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.

TIMPs Expression as A Maternal Cell Free Plasma Biomarker of Severe Preeclampsia: A Case-Control Study.

OBJECTIVE: Preeclampsia (PE) is a pregnancy related disorder with prevalence of 6-7%. Insufficient trophoblastic invasion leads to incomplete remodeling of spiral arteries and consequent decrease in feto-placental perfusion. Altered placental expression of tissue inhibitors of matrix metalloproteinase (TIMPs) is considered to be involved in this process while the balance between matrix metalloproteinases (MMPs) and TIMPs contributes to remodeling of the placenta and uterine arteries by degradation and refurbishing of extracellular matrix (ECM). Therefore, TIMPs, fetal expression pattern was evaluated with the aim of its potential to be used as a determinant for the (early) detection of PE. MATERIALS AND METHODS: In this case-control study, cell free fetal RNA (cffRNA) released by placenta into the maternal blood was used to determine expression patterns of TIMP1, 2, 3 and 4 in the severe preeclamptic women in comparison with the normal pregnant women. Whole blood from 20 preeclamptic and 20 normal pregnant women in their 28-32 weeks of gestational age was collected. The second control group consisted of 20 normal pregnant women in either 14 or 28 weeks of gestation (each 10). cffRNA was extracted from plasma and real-time polymerase chain reaction (PCR) was done to determine the expression levels of TIMP1, 2, 3 and 4 genes. RESULTS: Statistical analysis of the results showed significant higher expression of TIMP1-4 in the preeclamptic women in comparison with the control group (P=0.029, 0.037, 0.037 and 0.049, respectively). Also, an increased level of TIMPs expression was observed by comparing 14 to 28 weeks of gestational age in the normal pregnant women in the second control group. CONCLUSION: An increased cffRNA expression level of TIMPs may be correlated with the intensity of placental vascular defect and may be used as a determinant of complicated pregnancies with severe preeclampsia.

Disturbed progesterone signalling in an advanced preclinical model of endometriosis.

RESEARCH QUESTION: Do human endometriosis organoids recapitulate aberrant progesterone signalling in the disease to serve as advanced experimental models for uncovering epigenetic mechanisms involved in attenuated progesterone response in endometriosis? DESIGN: Initially, the organoids were established from acquired biopsies (women with and without endometriosis) and characterized by morphological, histological and immunostaining analyses. RESULTS: A panel of endometriosis-related genes showed a pattern of expressions in cytochrome c oxidase subunit II (COX2), matrix metalloproteinase 2 (MMP2), MMP9, tissue inhibitor of metalloproteinase-3 (TIMP3), transforming growth factor beta 1 (TGF-ß1), and zinc finger E-box binding homeobox 1 (ZEB1), and a contradictory expression pattern for cadherin (CDH1), POU class 5 homeobox 1 (POU5F1; also known as OCT4), and Nanog homeobox (NANOG) in the endometriosis organoids that is concordant with published research. These endometriosis organoids failed to upregulate 17ß-Hydroxysteroid dehydrogenase 2 (17HSDß2), progestogen associated endometrial protein (PAEP), secreted phosphoprotein 1 (SPP1), and leukaemia inhibitory factor (LIF) in response to progesterone at the level observed in control endometrium organoids. Progesterone receptor B (PRB) gene expression significantly decreased in both eutopic and ectopic organoids compared with control endometrium organoids. DNA hypermethylation, as an epigenetic mechanism for suppression of transcription, was detected at the PRB promoter in the eutopic, but not ectopic, organoids. Therefore, other epigenetic mechanisms, such as histone modifications and microRNAs, may be responsible for PRB downregulation in ectopic organoids. CONCLUSIONS: Endometriosis organoids are powerful preclinical models that can be used to investigate the molecular mechanisms involved in endometriosis-associated progesterone resistance.

KH domain containing 3 like (KHDC3L) frame-shift mutation causes both recurrent pregnancy loss and hydatidiform mole.

OBJECTIVE: Recurrent pregnancy loss (RPL) is a common infertility-related complication that affects approximately 1-3 % of women worldwide. Known causes of etiology are found in approximately half the cases but the other half remain unexplained. It is estimated that several thousands of genes contribute to reproductive success in mammals and the genetic causes of RPL cannot be fully addressed through targeted genetic tests. In recent years, massive parallel sequencing technologies has helped discovering many causal mutations in hereditary diseases such as RPL. STUDY DESIGN: Using whole-exome sequencing (WES), we studied a large multiplex consanguineous family with multiple cases of RPL and hydatidiform moles (HM). In addition, targeted Sanger sequencing was applied to 40 additional non-related individuals with RPL. RESULTS: The use of WES permitted to identify the pathogenic variant in KHDC3L (c.322_325delGACT) in related who experienced RPL with or without HM. Sanger sequencing confirmed the segregation of the mutation throughout the pedigree and permitted to establish this variant as the genetic cause responsible for RPL and HM in this family. CONCLUSION: KHDC3L is well established as a susceptibility gene for HM but we confirmed here that KHDC3L deleterious variants can also induce RPL. In addition, we observed a genotype-phenotype correlation, demonstrating that women with a truncating KHDC3L homozygous variant could not sustain a pregnancy and often had pregnancy losses mainly due to HM while those with the same heterozygous variant could have children but often endured RPL with no HM.

