Secretory Endometrium Research Articles

A Retrospective Cross-Sectional Study on Histopathological Findings of Abnormal Uterine Bleeding in Females Visiting a Paramilitary Tertiary Care Hospital

Introduction: Abnormal uterine bleeding (AUB) is a common gynaecological condition majority of which are diagnosed by sampling the endometrium through dilatation and curettage (D and C) and histopathological examination. The aim of this study was to analyse the different types of endometrial histopathology of patients with the abnormal uterine bleeding and associate between different clinical presentation and histopathological findings. Methods: This was a retrospective study carried out in the Department of Obstetrics and Gynaecology of Nepal Armed Police Force Hospital, Kathmandu over a period of one year from April 2022 to March 2023. All cases of AUB who underwent D and C procedure were included in the study. Data was entered and managed in Microsoft excel and descriptive and comparative analysis were performed using the EZR version 1.36 software. Result: Among the 118 patients, age group ranged from 29 to 72 years and most common age group presenting with AUB were perimenopausal (40- 50 years). Menorrhagia was present in 49 women(41.5% ), which was more common among reproductive age group, perimenopausal and multipara women. Secretory endometrium was major findings with 35 women(29.7%), which was more common among reproductive age group, perimenopausal and multipara women. Conclusion: Clinical presentation of AUB and the histopathological examination of the endometrium may be supportive in distinguishing normal from abnormal causes of AUB.

Immunohistochemical Analysis of GATA2 Expression in Endometrium and its Relationship with Hormone Receptor Expression in Benign and Premalignant Endometrial Disorders

The GATA gene family encodes highly conserved zinc-finger transcription factors that facilitate the development and function of multiple organ systems including the uterus. In the endometrium, GATA2 functions in a positive autoregulatory loop with the progesterone receptor (PGR) and colocalizes with PGR on chromatin to promote PGR transcriptional programs. GATA2 also has PGR-independent functions that maintain endometrial cell identity, and GATA2 transcripts reportedly are down-regulated in endometrial disorders including endometriosis. This event is accompanied by a reciprocal increase in GATA6. Here, we applied custom anti-GATA2 monoclonal antibodies and performed GATA2 immunohistochemistry (IHC) on patient endometrial tissues corresponding to proliferative, secretory, inactive, and hormone-treated endometrium, as well as endometriosis and endometrial atypical hyperplasia/endometrioid intraepithelial neoplasia (EAH/EIN). We also performed IHC for the estrogen receptor, PGR, and GATA6 in relevant groups. The results reveal a tight correlation between GATA2 and PGR expression in the glandular and stromal cells of benign endometrium. GATA2 expression is markedly reduced in stromal but not glandular cells in endometriosis and EAH/EIN. This reduction in GATA2 expression does not lead to a detectable increase in GATA6 expression in endometriosis. Although average glandular GATA2 expression was preserved in endometriosis and EAH/EIN cases, its expression was decoupled from PGR, implying that alternative pathways regulate GATA2 levels in these disorders. Our findings indicate that GATA2 dysregulation is a feature of endometriosis and EAH/EIN, and support a model whereby loss of stromal GATA2 in these disorders contributes to their progesterone insensitivity.

Endometrial pattern in women with abnormal uterine bleeding attending a tertiary care centre of Kathmandu

Background: Abnormal uterine bleeding is one of the most common problems among pre and postmenopausal women. Abnormal uterine bleeding is diagnosed when there is a substantial change in frequency, duration, or amountof bleeding during or between periodsin the absence of any organic pathology. The study was perform to assess the endometrial causes of abnormal uterine bleeding and their incidence in different age groups. Materials and methods: It was a cross sectional and descriptive study carried out at a Manmohan medical College & hospital, over a period of two year from 13th April, 2020 to 15th March, 2022. In the study all the endometrial tissue either obtained by endometrial curettage and biopsy or hysterectomy presenting with abnormal uterine bleeding for more than 3 months were included Results: A total of 88 cases were received during this period, out of which there were 78 endometrial tissue samples and 10 hysterectomy specimens. The patient’s age ranged from 20 years to 73years. The pattern of endometrial biopsy are Proliferative (37.5%) and secretory (20.45%) phase endometrium is the most common pattern seen, followed by disordered proliferative endometrium (13.63%), endometrial hyperplasia without atypia (9.09%), Change consistent with Exogenous Hormone (5.68%), Chronic Endometritis (5.68%), Endometrial Polyp(3.40%), Endometrial Hyperplasia with Atypia(2.27%) and Endometrial Carcinoma (2.27%). Conclusions: Patients with abnormal uterine bleeding show a varying spectrum of endometrial pattern. Cyclic endometrium is commonly seen in the reproductive age group whereas hyperplasia and carcinomas are seen in pre/postmenopausal age group. Endometrial biopsy with histopathological examination is a major diagnostic tool in the evaluation of abnormal uterine bleeding.

Integrated transcriptomic analysis reveals dysregulated immune infiltration and pro-inflammatory cytokines in the secretory endometrium of recurrent implantation failure patients.

