Endometrial Samples Research Articles

NLRC5 exerts anti-endometriosis effects through inhibiting ERβ-mediated inflammatory response

BackgroundEndometriosis is well known as a chronic inflammatory disease. The development of endometriosis is heavily influenced by the estrogen receptor β (ERβ), while NOD-like receptors (NLRs) family CARD domain-containing 5 (NLRC5) exhibits anti-inflammatory properties during endometriosis. However, whether NLRC5-mediated anti-inflammation is involved in the ERβ-mediated endometriosis is still uncertain. This study aimed to assess that relation.MethodsNine cases of eutopic endometrial tissue and ten cases of ectopic endometrial tissue were collected from patients with endometriosis, and endometrial samples from ten healthy fertile women were analyzed, and the expression levels of ERβ were quantified using immunohistochemistry (IHC). Subsequently, we constructed mouse model of endometriosis by intraperitoneal injection. We detected the expression of ERβ, NLRC5, tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, and IL-10 and measured the volume of ectopic lesions in mice with endometriosis. In vitro, human endometrial stromal cells (hESCs) were transfected respectively with ERβ-overexpressing and NLRC5-overexpressing plasmids. We then assessed the expression of ERβ and NLRC5 using quantitative real-time PCR (qRT-PCR) and western blot analysis. Furthermore, we measured the concentrations of TNF-α, IL-6, and IL-10 in the cell culture supernatant through enzyme-linked immunosorbent assay (ELISA). Additionally, we evaluated the migration and invasion ability of hESCs using transwell and wound healing assays.ResultsInhibition of NLRC5 expression promotes the development of ectopic lesions in mice with endometriosis, upregulates the expression of pro-inflammatory factors TNF-α and IL-6, and downregulates the expression of anti-inflammatory factor IL-10. The high expression of NLRC5 in endometriosis depended on the ERβ overexpression. And ERβ promoted the migration of hESCs partially depend on inflammatory microenvironment. Lastly, NLRC5 overexpression inhibited ERβ-mediated development and inflammatory response of endometriosis.ConclusionsOur results suggest that the innate immune molecule NLRC5-mediated anti-inflammation participates in ERβ-mediated endometriosis development, and partly clarifies the pathological mechanism of endometriosis, expanding our knowledge of the specific molecules related to the inflammatory response involved in endometriosis and potentially providing a new therapeutic target for endometriosis.

Immunohistochemical Investigation of P16 Expression in Curettage Biopsies

Our study aims to investigate the immunohistochemical expression of the P16 molecule, which is involved in the cell cycle and plays a role in developing endometrial cancer in normal epithelium, endometrial polyp, and precursor lesions. A total of 68 patients underwent endometrial sampling for various reasons at the Department of Obstetrics and Gynecology, Faculty of Medicine, Kafkas University, between 2020 and 2021 were included in the study. The selected cases were categorized into four groups: proliferative endometrium, endometrial hyperplasia without atypia, atypical hyperplasia / endometrioid intraepithelial neoplasia (AH / EIN) and endometrial polyp. There were no cases with a diagnosis of endometrial tumors in our study. All patients’ pathology samples were re-evaluated, and P16 immunohistochemistry was applied to tissue samples. Among patients diagnosed with atypical endometrial hyperplasia, 72.7% exhibited moderate P16 protein expression, 18.2% had low expression, and 9.1% had high protein expression. The number of patients diagnosed with AH / EIN had a very low frequency in the study population. Among patients diagnosed with endometrial polyps, 50.0% showed moderate P16 protein expression, 20.0% exhibited low protein expression, and 30.0% had high protein expression. High P16 expression has been reported to be significantly associated with endometrial cancer in the literature. P16 expression is significant in precancerous lesions and stages of cancer development. Larger-scale studies with more cases are needed in this regard.

Model Predicting the Risk of Endometrial Hyperplasia Developing into Endometrial Cancer.

