Estradiol versus tranexamic acid on the amount and duration of acute cyclic heavy menstrual bleeding

P-189 Tuesday, October 20, 2015 GENE EXPRESSIONS OF L-SELECTIN LIGANDS ARE DECREASED AT MID-SECRETORY PHASE IN WOMEN WITH ADENOMYOSIS. T. Lai,a,b,c F. Lee,a,b Q. Ling.c,d aObstetrics and Gynecology, Cathay General Hospital, Taipei, Taiwan; bSchool of Medicine, Fu Jen Catholic University, New Taipei, Taiwan; cInstitute of Systems Biology and Bioinformatics, National Central University, Taoyuan, Taiwan; dCathay Medical Research Institute, Cathay General Hospital, Taipei, Taiwan. OBJECTIVE: To evaluate the gene expression of L-selectin ligands’ subtypes during different phases of the menstrual cycles in women with adenomyosis. DESIGN: A prospective study on endometrial samples from reproductiveaged women with adenomyosis at various phases in the normal menstrual cycles. MATERIALS AND METHODS: The Institutional Review Board of Cathay General Hospital approved this study. Tissue samples of endometrium were collected from women (age 35-49 y) with adenomyosis undergoing hysterectomy. Forty-two endometrial biopsies included 12 at proliferative (day714), 10 at early secretory (day15-19), 9 at mid-secretory (day20 to 24) and 11 at late secretory phase (dayS 25). Immunohistochemistry was performed using the antibody, MECA-79, which recognizes the ligands for the L-selectin adhesion molecule. The mRNA expression profiles of L-selectin ligand’s genes were analyzed using RT-PCR for the 4 L-selectin ligand’s subtypes: podocalyxin, endomucin, GlyCAM-1 and CD34. Western blot were used to quantify protein levels. Kruskal-Wallis (K-W) analysis with multiple comparisons was performed to examine differences between different phases in the menstrual cycle. Findings with a two-sided P value < 0.05 were considered to indicate statistically significant differences between the phases. RESULTS: Mean age and BMI of the patients among 4 phases were no different (P>0.05). Immunohistochemistry showed L-selectin ligands were expressed in the luminal and glandular epithelium of the endometrium at all the phases. The intensity of L-selectin ligands was greatest at mid-secretory phase which was at the stage of implantation window. However, the gene expression profiles of L-selectin ligands including endomucin, GlyCAM-1 and CD34 revealed gradually decreased from proliferative to mid-secretory phase. Endomucin, GlyCAM-1 and CD34 had the lowest expression during mid-secretory phase. Interestingly, the highest gene expression of podocalyxin, endomucin, GlyCAM-1 and CD34 were found at late secretory phase (P<0.05). CONCLUSIONS: Decreased expression of L-selectin ligand’s genes at mid-secretory phase may be related to the defects in endometrial receptivity in women with adenomyosis. Further evaluation of the L-selectin system is necessary to determine its role in the process of implantation. Supported by: Cathay General Hospital, Taipei City, Taiwan. P-190 Tuesday, October 20, 2015 DYSREGULATION OF SGK1 IN THE ENDOMETRIUM CAUSES ENDOMETRIAL RECEPTIVITY DEFECTS AND EMBRYO IMPLANTATION FAILURE IN DIABETES. H. Xu,a J. Li,a Z. Ke,a H. Huang.b aDepartment of Reproductive Endocrinology, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou, China; bDepartment of Reproductive Endocrinology, School of Medicine, Zhejiang University, Hangzhou, China. OBJECTIVE: To investigate whether diabetes affects the natural events involved with successful implantation in female and to confirm the expression level of the serum- and glucocorticoid-inducible kinase SGK1 in the endometria of diabetics, furthermore to uncover the underlying mechanisms. DESIGN: Explore the morphological features of endometria from diabetics and control patients during the window of implantation, examine the JAr spheroid attachment rates in a model in vitro system of the human endometrial cancer (Ishikawa) cell line and detect the expression levels of SGK1 and other molecules associated with uterine receptivity and embryo implantation. MATERIALS AND METHODS: The endometrial samples were obtained from diabetics and euglycemia women with male factor infertility (control) during the window of implantation. Scanning electron microscope examinations were performed to assess morphological features of uterine receptivity. Ishikawa cells were grown in steroid-depleted media administered with 5mM glucose(control group) or 25mM glucose(high glucose group).The effect of high glucose on implantation was examined via attachment assay of JAr spheroids to Ishikawa cells. The expression levels of molecules associated with uterine receptivity and embryo implantation were detected by qRT-PCR and Western blot. Nuclear-cytoplasmic localization of SGK1 after high glucose treatment was detected by laser scanning confocal microscopy in Ishikawa cells.

