Endometrium Of Women Research Articles

Стан мікрофлори та ступінь вираженості місцевого імунного захисту в ендометрії жінок з безпліддям, що розвивається на тлі хронічного ендометриту поєднаного з бактеріальним вагінозом

The purpose of the study was to evaluate the taxonomic profile of the microbiota and the state of local immunity in the endometrium of women with infertility under conditions of chronic endometritis combined with bacterial vaginosis. Materials and methods. Diagnosis of chronic form of endometritis was performed on clinical symptoms, medical history, scraping, endoscopic and ultrasound examinations. All women were divided into three groups: Group I – 12 healthy women, who had no signs of chronic endometritis and formed a control group; Group II – patients with severe chronic endometritis (n = 16); Group III – patients with chronic endometritis developed on the background of bacterial vaginosis (n = 12). Diagnosis of bacterial vaginosis in patients was performed by detecting an increase in the pH of vaginal secretions >4.5, the presence of homogeneous vaginal discharge with an unpleasant odor, detection of vaginal epitheliocytes with adherent gram-variable microorganisms, detection of "fishy" odor. The study was performed in the second phase of the menstrual cycle (day 18-23). Endometrial aspirate for the study of microflora was selected using a device for obtaining endometrial cytology Endobrash Standard for Endometrial Cytology. Results and discussion. It was found that the development of chronic endometritis on the background of bacterial vaginosis increases the frequency of detection of opportunistic anaerobic microflora in the endometrium compared to women without bacterial vaginosis. It has been shown that a decrease in the level of T-helpers and an increase in the level of T-suppressors / killers in the endometrium are exacerbated in patients with chronic endometritis developed against the background of bacterial vaginosis. Thus, the level of CD4 + cells decreased by 1.6 times, and the level of CD8 + cells increased by 1.9 times compared to healthy women. Against this background, the level of B-lymphocytes and IgA in the endometrium of women with chronic endometritis and bacterial vaginosis decreased. Conclusion. The development of chronic endometritis is associated with an increase in the frequency of detection and quantitative content in the endometrium of Streptococcus spp., Staphylococcus spp. The development of bacterial vaginosis is accompanied by the detection in the endometrium of new species of microorganisms – Prevotella spp., Mobiluncus spp. The imbalance of the endometrial microflora is accompanied by significant violations of local cellular and humoral immune defense, which is expressed by a decrease in the level of T-helpers, B-lymphocytes and IgA with a simultaneous increase in T-suppressors / killers in women with chronic endometritis. The established changes amplify at a bacterial vaginosis

Number and function of uterine natural killer cells in recurrent miscarriage and implantation failure: a systematic review and meta-analysis.

BACKGROUNDUterine natural killer cells (uNK) are the most abundant lymphocytes found in the decidua during implantation and in first trimester pregnancy. They are important for early placental development, especially trophoblast invasion and transformation of the spiral arteries. However, inappropriate uNK function has been implicated in reproductive failure, such as recurrent miscarriage (RM) or recurrent implantation failure (RIF). Previous studies have mainly focussed on peripheral NK cells (pNK), despite the well-documented differences in pNK and uNK phenotype and function. In recent years, there has been an explosion of studies conducted on uNK, providing a more suitable representation of the immune environment at the maternal–foetal interface. Here, we summarize the evidence from studies published on uNK in women with RM/RIF compared with controls.OBJECTIVE AND RATIONALEThe objectives of this systematic review and meta-analysis are to evaluate: differences in uNK level in women with RM/RIF compared with controls; pregnancy outcome in women with RM/RIF stratified by high and normal uNK levels; correlation between uNK and pNK in women with RM/RIF; and differences in uNK activity in women with RM/RIF compared with controls.SEARCH METHODSMEDLINE, EMBASE, Web of Science and Cochrane Trials Registry were searched from inception up to December 2020 and studies were selected in accordance with PRISMA guidelines. Meta-analyses were performed for uNK level, pregnancy outcome and uNK/pNK correlation. Narrative synthesis was conducted for uNK activity. Risk of bias was assessed by ROBINS-I and publication bias by Egger’s test.OUTCOMESOur initial search yielded 4636 articles, of which 60 articles were included in our systematic review. Meta-analysis of CD56+ uNK level in women with RM compared with controls showed significantly higher levels in women with RM in subgroup analysis of endometrial samples (standardized mean difference (SMD) 0.49, CI 0.08, 0.90; P = 0.02; I2 88%; 1100 women). Meta-analysis of CD56+ uNK level in endometrium of women with RIF compared with controls showed significantly higher levels in women with RIF (SMD 0.49, CI 0.01, 0.98; P = 0.046; I2 84%; 604 women). There was no difference in pregnancy outcome in women with RM/RIF stratified by uNK level, and no significant correlation between pNK and uNK levels in women with RM/RIF. There was wide variation in studies conducted on uNK activity, which can be broadly divided into regulation and receptors, uNK cytotoxicity, cytokine secretion and effect of uNK on angiogenesis. These studies were largely equivocal in their results on cytokine secretion, but most studies found lower expression of inhibitory receptors and increased expression of angiogenic factors in women with RM.WIDER IMPLICATIONSThe observation of significantly increased uNK level in endometrium of women with RM and RIF may point to an underlying disturbance of the immune milieu culminating in implantation and/or placentation failure. Further research is warranted to elucidate the underlying pathophysiology. The evidence for measuring pNK as an indicator of uNK behaviour is sparse, and of limited clinical use. Measurement of uNK level/activity may be more useful as a diagnostic tool, however, a standardized reference range must be established before this can be of clinical use.

