Increased Gene Expression of LITAF, TNF-α and BCL6 in Endometrial Tissues of Women with Endometriosis: A Case-Control Study.

OBJECTIVE: Endometriosis, as a common inflammatory chronic disease is characterized by endometrial tissue growth outside the uterine cavity. It was reported that lipopolysaccharides (LPS) activate a transcription factor called LPSinduced tumor necrosis factor-alpha (LITAF) in macrophages, which induced transcription of cytokine genes such as tumor necrosis factor alpha (TNF-α). B-cell lymphoma 6 protein (BCL6) is a transcription factor which expression was increased in endometrial tissues of infertile women with endometriosis. In addition, it was shown that mRNA and protein of LITAF and BCL6 were inversely related in mature B lymphocytes and B-Cell lymphomas. Accordingly, we investigated gene expression of LITAF, BCL6 and ,TNF-α in eutopic and ectopic endometrial tissues of women with endometriosis compared to the controls. MATERIALS AND METHODS: In this case-control study, 10 women with endometriosis (endometriosis group) and 10 women without endometriosis (control group) enrolled after diagnostic laparoscopy. Real-time polymerase chain reaction (PCR) technique was used to quantitatively analyze gene expression. One-Way ANOVA was used for data analysis. RESULTS: This study showed that LITAF gene expression was significantly higher in ectopic endometrial tissues compared to the control samples. Expression level of BCL6 gene was significantly increased in the ectopic tissues of women with endometriosis compared to the control and eutopic samples. Although TNF-ɑ gene expression was increased in the ectopic lesions compared to the eutopic and control endometrial samples, these differences were not significant. CONCLUSION: The results suggested that over-expression of these inflammatory genes in ectopic endometrial lesions can be considered as a molecular scenario in pathophysiology of endometriosis by induction of inflammatory cascades and cellular proliferation.

Altered gene expression of VEGF, IGFs and H19 lncRNA and epigenetic profile of H19-DMR region in endometrial tissues of women with endometriosis.

BACKGROUND: Endometriosis, as chronic estrogen-dependent disease, is defined by the presence of endometrial-like tissue outside the uterus. Proliferation of endometrial tissue and neoangiogenesis are critical factors in development of endometriosis. Hence, vascular endothelial growth factor (VEGF) as well as insulin-like growth factor 1 and 2 (IGF1, 2) may be involved as inducers of cellular proliferation or neoangiogenesis. Imprinted long noncoding RNA H19 (lncRNA H19) has been suggested to be involved in pathogenesis of endometriosis via regulation of cellular proliferation and differentiation. Epigenetic aberrations appear to play an important role in its pathogenesis. The present study was designed to elucidate VEGF, IGF1, IGF2 and H19 lncRNA genes expression and epigenetic alterations of differentially methylated region (DMR) of H19 (H19-DMR) regulatory region in endometrial tissues of patients with endometriosis, in comparison with control women. METHODS: In this case-control study, 24 women with and without endometriosis were studied for the relative expression of VEGF, IGF1, IGF2 and H19 lncRNA genes using real-time polymerase chain reaction (PCR) technique. Occupancy of the MeCP2 on DMR region of H19 gene was assessed using chromatin immunoprecipitation (ChIP), followed by real-time PCR. RESULTS: Genes expression profile of H19, IGF1 and IGF2 was decreased in eutopic and ectopic endometrial tissues of endometriosis group, compared to the control tissues. Decreased expression of H19 in ectopic samples was significant in comparison with the controls (P < 0.05). Gene expression of VEGF was increased in eutopic tissues of endometriosis group, compared to control group. Whereas its expression level was lower in ectopic lesions versus eutopic and control endometrial samples. ChIP analysis revealed significant and nearly significant hypomethylation of H19-DMR region II in eutopic and ectopic samples, compared to the control group respectively. This epigenetic change was aligned with expression of IGF2. While methylation of H19-DMR region I was not significantly different between the eutopic, ectopic and control endometrial samples. CONCLUSION: These data showed that VEGF, IGF1, IGF2 and H19 lncRNA genes expression and epigenetic alterations of H19 lncRNA have dynamic role in the pathogenesis of endometriosis, specifically in the way that hypomethylation of H19-DMR region II can be involved in IGF2 dysregulation in endometriosis.

