Effects of cilostamide and forskolin on the meiotic resumption and embryonic development of immature human oocytes.

BACKGROUND: In an attempt to allow for acquisition of oocyte cytoplasmic maturation, PDE3 specific inhibitor, cilostamide and adenylate cyclase activator, forskolin were used to extend pre-maturation culture of immature human oocytes. METHODS: Cumulus-oocyte complexes retrieved from unstimulated ovaries were continuously cultured under 20 microM cilostamide or 50 microM forskolin, alone or in combination for 6, 12, 24 or 48 h, respectively. Levels of intercellular gap junction communication (GJC) and maturational status were examined at these designated time points. Metaphase II oocytes obtained following 54 h biphasic culture (with meiotic inhibitors from 0 to 24 h, no meiotic inhibitors from 24 to 54 h) were subject to intracytoplasmic sperm injection and embryos were cultured for five more days. RESULTS: Both cilostamide and forskolin delayed spontaneous meiotic progression after continuous culture with immature human oocytes. Combined treatment of cilostamide and forskolin significantly lowered the rates of germinal vesicle breakdown (GVBD) at 6, 12, 24 or 48 h after meiotic inhibitory culture, when compared with the control (all P < 0.05). A delay of 6 h for the loss of GJC was also observed under the combined treatment of cilostamide and forskolin. The fertilization rate was significantly higher under the combined treatment of cilostamide and forskolin than that of the control. Although the rates of oocyte maturation and embryo cleavage were similar among groups, there was a slight but non-significant increase in blastocyst formation rate with the treatment of cilostamide and forskolin. CONCLUSIONS: Combined treatment of cilostamide and forskolin positively influences oocyte developmental competence by exhibiting a synergistic effect on the prevention of GJC loss and resumption of meiosis.

Predictive value of excision repair cross-complementing rodent repair deficiency complementation group 1 and ovarian cancer risk.

OBJECTIVE: We aimed to analyze the association between excision repair cross-complementing rodent repair deficiency complementation group 1 (XRCC1) and ovarian cancer risk. METHODS: We performed a hospital-based case-control study with 155 cases and 313 controls in China. All Chinese cases with newly diagnosed primary ovarian cancer between May 2005 to May 2010 in our hospital were invited to participate within 2 months of diagnosis. Controls were randomly selected from people who requested general health examinations in the same hospital during the same period. SNPs in EXCC1, ERCC1 C8092A and ERCC1 T19007C, were analyzed by PCR-RFLP method. RESULTS: We observed a non-significantly increased risk of ovarian cancer among individuals with ERCC1 8092TT compared with those with the 8092CC genotype (adjusted OR=1.55, 95% CI%=0.74-2.97). Moreover, 19007TT genotype carriers also showed a non-significant increased risk of ovarian cancer over those with the 19007CC genotype (adjusted OR=1.78, 95% CI%=0.91-3.64). CONCLUSION: Our firstly investigation of links between polymorphisms in the ERCC1 gene and the risk of ovarian cancer in Chinese population demonstrated no significant association. Further large sample studies in Chinese populations are needed.

[Effect of levonorgestrel-releasing intrauterine system combined with GnRH analogue for treatment of large adenomyosis].

OBJECTIVE: To evaluate the effects of levonorgestrel-releasing intrauterine system (LNG-IUS) combined with GnRH analogue (GnRH-a) in the treatment of adenomyosis with uterine body enlargement. METHODS: Twelve women (mane age 40.3 years) with adenomyosis and uterine cavity depth over 11 cm received injections of GnRH-a every 4 weeks, and after the uterine cavity depth was reduced to below 10 cm, LNG-IUS was deployed. VAS pain score, PBAC bleeding score, uterine volume, and hemoglobin levels of the women were measured before the treatment and at 6 and 12 months after LNG-IUS placement. RESULTS: The VAS pain score was significantly lowered at 6 and 12 month after LNG-IUS placement (P<0.05), and the PBAC bleeding score also showed significant reductions (P<0.05). The uterine volume decreased significantly at 6 and 12 months after LNG-IUS placement as compared with that before the treatment, but was significantly greater at 6 month in comparison with that at the time of LNG-IUS placement (P<0.05). Serum hemoglobin levels underwent significant increments after LNG-IUS placement (P<0.05). CONCLUSION: LNG-IUS combined with GnRH analogue injection can be effective in the treatment of adenomyosis with dysmenorrhea and hypermenorrhea.

[Expression and methylation of adenomatous polyposis coli gene in endometrioid adenocarcinoma].

BACKGROUND & OBJECTIVE: Endometrial carcinoma is a common malignant tumor of female reproductive system, with an increasing incidence in China. Adenomatous polyposis coli (APC) gene, a tumor suppressor gene, is expressed in many tissues, and has a certain relationship with ovarian cancer. This study was to observe the expression and DNA methylation of APC gene in endometrioid adenocarcinoma, and explore its correlations to the occurrence and development of this disease. METHODS: The methylation, mRNA and protein expression of APC gene were detected by methylation-specific polymerase chain reaction (PCR), reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry in 30 specimens of normal proliferative endometrium, 30 specimens of atypical hyperplastic endometrium and 60 specimens of endometrioid adenocarcinoma. RESULTS: The methylation rate of APC gene was significantly higher, the positive rates of APC mRNA and protein were significantly lower in endometrioid adenocarcinoma than in atypical hyperplastic endometrium and normal proliferative endometrium (65.0% vs. 33.3% and 23.3%, 33.3% vs. 63.3% and 73.3%, 30.0% vs. 50.0% and 66.7%,P<0.05). There was no significant difference between atypical hyperplastic endometrium and normal endometrium (P>0.05). APC methylation was positively correlated to APC mRNA expression. CONCLUSION: The expression and DNA methylation of APC gene are certainly related with the occurrence and development of endometrioid adenocarcinoma.