The mid-secretory endometrial transcriptomic landscape in endometriosis: a meta-analysis.

STUDY QUESTIONDo women with endometriosis have a different endometrial gene expression profile at the time of embryo implantation than women without endometriosis?SUMMARY ANSWERThe endometrial gene expression profile of women with endometriosis differs from that of women without endometriosis at the mid-secretory phase, although the differences are small.WHAT IS KNOWN ALREADYAbout 50% of women with endometriosis suffer infertility. Several molecular studies have suggested impaired endometrial receptivity in women with endometriosis, while others have detected no dysregulation of endometrial receptivity. Nevertheless, the previous endometrial transcriptome studies comparing women with and without endometriosis have been performed in small sample size with limited statistical power. We set out to systematically search and compile data of endometrial gene expression signatures at the receptive phase in women with endometriosis versus control women. Based on the obtained data, we conducted a meta-analysis of differentially expressed genes in order to raise the power of the analysis for identifying the molecular profiles of receptive phase endometria in endometriosis.STUDY DESIGN, SIZE, DURATIONA systematic literature search was conducted up to February 2022 following PRISMA criteria and included PubMed, Cochrane and Web of Science databases. For the systematic search, the term ‘endometriosis’ was paired with the terms ‘transcriptomics’, ‘transcriptome’, ‘gene expression’, ‘RNA-seq’, ‘sequencing’ and ‘array’, by using the Boolean operator ‘AND’ to connect them. Articles written in English were screened and interrogated for data extraction.PARTICIPANTS/MATERIALS, SETTING, METHODSA meta-analysis was performed on the selected studies to extract the differentially expressed genes described at the mid-secretory phase in women with endometriosis versus women without endometriosis in natural cycles, using the robust rank aggregation method. In total, transcriptome data of 125 women (78 patients and 47 controls) were meta-analysed, with a special focus on endometrial receptivity-specific genes based on commercial endometrial receptivity tests.MAIN RESULTS AND THE ROLE OF CHANCEIn total, 8 studies were eligible for the quantitative meta-analysis, gathering transcriptome data from the mid-secretory phase endometria of 125 women. A total of 7779 differentially expressed transcripts between the study groups were retrieved (3496 up-regulated and 4283 down-regulated) and were meta-analysed. After stringent multiple correction, there was no differential expression of any single molecule in the endometrium of women with endometriosis versus controls, while enrichment analysis detected that the pathways of chemotaxis and locomotion are dysregulated in endometriosis. Further analysis of endometrial receptivity-specific genes highlighted dysregulation of C4BPA, MAOA and PAEP and enrichment of immune and defence pathways in women with endometriosis.LIMITATIONS, REASONS FOR CAUTIONMost of the studies included into the meta-analysis were relatively small and had different study designs, which might have contributed to a bias.WIDER IMPLICATIONS OF THE FINDINGSThe current meta-analysis supports the hypothesis that endometrial receptivity is altered in women with endometriosis, although the changes are small. The molecules and pathways identified could serve as future biomarkers and therapeutical targets in detecting and treating endometriosis-associated infertility.STUDY FUNDING/COMPETING INTEREST(S)The authors declare no competing interests. This work was supported by the Spanish Ministry of Education, Culture and Sport [grant FPU15/01193] and the Margarita Salas program for the Requalification of the Spanish University system [grant UJAR01MS]; Spanish Ministry of Economy, Industry and Competitiveness (MINECO) and European Regional Development Fund (FEDER): grants RYC-2016-21199 and ENDORE SAF2017-87526-R; Programa Operativo FEDER Andalucía (B-CTS-500-UGR18; A-CTS-614-UGR20); the Junta de Andalucía [BIO-302; and PAIDI P20_00158]; the University of Jaén [PAIUJA-EI_CTS02_2017]; the University of Granada, Plan Propio de Investigación 2016, Excellence actions: Units of Excellence; Unit of Excellence on Exercise and Health (UCEES), and by the Junta de Andalucía, Consejería de Conocimiento, Investigación y Universidades and European Regional Development Fund (ERDF), ref. SOMM17/6107/UGR; the Estonian Research Council (grant PRG1076); Horizon 2020 innovation (ERIN, grant no. EU952516) of the European Commission and Enterprise Estonia (grant EU48695).TRIAL REGISTRATION NUMBERThe systematic review was registered at PROSPERO (identifier: CRD42020122054).

