VDR and TNFSF11 polymorphisms are associated with osteoporosis in Thai patients.

Determining molecular markers for osteoporosis may be valuable for improving the quality of life of affected elderly patients by aiding in early detection and disease management. In the present study, the association between single nucleotide polymorphisms (SNPs) of the vitamin D receptor (VDR) and tumour necrosis factor superfamily number 11 (TNFSF11) genes and the susceptibility of developing osteoporosis was investigated in a Thai female cohort. The study group consisted of 105 Thai postmenopausal patients diagnosed with osteoporosis and 132 healthy Thai postmenopausal female volunteers. DNA extracted from blood samples was used to genotype the VDR and TNFSF11 genes using polymerase chain reaction-restriction fragment length polymorphism and sequencing analysis. For VDR, the frequencies of the genotypes TT, CT and CC for the TaqI SNP (rs731236) were 87.88, 11.36 and 0.76%, respectively, in the control group, while in the osteoporosis cohort were 92.38, 5.71 and 1.91%, respectively. For the FokI SNP (rs2228570), the frequencies of the genotypes CC, CT and TT were 31.06, 55.30 and 13.64%, respectively, in the control group, and in the osteoporosis group were 29.52, 43.81 and 26.67%, respectively. For BsmI SNP (rs1544410), the frequencies of the genotypes GG, GA and AA were 78.03, 18.94 and 3.03%, respectively, in control group, and in the osteoporosis group were 80.95, 18.10 and 0.95%, respectively. The significant risk of osteoporosis associated with the FokI SNP was determined. The odds ratio (95% confidence interval) was 2.30 (1.14-4.69; P=0.01) among patients with osteoporosis with TT as the susceptibility genotype. For TNFSF11, the frequencies of the genotypes TT, CT and CC for the -290C>T SNP (rs9525641) in the control group were 36.36, 50.76 and 12.88%, respectively, while in the osteoporosis group were 31.43, 56.19 and 12.38%, respectively. For the -643C>T SNP (rs9533156), the frequencies of the genotypes TT, CT and CC in the control group were 35.61, 48.48 and 15.91%, respectively, while in the osteoporosis group were 32.38, 55.24 and 12.38%, respectively. For the -693G>C SNP (rs9533155), the frequencies of the genotypes CC, CG, and GG in the control group were 39.39, 46.97 and 13.64%, respectively, and in the osteoporosis group were 36.19, 53.33 and 10.48%, respectively. No significant associations of the TNFSF11 SNPs with osteoporosis were determined; however, it was notable that the GCT haplotype of TNFSF11 may be a protective haplotype for osteoporosis. Therefore, it was concluded that the SNP FokI of VDR may be a potential molecular biomarker for the development of osteoporosis in Thai females.

Differential expressions of estrogen and progesterone receptors in endometria and cyst walls of ovarian endometrioma from women with endometriosis and their responses to depo-medroxyprogesterone acetate treatment.

BACKGROUND: Depo-medroxyprogesterone acetate (DMPA) is an injectable progestin contraceptive that provides a highly effective reduction of pelvic pain in women with endometriosis. Despite its wide use to treat pain associated with endometriosis, its precise mechanisms of action remain unclear. The aims of this study were to investigate the differential expressions of estrogen receptors (ERs), and progesterone receptors (PRs) in endometria and ovarian endometrioma cyst walls of women with endometriosis with and without DMPA treatment. METHODS: Endometria and cyst walls of endometrioma were obtained from 25 to 45 year-old women who suffered from endometriosis and had ovarian endometrioma with the size ≥3 cm. The expression levels of ERs and PRs and the numbers of ER- and PR-positive cells before and after treatment with DMPA were evaluated by Western blot, real-time PCR, and immunohistochemistry. RESULTS: The levels of ERα and ERß expression, their corresponding mRNAs, and numbers of ERα- and ERß-immunoreactive cells in stroma and glands of endometria of the DMPA group were significantly decreased when compared with those of the untreated groups (p < 0.05). In contrast, the levels of PRA/B expression and numbers of PRA/B positive cells in stroma and number of PRB positive cells in stroma and endometrial glands were significantly increased in endometria of the DMPA group when compared with those of the untreated groups. However, in cyst wall the expression levels of these proteins, their corresponding mRNAs, and immonoractive cells were low compared to those in endometria, and DMPA-treatment did not cause any significant changes in these parameters. CONCLUSION: These data indicated that DMPA could upregulate the expressions of PRA/B and down-regulate ERα and ERß in endometria but not in cyst walls from women with endometriosis.

EGFL7 and RASSF1 promoter hypermethylation in epithelial ovarian cancer.

DNA methylation is one of the epigenetic mechanisms associated with gene expression and plays a key role as in activation and deactivation of oncogenes and tumor suppressor genes, respectively. This study employed DNA methylation array to identify methylated genes which are highly correlated with various phenotypes of epithelial ovarian cancer (EOC) in Thai patients and to quantify promoter CpG-island methylation of candidate genes. Tissues from patients with serous and non-serous EOC showed significantly higher promoter methylation of EGFL7 and RASSF1 compared to benign cases. These results indicate the potential of investigating promoter CpG-island methylation of cancer-associated genes as biomarkers of disease progression and even possibly of early detection.