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RESULTS: There was a reduced number of mature uterodomes in the endometria of diabetics compared to control, shown as patches of luminal surface dominated by underdeveloped or immature microvilli. Treating Ishikawa cells with high glucose decreased JAr spheroid attachment rate, up-regulated the expression levels of SGK1 and ENACa, but significantly down-regulated the expression levels of LIF,HOXA10 and HBEGF. Furthermore high glucose treatment can also activate TGFb1/Smad2 signaling pathway, which may lead to the increased expression of SGK1 directly. Those effects can be reversed by the TGFbR1 inhibitor SB431542. High glucose treatment also promoted the nuclear translocation of SGK1 in Ishikawa cells, and the activated SGK1 regulated the expression of a subset of genes that were involved in uterine receptivity, including ENACa, LIF, HOXA10,HBEGF. CONCLUSIONS: Diabetes may cause endometrial receptivity impairment and embryo implantation failure via increasing the expression level of SGK1 and promoting its activation, which may be due to a high glucose microenvironment in endometrium during the window of implantation. P-191 Tuesday, October 20, 2015 ESTRADIOL VERSUS TRANEXAMIC ACID ON THE AMOUNT AND DURATION OF ACUTE CYCLIC HEAVY MENSTRUAL BLEEDING. K. Wauda, D. F. Archer.b aJones Institute for Reproductive Medicine, Norfolk, VA; bEastern Virginia Medical School, Norfolk, VA. OBJECTIVE: To document and quantify the amount and duration of menstrual bleeding during treatment with estradiol or tranexamic acid, and compare the efficacy of estradiol with that of tranexamic acid. DESIGN: This randomized controlled trial, registered as NCT01659008 with clinicaltrilas.gov, is a comparative study with parallel design comparing efficacy of estradiol 1.0 mg three times a day to tranexamic acid 1300 mg three times a day in reducing acute menstrual blood loss. Sample size is calculated based on detecting less than 30 ml difference between the mean menstrual blood loss of the two treatment groups. Power of 80% with confidence interval of 95% is used. Student’s t-test is used to compare mean blood loss during first 24 hours with blood loss at 24 to 48 hrs. MATERIALS AND METHODS: Subjects with acute, cyclical menstrual bleeding are recruited from local gynecology clinics and emergency departments. Subjects start treatment within 48hours of initiation of their cycles. All pads and tampons used during the treatment are collected. Blood loss is calculated using modified alkaline hematin method. RESULTS: Overall average age of subjects is 34. During treatment with estradiol or tranexamic acid, mean blood loss decreased from 28.4mL ( 95% CI,18.2-38.6) during first 24 hours to 18.7mL (95% CI, 9.2-27.2) by 48 hours. All subjects reported that the treatment resulted in a meaningful decrease in blood loss during the cycle. CONCLUSIONS: Blood loss is decreased after 24 hours of initiation of treatment with estradiol or tranexamic acid. References: 1. Magnay JL, Nevatte TM, Dhingra V, O’Brien S. Menstrual blood loss measurement: validation of the alkaline hematin technique for feminine hygiene products containing superabsorbent polymers. Fertil Steril 2010;94:2742-6. 2. Malik S, Day K, Perrault I, Charnock-Jones DS, Smith SK. Reduced levels of VEGF-A and MMP-2 and MMP-9 activity and increased TNF-alpha in menstrual endometrium and effluent in women with menorrhagia. Hum Reprod 2006;21:2158-66. 3. Gannon MJ, Day P, Hammadieh N, Johnson N. A new method for measuring menstrual blood loss and its use in screening women before endometrial ablation. British journal of obstetrics and gynaecology 1996;103:1029-33. 4. Gleeson NC. Cyclic changes in endometrial tissue plasminogen activator and plasminogen activator inhibitor type 1 in women with normal menstruation and essential menorrhagia. American journal of obstetrics and gynecology 1994;171:178-83. Supported by: ACOG/ Ferring Pharmaceuticals Inc. Research Award on Menstrual Bleeding Disorders. P-192 Tuesday, October 20, 2015 EXOGENOUS HUMAN CHORIONIC GONADOTROPIN EXPOSURE FROM THE OVULATORY TRIGGER DOES NOT IMPAIR ENDOMETRIAL RECEPTIVITY. B. S. Shapiro, S. Daneshmand, F. Garner, M. Aguirre, C. Hudson. Fertility Center of Las Vegas, Las Vegas, NV.

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