Hysteroscopic and morphological evaluation of endometrium in reproductive age women with comorbid polycystic ovarian syndrome and chronic endometritis in the protocols of in vitro fertilization

Summary. Today, infertility, which is associated with polycystic ovary syndrome, is a pressing problem in obstetrics, gynecology and reproductive medicine. Polycystic ovary syndrome (PCOS) is a common endocrine disease that affects about 6 to 20 % of women of childbearing potential. Its main signs are ovulatory and menstrual dysfunction (including anovulation), increased levels of gonadotropin-releasing hormone, hypersecretion of luteinizing hormone, hyperandrogenemia, metabolic disorders and morphological changes in the endometrium and ovaries. The aim of the study – to evaluate hysteroscopic and morphological changes in the endometrium of women with infertility against the background of PCOS and chronic endometritis and to analyze the effect of a modified treatment regimen in the protocols of in vitro fertilization on the manifestations of chronic endometritis. Materials and Methods. We examined 140 women with endocrine infertility against the background of PCOS and chronic endometritis, which were divided into 2 study groups, depending on the cause and treatment approach for infertility, as well as 35 women with male-caused infertility who comprised the control group. All examined patients underwent hysteroscopy, targeted biopsy and immunohistochemical study of the endometrium. Histological study of the endometrium was performed by staining the tissue with hematoxylin and eosin according to a standard protocol in order to understand the structural changes of the endometrium in women of the study groups. Immunohistochemical testing involved the detection of CD138 using standard Daco antibody kits (Denmark). Results. Patients with PCOS and chronic endometritis used standard anti-inflammatory treatment which followed our modified protocol. This resulted in significantly reduced manifestations of chronic endometritis. The modified protocol includes inositol and alpha-lipoic acid in pre-pregnancy preparation; PRP therapy of the uterine cavity; and lipofundin infusions on day 2–3 of the menstrual cycle and on the day of embryo transfer. Conclusions. Histological examination of the endometrium with manifestations of stromal-adenomatous polyps revealed reduced signs of cytogenic stroma proliferation, decreased perivascular edema and the number of erythrocyte diapedesis. The number of glandular elements and the size of the glands also decreased

Altered endometrial oestrogen-responsiveness and recurrent reproductive failure.

Graphical abstract Recurrent reproductive failure (RRF) encompasses recurrent implantation failure (RIF) and recurrent pregnancy loss (RPL). These highly prevalent, distressing conditions have many unanswered questions regarding aetiology and management. Oestrogen receptor beta (ERβ) is the predominant oestrogen receptor expressed in the vascular endothelium of the endometrium during the window of implantation (WOI). The establishment of normal endometrial receptivity is integrally associated with progesterone receptor (PR). Therefore, we aimed to investigate whether women with RRF have clinical, type-specific endometrial aberrations of ERβ, PR and Ki-67 expression during the WOI. Thirty-eight endometrial biopsies were collected; 29 RRF (10 RIF, 9 recurrent loss of early pregnancy (RLEP) and 10 recurrent fetal loss (RFL)) and 9 fertile controls (FC). Within RIF, RLEP and RFL groups, the perivascular compartment showed significantly lower levels of ERβ vs FC (P = 0.02, P = 0.03 and P = 0.01, respectively). Vascular endothelium also displayed significantly lower levels of ERβ within RIF and RFL cohorts vs FC (P = 0.03 and P = 0.003). The expression of Ki-67 was significantly lower within vascular endothelium of all RRF; RIF (P = 0.02), RLEP (P = 0.02) and RFL (P <0.01). PR was significantly reduced (P <0.001) in the perivascular area of women with RIF. These findings provide novel insights into biological correlates of clinical subtypes of RRF. The endometrium of women with RRF display significantly altered levels of ERβ, PR and Ki-67 during the WOI, furthering our understanding of the defective endometrial phenotype of women suffering from RRF, with possible impaired glandular function, angiogenesis and decidualisation.Lay summaryRecurrent reproductive failure (RRF) refers to a group of devastating conditions with many unanswered questions regarding their causes and treatment options. The lining of the womb, the endometrium, is primed and suitable for successful embryo implantation for a short time during the menstrual cycle; the window of implantation (WOI). Oestrogen is a key hormone that plays an important role in regulating the endometrium and its effects are exerted via two oestrogen receptor subtypes. Oestrogen receptor beta (ERβ) is the main oestrogen receptor present during the WOI. Progesterone receptor allows the other main hormone, progesterone, to influence the endometrial activity and Ki-67 reflects the proliferative activity of the cells within the endometrium. We investigated the expression of these markers in endometrial samples collected from women with RRF and proven fertility. We found that the endometrium of women with RRF has significantly lower levels of ERβ and Ki-67 during the WOI, possibly leading to unsuccessful pregnancies.

Receptivity at different endometrial thickness in women of reproductive age (literature review)

Overcoming infertility and reducing miscarriage are important challenges in obstetrics and gynecology. In the Russian Federation, the frequency of infertile couples reaches 24%; the percentage of miscarriage does not decrease, accounting for up to 20% of all desired pregnancies. The endometrial factor plays an important role in the causes of gravidar losses. Inadequate phase transformation of the endometrium is a predictor of fetal egg implantation failures. An integral part of endometrial cyclic transformations is the full proliferation of the mucous membrane of the uterine body. However, there is a cohort of women who, due to the insufficiency of proliferative processes in the endometrium, have a syndrome of thin endometrium. The pathogenesis of the formation of a hypoplastic endometrium in women with normal characteristics of hormonal factors is still poorly understood. Particular attention is paid to determining the function of proteomic markers (leukemia inhibiting factor, Forkhead box-protein family, etc.) in the genesis of the hypoplastic endometrium. This review presents an analysis of modern Russian and foreign sources on the receptivity of the endometrium, including the syndrome of thin endometrium in women of reproductive age.

Differences between proximal and distal endometrial monolayer thickness in women aged over 50 years undergoing saline infusion sonohysterography.