Recurrent implantation failure (RIF) is a leading impediment to assisted reproductive technology, yet the underlying pathogenesis of RIF remains elusive. Recent studies have sought to uncover novel biomarkers and etiological factors of RIF by profiling transcriptomes of endometrial samples. Nonetheless, the inherent heterogeneity among published studies and a scarcity of experimental validations hinder the identification of robust markers of RIF. Hence, we integrated six publicly accessible datasets with 209 samples, including microarray profiles of endometrial samples in the secretory phase. After removing batch effects, we identified 175 differentially expressed genes. Gene set enrichment analysis identified dysregulation of immunological pathways in RIF. We also observed altered immune infiltration and pro-inflammatory cytokines in RIF. Protein-protein interaction network analysis identified ten hub genes, representing two co-expression modules significantly related to RIF. Knockdown of ENTPD3, one of the hub genes, promoted the epithelial-mesenchymal transition process and resulted in elevated levels of pro-inflammatory cytokines. Collectively, our study reveals abnormal gene expressions involving the regulation of epithelial-mesenchymal transition and immune status in RIF, providing valuable insights into its pathogenesis.

Limiting pre-menstrual endometrial Hypoxia Inducible Factor 2 Alpha may fine-tune endometrial function at menstruation.

Heavy menstrual bleeding (HMB) is common and debilitating, but the precise endometrial mechanisms causing increased menstrual blood loss (MBL) remain undefined. We have previously identified a role for hypoxia in endometrial repair following progesterone withdrawal. As hypoxia inducible factor 2 alpha (HIF2A) is known to alter vascular function in other tissues, we hypothesised that endometrial HIF2A is involved in pre-menstrual optimisation of endometrial function during the secretory phase to limit MBL. Women with objective HMB had higher endometrial HIF2A during the mid-secretory phase when compared to those with normal MBL (p=0.0269). In a mouse model of simulated menses, genetic or pharmacological manipulation of HIF2A did not significantly affect endometrial breakdown/repair, volume of MBL or endometrial hypoxia. However, 88% of Hif2a heterozygote mice reached early-full repair by 24h versus only 65% of wild-type mice. Mean MBL was 0.39 μl (±0.67) in Hif2a heterozygote mice versus 0.98 μl (±0.79) in wild-type mice. Conversely, when we increased HIF2A pre-menstrually, 11% reached early repair at by 8h versus 30% of vehicle-treated mice. Mean MBL was 2.61 μl (±1.10) in mice with HIF2A stabilisation and 2.24 μl (±1.14) in vehicle-treated mice. These non-significant but consistent trends indicate that increased endometrial HIF2A may contribute to delayed endometrial repair and HMB. Increased HIF2A in the secretory endometrium is unlikely to be sufficient to account for the phenotype of HMB, but limitation of HIF2 levels may optimise endometrial function at menstruation.

Assessing the Role of Transvaginal Sonography in Post-menopausal Bleeding: A Less Invasive Approach to Identify Endometrial Carcinoma.

Background Post-menopausal bleeding (PMB) is a very common complaint seen in current practice. Endometrial carcinoma (EC) commonly presents with PMB. Endometrial biopsy is the tool for definitive diagnosis, but it is invasive. Transvaginal sonography (TVS) is a non-invasive tool that can help us in the initial evaluation of such patients. Methods A prospective observational study was conducted on 76 women with PMB. TVS and histopathological study, along with basic evaluation and investigations, were performed on all participants, followed by necessary treatment and follow-up. Data collected were studied and statistically analyzed. Results A maximum of 27.63% (n=21) of patients had endometrial atrophy causing their PMB. Proliferative endometrium was observed in 21.06% (n=16) of cases, 13.15% (n=10) of women had secretory endometrium, 23.68% (n=18) had simple endometrial hyperplasia, 3.94% (n=3) had complex endometrial hyperplasia without atypia, and another 3.94% (n=3)had complex endometrial hyperplasia with atypia. Further classifying, women with benign hyperplasia included 27.63% (n=21) and those with atypical hyperplasia included 3.94% (n=3). Out of the 5.26% (n=4) patients diagnosed with EC on histopathology, TVS identified carcinoma in75% (n=3) cases. This indicates that the sensitivity and specificity of TVS in detecting EC are 75% and 100%, respectively. The positive predictive value (PPV) is 100%, the negative predictive value (NPV) is 98.63%, and the accuracy is 98.68%. Conclusion If the cut-off for endometrial thickness is set at 4 mm, then TVS proves to be an effective and reliable tool for screening and diagnosing EC. It can thus serve as a safe threshold to screen patients with PMB using TVS.

Comparative Assessment of miR-185-5p and miR-191-5p Expression: From Normal Endometrium to High-Grade Endometrial Cancer.