This study retrospectively analyzed the medical records of 200 patients with endometrial hyperplasia to predict the risk of concurrent endometrial cancer. Patients were categorized into either the endometrial cancer group or the endometrial hyperplasia group based on post-hysterectomy pathology. The investigation compared general information, tumor indices, fertility history, preoperative endometrial sampling methods, comorbidities, and clinical symptoms between the groups to identify risk factors for endometrial hyperplasia complicating endometrial cancer. (1) Of the 200 patients, 68 (34.0%) were diagnosed with concurrent endometrial cancer post-hysterectomy. Among these, 60 (88.24%) had endometrioid adenocarcinoma, while 8 (11.76%) had other types. Stage I was identified in 58 patients (85.29%) and Stage II in 10 patients (14.71%). High differentiation was observed in 57 cases (83.82%), moderate differentiation in 7 cases (10.29%), and poor differentiation in 4 cases (5.89%), indicating that most endometrial cancers complicated by hyperplasia were early-stage, well-differentiated endometrioid carcinomas; (2) Univariate analysis revealed statistically significant differences in age, menopausal status, length of menopause, and preoperative endometrial pathology of severe atypical hyperplasia between the groups; (3) Multivariate analysis indicated significant differences for age ≥ 53.5 years (OR: 4.307, 95% CI: 2.018-9.192, p < 0.05), menopausal status (OR: 5.250, 95% CI: 2.449-11.252, p < 0.05), and severe atypical endometrial hyperplasia (OR: 4.817, 95% CI: 1.260-18.419, p < 0.05); (4) Significant differences were observed among patients with endometrial hyperplasia when stratified by the presence of zero, one, two, or three high-risk factors. In conclusion, patients aged ≥ 53.5 years, those who are menopausal, and those with severe atypical endometrial hyperplasia preoperatively are at higher risk for endometrial cancer. The risk increases with the number of high-risk factors present in patients with atypical endometrial hyperplasia.

Uterine Cancer: A Nine-year Review from a Tertiary Hospital in Tamil Nadu

Background: With increasing uterine cancer in developing nations, there is a need for timely determination of the diagnosis, prognosis, and management options to reduce morbidity and mortality. Objective: To analyze the socio-demographic, etio-pathological features and management of uterine cancer and evaluate its correlation with grading/staging in our population. Methods: This retrospective descriptive study analyzed data from 97 histologically proven uterine cancer cases. Age, parity, symptoms, co-morbidities, body mass index (BMI), ultrasound features, histopathology type, stage and grade of the tumor, type of hysterectomy done, complications and mortality were analysed. Statistical analysis was done using ANOVA and chi-square test, and a p-value&lt;0.05 indicated statistical significance. Results: The mean age of diagnosis was 57.91 years, and the mean BMI was 29.32 Kg/m2. Majority of the patients were multiparous (42.27%), and only 10% were nulliparous. The disease was detected at an earlier age in nulliparous and obese women. Diabetes and hypertension were found in 75.25%. Most of the patients were detected with stage I cancer (80.6%). Patients diagnosed with uterine cancer on biopsy were treated with total abdominal hysterectomy with bilateral salpingo-oophorectomy andbilateral pelvic lymph node dissection (55.8%). Over 36% of patients received postop radiotherapy and/or chemotherapy. 21% patients were lost to follow-up and 12.37% died. Also, 24 cases had postoperative complications (wound infection). Conclusion: Uterine cancer is common among obese women with diabetes and hypertension. In nulliparous and the obese, the cancer was detected at an earlier age. Most of our patients had stage 1 disease, and 90% was endometroid cancer. The study highlights the importance of endometrial sampling before hysterectomy in perimenopausal women to avoid suboptimal surgery in patients diagnosed with uterine cancer after a simple hysterectomy.

Exploring Morphometry of Endometrial Glands and Blood Vessels in Reproductive Age Women With Abnormal Uterine Bleeding.

Background Abnormal uterine bleeding (AUB) is one of the most common problems encountered in gynaecological practice. Defective endometrial angiogenesis has been implicated in various benign and malignant disorders of the endometrium. Aim This study aims to assess morphometry of endometrial glands and blood vessels in patients with AUB. Material and methods This is a one year retrospective cross sectional analysis of endometrial samples received with the diagnosis of AUB in reproductive age group. All samples were routinely processed and stained. Sections were analysed for morphometry of blood vessels and endometrial glands. Results A total of 374 cases were included. Most common histological group was proliferative followed by secretory phase. A significant difference was noted in mean vascular density, diameter, mitotic scores and height of glandular epithelium in different benign and malignant groups. Conclusion This study highlights the fact that glandular and vascular morphometry can be used to differentiate between various proliferative disorders of the endometrium. In the current era of new anti-angiogenic therapies, endometrial angiogenesis and changes in vascular morphology can be targeted, thus improving treatment modalities and patient care.