Serum estradiol level on trigger day impacts clinical pregnancy rate in modified natural frozen embryo transfer cycles.

OBJECTIVE: To examine the predictive value of serum estradiol and progesterone on the day of human chorionic gonadotropin (hCG) administration and embryo transfer for clinical pregnancy rate in modified natural-cycle frozen embryo transfer (NC-FET). METHODS: In a longitudinal prospective study, all eligible women who underwent NC-FET cycles with hCG triggering in Royan Institute, Tehran, Iran, from June 1, 2015, to December 31, 2016, were evaluated. Serum estradiol and progesterone levels were measured at menstrual cycle initiation, on day of trigger with hCG, on day of embryo transfer, and in pregnant women every 7 days until the observation of a gestational sac with embryonic heartbeat. RESULTS: In total, 101 modified natural FET cycles were assessed, and the clinical pregnancy and live birth rates achieved were 34 (33.6%) and 32 (31.6%), respectively. The changes in estradiol level during early pregnancy showed an increase by an average of 200 pg/mL per week. Multivariable logistic regression analysis showed that only the estradiol level on the hCG day was a significant predictive variable for clinical pregnancy following NC-FET (P=0.04). CONCLUSION: Estradiol level on the day of hCG trigger predicted the clinical pregnancy rates after modified NC-FET; this likely mirrored the developmental competence of the corpus luteum and an appropriate luteal structure-function.

Live birth rates after different endometrial preparation methods in frozen cleavage-stage embryo transfer cycles: a randomized controlled trial.

PURPOSE: This study aimed to compare the clinical outcomes in different endometrial preparation methods prior to frozen embryo transfer (FET) in women with normal menstrual cycles. METHODS: A total of 471 eligible patients were randomly allocated into four groups of endometrial preparation prior to FET: natural cycle with spontaneous ovulation (n = 120), natural cycle with human chorionic gonadotropin (hCG) for ovulation induction (n = 117), hormone replacement cycle (HRC) (n = 113) and HRC with pre-treatment with GnRH-a (n = 121). Natural cycle with hCG also received hCG in luteal phase. The primary outcome was live birth rate. The secondary outcomes included implantation, biochemical and clinical pregnancy, ongoing pregnancy, and late miscarriage rates. Data analysis included t test, ANOVA and χ2. RESULTS: There were no statistically significant differences in the mean age (p = 0.31), duration (p = 0.43) and cause of infertility (p = 0.77) and the number (p = 0.33) and quality (p = 0.21) of embryos transferred between the groups. No significant differences regarding the implantation rates per embryo transfer (p = 0.97) and biochemical pregnancy rates (p = 0.90) were observed between the groups. The rates of clinical pregnancy were 34.2%, 32.5%, 31% and 36.4% in the natural cycle, natural with hCG, HRC and HRC with GnRH-a groups, respectively (p = 0.83). Ongoing pregnancy (p = 0.89) and miscarriage (p = 0.33) rates were comparable between groups. The rate of live birth was 30.8% in the natural group, 30% in the natural with hCG, 27.4% in the HRC and 31.4% in the HRC with GnRH-a groups (p = 0.91). CONCLUSION: Four different types of endometrial preparation methods for FET cycles appear to be equally effective in terms of implantation, pregnancy, miscarriage and live birth rates in women with normal menstrual cycles. CLINICAL TRIAL REGISTRATION NUMBER: NCT02251925.

Interstitial ectopic pregnancy diagnosis by three-dimensional ultrasound and its laparoscopic management: A case report.

BACKGROUND: Interstitial Ectopic Pregnancy (IEP) is an uncommon type of ectopic pregnancy with the risk of rupturing and bleeding. The incidence of IEP is about 2-4% of all EPs. The diagnosis and management are challenging. We present a well-timed and managed case of IEP. CASE: The case was a 37-yr-old woman presented at the Royan Institute with a chief complain of sudden onset of pelvic pain and moderate vaginal bleeding, three weeks after her positive pregnancy test. She had got pregnant with in-vitro fertilization procedure. She was admitted for a two-dimensional ultrasound (2DUS). The 2DUS findings showed a gestational sac with live embryo and yolk sac which was located high in the fundus and eccentric to the endometrium. The suspicion of IEP rose after the 2DUS findings, the confirmation of further diagnosis was then done by three-dimensional ultrasound, and the treatment was done by laparoscopy. The patient underwent laparoscopic left corneal resection. She was discharged after two days and her ß-hCG achieved complete resolution ( < 5 mIU/mL) after two weeks' follow-up. CONCLUSION: According to the life-threatening complications that are associated with IEP, acquaintance and suspicion about IEP is important. Specified information that obtained by three-dimensional ultrasound could be useful for exact locating and detection.