Endometrial cancer (EC) is a significant cause of cancer-related deaths in women. MicroRNAs (miRs) play a role in cancer development, acting as oncogenes or tumor suppressors. This study evaluated the diagnostic potential of hsa-miR-185-5p and hsa-miR-191-5p in EC and their correlation with clinical and histopathological features. A cross-sectional study analyzed formalin-fixed, paraffin-embedded tissue samples from 59 patients: 18 with EC, 21 with endometrial hyperplasia (EH), 17 with normal endometrium (NE), and 3 with endometrial polyps (EPs). Quantitative reverse transcription-polymerase chain reaction and TaqMan probes were used for miR expression analysis. The Shapiro-Wilk test was used to analyze the normal distribution of the data. Subsequently, parametric or non-parametric tests were used to evaluate the associations between the expression levels of each miR and clinical parameters. Both miRs were underexpressed in some precursor and malignant lesions compared to certain NE subtypes and benign lesions. Specifically, hsa-miR-185-5p showed underexpression in grade 3 EC compared to some NE and EH subtypes (FC: -57.9 to -8.5, p < 0.05), and hsa-miR-191-5p was underexpressed in EH and EC compared to secretory endometrium and EPs (FC: -4.2 to -32.8, p < 0.05). SETD1B, TJP1, and MSI1 were common predicted target genes. In conclusion, hsa-miR-185-5p and hsa-miR-191-5p are underexpressed in EC tissues, correlating with histopathological grades, highlighting their potential as diagnostic biomarkers and their role as tumor suppressors in EC.

Application of strain elastography ultrasound to the endometrium of normal women

BackgroundWhile there is a scarcity of studies utilizing strain elastography (SE) for the endometrium, commonly used gynecologic ultrasound instruments are equipped with built-in elastography modalities, primarily SE. With the objective of facilitating comprehensive examinations for gynecologic patients on a single ultrasound instrument, we undertook this study. Therefore, our aim was to study the value of SE ultrasonography in the assessment of endometrial elasticity in normal women.MethodsThree hundred and twenty normal women were recruited at our hospitals from November 2021 to December 2022. Each volunteer underwent a transvaginal two-dimensional (2D) and SE ultrasound during either the endometrial proliferative or secretory phase. The 2D ultrasound indices obtained included endometrial thickness, echo type (type A, B, and C), and blood flow grading (grades 0, 1, 2, and 3). SE indices obtained included endometrial strain values, myometrial strain values, and endometrial strain ratios. Differences in endometrial ultrasound indices between different menstrual cycles and different age groups were compared.ResultsComparison of 2D ultrasound parameters revealed that endometrial thickness in the proliferative phase endometrium group was smaller than that in the secretory phase endometrium group, with a statistically significant difference. Additionally, there was a statistically significant difference in endometrial echo types between the two groups, while the disparity in endometrial blood flow grading was not significant. Regarding SE parameters, the median and mean values of endometrial strain ratio in the proliferative phase endometrium group were smaller than those in the secretory phase endometrium group, showing a statistically significant difference. However, there were no significant differences observed between the two groups in endometrial strain and myometrial strain in the fundus. Furthermore, there were no significant differences in any of the endometrial ultrasound indices among the different age groups.ConclusionsSE can reflect changes in endometrial stiffness in different menstrual cycles and is an important tool for assessing endometrial softness.

METTL3-dependent m6A modification facilitates decreased endometrial decidualization via attenuation of MET in endometriosis.

Failure to induce mesenchymal-epithelial transition (MET) during stromal cell decidualization can lead to consequences such as impaired fertility in patients with endometriosis. METTL3-mediated m6A modification plays an important role in attenuating MET and defective decidualization of endometrial stromal cells and contributes to the development of reduced endometrial receptivity in endometriosis. Mesenchymal-epithelial transition (MET)-mediated endometrial decidualization is pivotal for achieving endometrial receptivity and successful pregnancy. We observed blockade of MET in the eutopic secretory endometrium of patients with endometriosis, but the underlying mechanism is unknown. In this study, real-time PCR was used to detect PRL and IGFBP1 expression, whereas western blotting was used to detect the expression of MET markers and METTL3. Phalloidin staining was used to identify changes in cell morphology. M6A levels were quantified using a colorimetric method and m6A dot blots, and functional analysis was performed using spheroid adhesion assays. We first found that increased E-cadherin expression was accompanied by decreased vimentin and Slug expression in the eutopic secretory endometrium of individuals with endometriosis. We also detected a significant increase in both the m6A level and the expression of the related methyltransferase METTL3. Finally, METTL3 expression was negatively correlated with PRL, IGFBP1, and MET markers expression. Collectively, our findings suggest that METTL3 mediates m6A modification, thereby inhibiting MET formation within the eutopic secretory endometrium of patients with endometriosis. Increased METTL3-mediated m6A modification plays a crucial role in attenuating MET formation and decidualization impairment in endometrial stromal cells, ultimately contributing to compromised endometrial receptivity in individuals with endometriosis. These insights could lead to the identification of potential therapeutic targets for improving both endometrial receptivity and pregnancy rate among individuals affected by endometriosis.

Endometrial Proliferative Phase-Centered View of Transcriptome Dynamics across the Menstrual Cycle.