Differential expression of ion channel coding genes in the endometrium of women experiencing recurrent implantation failures

Our study probed the differences in ion channel gene expression in the endometrium of women with Recurrent Implantation Failure (RIF) compared to fertile women. We analyzed the relative expression of genes coding for T-type Ca2+, ENaC, CFTR, and KCNQ1 channels in endometrial samples from 20 RIF-affected and 10 control women, aged 22–35, via microarray analysis and quantitative real-time PCR. Additionally, we examined DNA methylation in the regulatory region of KCNQ1 using ChIP real-time PCR. The bioinformatics component of our research included Gene Ontology analysis, protein–protein interaction networks, and signaling pathway mapping to identify key biological processes and pathways implicated in RIF. This led to the discovery of significant alterations in the expression of ion channel genes in RIF women’s endometrium, most notably an overexpression of CFTR and reduced expression of SCNN1A, SCNN1B, SCNN1G, CACNA1H, and KCNQ1. A higher DNA methylation level of KCNQ1’s regulatory region was also observed in RIF patients. Gene-set enrichment analysis highlighted a significant presence of genes involved with ion transport and membrane potential regulation, particularly in sodium and calcium channel complexes, which are vital for cation movement across cell membranes. Genes were also enriched in broader ion channel and transmembrane transporter complexes, underscoring their potential extensive role in cellular ion homeostasis and signaling. These findings suggest a potential involvement of ion channels in the pathology of implantation failure, offering new insights into the mechanisms behind RIF and possible therapeutic targets.

Progressively Diminished Prostaglandin E2 Signaling in Concordance with Increasing Fibrosis in Ectopic Endometrium.

The prostaglandin E2 (PGE2) signaling has traditionally been viewed to play a pivotal role in endometriosis, linking inflammation and hyperestrogenism. We have previously reported that asectopic endometrium becomes more fibrotic, the expression of both COX-2 and PGE2 receptors (EP2 and EP4) are reduced. This study further investigatedwhether the expression levels of genes involved in the biosynthesis and metabolism of PGE2in ectopic endometrium diminish in concordance with increasing lesional fibrosis. We performed immunohistochemistry analyses of COX-2, mPGES-1, mPGES-2, cPGES, 15-PGDH, EP2 and EP4 and Masson trichrome staining for ovarian endometrioma (OE), adenomyosis (AD), and deep endometriosis (DE) tissue samples and control endometrial tissue samples (CT). Gene and protein expression analyses were performed by real-time RT-PCR and Western blotting, respectively. We found that as the extent of lesional fibrosis increased, immunoexpression of COX-2, mPGES-1/2, cPGES, EP2 and EP4 in OE lesions was increased but no change in these genes/proteins in DE lesions as compared with CT. Immunoexpression of COX-2 was found to be reduced while that of 15-PGDH was found to be elevated in DE lesions. In AD lesions, only EP2 and COX-2 were overexpressed. Thus, our data indicate that when the extent of lesional fibrosis is high, the PGE2 signaling pathway is depressed, manifesting as reduced COX-2 expression and elevated expression of 15-PGDH. They underscore the fact that not all ectopic endometria are the same and equal, and highlight the importance of the extracellular matrix in shaping the lesional behavior and response to drug treatment.