Thymic stromal lymphopoietin (TSLP) is a potent pro-inflammatory mediator which is epigenetically deregulated in endometriosis.

OBJECTIVE: Endometriosis is an estrogen-dependent disease characterized by the presence of endometriotic tissue outside the uterine cavity, the condition that immunological factors play important roles in its pathogenesis. Thymic stromal lymphopoietin (TSLP) is an interleukin 7-like cytokine that triggers dendritic cell-mediated T helper2 inflammatory responses. TSLP receptor, or cytokine receptor- like factor 2 (CRLF2), forms a functional heterodimeric complex with IL-7 receptor alpha (IL-7Rα) to bind with TSLP. The present study aimed to elucidate the expression and epigenetic alterations of TSLP gene parallel to TSLP receptor and IL-10 genes expression in endometrial tissues of patients with endometriosis compared to controls. MATERIALS & METHODS: In this case-control study, 45 women with and without endometriosis was enrolled. The relative expression of TSLP, TSLPR and IL-10 genes were examined using qPCR. Chromatin Immunoprecipitation (ChIP) was also used to monitor epigenetic marks of methylation and acetylation on lysine 9 of histone H3 (H3K9me/ac) and DNA methylation in TSLP promoter. RESULTS AND CONCLUSION: TSLP, TSLPR and IL-10 genes were overexpressed in ectopic endometriotic lesions compared to controls. In ectopic samples, significant H3K9 hyper-acetylation and hypo-methylation parallel to DNA hypo-methylation were detected in TSLP promoter compared to eutopic and control groups (p < 0.05). These epigenetic changes were aligned with TSLP gene expression profile. These data collectively identify TSLP and TSLPR as candidate genes critically involved in development of endometriosis beyond their role in promoting Th2 immune responses. In addition, acetylation and methylation of H3K9 may have effective roles in TSLP dysregulation in endometriosis.

Differential expression of innate/adaptive immunity genes induced by endometrial scratching as a hopeful approach for implantation boosting in unexplained, repeated implantation failure: An RCT.

BACKGROUND: Endometrial scratching (ES) has been proposed as a potential treatment for implantation improvement in unexplained repeated implantation failure (uRIF) patients, however, little is known about its exact molecular mechanisms. OBJECTIVE: This randomized controlled trial (RCT) was conducted on twenty uRIF patients to investigate the expression of innate and adaptive immune signaling genes after ES. METHODS: Ten uRIF patients in the intervention (twice endometrial sampling in follicular and luteal phases) and 10 uRIF patients in the control group (only luteal phase sampling) were randomly enrolled. Gene expression analysis with innate and adaptive immune response PCR-array kit between intervention and control groups were performed. RESULTS: Among innate immune-associated genes, a significant decrease was observed in the expression of APCS, CPR, CCL2, NLRP3, HLA-A, TLR3 and TLR4 in the intervention group. In adaptive immune-related genes, the expression level of CD80, CD86, CXCR3, IFNγ, IFNα1, IFNß, MBL2, CCR6, CCR8 and IL17A were decreased and CSF2, GATA3, and IL4 increased significantly in the intervention group (P < 0.05). Of 14 uRIF patients, five live birth (35.71 %) was achieved. CONCLUSION: ES in uRIF patients may exert positive effects on the endometrial preparation which increases its receptivity for embryo implantation by modulating the expression of an array of immune signaling pathway genes.

Detailed Investigation of Downstream TLR Signaling in the Follicular Cells of Women with Endometriosis.