The endometrium, the inner mucosal lining of the uterus, undergoes complex molecular and cellular changes across the menstrual cycle in preparation for embryo implantation. Transcriptome-wide analyses have mainly been utilized to study endometrial receptivity, the prerequisite for successful implantation, with most studies, so far, comparing the endometrial transcriptomes between (i) secretory and proliferative endometrium or (ii) mid-secretory and early secretory endometrium. In the current study, we provide a complete transcriptome description of the endometrium across the entire menstrual cycle and, for the first time, comprehensively characterize the proliferative phase of the endometrium. Our temporal transcriptome analysis includes five time points including the mid-proliferative, late proliferative (peri-ovulatory phase), early secretory, mid-secretory, and late secretory phases. Thus, we unveil exhaustively the transitions between the consecutive proliferative and secretory phases, highlighting their unique gene expression profiles and possible distinct biological functions. The transcriptome analysis reveals many differentially expressed genes (DEGs) across the menstrual cycle, most of which are phase-specific. As an example of coordinated gene activity, the expression profile of histone-encoding genes within the HIST cluster on chromosome 6 shows an increase in cluster activity during the late proliferative and a decline during the mid-secretory phase. Moreover, numerous DEGs are shared among all phases. In conclusion, in the current study, we delineate the endometrial proliferative phase-centered view of transcriptome dynamics across the menstrual cycle. Our data analysis highlights significant transcriptomic and functional changes occurring during the late proliferative phase-an essential transition point from the proliferative phase to the secretory phase. Future studies should explore how the biology of the late proliferative phase endometrium impacts the achievement of mid-secretory endometrial receptivity or contributes to molecular aberrations leading to embryo implantation failure.

A prospective study of endometrial histopathology in post-menopausal women in Jharkhand.

Postmenopausal bleeding (PMB) refers to any uterine bleeding in a menopausal women. In the early menopausal years, endometrial hyperplasia, polyps and submucosal fibroids are common etiologies of post menopausal bleeding. The most common cause of postmenopausal bleeding is endometrial atrophy, comprises of 60-80%, while endometrial hyperplasia and endometrial cancer contribute to only 11% of Post menopausal bleeding. The aim of study is to analyses histomorphological pattern of endometrium in patients presenting with post-menopausal bleeding in Jharkhand. 103 postmenopausal women presenting to tertiary center of Jharkhand in 2020-22 with bleeding were subjected to endometrial curettage for histopathology. Analysis is based on morphological criteria to assess endometrium. Endometrial histology is of four categories: Proliferative, Secretory, premalignant and carcinoma. The highest incidence of postmenopausal bleeding was noticed in age group of < 60 years and incidence of malignancy was higher after 57 years of age. The majority of patients had parity between 1 and 3 (78.6%). Malignant & premalignant lesions comprises about 22.3% among that 77.7% were due to benign causes. Among the benign causes of postmenopausal bleeding, proliferative endometrium was the commonest finding. Types of hyperplasia encountered were simple hyperplasia without atypia (6.8%), Complex hyperplasia without atypia (3.9%),Complex hyperplasia with atypia (4.8%) and Simple hyperplasia with atypia (4.8%). 21.4% of cases of postmenopausal bleeding were associated with atrophic endometrium. Secretory endometrium seen in 17.5% of women. Endometrial carcinoma accounted for 12.6% of cases of postmenopausal bleeding. Out of these 69.2% were of endometroid type of endometrial carcinoma, 15.3% were of papillary serous carcinoma and 15.3% had clear cell carcinoma. The mean age of patients with endometrium carcinoma was 62.3 years. All cases of endometrial carcinoma were associated with 1 or more risk factor like diabetes/hypertension/Nulligravida. Proliferative Endometrium was a major cause of postmenopausal bleeding. Among the malignant causes, endometrial adenocarcinoma of endometroid type was most frequent with a lower mean age at presentation than other high grade cancers like papillary serous carcinoma & clear cell carcinoma.

Laparoscopic Management of Short-Interval Interstitial Pregnancy After Recent Tubal Ectopic: A Case Report [ID 2683561

INTRODUCTION: A 24-year-old G6P2032 presents with interstitial pregnancy 3 months after ipsilateral laparoscopic salpingectomy for ruptured tubal ectopic pregnancy. This case report demonstrates the management of short-interval interstitial pregnancy after recent ipsilateral laparoscopic salpingectomy. METHODS: The patient initially presented with a right-sided ruptured tubal ectopic pregnancy. Salpingectomy was performed to completely remove the fallopian tube and desiccate it at the cornua, leaving no residual visible tubal remnants. The patient presented with ipsilateral interstitial pregnancy 3 months later and initially underwent systemic methotrexate injection, which failed to resolve the pregnancy. She then underwent laparoscopic cornuostomy and dilation and curettage (D&amp;C). Cornuostomy was performed with injection of dilute vasopressin around the gestational sac to help minimize blood loss followed by hydrodissection and sharp dissection to remove the pregnancy. The myometrium and serosa were closed in layers. There was attention to preserve uterine myometrial tissue integrity at the cornua. The patient did well postoperatively and was discharged on postoperative day 0 (POD0). Her β-hCG trended from 11,902 mIU/mL pre-surgery to 7,726 on POD0, and 289 at 1 week. Pathology showed fragments of chorionic villi, and the D&amp;C showed decidualized secretory endometrium. CONCLUSION: Short-interval interstitial ectopic pregnancies after prior salpingectomy for tubal ectopic are extremely rare. We demonstrated successful management with a minimally invasive laparoscopic cornuostomy after failed methotrexate. Patients should be counseled carefully about the risks of short-interval pregnancy after tubal ectopic even after care is taken to remove all visible portions of the fallopian tube.