Saline infusion sonohysterography (SIS) is a procedure performed to evaluate the endometrium in women with postmenopausal bleeding. To investigate differences in endometrial monolayer measurements in women aged >50 years undergoing SIS. Retrospective study of women aged >50 undergoing SIS. Endometrial echo (EE) was measured according to the International Endometrial Tumor Analysis (IETA) guidelines. Monolayer thickness was compared between anterior and posterior uterine walls and between the monolayer that was proximal or distal to the ultrasound probe. Presence and location of focal thickening and polyps on each of the monolayers were assessed. SIS was performed in 608 patients. Of them, 485 (79.8%) had anteverted, 85 (14%) retroverted, and 38 (6.2%) a midposition uterus. The mean posterior monolayer was thicker than the anterior monolayer (2.14 mm vs. 1.88 mm; P = 0.002). The distal monolayer was thicker than the proximal layer in both anteverted and retroverted uteri (2.18 mm vs. 1.84 mm; P < 0.0001). In 16% of women, the difference between distal and proximal monolayers was ≥1 mm. Focal thickening was seen 3.3 times more frequently in the distal endometrium. Among women with a double layer EE >4 mm, 18.8% had a proximal layer of <2 mm while only 4.6% had a distal EE <2 mm. Distal endometrium measures thicker than the proximal endometrium in most SIS cases and in one out of six women, the difference is >1 mm. The distal layer is three times more likely to contain focal thickening. Sonologists should be conscious of possible enhancement artifact when measuring the EE during SIS.

Whole-Tissue Deconvolution and scRNAseq Analysis Identify Altered Endometrial Cellular Compositions and Functionality Associated With Endometriosis.

The uterine lining (endometrium) exhibits a pro-inflammatory phenotype in women with endometriosis, resulting in pain, infertility, and poor pregnancy outcomes. The full complement of cell types contributing to this phenotype has yet to be identified, as most studies have focused on bulk tissue or select cell populations. Herein, through integrating whole-tissue deconvolution and single-cell RNAseq, we comprehensively characterized immune and nonimmune cell types in the endometrium of women with or without disease and their dynamic changes across the menstrual cycle. We designed metrics to evaluate specificity of deconvolution signatures that resulted in single-cell identification of 13 novel signatures for immune cell subtypes in healthy endometrium. Guided by statistical metrics, we identified contributions of endometrial epithelial, endothelial, plasmacytoid dendritic cells, classical dendritic cells, monocytes, macrophages, and granulocytes to the endometrial pro-inflammatory phenotype, underscoring roles for nonimmune as well as immune cells to the dysfunctionality of this tissue.

Transcriptomics of receptive endometrium in women with sonographic features of adenomyosis

BackgroundWomen with uterine adenomyosis seeking assisted reproduction have been associated with compromised endometrial receptivity to embryo implantation. To understand the mechanisms involved in this process, we aimed to compare endometrial transcriptome profiles during the window of implantation (WOI) between women with and without adenomyosis.MethodsWe obtained endometrial biopsies LH-timed to the WOI from women with sonographic features of adenomyosis (n=10) and controls (n=10). Isolated RNA samples were subjected to RNA sequencing (RNA-seq) by the Illumina NovaSeq 6000 platform and endometrial receptivity classification with a molecular tool for menstrual cycle phase dating (beREADY®, CCHT). The program language R and Bioconductor packages were applied to analyse RNA-seq data in the setting of the result of accurate endometrial dating. To suggest robust candidate pathways, the identified differentially expressed genes (DEGs) associated with the adenomyosis group in the receptive phase were further integrated with 151, 173 and 42 extracted genes from published studies that were related to endometrial receptivity in healthy uterus, endometriosis and adenomyosis, respectively. Enrichment analyses were performed using Cytoscape ClueGO and CluePedia apps.ResultsOut of 20 endometrial samples, 2 were dated to the early receptive phase, 13 to the receptive phase and 5 to the late receptive phase. Comparison of the transcriptomics data from all 20 samples provided 909 DEGs (p<0.05; nonsignificant after adjusted p value) in the adenomyosis group but only 4 enriched pathways (Bonferroni p value < 0.05). The analysis of 13 samples only dated to the receptive phase provided suggestive 382 DEGs (p<0.05; nonsignificant after adjusted p value) in the adenomyosis group, leading to 33 enriched pathways (Bonferroni p value < 0.05). These included pathways were already associated with endometrial biology, such as “Expression of interferon (IFN)-induced genes” and “Response to IFN-alpha”. Data integration revealed pathways indicating a unique effect of adenomyosis on endometrial molecular organization (e.g., “Expression of IFN-induced genes”) and its interference with endometrial receptivity establishment (e.g., “Extracellular matrix organization” and “Tumour necrosis factor production”).ConclusionsAccurate endometrial dating and RNA-seq analysis resulted in the identification of altered response to IFN signalling as the most promising candidate of impaired uterine receptivity in adenomyosis.