Value of Endometrial Echopattern at HCG Administration Day in Predicting IVF Outcome.

BACKGROUND: Assisted reproductive  treatment  (ART)  cycle  is an  expensive  procedure with low implantation and pregnancy rate; therefore, it is necessary to evaluate the predictors of success in these patients. OBJECTIVE: The relationship between endometrial echopattern and pregnancy rate was evaluated. METHODS: The endometrial  echopattern  was  analyzed  prospectively  on  the  day  of  human chorionic  gonadotropin  (HCG) administration  in  280  in  vitro  fertilization/intracytoplasmic  sperm  injection (IVF/ICSI) cycles with 8-14 mm endometrial thickness at Royan institute in 2013-2014. Based on echopattern, three groups were developed: pattern A (Triple line), pattern B (Heterogeneous-echogen) and pattern C (Homogeneous-echogen). Pregnancy rate (PR) was compared between all groups. Data was analyzed using SPSS (v.18, Chicago, IL, USA), and descriptive tests such as Chi-square and analytical tests like logistic regression, for controlling confounder variables like age. RESULTS: Among 280 patients finally evaluable, the distribution of endometrial echopattern on the day of HCG administration was 127 patients (45.4%) in group A, 98 patients (35%) in group B and 55 patients (19.6%) in group C. The highest PR per transfer pertained to group A (49.6%), and the lowest to group B (32.7%). CONCLUSION: The presence of pattern A (Triple line) appears more likely to favor pregnancy. Therefore, ultrasonographic evaluation of endometrial echopattern on the day of HCG administration has prognostic value in clinical settings for predicting implantation in ART cycle.

Metabolic syndrome in infertile women with polycystic ovarian syndrome

ABSTRACT Objective The aim of the present study was to determine the prevalence of metabolic syndrome (MS) in infertile Iranian women with polycystic ovary syndrome (PCOS) using the ATPIII criteria. Subjects and methods In this cross-sectional study, 624 women with PCOS were enrolled at a tertiary referral center in Tehran, Iran, between April, 2012 and March, 2013. Diagnosis of MS was according to ATPIII criteria. Also, we divided PCOS patients into following two main groups: (i) with MS (n = 123) and (ii) without MS (n = 501), and then compared variables between two groups. Results The mean age, body mass index (BMI) and waist circumference were 28.6 ± 4.3 years, 26.7 ± 3.7 kg/m2 and 85.2 ± 8.7 cm, respectively. The prevalence of MS was 19.7%. Our findings showed that age, BMI, waist circumference and all metabolic parameters were higher in PCOS women with MS than related values in those without MS. The most and least prevalent forms of MS were low level of high density lipoprotein-cholesterol (HDL-C) and hypertension, respectively. Conclusion It seems the prevalence of metabolic syndrome in our country isn’t as high as western countries. The prevalence rate of MS increased with age and BMI. One of the major cardiovascular risk factors, low level of HDL-C, is the most prevalent metabolic abnormality in our participants.

Relationships between Serum Luteinizing Hormone Level, Endometrial Thickness and Body Mass Index in Polycystic Ovary Syndrome Patients with and without Endometrial Hyperplasia.

BACKGROUND: The endometrial hyperplasia measured by ultrasound in polycystic ovary syndrome (PCOS) women is strongly related to pathologic endometrial thickness, but there is no consensus on the relation between serum luteinizing hormone (LH) and either of these factors: pathologic endometrial hyperplasia and body mass index (BMI). MATERIALS AND METHODS: In this observational cross-sectional study, three hundred fifty infertile PCOS women were involved in this research. An endometrial biopsy was taken by using a pipelle instrument, regardless of menstrual cycle's day and all samples were reported by the same pathologist. Basal serum LH level was compared between two subgroups (hyperplasia and non-hyperplasia). The intended population was divided into three groups according to BMI and basal serum LH, later on the comparison was made in three groups. Chi-square test was applied to compare nominal variables between groups. Mann-Whitney U, and one way ANOVA tests were used to compare means on the basis of the result of normality test. RESULTS: The frequency of endometrial hyperplasia was 2.6%. Endometrial thickness in the patients with endometrial hyperplasia was significantly higher than that of a normal endometrium (10.78 ± 3.70 vs. 7.90 ± 2.86 respectively, P=0.020). There was no relation between endometrial hyperplasia and serum LH (P=0.600). The ANOVA test showed serum LH levels were not the same among three BMI groups (P=0.007). Post hoc test was also performed. It showed that the LH level in normal BMI group was significantly higher than those of other groups (P=0.005 and P=0.004), but there was no statistical difference between overweight and obese groups (P=0.8). We found no relationship between BMI and endometrial thickness in PCOS patients (P=0.6). CONCLUSION: Sonographic endometrial stripe thickness is predictive for endometrial hyperplasia in PCOS women. We could not find out any relationship between serum LH level and BMI with endometrial thickness in PCOS patients. However, our study confirmed a diverse relationship between serum LH level and BMI in PCOS patients.