BACKGROUND: Inflammatory responses within the peritoneal cavity may result in endometrial dysfunction in women with endometriosis. The true causes of this disease remain poorly understood. It is hypothesized that downstream toll-like receptors (TLRs) inflammatory cytokines in response to pathogens may be associated with endometriosis. So, this study was aimed at evaluating the expression of TLRs signaling and endometriosis-associated inflammatory responses. METHODS: Totally, 20 infertile endometriosis patients and 20 normal women undergoing controlled ovarian stimulation were enrolled. The cellular pellet and supernatant were obtained by centrifugation of follicular fluid (FF). Evaluation of TLRs and their signaling pathway gene expression was performed on cellular pellets using quantitative-PCR. The supernatant was used for determination of cytokine protein expression by ELISA. The results are expressed as mean±SEM and a p<0.05 was considered statistically significant. RESULTS: Quantitative-PCR analysis suggested that TLR1, 5, 6, 7, 8, 10, MYD88, NF-ĸB, IL-10 and TGF-ß genes expression significantly increased in patients compared to the control group (p<0.05). TLR3, 9, INF-ß genes expression was significantly lower in endometriosis than control group (p<0.05). There was no significant difference in the expression of TLR2, TLR4, TIRAP, TRIF, TRAM, and IRF3 between two groups. Also, significant increase in the levels of IL-6, IL-8 and MIF protein in FF of endometriosis group was detected in comparison with normal women (p<0.05). CONCLUSION: The expression of TLR downstream signaling in the follicular cells can initiate inflammatory responses and changes in the FF cytokine profile which in turn may induce endometriosis and infertility disorder.

A Comparison of Postpartum Depression in Mothers Conceived by Assisted Reproductive Technology and Those Naturally Conceived.

BACKGROUND: It is thought that mothers who conceive via assisted reproductive technology (ART) may be at greater risk of postpartum depression (PPD) because of the problems and psychological stresses associated with ART treatment. The aim of the present study is to determine the occurrence of PPD among mothers who conceive by ART in comparison with those who naturally conceive. The Edinburgh Postnatal Depression Scale (EPDS) was used to assess PPD. MATERIALS AND METHODS: This historical cohort study investigated 406 mothers with infants aged 3-9 months. Three hundred and eight women with natural pregnancies were selected as the control group from mothers who referred to Tehran healthcare centres for infant vaccinations. The ART group consisted of 98 women who conceived via ART at Royan Institute. Participants completed a general questionnaire that asked about education, occupation, number of children, delivery method, history of infant hospitalization, breastfeeding, mothers' and infants' ages, cause of infertility (ART group), and history of depression. A validated Persian version of the EPDS was used to measure depressive symptoms. RESULTS: The mean EPDS score in mothers who naturally conceived was 8.38 ± 0.35 in comparison with mothers who conceived via ART (7.59 ± 0.63). The proportions of women who reported PPD were 26.0% for the control group and 20.4% for the ART group. There was no statistically significant difference in PPD between the control and ART groups (P=0.26). CONCLUSION: The occurrence of PPD in mothers who conceived via ART was similar to those who conceived naturally.

Sex hormones alter the response of Toll-like receptor 3 to its specific ligand in fallopian tube epithelial cells.

OBJECTIVE: The fallopian tubes play a critical role in the early events of fertilization. The rapid innate immune defense is an important part of the fallopian tubes. Toll-like receptor 3 (TLR3), as a part of the innate immune system, plays an important role in detecting viral infections. In this basic and experimental study, the effect of sex hormones on the function of TLR3 in the OE-E6/E7 cell line was investigated. METHODS: The functionality of TLR3 in this cell line was evaluated by cytokine measurements (interleukin [IL]-6 and IL-1b) and the effects of sex hormones on TLR3 were tested by an enzyme-linked immunosorbent assay kit. Additionally, TLR3 small interfering RNA (siRNA) and a TLR3 function-blocking antibody were used to confirm our findings. RESULTS: The production of IL-6 significantly increased in the presence of polyinosinic-polycytidylic acid (poly(I:C)) as the TLR3 ligand. Using a TLR3-siRNA-ransfected OE-E6/E7 cell line and function-blocking antibody confirmed that cytokine production was due to TLR3. In addition, 17-ß estradiol and progesterone suppressed the production of IL-6 in the presence and absence of poly(I:C). CONCLUSION: These results imply that sex hormones exerted a suppressive effect on the function of TLR3 in the fallopian tube cell line when different concentrations of sex hormones were present. The current results also suggest that estrogen receptor beta and nuclear progesterone receptor B are likely to mediate the hormonal regulation of TLR3, as these two receptors are the main estrogen and progesterone receptors in OE-E6/E7 cell line.