Clock Genes - A New Perspective on Pregnancy Complications-

The circadian rhythm is controlled by clock genes, which is crucial for development, reproduction and pregnancy. The precise timing of kisspeptin, gonadotropin-releasing hormone, and gonadotropin secretion in the hypothalamic-pituitary-gonadal axis determines fertility. Additionally, the mouse uterus demonstrated the circadian rhythm regulated by circadian clock genes. The expression level of aryl hydrocarbon receptor nuclear translocator-like protein (Bmal1), a core component of circadian oscillation, was shown to be lower in the endometrium of women who experienced recurrent miscarriages. Bmal1 participates in a variety of physiological processes. It is known that Bmal1-deficient mice were sterile and had damaged gametes and reproductive systems. In addition, hormone secretion is disrupted in Bmal1-deficient mice. Recently, we discovered that uterus-specific Bmal1-deficient mice had impaired placental development and were unable to sustain pregnancy. Additionally, we revealed that time-restricted feeding controlled the uterine clock's circadian rhythm. In humans, we found a higher incidence of hypertensive disorders of pregnancy in pregnant women who had dysmenorrhea in the past at a younger age. These findings suggest that the uterine clock system is a candidate for the new perspective on pregnancy complications.

MORPHOLOGICAL AND IMMUNOHISTOCHEMICAL CHANGES OF THE ENDOMETRIUM IN WOMEN WITH HYPERPROLIFERATIVE PATHOLOGY OF THE UTERUS AND REPRODUCTIVE DYSFUNCTION

Hyperproliferative pathology of the uterus (HPЕ) occupies one of the leading places in the structure of female infertility, leading to women’s generative dysfunction in 80% of cases. The aim of the study: to assess the immunohistochemical and morphological features of the endometrium in patients with hyperproliferative pathology of the endometrium and infertility. Materials and methods. Forty endometrial samples from women with infertility and hyperproliferative pathology of the endometrium were examined: 20 samples of endometrium obtained from women with infertility and endometrial polyps (1 group) and 20 samples obtained from women with infertility and endometrial hyperplasia without atypia. The endometrium of 20 healthy women was examined as the control group. Pipelle biopsy of the endometrium was performed in the luteal phase of the menstrual cycle in women of the main and control groups to obtain the material. The morphological and immunohistochemical features of the endometrium in women with infertility and hyperproliferative pathology of the endometrium were determined. Histogram sections were stained with hematoxylin and eosin, and picrofuxin (Van Gizon’s stain). An immunohistochemical study was performed with the polymer detection method of antigen application using the UltraVisionQuanto peroxidase polymer and DAB plus chromogen detection system to determine the expression of monoclonal antibodies to estrogen and progesterone receptors, natural killers СD-56 and СD-138. The results of the investigation. Morphological examination of the endometrium of women with infertility and hyperproliferative pathology of the endometrium revealed a number of pathological changes: glandular and glandular-fibrous polyps of the endometrium (70–80%), atypical endometrial hyperplasia (10–20%), dyschronosis of endometrial receptivity (40–50%), morphological signs of chronic endometritis, positive expression of CD-138 (10%), as well as their combination. Conclusions. Women with infertility and hyperproliferative pathology of the uterus and unsuccessful attempts at ART have expressed dyshronosis of receptivity of the endometrium on the background of reduced expression of endometrial natural killers and positive expression of CD-138. It is indicative of the dysregulation of immunological homeostasis on the local level, which may be the cause of disruption of blastocyst implantation processes, unsuccessful attempts of ART and lead to early pregnancy loss.

Ультразвукове дослідження органів малого таза в жінок із невдалими спробами ДРТ на тлі хронічного ендометриту

The subject of discussion is the effectiveness of sonographic examination in the diagnosis of chronic endometritis, despite the high availability of this method of research and ultrasound criteria of chronic endometritis. The validity of the ultrasound diagnosis of chronic endometritis is denied by many experts, but there is a consensus on the high prognostic detectable values of the signs for further verification of this nosology. Purpose — to determine the diagnostic value of pelvic ultrasound to assess the condition of the endometrium in women with unsuccessful attempts at ART in the history of chronic endometritis. Materials and methods. The prospective study of women with chronic endometritis and unsuccessful attempts at ART, based on MC LLC «ISIDA-IVF», for the period from 2019 to 2021. I and II groups were 110 women of childbearing age with chronic endometritis and repeated unsuccessful attempts at ART, and 40 women of reproductive age without impaired fertility, who formed a control group. Ultrasound of the pelvic organs was performed in all patients of the studied groups on days 5–7, 12–15, 22–24 of the menstrual cycle. The threshold value of static reliability is p&lt;0.05. Results. In women with infertility on the background of chronic endometritis in history, the average thickness of the median M-echo in the periovulatory period in group I (with pre-conception preparation) was 9.1±0.08 mm, in group II (without pre-conception preparation) — 7.6±0.9 mm, in the CG — 9.3±0.1 mm and had statistically significant differences (p1,2&lt;0.05, p1,3&gt;0.05, p2,3&lt;0.05). Thin endometrium was detected in patients of group I with pre-conception preparation in 16.3% and twice as often in women in group II without pre-conception preparation — 30.9% (p1,2&lt;0.05). The inconsistency of endometriality was approximately the same in the number of patients in the 1st and 2nd group, which was seen during the day in the cohort of the control group. In case of hyperplastic chronic endometritis macrotype in both groups, polypendometry was tested, as in 2.5 times it was diagnosed in patients of the II group — 9.6% versus 3.6% in women of the I group (p1,2&lt;0.05). The incidence of diagnosed hyperplasia endometrial in pre-existing groups is not small statistically significant differences — 1.8% and 5.4% (p1,2&gt;0.05) apparently. In case of hypoplastic chronic endometritis macrotype, thin endometrium is 2 times and more often in the II group. Conclusions. Transvaginal echography has high specificity (96.6%), sensitivity (55.2%) and diagnostic accuracy (68.2%). Criterias for the active readiness of women to the vagina for pre-conception health improvement are updating the photographic picture of endometries with values of M-exo&gt;7 mm; normalization of uterine hemodynamics (RI&lt;0.9, PI&lt;3.0). The research was carried out in accordance with the principles of the Helsinki declaration. The study protocol was approved by the Local ethics committee of all participating institutions. The informed consent of the patient was obtained for conducting the studies. No conflict of interest was declared by the authors. Key words: ultrasound examination, assisted reproductive technologies, chronic endometritis.