Correlation of Hysteroscopy With Histopathological Findings of the Endometrium in Women With Polycystic Ovary Syndrome (PCOS)-Related Infertility.

Introduction There have been numerous studies on the anovulatory factor, leading to infertility inwomen with polycystic ovary syndrome (PCOS); however, studies on the endometrium factor causing infertility in PCOS women are scarce. While hysteroscopy can accurately diagnose endometrial disorders such as endometrial polyps, it may be ineffective in detecting probable endometrial pathologies due to different hormonal habitats in these patients. Materials and methods Sixty patients with PCOS-related infertility were included in the study. All participants underwent hysteroscopic examination followed by endometrial biopsy and histopathological examination. The clinical and hormonal profiles of two main subgroups, that is, (a) normal endometrium (N), which included proliferative endometriumand secretory endometrium on histology, and (b) disordered endometrium (D), which included disordered endometrium on histology, were compared. Results There was no correlation between hysteroscopic and histopathological findings of PCOS infertile women. In the subgroup analysis of the two main histological types, that is, normal (proliferative and secretory) and disordered (disordered endometrium), age (28.70±4.66 vs. 32.9±5.61, p=0.012) and duration of amenorrhea (5.49±2.43 vs. 7.82±2.93, p=0.008) were significantly higher in the disordered group. There was a statistically nonsignificant higher BMI in the patients of the disordered endometrium group. Conclusion These findings suggest that endometrial biopsy and histopathological evaluation along with hysteroscopy should be desired in women with PCOS-related infertility, especially if they are in the late reproductive age group and have a longer duration of amenorrhea, regardless of endometrial thickening. This approach is essential to diagnose and treat endometrial disorder, which can be an additional cause of infertility, recurrent implantation failure, and recurrent pregnancy loss, in addition to ovulatory dysfunction.

Pre-pregnancy preparation of women with hypoandrogenism and female sexual dysfunction

The onset of pregnancy in the natural cycle is a task that requires the coordinated work of both central and peripheral levels of regulation of hypothalamic‑pituitary hormones. An analysis of the reproductive health of women with a menstrual cycle disorder against the background of hypoandrogenism indicates an unsatisfactory course of the early stages of the pregnancy process with the subsequent formation of infertility against the background of a reduced ovarian reserve. Hypolubrication is considered a clinical marker of hypoandrogenism in women of reproductive age.&#x0D; Objective — Optimization of pre‑pregnancy training in women with hypoandrogenism, menstrual cycle disorders and early pregnancy losses in the anamnesis.&#x0D; Materials and methods. A comprehensive clinical, laboratory and instrumental examination involved 90 women with menstrual cycle disorders and hypoandrogenism, from them 45 women received the proposed therapy (group Ia), and 45 patients traditional therapy (group IIa). The control group consisted of 30 healthy women at the stage of pre‑gravid examination. The age of women was 18 to 35 years. Hormonal reproductive background was determined on the 5 ‑ 7th day of the menstrual cycle in all examined women, as well as levels of thyroid‑stimulating hormone (TSH), thyroid peroxidase antibody, vitamin D, homocysteine. The proposed algorithm of pre‑gravid measures included: 1) optimization of thyroid gland function (after consultation with an endocrinologist); 2) restoration of pituitary‑ovarian relationships in case of functional central hypogonadism; 3) therapy aimed at optimizing the prolactin level (target value 12.5ng/ml); 4) reduction of homocysteine level (target level 5.6 mmol/l); 5) normalization of vitamin D level (norm is 75—125 nM/l, insufficiency defined as 50—75 nM/l, and deficiency &lt;50 nM/l). The target control parameters included restoration of lubrication, ovulatory menstrual cycle, onset and progression of pregnancy. IBM SPSS Statistics version 25.0 software was used for statistical processing of the obtained data. The control periods of therapy were 3, 6 and 9 months.&#x0D; Results. In women with proposed treatment algorithm, therapy resulted in the recovery of vitamin D levels, starting from the 6th month of therapy, and complete normalization was recorded at the 9th month of treatment. This indicates the need for long‑term supplementation of vitamin D in women with hypoandrogenism. A decrease in the homocysteine levels was also noted on the 6th month of treatment, and its normalization after 9 months of treatment. Elevated homocysteine levels and vitamin D insufficiency/deficiency were observed throughout the follow‑up period in the conventional therapy group. In women with the proposed therapy, the levels of follicle‑stimulating hormone (FSH), TSH, luteinizing hormone (LH) and prolactin tend to normalize already from the 3rd month of therapy, while the highest positive increase was noted in the LH indicator on the 3rd‑6th month of treatment. Ovarian steroidogenesis approached the indicators of the control group after 6—9 months of therapy. Proandrogenic steroidogenesis began to increase on the 3rd month of therapy, reaching maximum on the 9th month. Unsatisfactory indicators of changes in free testosterone and dehydroepiandrosterone sulfate (DHEA‑C) were noted in women from the group of the traditional pre‑pregnancy preparation even after 9 months.&#x0D; The specified features of steroid support of the hormonal background of women’s reproductive health demonstrated the following: signs of female sexual dysfunction in the group with the proposed therapy completely disappeared after the 9th month of treatment, and in the group of traditional therapy they continued to bother 91.1% of women. At the end of the observation period, the regular menstrual cycle was gained in 40.0% of women with the proposed therapy and in 6.7% with the traditional one. Progressive pregnancies in the group with the proposed algorithm were achieved in 37.5% of women (successfully overcame the luteoplacental barrier), and in 8.8% in the group of the traditional approach (ended by early termination of pregnancy).&#x0D; Conclusions. The period of formation of the luteal phase insufficiency is the specific feature of the restoration of menstrual cycle fullness in women with hypoandrogenism, which requires additional therapy to ensure the conditions for a full‑fledged secretory endometrium transformation. The pregnancy process takes place in unfavorable conditions, which can lead to early pregnancy losses. The period of pre‑pregnancy measures in this group of patients should last at least 6—9 months. Traditional pre‑pregnancy preparation does not contribute to the restoration of the central mechanisms of menstrual cycle regulation in women with hypoandrogenism, nor does it contribute to the restoration of full‑fledged ovarian steroidogenesis, it is accompanied by anovulation and insufficiency of the luteal phase, early pregnancy losses. Symptoms of female sexual dysfunction do not disappear under the influence of traditional treatment.