The Association between ABO and Rh Blood Groups and Risk of Endometriosis in Iranian Women.

BACHGROUND: Endometriosis is a common gynaecological disease that affects quality of life for women. Several studies have revealed that both environmental and genetic factors contribute to the development of endometriosis. The aim of this study was to investigate the distribution of ABO and Rh blood groups in Iranian women with endometriosis who presented to two referral infertility centers in Tehran, Iran. MATERIALS AND METHODS: In this case-control study, women who referred to Royan Institute and Arash Women's Hospital for diagnostic laparoscopy between 2013 and 2014 were assessed. Based on the laparoscopy findings, we categorized the women into two groups: endometriosis and control (women without endometriosis and normal pelvis). Chi-square and logistic regression tests were used for data analysis. RESULTS: In this study, we assessed 433 women, of which 213 patients were assigned to the endometriosis group while the remaining 220 subjects comprised the control group. The most frequent ABO blood group was O (40.6%). The least frequent blood group was AB (4.8%). In terms of Rh blood group, Rh+ (90.1%) was more frequent than Rh- (9.9%). There was no significant correlation between ABO (P=0.091) and Rh (P=0.55) blood groups and risk of endometriosis. Also, there was no significant difference between the two groups with regards to the stage of endometriosis and distribution of ABO and Rh blood groups (P>0.05). CONCLUSION: Although the O blood group was less dominant in Iranian women with endometriosis, we observed no significant correlation between the risk of endometriosis and the ABO and Rh blood groups. Endometriosis severity was not correlated to any of these blood groups.

The effects of superovulation with gonadotropins on autoantibody levels in patients undergoing assisted reproductive cycles.

PURPOSE: To evaluate the effect of controlled ovarian stimulation (COH) with gonadotropins on the serum levels of autoantibodies in the women who underwent in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles and to compare basal levels of these autoantibodies between groups according to history of COH. METHODS: This prospective cohort study was performed from October 2014 to March 2016 in the Royan Institute. The volunteered infertile women with regard to the inclusion criteria, who underwent IVF/ICSI cycles, were recruited. The COH was performed according to standard long GnRH agonist protocol. The mean levels of the autoantibodies including anti-nuclear, anti-smooth muscle, anti-ovarian, anti-mitochondrial, anti ß2-glycoprotein I, anti-parietal cell and anti-follicle-stimulating hormone antibodies were measured at three time points: on the 3-5 days of the menstrual cycle, 1 week after starting of COH and the ovum pick-up (OPU) day. RESULTS: Of all participants (n = 189), 73 women had history of COH (group B) and 116 women did not have such history (group A). The analysis indicated that the autoantibodies changes during COH were similar in both groups. COH has no significant impact on the level of autoantibodies during the stimulation cycle. Multiple logistic regression analysis showed that the serum levels of anti-smooth muscle antibody on OPU day was the positive predictive factors for live birth following ART cycles in the studied population. CONCLUSION: No significant effect of COH on the studied autoantibodies by the time of OPU was found but further studies are required to interpret these results.

mRNA expression of VEGF and its receptors in fallopian tubes of women with ectopic pregnancies.

