cell proliferation and differentiation compared with other types of MSCs identified to date. However, MSCs in prolonged culture have been found to exhibit defects in genetic stability and differentiation capacity. Epigenetic anomalies have been hypothesized to be a cause of these defects. Here, we investigated the genomic methylation and genetic imprinting in AFSs during prolonged in vitro culture. Four human imprinted genes, insulin-like growth factor 2 (IGF2), H19, small nuclear ribonucleoprotein polypeptide N gene (SNRPN), and mesoderm-specific transcript (MEST), were evaluated for their expression levels and methylation statuses in AFS lines. The data revealed epigenetic instability in high passage number AFS cultures. The real-time polymerase chain reaction analysis showed that the expression levels of the imprinted genes gradually increased with increased time in culture. The loss of parental allele-specific imprinting for at least 1 gene among IGF2, H19, and SNRPN was observed in every AFS line after passage 8 using allelic expression analysis. The imprinting control regions (ICRs) of the IGF2 and H19 genes were assayed for site-specific methylation using bisulfite sequencing. This assay revealed a variable level of methylated CpG sites in the ICRs of IGF2 and H19. This variable level of CpG methylation is related to the aberrant expression of the IGF2 and H19 genes in late-passage AFSs. Our results did not reveal any irregularity in the epigenetic control system in the early-passage AFSs, indicating that the standard in vitro culturing of AFSs used in medical treatments should be limited to 8 passages.

Efficacy of mefenamic acid and hyoscine for pain relief during saline infusion sonohysterography in infertile women: a double blind randomized controlled trial.

OBJECTIVES: To evaluate the efficacy and level of satisfaction from mefenamic acid and hyoscine when used for pain relief during saline infusion sonohysterography. STUDY DESIGN: In this double blind randomized controlled trial, 141 nulliparous women were allocated to receive 500 mg of mefenamic acid, 10mg of hyoscine or a placebo, which was packed in the same outer capsule. Saline infusion sonohysterography (SIS) was performed 30 min later by one operator. Pain and satisfaction scores were evaluated using a 10 cm visual analog scale. Baseline characteristics, pain and satisfaction scores were compared among the three groups. Pain scores were recorded before, after catheter insertion, during, immediately after, and 30 min after the procedure. RESULTS: No statistically significant differences were found in baseline characteristics, pain and satisfaction scores among the three groups. Maximum pain during SIS was 4.40 ± 3.34, 4.67 ± 3.14 and 4.85 ± 3.19 in the mefenamic acid, hyoscine and placebo groups respectively. There was a 31.1% prevalence of intrauterine abnormality and the most frequent finding was endometrial polyp. CONCLUSION: There is no benefit in using mefenamic acid and hyoscine in the prevention of pain occurring from SIS.