The involvement of RNA N6-methyladenosine and histone methylation modification in decidualization and endometriosis-associated infertility.

Defective decidualization of endometrial stromal cells (ESCs) in endometriosis (EM) patients leads to inadequate endometrial receptivity and EM-associated infertility. Hypoxia is an inevitable pathological process of EM and participates in deficient decidualization of the eutopic secretory endometrium. Enhancer of zeste homology 2 (EZH2) is a methyltransferase which catalyses H3K27Me3, leading to decreased expression levels of target genes. Although EZH2 expression is low under normal decidualization, it is abundantly increased in the eutopic secretory endometrium of EM and is induced by hypoxia. Chromatin immunoprecipitation-PCR results revealed that decidua marker IGFBP1 is a direct target of EZH2, partially explaining the increased levels of histone methylation modification in defected decidualization of EM. To mechanism controlling this, we examined the effects of hypoxia on EZH2 and decidualization. EZH2 mRNA showed decreased m6 A modification and increased expression levels under hypoxia and decidualization combined treatment. Increased EZH2 expression was due to the increased expression of m6 A demethylase ALKBH5 and decreased expression of the m6 A reader protein YTHDF2. YTHDF2 directly bind to the m6 A modification site of EZH2 to promote EZH2 mRNA degradation in ESCs. Moreover, selective Ezh2 depletion in mouse ESCs increased endometrial receptivity and improved mouse fertility by up-regulating decidua marker IGFBP1 expression. This is the first report showing that YTHDF2 can act as a m6 A reader to promote decidualization by decreasing the stability of EZH2 mRNA and further increasing the expression of IGFBP1 in ESCs. Taken together, our findings highlight the critical role of EZH2/H3K27Me3 in decidualization and reveal a novel epigenetic mechanism by which hypoxia can suppress EM decidualization by decreasing the m6 A modification of EZH2 mRNA.

Histopathological profiling of endometrial biopsies in cases of abnormal uterine bleeding: Insights from a tertiary care center in India

ABSTRACT Introduction: Abnormal uterine bleeding (AUB), characterized by excessive, irregular, or prolonged uterine bleeding, is a prevalent gynecological concern, accounting for more than 30% of outpatient gynecological visits. Aim and Objective: This study aimed to assess: (A) The histomorphological patterns of the endometrium in cases of AUB. (B) The distribution of various endometrial pathologies across different age groups. (C) The occurrence of endometrial carcinoma among patients experiencing AUB. Materials and Methods: A prospective investigation was conducted among patients seeking care for AUB and infertility at a tertiary care center in northern India. A total of 130 endometrial specimens (collected through endometrial curettage, biopsy, or hysterectomy) were examined over a 2-year period. Tissue samples were preserved in 10% neutral buffered formalin, processed routinely, and examined using light microscopy. Results: The largest proportion of cases (36.2%) fell within the 31–40-year age group. The predominant histopathological pattern observed was secretory endometrium in 47 patients (36.2%), followed by proliferative endometrium in 44 cases (33.8%). Hyperplasia with atypia was more frequently noted in the perimenopausal age group; however, the incidence of malignancy in this subset of the population was relatively low. Conclusion: Histomorphological examination in cases of AUB contributes valuable insights for the diagnosis and management of AUB.