DMPA Suppresses Cell Proliferation and Enhances Cell Apoptosis of Eutopic Endometrium in Women with Endometriosis: A Randomized Controlled Study.

Background: Although Depo-medroxyprogesterone acetate (DMPA), an injectable contraceptive progestin, is very effective for pain relief and prevention of recurrence in women with endometriosis, there is no report on the mechanism of this medication about cell proliferation and apoptosis. Objective: To investigate the effects of DMPA on cell proliferation and apoptosis in the eutopic endometrium of women with endometriosis. Material and Method: A randomized controlled study was conducted in 28 women with endometriosis. The DMPA-treated group included 14 women who were scheduled to undergo laparoscopic surgery after 150 mg of DMPA injections. The control group included 14 women who were scheduled to undergo the surgery without DMPA injection. The endometrial tissue was obtained from each woman by endometrial aspiration before surgery. The ELISA formats of PCNA and the quantitative colorimetric analysis of TUNEL were used for estimating cell proliferation and apoptosis of the eutopic endometrium. Results: There were no differences in the women characteristics between the two groups. The relative level of cell proliferation was significantly less in the DMPA than the control groups (1.08±0.57 vs. 1.73±0.50, p = 0.014). Whereas the relative level of cell apoptosis was greater in the DMPA group than that in the control group (1.12±0.36 vs. 0.82±0.39, p = 0.034). Conclusion: Three months of 150 mg DMPA treatment could suppress cell proliferation and enhance cell apoptosis of the eutopic endometrium of women with endometriosis.

Comparison of Cryopreserved Human Sperm between Ultra Rapid Freezing and Slow Programmable Freezing: Effect on Motility, Morphology and DNA Integrity.

BACKGROUND: Cryopreservation of sperm is common methods to preserve male fertility. Sperm freezing, suggest slow programmable freezing caused lower change of sperm morphology than sperm freezing in vapor of liquid nitrogen. Ultra rapid freezing is easy to be worked on, less time, low cost and does not need high experience. OBJECTIVE: To compare the effect on sperm motility, morphology and DNA integrity of post-thawed sperm after ultra rapid freezing and slow programmable freezing methods. MATERIAL AND METHOD: Experimental study at laboratory of infertility unit, Department of Obstetrics and Gynecology, Faculty of Medicine Ramathibodi Hospital. Thirty-seven semen samples with normal semen analysis according to World Health Organization (WHO) 1999 [normal sperm volume ( 2 ml) and normal sperm concentration (≥ 20 x10(6)/ml) and sperm motility (≥ 50%)]. Semen samples were washed. Then each semen sample was divided into six cryovials. Two cryovials, 0.5 ml each, were cryopreserved by slow programmable freezing. Four 0.25 ml containing cryovials, were cryopreserved by ultra rapidfreezing method. After cryopreservationfor 1 month, thawedprocess was carried out at room temperature. Main outcomes are sperm motility was determined by Computer-Assisted Semen Analysis (CASA), sperm morphology was determined by eosin-methylene blue staining and sperm DNA integrity was assessed by TUNEL assay. RESULTS: Sperm motility was reduced significantly by both methods, from 70.4 (9.0)% to 29.1 (12.3)% in slowprogrammable freezing and to 19.7 (9.8)% in ultra rapid freezing (p < 0.05). Sperm motility decreased significantly more by ultra rapid freezing (p < 0.001). The percentage of normal sperm morphology and DNA integrity were also reduced significantly by both methods. However, no significant difference between the two methods was found (p > 0.05). CONCLUSION: Cryopreservation of human sperm for 1 month significantly decreased sperm motility, morphology and DNA integrity in both methods. However sperm motility was decreased more by ultra rapid freezing.

A comparison of the effects of clomiphene citrate and the aromatase inhibitor letrozole on superovulation in Asian women with normal ovulatory cycles.

OBJECTIVE: To compare the effect of the aromatase inhibitor letrozole and clomiphene citrate (CC) on superovulation in women with normal ovulation. METHODS: A cross-over randomized study of 22 women with normal ovulation, divided randomly into two equal cohorts, was carried out. Each group of 11 women was randomly allocated to take letrozole or CC for one cycle. After washing out for one cycle, the alternative drug was administered in the subsequent cycle. The number and size of mature follicles, endometrial thickness, and estradiol and progesterone levels were monitored. RESULTS: The number of mature follicles and estradiol levels on ovulation day were significantly lower in the letrozole group than the CC group (p < 0.05 for both). However, no differences between the two groups in endometrial thickness and pattern were observed. Progesterone levels showed ovulation in all cycles. CONCLUSIONS: The administration of 50 mg CC on days 3-5 was superior to 2.5 mg letrozole for superovulation induction in women with normal ovulation.