Endometrial pathology in abnormal uterine bleeding (AUB) &amp; role of different techniques of endometrial sampling in evaluation of AUB: A cross-sectional study

: Abnormal uterine bleeding (AUB), traditionally defined as uterine bleeding that is abnormal in volume, regularity, and/or timing. It affects 14–25% of women of reproductive age. AUB has a major impact on quality of life of women. Aetiology of AUB may be structural and functional and varies with the age group. Histology of endometrium remains the mainstay in diagnosis of pathology and management of AUB. Various techniques such as endometrial sampling by pipelle, dilatation and curettage (D &amp; C) and hysteroscopy are used to obtain endometrium for histological diagnosis. A single center prospective cross-sectional study at teaching institute was carried out to include 150 females from 18-70 years presenting with abnormal uterine bleeding. Objective of study was to analyse prevalence of various endometrial pathologies in AUB, symptomatology as well as age wise distribution of various endometrial pathologies in AUB and compare feasibility of different techniques of collecting endometrial sample. Endometrial sample was obtained either by OPD Pipelle brush or Surgical Dilatation and Curettage method or hysteroscopy. Statistical Analysis was done by using Epi-Info-7 software.Commonest age group presenting with AUB was 41-50 (40%) years, 64% were para 3 or more, Proliferative and secretary changes, 53(35.3%) and 24(16%) cases while endometrial hyperplasia without atypia 18 (12%) cases and with atypia 7 cases (4.7%), endometrial carcinoma 5 cases (3.3%) while endometrial polyp was seen in 8 cases (5.3%). Heavy menstrual bleeding with normal duration was the commonest presentation in 64%. Endometrial sampling technical failure was seen in 2 cases with pipelle biopsy while in 7 cases inadequate sample was obtained with pipelle and dilatation and curettage. Uterine perforation was seen in one case which was managed conservatively.Technical failure and inadequate sample needs to be addressed considering background high risk factors, TVS imaging, repeat sample and use of hysteroscopy may be offered case to case basis.

Factors affecting pregnancy rates in mares bred with cryopreserved semen

Historically, 8 × 0.5 ml straws, containing approximately 800 million sperm and 250 million progressively motile sperm were provided as a single ‘breeding dose’ of cryopreserved stallion semen. With the use of deep horn artificial insemination, there is a trend to reduce the number of 0.5 ml straws sold as a breeding dose, sometimes down to as little as one straw. Our aims were to determine if the number of straws provided as a breeding dose, as well as other mare, stallion and management factors, have an impact on pregnancy outcome in mares inseminated with cryopreserved semen. Unexpectedly, we identified no effect of the number of 0.5 ml straws on pregnancy outcome. We also identified no difference in pregnancy outcome for those mares inseminated once post-ovulation compared to mares inseminated once pre- and once post- ovulation. Additionally, for mares inseminated once post-ovulation, we identified no benefit of breeding 0-3 hours post-ovulation vs. breeding 0-6 hours post-ovulation. Other factors not associated with pregnancy outcome included: whether an endometrial sample was obtained for bacteriologic culture, whether the endometrial sample produced bacterial growth, whether a mare developed fluid after breeding, whether a mare was treated for bacterial endometritis and/or uterine fluid, and post-thaw progressive sperm motility. These results suggest the existence of an effective industry self-selection process in which only semen from the most fertile stallions is marketed in these ‘ultra-low’ doses and that breeding mares within 3 hours post- ovulation provides no benefit to pregnancy outcome compared to breeding mares within 6 hours post-ovulation.

In Situ Colorimetric LAMP Based on One-Step Modified Filter Paper to Screen Human Papillomavirus (HPV)16/18 from Clinical Samples.

Cervical cancer is among the most common malignant tumors in women. The development of rapid screening techniques plays an important role in early screening for cancer treatment. We have developed an HPV screening method, which effectively combines the high-efficiency nucleic acid enrichment of chitosan-modified filter paper and the rapid visual detectability of colorimetric LAMP, along with the enhancement of the tolerance ability of the pH-sensitive LAMP reagent to acidic original samples, making the detection of HPV 16/18 easy to carry out and reliable, which is helpful for the epidemiological prevention and control strategies of HPV-induced cancer. This technique can simultaneously exhibit the "in situ amplification" capability of chitosan-modified filter paper and the nontemperature cycle dependence of visual LAMP detection. Therefore, DNA extraction and amplification can be performed efficiently and quickly within a single reaction where all DNA is concentrated in the QF paper disc. By embedding amino-modified filter paper into the plastic chip, a simple and reliable disposable chip was prepared for rapid HPV16 and HPV18 detection from clinical endometrial samples, and the results were 100% consistent with clinical diagnosis. More importantly, even after the sample was diluted 100-fold, HPV16/18-infected cells could be accurately identified, showing the advantages of the system in early cancer screening. Moreover, for endometrial samples containing plenty of cells, the filter paper could be used to enrich cells by filtration, preventing the acidic fluid from impacting pH-induced colorimetric LAMP detection and realizing direct amplification for HPV identification without nucleic acid extraction. This easy-to-operate system that can analyze a wide range of samples will be suitable for routine on-site HPV screening, dramatically extending the applications and utility for rapid, near-patient nucleic acid testing.