Influence of parity and Body Mass Index (BMI) on endometry thickness variation in women of Lokossa, Benin

The thickness of the endometrium is also an important parameter in the clinical management of women. It is often influenced by several parameters which are important to determine. This is why the present study looked at the factors that influence the variation in the thickness of the endometrium of women in Lokossa in the Republic of Benin. This descriptive, prospective and cross-sectional study concerns 166 women aged 15 to 40 in Lokossa. Ultrasound examination measured the thickness of the endometrium on the longitudinal section through the suprapubic route. Anthropometric parameters were taken by other equipment from each woman. Analysis of one-way proc glm ANOVA variants and Student-Newman-Keuls (SNK) tests identified factors associated with variation in endometrial thickness in women. At the end of the study, the minimum, average and maximum values ​​of the thickness of the endometrium obtained are respectively Vm = 3.50 mm, VMoy = 6.73 ± 1.38 mm, MV = 10.00 mm during the pre-ovulatory phase, Vm = 7.30 mm, MV avg = 10.75 ± 2.11 mm, MV = 16.00 mm during the post-ovulatory period. There is an association between the thickness of the endometrium and parameters such as age, weight, Body Mass Index, parity. BMI has been found to be associated with changes in endometrial thickness. And this variation in the thickness of the endometrium of the woman's uterus is related to the number of deliveries. Thus, the higher the number of childbirth, the greater the thickness of the endometrium deprived.

Expression of estrogen and progesterone receptors in the endometrium in women with impaired reproductive function with different functional activity of the corpus luteum of the ovary

Aim. To assess the expression of estrogen (ER) and progesterone receptors (PR) in the endometrium during the implantation "window" in women with impaired reproductive function with low (16.1P30 nmol/l) and high (P30 nmol/l)) functional activity of the corpus luteum of the ovary.&#x0D; Materials and methods. A prospective single-center cohort comparative study. The main group included patients with infertility (n=93) and miscarriage (n=47) of unclear reason in the anamnesis, the control group 16 healthy fertile women. In all cases, vacuum-aspiration endometrial biopsy was performed on the 68 days after ovulation with simultaneous collection of a peripheral blood sample to determine the levels of estradiol and progesterone. Endometrial samples were studied by histological and immunohistochemical (ER, PR expression) methods.&#x0D; Results. Women of the main (1) and control (2) groups had an ovulatory menstrual cycle. The serum estradiol (E2) level was 691.326.5 pmol/l (1) and 707.466.1 pmol/l (2); progesterone (P) 46.11.8 nmol/l (2) and 39.14.9 nmol/l (2). Mid-secretory endometrium (results of histological examination) was determined in all healthy women (n=16) and in 46% (n=64) of the patients of the main group. All women in the control group had adequate hormone-receptor characteristics of the endometrium, in women in the main group in 45% (n=63). Comparative analysis of the expression of ER, PR in the endometrium in patients with different functional activity of the corpus luteum of the ovary was carried out in the groups 1 and 2, in women with different reproductive dysfunctions in the anamnesis (without significant differences; p0.05); subgroup analysis was performed in cohorts of women with normal (63 women in group 1 and 16 in group 2) and inadequate (77 patients in group 1) hormone-receptor status of the endometrium. There were no significant differences in the expression of ER, PR in the endometrium with low/high functional activity of the corpus luteum of the ovary with similar morphological characteristics of the endometrium (p0.05). Correlation analysis did not reveal significant relationships between the level of P in the blood and indicators of endometrial expression ER, PR (p0.05).&#x0D; Conclusion. In the studied cohorts, the prognostic significance of the functional activity of the corpus luteum of the ovary for the adequate expression of estrogen and progesterone receptors has not been established. To clarify the presence/absence of endometrial dysfunction in women with a history of reproductive failures, a morphological examination of the endometrium of the middle secretion phase should be carried out.

Microbiological Study of Vaginal Microbiota and Endometrium in Women with Chronic Endometritis