BACKGROUND: Establishment of viable pregnancy requires embryo implantation and placentation. Ectopic pregnancy (EP) is a pregnancy complication which occurs when an embryo implants outside of the uterine cavity, most often in a fallopian tube. On the other hand, an important aspect of successful implantation is angiogenesis. Vascular endothelial growth factor (VEGF) is a potent angiogenic factor responsible for vascular development that acts through its receptors, VEGF receptor 1 (VEGFR1) and VEGFR2. This study aims to investigate mRNA expression of VEGF and its receptors in fallopian tubes of women who have EP compared with fallopian tubes of pseudo-pregnant women. We hypothesize that expression of VEGF and its receptors in human fallopian tubes may change during EP. MATERIALS AND METHODS: This was a case-control study. The case group consisted of women who underwent salpingectomy because of EP. The control group consisted of women with normal fallopian tubes that underwent hysterectomy. Prior to tubal sampling, each control subject received an injection of human chorionic gonadotropin (hCG) to produce a state of pseudo-pregnancy. Fallopian tubes from both groups were procured. We investigated VEGF, VEGFR1 and VEGFR2 mRNA expressions in different sections of these tubes (infundibulum, ampulla and isthmus) by reverse transcription polymerase chain reaction (RT-PCR) and quantitative PCR (Q-PCR). RESULTS: RT-PCR showed expressions of these genes in all sections of the fallopian tubes in both groups. Q-PCR analysis revealed that expressions of VEGF, VEGFR1 and VEGFR2 were lower in all sections of the fallopian tubes from the case group compared to the controls. Only VEGFR2 had higher expression in the ampulla of the case group. CONCLUSION: Decreased expressions of VEGF, VEGFR1 and VEGFR2 in the EP group may have a role in the pathogenesis of embryo implantation in fallopian tubes.

Gene expression analysis of VEGF and its receptors and assessment of its serum level in unexplained recurrent spontaneous abortion.

OBJECTIVE: Unexplained recurrent spontaneous abortion (URSA) is one of the main complications of pregnancy which is usually defined as three or more consecutive pregnancy losses before the 20(th) week of gestation without a known cause. Vascular endothelial growth factor (VEGF) is a potent angiogenic factor and shown, along with its receptors (VEGFR1, 2), to play important roles in several physiologic processes including reproduction. The aim of the present study was to analyze gene expression of VEGF and VEGF receptors in endometrium of patients with a history of URSA compared with normal fertile women. In addition, serum VEGF concentration was assessed and compared between the two groups at the same time. MATERIALS AND METHODS: In this case control study, endometrial and blood samples were obtained between day 19(th)and 24(th) of menstrual cycle (window of implantation) from 10 women with a history of URSA (case group) and 6 fertile women who had at least one successful pregnancy (control group). Expression of VEGF and VEGFRs was studied by reverse transcription- polymerase chain reaction (RT-PCR) and then quantified by real time PCR. Normalization of expression levels was done by comparison with beta-actin expression level as an internal control. Relative VEGF, VEGFR1 and VEGFR2 expression quantities were compared between the two groups. Enzyme linked immunosorbent assay (ELISA) was used for serum VEGF assay. RESULTS: VEGF, VEGFR1 and VEGFR2 gene expression was detected in endometrial samples of both groups. The mean relative expression of VEGF gene was lower in the case group compared with control women, however, both VEGF receptors were expressed higher in endometrium of the case group. In addition, the serum level of VEGF was significantly higher in the case group compared with the controls. CONCLUSION: Alteration in gene expression of VEGF and its receptors in endometrium and changes of serum VEGF might play important roles in pathogenesis of unexplained RSA.

Risk factors associated with endometriosis among infertile Iranian women.

INTRODUCTION: Endometriosis is defined as overgrowth of endometrial tissue outside the uterine cavity. Endometriosis may be asymptomatic or associated with dysmenorrheal symptoms, dyspareunia, pelvic pain, abnormal uterine bleeding and infertility. The aim of this study was to explore the risk factors related to endometriosis among infertile Iranian women. MATERIAL AND METHODS: In this case control study, infertile women referred for laparoscopy and infertility workup to two referral infertility clinics in Tehran, Iran were studied. According to the laparoscopy findings, women were divided into case (women who had pelvic endometriosis) and control (women with normal pelvis) groups. The case group was divided into two subgroups: stage I and II of endometriosis were considered as mild while stage III and IV were categorized as severe endometriosis. A questionnaire was completed for each patient. RESULTS: Logistic regression showed that age, duration of infertility, body mass index (BMI), duration of menstrual cycle, abortion history, dyspareunia, pelvic pain and family history of endometriosis are independent predictive factors for any type of endometriosis. In addition, it was shown that education, duration of infertility, BMI, amount and duration of menstrual bleeding, menstrual pattern, dyspareunia, pelvic pain and family history of endometriosis are independent predictive factors of severe endometriosis. The AUCs for these models were 0.781 (0.735-0.827) and 0.855 (0.810-0.901) for any type of endometriosis and severe endometriosis, respectively. CONCLUSIONS: It seems that any type of endometriosis and severe ones could be predicted according to demographic, menstrual and reproductive characteristics of infertile women.