Aging promotes accumulation of senescent and multiciliated cells in human endometrial epithelium.

What changes occur in the endometrium during aging, and do they impact fertility? Both the transcriptome and cellular composition of endometrial samples from women of advanced maternal age (AMA) are significantly different from that of samples from young women, suggesting specific changes in epithelial cells that may affect endometrial receptivity. Aging is associated with the accumulation of senescent cells in aging tissues. Reproductive aging is mostly attributed to the decline in ovarian reserve and oocyte quality, whereas the endometrium is a unique complex tissue that is monthly renewed under hormonal regulation. Several clinical studies have reported lower implantation and pregnancy rates in oocyte recipients of AMA during IVF. Molecular studies have indicated the presence of specific mutations within the epithelial cells of AMA endometrium, along with altered gene expression of bulk endometrial tissue. Endometrial transcriptome profiling was performed for 44 women undergoing HRT during the assessment of endometrial receptivity before IVF. Patients younger than 28 years were considered as the young maternal age (YMA) group (age 23-27 years) and women older than 45 years were considered as the AMA group (age 47-50 years). Endometrial biopsies were obtained on Day 5 of progesterone treatment and RNA was extracted. All endometrial samples were evaluated as being receptive based on the expression of 68 common endometrial receptivity markers. Endometrial samples from another 24 women classified into four age groups (YMA, intermediate age group 1 (IMA1, age 29-35), intermediate age group 2 (IMA2, age 36-44), and AMA) were obtained in the mid-secretory stage of a natural cycle (NC) and used for validation studies across the reproductive lifespan. A total of 24 HRT samples (12 YMA and 12 AMA) were subject to RNA sequencing (RNA-seq) and differential gene expression analysis, 20 samples (10 YMA and 10 AMA) were used for qPCR validation, and 24 NC samples (6 YMA, 6 IMA1, 6 IMA2 and 6AMA) were used for RNA-seq validation of AMA genes across the woman's reproductive lifespan. Immunohistochemistry (IHC) was used to confirm some expression changes at the protein level. Computational deconvolution using six endometrial cell type-specific transcriptomic profiles was conducted to compare the cellular composition between the groups. Comparisons between YMA and AMA samples identified a lower proportion of receptive endometria in the AMA group (P = 0.007). Gene expression profiling identified 491 differentially expressed age-sensitive genes (P adj < 0.05) that revealed the effects of age on endometrial epithelial growth and receptivity, likely contributing to decreased reproductive performance. Our results indicate that changes in the expression of the cellular senescence marker p16INK4a and genes associated with metabolism, inflammation, and hormone response are involved in endometrial aging. Importantly, we demonstrate that the proportion of multi-ciliated cells, as discovered based on RNA-seq data deconvolution and tissue IHC results, is affected by endometrial aging, and propose a putative onset of age-related changes. Furthermore, we propose that aging has an impact on the transcriptomic profile of endometrial tissue in the context of endometrial receptivity. The raw sequencing data reported in this article are deposited at the Gene Expression Omnibus under accession code GSE236128. This retrospective study identified changes in the endometrium of patients undergoing hormonal replacement and validated these changes using samples obtained during a NC. However, future studies must clarify the importance of these findings on the clinical outcomes of assisted reproduction. The findings reported in this study have important implications for devising future strategies aimed at improving fertility management in women of advanced reproductive age. This research was funded by the Estonian Research Council (grant no. PRG1076), Horizon 2020 innovation grant (ERIN, grant no. EU952516), Enterprise Estonia (grant no. EU48695), MSCA-RISE-2020 project TRENDO (grant no. 101008193), EU 874867 project HUTER, the Horizon Europe NESTOR grant (grant no. 101120075) of the European Commission, the EVA specialty program (grant no. KP111513) of the Maastricht University Medical Center (MUMC+), MICIU/AEI/10.13039/501100011033 and FEDER, EU projects Endo-Map (grant no. PID2021-12728OB-100), ROSY (grant no. CNS2022-135999), and the National Science Fund of Bulgaria (grant no. KII-06 H31/2). The authors declare no competing interests.