The research objective was to study the spectrum of the vagina and endometrium microorganisms in women with chronic endometritis (CE) in order to take adequate therapeutic measures. Methods and Results: We did a cross-sectional study in 47 women (average age of 35.38±5.19 years) with histologically confirmed CE. The vaginal microbiota and endometrial biopsies were assessed using microbiological research methods in accordance with the medical technology "Integral assessment of the state of the vaginal microbiota." To identify the share of different types of microorganisms in the structure of the biocenosis, the coefficient of constancy of the species (C) was used. Only 19% of patients had a titer of Lactobacillus spp. within the age norm, while the deficit was observed in 80% of women. Among the representatives of Enterobacteriaecae, Escherichia coli and Klebsiella aerogenes were sown, which are considered to be random species (C=11% and C=2.1%, respectively). The average titer for E. coli was 3.6±1.3 lg CFU/swab and for K. aerogenes - 2.14 lg CFU/swab. An atypical variant of E. coli with hemolytic properties was found in only one sample. All isolates of the genus Staphylococcus were also random species (C did not exceed 25%). Coagulase-negative staphylococci (CoNS) were detected in 7 patients (C=15%), while the average titer was 2.1±0.4 lg CFU/swab. S. aureus was isolated from only one patient at a titer of 5 lg CFU/swab. Corynebacterium spp. were isolated in 11% of cases (C = 11% - random species), in a titer of 3.2±0.8 lg CFU/swab. Enterococcus spp. also belonged to random species (C=23.4%). At the same time, E. faecalis was inoculated in 19% of cases and E. faecium was sown in 4.3%, the average titer of which was 3.1±0.9 and 5 lg CFU/swab. Streptococcus spp. were recorded in only one case at a concentration of 5 lg CFU/swab. Fungi of the Candida were isolated as a random species in 8.5% of cases. The growth of microorganisms in endometrial samples was obtained only in 3 examined women with CE (6.4% of cases). The endometrial microbiota were represented only by random species, for which the C index ranged from 2.1% to 4.3%. Conclusion: The microbiological study of the microbiota of vaginal discharge showed the presence of dysbiotic disorders with a significant deficiency of lactobacilli (80%) without the dominance of representatives of the Lactobacillus spp. In the structure of opportunistic microflora, Escherichia coli, coagulase-negative staphylococcus, Enterococcus spp., and E. faecalis prevailed as random species. Representatives of the microbiota in endometrial biopsies were identified only in 6.4% of cases, and are represented by random species.

Increased METTL3-mediated m6A methylation inhibits embryo implantation by repressing HOXA10 expression in recurrent implantation failure

BackgroundRecurrent implantation failure (RIF) is a major limitation of assisted reproductive technology, which is associated with impaired endometrial receptivity. Although N6-methyladenosine (m6A) has been demonstrated to be involved in various biological processes, its potential role in the endometrium of women with RIF has been poorly studied.MethodsGlobal m6A levels and major m6A methyltransferases/demethylases mRNA levels in mid-secretory endometrium from normal and RIF women were examined by colorimetric m6A quantification strategy and quantitative real-time PCR, respectively. The effects of METTL3-mediated m6A modification on embryo attachment were evaluated by an vitro model of a confluent monolayer of Ishikawa cells co-cultured with BeWo spheroids, and the expression levels of homeo box A10 (HOXA10, a well-characterized marker of endometrial receptivity) and its downstream targets were evaluated by quantitative real-time PCR and Western blotting in METTL3-overexpressing Ishikawa cells. The molecular mechanism for METTL3 regulating HOXA10 expression was determined by methylated RNA immunoprecipitation assay and transcription inhibition assay.ResultsGlobal m6A methylation and METTL3 expression were significantly increased in the endometrial tissues from women with RIF compared with the controls. Overexpression of METTL3 in Ishikawa cells significantly decreased the ration of BeWo spheroid attachment, and inhibited HOXA10 expression with downstream decreased β3-integrin and increased empty spiracles homeobox 2 expression. METTL3 catalyzed the m6A methylation of HOXA10 mRNA and contributed to its decay with shortened half-life. Enforced expression of HOXA10 in Ishikawa cells effectively rescued the impairment of METTL3 on the embryo attachment in vitro.ConclusionIncreased METTL3-mediated m6A modification represents an adverse impact on embryo implantation by inhibiting HOXA10 expression, contributing to the pathogenesis of RIF.

Interleukin 22 Expression During the Implantation Window in the Endometrium of Women with Unexplained Recurrent Pregnancy Loss and Unexplained Infertility Compared to Healthy Parturient Individuals.

We evaluated the expression of interleukin-22 (IL-22) in the endometrium of women with unexplained recurrent pregnancy loss (uRPL) and unexplained infertility (UI) compared to the women with normal pregnancies. Endometrial tissues were collected from 20 women with UI, 20 women with uRPL, and 24 healthy women as a control group. Immunohistochemical expression and gene expression of IL-22 were analyzed by immunohistochemistry (IHC) and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) methods. The controls showed lower IL-22 expression than the uRPL group (P > 0.05) using PCR. It was also found that patients with UI had lower levels of IL-22 expression compared to the uRPL group (P > 0.05). Although IL-22 expression in the endometrium of patients with UI was higher than the control group, this difference was not statistically significant (P < 0.05). IL-22 immunoreactivity was observed in the endometrial glands and stromal tissues using IHC. We found the lowest IL-22 expression in the control group and the highest in uRPL samples (P < 0.05). Our findings suggest that a significant increase in IL-22 expression in uRPL patients may affect fertility and pregnancy outcomes or even have a considerable impact on immune function deficits. Further studies on the critical function of IL-22 during pregnancy are suggested.

Disrupted PGR-B and ESR1 signaling underlies defective decidualization linked to severe preeclampsia.