The role of toll like receptors in pregnancy.

For many years, the innate immunity was of less interest than the adaptive immunity because it was perceived to have secondary importance in the functionality of the immune system. During the past decades, with the advancement of knowledge about innate immune system, interest in innate immunity has grown dramatically and thus its function has been extensively studied. Innate immunity plays fundamental roles in the initiation and induction of adaptive immune responses. It consists of several cells and receptors including natural killer (NK) cells, macrophages (MQs), dendritic cells (DCs) and pattern recognition receptors (PRRs). Two decades ago, Toll like receptors (TLRs) family was known as one of the important PRRs with unique functions especially in protection against invading pathogens. Since the female reproductive tract has access to the outside environment and has a unique interaction with different pathogens whether invading microorganisms or normal flora, allogenic sperm and semi allogenic fetus, it has an essential need for effective immune responses. It has therefore been suggested that TLRs may play important roles in the immune regulation of the female reproductive tract. In addition, it has been demonstrated that immune disturbance may be responsible for some adverse pregnancy outcomes such as preeclampsia (PE), recurrent spontaneous abortion (RSA) and intrauterine growth restriction (IUGR). Our focus in this review is to show the importance of TLRs in pregnancy with emphasis on the expression of these receptors in different tissues related to pregnancy.

The expression of Toll-Like Receptors (TLRs) in testicular cancer: A case control study.

BACKGROUND: It has been suggested that malfunction of immune system may causes testicular cancer. Recently, our understanding of innate immune system has been expanded, by discovery of "Toll-Like Receptors" (TLRs). Some studies have shown that polymorphisms of TLR2 and 4 may affect on the risk of cancer. Also, the role of TLRs 3 and 9 have been shown in apoptosis and metastasis of cancer cells in animal models. OBJECTIVE: Little information is available about the influence of innate immunity on testicular malignancy. Therefore, expression of TLRs 2, 3, 4 and 9 as main components of innate immunity has been investigated in this study. MATERIALS AND METHODS: In this case control study, TLRs gene expression was examined by RT-PCR in normal testis and testicular cancer tissues. Real time quantitative PCR (Q-PCR) analysis was used to compare the relative expression of TLRs between the samples. RESULTS: mRNAs of TLR 2, 3, 4 and 9 were expressed in all normal and cancer samples. Q-PCR reveals that cancer samples had stronger expression of these genes compared with normal ones. CONCLUSION: It seems that the different TLRs expression in testicular cancer cells may contribute to extensive signaling pathways involved in carcinogenesis.

The effect of adding oral oestradiol to progesterone as luteal phase support in ART cycles - a randomized controlled study.

INTRODUCTION: Luteal phase support in assisted reproductive technology (ART) cycles is still controversial. The present study was conducted to evaluate the effect of adding oral oestradiol to progesterone during ART cycles. MATERIAL AND METHODS: In this prospective case control study, infertile women under 35 years old who were candidates for IVF/ICSI cycles in Royan Institute were enrolled. A long gonadotropin-releasing hormone (GnRH) agonist protocol was used for ovarian stimulation. Patients were randomly divided into two groups for luteal phase support: the control group received vaginal administration of progesterone supplementation alone starting on the day after oocyte retrieval and continued until the tenth week if the chemical pregnancy test was positive. In the oestradiol group, 2 mg of oestradiol valerate was initiated orally with progesterone. The control group received a placebo instead of oestradiol. RESULTS: Ninety-eight women were studied as oestradiol (N = 47) and control groups (N= 51). There were no significant differences in the mean number of retrieved oocytes, number of transferred embryos, or chemical and clinical pregnancy rates between the two groups. Although the serum progesterone concentration was higher in the oestradiol group in comparison to the control group on day 7, 10 and 12 after embryo transfer, these differences were not statistically significant. CONCLUSIONS: The results suggested that adding oral oestradiol to vaginal progesterone supplementation does not improve the chemical and clinical pregnancy rates of IVF/ICSI cycles.