Evaluation of Endometrial Thickness by Transvaginal Sonography in Postmenopausal Women with Bleeding: A Cross-sectional Study

Introduction: Abnormal Uterine Bleeding (AUB), occurring atleast one year after menopause, is one of the most concerning complaints among gynaecological patients. Endometrial atrophy is the most common endometrial finding in women with Postmenopausal Bleeding (PMB), approximately 10-15% of whom have endometrial carcinoma. Earlier studies suggested that obesity, Type 2 diabetes mellitus, and hypertension are risk factors for endometrial hyperplasia. Diabetes mellitus, obesity, and hypertension form the triad of endometrial cancer. Aim: To assess endometrial thickness by Transvaginal Sonography (TVS) and compare it with histopathological findings in women with PMB, evaluating whether an endometrial thickness of &lt;4 mm on TVS is more suggestive of malignancy. Materials and Methods: The present cross-sectional study was conducted in the Department of Obstetrics and Gynaecology at King George Hospital, Vishakapatnam, Andhra Pradesh, India, from January 2022 to December 2022, comprising n=100 subjects. After routine necessary investigations for PMB, a transvaginal ultrasound examination was carried out to calculate endometrial thickness and compare it with the results of histopathological diagnosis of the endometrium. The parameters assessed were endometrial thickness and appearance, Type 2 Diabetes Mellitus (T2DM), hypertension, and Body Mass Index (BMI). The data was compiled and compared using the Chi-square test, with a p-value of ≤0.05 considered highly significant. Results: The results showed normal atrophic and thickened endometrium in 36% of subjects each, and abnormal endometrium such as hyperplasia, carcinoma, and polyps were recorded in 14%, 8%, and 6% (n=6) of cases, respectively. The histopathological results showed normal endometrium such as atrophic, secretory, and proliferative endometrium in 49%, 6%, and 19% of subjects, respectively. Abnormal endometrial findings such as endometrial hyperplasia, carcinoma, and polyps were observed in 14%, 7%, and 5% of cases, respectively. The majority of cases (58%) had endometrial thickness ≤4 mm, followed by 36% of cases with 5-10 mm and 6% of cases &gt;11 mm, with mean and Standard Deviation (SD) values of 5.8±3. mm Statistical analysis data on the association between TVS findings and Histopathological Examination (HPE) findings was observed to be statistically significant (p&lt;0.001). A statistically significant association was observed between T2DM, and BMI &gt;25 kg/m2 together, DM+hypertension+BMI (Triad) and endometrial thickness, respectively. Conclusion: The TVS is easy, simple, non invasive, and involves no complications. Therefore, it can be used as the first diagnostic step in the investigations of women with PMB, combined with histopathological assessment.

MORPHOLOGICAL AND IMMUNOHISTOCHEMICAL CHARACTERISTICS OF ENDOMETRIAL LAYERS IN WOMEN OF REPRODUCTIVE AGE WITH ENDOMETRIAL HYPERPLASIA WITHOUT ATYPIA

Immunohistochemical staining may be useful for the differential diagnosis of endometrial hyperplasia (EH) without/with atypia and carcinoma. Several immunohistochemical biomarkers have already been investigated for use as diagnostic adjuncts in the diagnosis and classification of EH and may also predict progression from EH to carcinoma. However, the optimal molecular biomarker would be one that could reliably distinguish between hyperplastic benign with/without risk of recurrence, precancerous (hyperplastic atypical) and malignant endometrium, and indicate/predict transition between these three groups. To date, no candidate has been found to fully fill this role, so the search is ongoing.&#x0D; Aim. To study the morphological and immunohistochemically features of the structure of all layers of the endometrium, divided into functional and non-functional zones, which may have an impact on the development of endometrial hyperplasia&#x0D; Materials and methods. The study was performed on endometrial morphological material obtained by diagnostic biopsy from 21 women with abnormal uterine bleeding (AUB) in the gynecological department of the Dnipro Clinical Hospital No. 9 in Dnipro during 2022-2023. The study investigated the expression of the biomarkers ER, PgR, Ki67, CK7, and CK8 in women with endometrial hyperplasia without atypia and secretory endometrium.&#x0D; Results. In women with EH without atypia, the expression of ER in the stroma spontaneously increases, while epithelial cells do not show sensitivity to estrogen. The expression of PgR in the glands spontaneously increases, while the expression of PgR in the stroma is low. This may indicate that stromal cells may be less sensitive to progesterone. High expression of Ki67 in endometrial hyperplasia causes active processes of cell proliferation, while low expression of Ki67 in normal endometrium indicates its physiological state. These data can be useful for diagnosis and detection of pathological changes of the endometrium. SK7 is expressed in epithelial cells, even with increased numbers. Normally, during functional observations of the endometrium, the expression of SK7 in epithelial cells is also observed. The functional zone is a layer of the endometrium that undergoes cyclical changes depending on the menstrual cycle of a woman. In this case, there is a regular renewal of cells expressing CK7, which remains one of the typical markers of epithelial cells. The analysis of SK7 expression allows us to detect differences in the processes of cell proliferation and differentiation between endometrial hyperplasia without atypia and normal endometrium. These data coincide with the findings of other authors, are important for a deeper identification of pathophysiological mechanisms associated with endometrial hyperplasia, and also include additional markers for the diagnosis and prognosis of this condition.&#x0D; Conclusion The study results suggest that the expression of ER, PgR, Ki67, CK7, and CK8 may be associated with the development of EH without atypia. Further studies are needed to confirm these findings and to investigate the role of other biomarkers in the pathogenesis and diagnosis of EH.