Decidualization of the uterine mucosa drives the maternal adaptation to invasion by the placenta. Appropriate depth of placental invasion is needed to support a healthy pregnancy; shallow invasion is associated with the development of severe preeclampsia (sPE). Maternal contribution to sPE through failed decidualization is an important determinant of placental phenotype. However, the molecular mechanism underlying the in vivo defect linking decidualization to sPE is unknown. Global RNA sequencing was applied to obtain the transcriptomic profile of endometrial biopsies collected from nonpregnant women who suffer sPE in a previous pregnancy and women who did not develop this condition. Samples were randomized in two cohorts, the training and the test set, to identify the fingerprinting encoding defective decidualization in sPE and its subsequent validation. Gene Ontology enrichment and an interaction network were performed to deepen in pathways impaired by genetic dysregulation in sPE. Finally, the main modulators of decidualization, estrogen receptor 1 (ESR1) and progesterone receptor B (PGR-B), were assessed at the level of gene expression and protein abundance. Here, we discover the footprint encoding this decidualization defect comprising 120 genes-using global gene expression profiling in decidua from women who developed sPE in a previous pregnancy. This signature allowed us to effectively segregate samples into sPE and control groups. ESR1 and PGR were highly interconnected with the dynamic network of the defective decidualization fingerprint. ESR1 and PGR-B gene expression and protein abundance were remarkably disrupted in sPE. Thus, the transcriptomic signature of impaired decidualization implicates dysregulated hormonal signaling in the decidual endometria in women who developed sPE. These findings reveal a potential footprint that could be leveraged for a preconception or early prenatal screening of sPE risk, thus improving prevention and early treatments. This work has been supported by the grant PI19/01659 (MCIU/AEI/FEDER, UE) from the Spanish Carlos III Institute awarded to TGG. NCM was supported by the PhD program FDGENT/2019/008 from the Spanish Generalitat Valenciana. IMB was supported by the PhD program PRE2019-090770 and funding was provided by the grant RTI2018-094946-B-100 (MCIU/AEI/FEDER, UE) from the Spanish Ministry of Science and Innovation with CS as principal investigator. This research was funded partially by Igenomix S.L.

Reduced innate lymphoid cells in the endometrium of women with endometriosis.

Innate lymphoid cells (ILCs), a recently discovered family of innate immune cells, are responsible for the early immune response, and control both innate and adapted immune system via cytokine secretion. The role of ILCs in endometriosis has not been investigated; therefore, here, we aimed to investigate how the proportion of ILCs changes in endometriosis. The percentage of each ILC group in CD45+ cells was examined in the peripheral blood, peritoneal fluid, endometrium, and ovarian endometrioma obtained from women with and without endometriosis (ERB-C-1216) using flow cytometry. Specimens were obtained from 19 women with endometriosis and 15 without endometriosis. In the endometrium, patients with endometriosis had lower proportion of ILC2 and 3 compared to control specimens (ILC2: .02±.01% vs .07±.03%; P<.05, ILC3: .31±.14% vs 1.10±.93%; P<.05). There was no significant change in the peripheral blood or the peritoneal fluid between the two groups. Additionally, ovarian endometrioma increased the proportion of ILCs (ILC1: .92±1.12%, ILC2: .08±.08%, ILC3: .70±.39%) compared to the endometrium samples of patients with endometriosis each with P<.05. Immunohistochemistry of IL-1β and IL-23, which are ILC3-inducing factors, showed no significant change in the H-score of the epithelium of the two groups, but a significant increase was found in ovarian endometrioma. The proportion of ILC2 and 3 was reduced in the endometrium of patients with endometriosis, and ILCs were increased in ovarian endometrioma. Our findings may indicate a new immunological approach to understand the pathophysiology of endometriosis.

Histological and Immunohistochemical Assessment of Ovarian Tissue and Endometrium in Patients with Diminished Ovarian Reserve

<h3>Study Objective</h3> To analyze the histological and immunohistochemical features of ovarian tissue and endometrium in women with diminished ovarian reserve and to investigate the correlation between these parameters and the effectiveness of infertility treatment <h3>Design</h3> Prospective study, Level II. <h3>Setting</h3> National Medical Research Center. <h3>Patients or Participants</h3> 70 patients with diminished ovarian reserve who underwent drug-free in vitro activation using a modified surgical technique. Two bilateral ovarian cortical tissue samples and one endometrial sample were obtained from each patient during the surgery. 40 patients out of 70 underwent ovarian stimulation after at least a one-month postoperative period. <h3>Interventions</h3> Bilateral ovarian cortex biopsy via laparoscopy, ovarian tissue fragmentation in aliquots of 1 mm², and hysteroscopy with endometrial biopsy were performed in all patients, followed by histological and immunohistochemical analysis (expression of proteins of the Hippo signaling pathway (YAP, MST-1, LATS-1) and PTEN (inhibitor of PI3K signaling pathway). The Human Protein Atlas datasets were used as a comparison group. <h3>Measurements and Main Results</h3> We revealed an impaired functioning of intracellular signaling pathways (primarily YAP- and PTEN-dependent) both in ovarian tissue and endometrial samples. Moreover, the expression of the proteins targeted in the study was significantly reduced in patients who responded to stimulation (n=31). Histological analysis of tissue samples of patients with diminished ovarian reserve indicated profound changes in various structures of the ovaries: surface epithelium, vascular component, stroma of ovarian cortex. In 25% of patients, chronic endometritis was also determined. <h3>Conclusion</h3> Specific histological and immunohistochemical features of ovarian tissue and endometrium associated with diminished ovarian reserve and the outcome of ovarian stimulation were shown.