Half-dose, long-acting gonadotropin-releasing hormone agonist (Diphereline) is comparable with daily injections of short-acting gonadotropin-releasing hormone agonist (Suprefact) in IVF/ICSI cycles.

INTRODUCTION: The aim of the study was to compare the efficacy of half-dose, long-acting GnRH analogue (Diphereline) with Suprefact in IVF/ICSI (in vitro fertilization/intracytoplasmic sperm injection) cycles. MATERIAL AND METHODS: In this randomized clinical trial performed in Royan Institute, 126 infertile women who were first time candidates for IVF/ICSI were enrolled. Patients were randomly divided into two groups by using a random number table. In one group, 62 patients received a single half dose, 1.87 mg Diphereline, in mid-luteal phase. In the other group, 64 cases were treated with buserelin from the previous mid-luteal phase. P value less than 0.05 was considered significant. RESULTS: The mean age of patients in the Diphereline and Suprefact groups was 27.9 ±3.6 and 29.6 ±3.5 years, respectively (p = 0.01). In the Diphereline group, the mean number of used gonadotropins was 25.6 ±12.1 ampoules, while in the second group it was 25.9 ±8.5 ampoules. Numbers of retrieved and MII oocytes were significantly higher in the Diphereline group (12.1 ±6.3 and 9.6 ±5.5) in comparison to the Suprefact group (9.4 ±6.4 and 7.2 ±5.1). Although the number of developed embryos in the Diphereline group was statistically higher than in the Suprefact group (6.1 ±3.9 vs. 4.7 ±3.4, p = 0.04) there was no significant difference in pregnancy rate (37.1%, 95% CI [26.16-49.54] vs. 37.5%, 95% CI [26.67-49.75]). CONCLUSIONS: A half-dose, long-acting GnRH agonist can be successfully used in ovarian stimulation and produces a higher number of MII oocytes and embryos. The pregnancy rates with this method are acceptable.

Comparison of the efficacy of the aromatase inhibitor letrozole and clomiphen citrate gonadotropins in controlled ovarian hyperstimulation: a prospective, simply randomized, clinical trial.

OBJECTIVE: To study the efficacy of the aromatase inhibitor letrozole in controlled ovarian hyperstimulation (COH). MATERIAL AND METHODS: In this prospective simply randomized clinical trial, one hundred forty patients with unexplained infertility undergoing intrauterine insemination (IUI) therapy were randomized to receive either letrozole or clomiphene citrate (CC)-gonadotropin. The patients were selected among patients referred to one university hospital and one private infertility clinic. A letrozole dose of 5 mg/day (n = 70) was given on days 3-7 of the menstrual cycles. Clomiphen citrate a dose of 100 mg/day was given like letrozole but combined with human menopausal gonadotropin (hMG) dose of 75 IU/ml administered every day starting on day 6. Ovulation was triggered with urinary hCG (10,000 IU) when the leading follicle(s) reached 18 mm in diameter. A single IUI was performed 36 hours later. The luteal phase was supplemented with micronized progesterone vaginally. Ovarian stimulation response (E2 levels and number of follicles) was primary outcome. RESULTS: There were no differences in demographic characteristics between groups. The number of mature follicles (1.8 +/- 0.7 vs. 2.46 +/- 2.3; P = 0.042) and serum E2 level on the day of hCG (310 +/- 135.4 vs. 1,670.7 +/- 1021.8 pg/ml, respectively; P < 0.0001) were significantly lower in letrozole group. A significantly higher endometrial thickness was observed at the time of hCG administration in patients that received letrozole (9.7 +/- 1.6 mm vs. 7.8 +/- 2 mm; P < 0.001). Clinical pregnancy rates also were significantly higher in letrozole group (32.8% vs. 14.3%, respectively; P < 0.01). CONCLUSION: The aromatase inhibitor letrozole appears to constitute a good alternative to CC-gonadotropin in patients with unexplained infertility undergoing COH cycles combined with IUI therapy.