OR09-05 GPBAR1- Understanding Its Function In The Secretory Endometrium

Abstract Disclosure: M. Saad-Naguib: None. P. Timmins: None. L. George: None. C. Samuels: None. N. Douglas: None. A.V. Babwah: None. Every successful pregnancy relies on implantation of an embryo into a receptive uterine endometrium. While implantation failure accounts for 50% of pregnancy failures, it remains the least understood aspect of pregnancy. In the mid-secretory (MS) phase of the menstrual cycle, under the influence of progesterone and cAMP, fibroblasts decidualize, a critical requirement for implantation. Decidualization involves the differentiation of stromal fibroblasts into secretory epithelioid cells that contribute to the formation of the decidua, the support of the pregnancy prior to placentation. A defective decidua can lead to implantation failure, but if implantation occurs, it could compromise placenta formation and function. Several studies have demonstrated the importance of G protein-coupled receptors (GPCRs) in mouse endometrial receptivity and decidualization. GPCRs are transmembrane signaling molecules that are ubiquitously expressed in the body and regulate almost every cellular and physiological process. They are major pharmaceutical targets, with 35% of all the drugs approved by the US FDA targeting GPCRs. To identify novel GPCR regulators of embryo implantation, we conducted bulk RNA sequencing of the human endometrium at the periovulatory (PO) and mid-secretory (MS) phase in natural menstrual cycles in healthy women of reproductive age and observed that genes encoding 11 GPCRs, not previously reported to regulate implantation, were differentially expressed between these two time points. Among these was the gene encoding the G protein-coupled bile acid receptor 1 (GPBAR1), and this was upregulated 4.6-fold in the MS vs PO phase. Single cell RNA sequence analysis showed that within the human endometrium, GPBAR1 mRNA expression is mostly and strongly localized to stromal fibroblasts. Upon activation by secondary bile acids, GPBAR1 couples to Gαs leading to an increase in intracellular cAMP levels. We hypothesize that in the MS endometrium, upon ligand binding, GPBAR1 produces cAMP and induces human endometrial stromal cell (HESC) decidualization. This hypothesis is being tested in vitro using primary HESCs. Interestingly, in the pregnant mouse, GPBAR1 mRNA is barely expressed at the implantation site on E7.5, and therefore unlikely plays a role in stromal cell decidualization in the mouse. Whether Gpbar1 is expressed in the pregnant mouse endometrium prior to implantation and plays a role in regulating endometrial receptivity is being investigated. Overall, our studies have uncovered a GPCR, that to our knowledge, has not been previously described in the human endometrium. Our data show that the expression of this GPCR is upregulated at the time of endometrial stromal cell decidualization, and therefore it might regulate decidualization and/or be required for decidual function in women. These possibilities are being examined. Presentation: Friday, June 16, 2023

Endometrial protein expression and phosphorylation landscape decipher aberrant insulin and mTOR signalling in patients with recurrent pregnancy loss

What are the proteomic and phosphoproteomic differences between the endometrium of women with recurrent pregnancy loss (RPL) and the endometrium of healthy control women during the proliferative and secretory phases of the menstrual cycle? In total, 54 endometrial samples were collected during the proliferative and secretory phases from women with RPL (n = 28) and healthy controls (n = 26). Comprehensive proteomic and phosphoproteomic analyses were conducted using label-free liquid chromatography-tandem mass spectrometry (n = 44), and verified through Western blotting (n = 10). Three comparison groups were established: total RPL endometrium versus total control endometrium; RPL proliferative endometrium versus control proliferative endometrium; and RPL secretory endometrium versus control secretory endometrium. Differentially expressed proteins and differentially phosphorylated proteins were identified in the three comparison groups. Combining pathway enrichment, network analysis and soft clustering analysis, the insulin/cyclic nucleotide signalling pathway and AMPK/mTOR signalling pathway were identified as the major contributors to the aberration of RPL endometrium. Western blotting verified altered expression of four proteins: cAMP-dependent protein kinase type I-β regulatory subunit, adenylate cyclase type 3, 5'-AMP-activated protein kinase catalytic subunit α-2 and phosphatidate phosphatase LPIN2. This exploratory study provides insights into the differentiated protein expression and phosphorylation profiles of the endometrium of women with RPL in both the proliferative and sectretory phases of the menstrual cycle. The results highlight potential proteins associated with the pathogenesis of RPL that may serve as potential indicators for RPL. The findings contribute to the identification of potential targets for RPL treatment as well as its pathogenesis.