Assisted reproductive outcomes in women with different polycystic ovary syndrome phenotypes: the predictive value of anti-Müllerian hormone.

This cross-sectional study aimed to evaluate IVF/intracytoplasmic sperm injection (ICSI) outcomes in different polycystic ovary syndrome (PCOS) phenotypes (A, B, C and D) compared with a control group and the predictive values of serum anti-Müllerian hormone (AMH) in PCOS phenotypes for main outcomes. This study evaluated 386 PCOS women and 350 patients with male factor infertility. Women with phenotypes A and C had significantly higher concentrations of AMH than those with phenotype B (P < 0.001). Clinical pregnancy rate (CPR) in the phenotype D group (53.3%) was higher than other groups (32.5%, 26.4% and 36.8%, respectively, in phenotypes A, B and C), but not to a significant level. Multivariable regression analysis, after adjusting for women's age and body mass index, revealed that PCOS phenotypes A and B were associated with a decreased CPR compared with the control group (odds ratio [OR]: 0.46, confidence interval [CI]: 0.26-0.8, P = 0.007 and OR: 0.34, CI: 0.18-0.62, P = 0.001, respectively). It seems a combination of hyperandrogenism and chronic anovulation is associated with a negative impact on the CPR in these patients. These results demonstrated that AMH concentration is related to PCO morphology but not predictive for CPR and live birth rate.

Metabolic syndrome in infertile women with polycystic ovarian syndrome.

OBJECTIVE: The aim of the present study was to determine the prevalence of metabolic syndrome (MS) in infertile Iranian women with polycystic ovary syndrome (PCOS) using the ATPIII criteria. SUBJECTS AND METHODS: In this cross-sectional study, 624 women with PCOS were enrolled at a tertiary referral center in Tehran, Iran, between April, 2012 and March, 2013. Diagnosis of MS was according to ATPIII criteria. Also, we divided PCOS patients into following two main groups: (i) with MS (n = 123) and (ii) without MS (n = 501), and then compared variables between two groups. RESULTS: The mean age, body mass index (BMI) and waist circumference were 28.6 ± 4.3 years, 26.7 ± 3.7 kg/m2 and 85.2 ± 8.7 cm, respectively. The prevalence of MS was 19.7%. Our findings showed that age, BMI, waist circumference and all metabolic parameters were higher in PCOS women with MS than related values in those without MS. The most and least prevalent forms of MS were low level of high density lipoprotein-cholesterol (HDL-C) and hypertension, respectively. CONCLUSION: It seems the prevalence of metabolic syndrome in our country isn't as high as western countries. The prevalence rate of MS increased with age and BMI. One of the major cardiovascular risk factors, low level of HDL-C, is the most prevalent metabolic abnormality in our participants.

Clinical features, diagnostic approach, and therapeutic outcome in myasthenia gravis patients with thymectomy.

PURPOSE: Thymectomy has been widely employed in the treatment of myasthenia gravis (MG). However, little data exist in Iran demonstrating the efficacy and morbidity of thymectomy. The aim of this study was to determine the clinical features, diagnostic approach, and therapeutic outcome in patients with MG who underwent thymectomy. METHODS: This historical cohort study was conducted in 3 university hospitals in Tehran. Preoperative and operative indices of 61 patients with MG who had been treated with thymectomy in these hospitals from September 2000 to July 2005 were reviewed. Among them, 20 patients were followed during one year after operation for determination of postoperative complications and one year mortality rate. RESULTS: The most common manifestations of MG were ptosis (77.0%) and upper limbs weakness (70.4%). CT scans of the thymus showed thymus enlargement, thymoma and thymus hyperplasia in 51.5% (22/43), 11.6% (5/43) and 2.32% (1/43) of patients, respectively. The postoperative complications were found in 13.1% of patients and one year mortality rate of thymectomy was 6.6%. CONCLUSION: Regarding to high one year mortality rate of thymectomy in patients of MG in this study, the assessment of the factors related to the mortality and outcome of patients who underwent thymectomy in Iran are necessary.