The role of inhibin B in prediction of in vitro fertilization or intracytoplasmic sperm injection cycles' outcome.

OBJECTIVE: To investigate the usefulness of inhibin B concentrations obtained on the fifth day in predicting ovarian response and assisted reproductive technologies outcome. METHODS: In this prospective multi-center study, infertile women who were candidate for in vitro fertilization or intracytoplasmic sperm injection for the first time were enrolled. These patients were referred to the Royan Institute, Vali-Asr Hospital and Alvand Hospital, Tehran, Iran between 2003 and 2004. The inclusion criteria were female age (20-35 years), body mass index (BMI) of 20-28 Kg/m2, duration of infertility>2 years, a normal menstrual cycle and a normal day 3 follicle stimulating hormone level of <8.5 IU/l. All patients underwent long standard gonadotrophin releasing hormone agonist protocol. Plasma level of inhibin B was checked on the fifth day of menstrual cycle. The diagnostic accuracy of inhibin B, were assessed by the area under the receiver operating characteristic (ROC) curve. RESULTS: In this study, 107 infertile patients were studied. Using the value of 283 pg/ml for inhibin B as the cut-off point, day 5 inhibin B had 77% sensitivity, 30% specificity, 31.2% positive predictive values (PPV) and 76.7% negative predictive values (NPV) for poor ovarian response. There were statistically significant correlation among day 5 inhibin B concentration and BMI, number of mature follicles, retrieved oocytes, developed and transferred embryos, chemical pregnancy, ovarian hyperstimulation syndrome (OHSS) and poor responder. CONCLUSION: Although the chemical pregnancy, number of retrieved oocytes, developed and transferred embryos were higher in patients with higher day 5 inhibin B concentration but considering its sensitivity, specifity, PPV and NPV, it cannot be used as a strong test for prediction of cancellation, pregnancy, poor responses and OHSS.

Dermopathy and retinopathy in diabetes: is there an association?

BACKGROUND/AIMS: Diabetic dermopathy is the most common cutaneous marker of diabetes mellitus presenting as single or multiple well-demarcated brown atrophic macules, predominantly on the shins. Although diabetic dermopathy and diabetic retinopathy are both considered by some authors as manifestations of diabetic microangiopathy, only a few studies are published about their possible association. Our purpose was to investigate the association of diabetic dermopathy and diabetic retinopathy. METHODS: We conducted a cross-sectional study in an outpatient diabetes clinic during a 6-month period. One-hundred and eighty-one consecutive patients (8 cases of insulin-dependent diabetes mellitus and 173 cases of non-insulin-dependent diabetes mellitus) were examined for the presence of diabetic dermopathy and diabetic retinopathy. RESULTS: Forty-seven (26%) showed diabetic dermopathy and 68 patients (37.6%) suffered from diabetic retinopathy. The frequency of retinopathy in patients with diabetic dermopathy (44%; 30 cases) was significantly greater than in patients without dermopathy (15%; 17 cases; p < 0.0001). Retinopathy showed a statistically significant association with dermopathy [odds ratio (OR): 3.60; 95% confidence interval (CI): 1.53-8.44; p = 0.003] and diabetes duration (OR: 3.36; 95% CI: 1.67-6.77; p = 0.001). CONCLUSION: Our study further supports that diabetic dermopathy might be used as a telltale sign of diabetic retinopathy, necessitating more intensive ophthalmologic care, especially in long-lasting diabetes.