If CE affects infertile women’s outcomes after surgery?

ObjectiveTo investigate the prevalence of chronic endometritis (CE) in infertile patients and whether it affects spontaneous pregnancy after reproductive surgery in infertile patients. Materials and methodsIn this study, we collected clinical information on infertility patients who underwent reproductive surgery at the Reproductive Medicine Centre of the Second Hospital of Lanzhou University from 2021.1 to 2022.8. All patients underwent laparoscopic and hysteroscopic surgery. Tubal lubrication was performed concurrently with endometrial sample collection and pathological examination. The specimens were immunohistochemically stained with CD38 and CD138, and those who tested positive at the same time were diagnosed with chronic endometritis. As of 2023.9, the patients were followed up by telephone to determine whether chronic endometritis impacted postoperative pregnancy. OutcomeA total of 81 patients were finally included in the study. Of these, 25 were in the chronic endometritis group, and 56 were in the non-chronic endometritis group. There were no appreciable differences between the two groups’ demographic statistics. Furthermore, neither the bilateral appendages nor the uterus’s intraoperative conditions showed a statistically significant difference. Patients in the chronic endometritis group had a longer time to conception from the time of surgery (7 (6.00–11.75) vs. 10 (6.50–16.00), p = 0.467) and a lower rate of spontaneous pregnancies (8/25 = 32.00 % vs. 28/55 = 50.00 %, p = 0.132) than patients with non-chronic endometritis. Among the patients who had successful spontaneous pregnancies after surgery, approximately 77.14 % had live births and 22.86 % had miscarriages, and the live birth rate between the two groups was not significantly different. (21/28 = 75.00 % vs 7/8 = 87.50 %, p = 0.651) ConclusionChronic endometritis affects approximately 31.82% of infertile patients, and following reproductive surgery, it has no discernible impact on spontaneous pregnancy.

Reproductive and transcriptome characteristics of endometrium at the window of im-plantation in patients after fertility-sparing treatment of endometrial intraepithelial neoplasm or cancer

BackgroundProgestin therapy is an option for patients with endometrial carcinoma (EC) or endometrial intraepithelial neoplasm (EIN) who fit specific criteria of fertility-sparing treatment. However, the implantation rate remains low among females receiving in vitro fertilization (IVF) even after the complete reversal of endometrial lesions. MethodsHere, ten patients with EC/EIN achieved complete regression (CR) in histology. Their relevant metabolic and IVF parameters were collected. An endometrial sampling at the window of implantation (WOI) and transcriptome analysis were conducted among them, and four healthy controls were analyzed to analyze endometrial receptivity. ResultsOn average, it took ten patients five months to achieve CR after four curettage procedures. The interquartile range of endometrium thickness on trigger day was between 8.8 and 10.0 mm, while the range was 15.2–18.5 mm for controls. Five patients got pregnant after a frozen-embryo transfer. According to ERA analysis, the endometrial sampling at WOI showed pre-receptive status in four cases. In total, 1458 differential expression genes were identified, and 70 belonged to the ERA genes. ImmuneScore indicated decreased NK cells in the endometrium, affecting endometrial receptivity. ConclusionsEven after EC/EIN reversal in histology, endometrial receptivity has already been compromised regarding altered WOI and immune microenvironment, leading to a low pregnancy rate.

Performance of the Idylla microsatellite instability test in endometrial cancer

ContextDNA mismatch repair (MMR) deficiency (dMMR) testing is now recommended in endometrial cancer. Defect identification in the molecules participating in this pathway, or the presence of microsatellite instability, are commonly employed for this purpose. Novel methods are continuously evolving to report dMMR/microsatellite instability and to easily perform routine diagnoses. ObjectiveThe main aim of this study was to compare the concordance of the Idylla microsatellite instability test for the identification of dMMR endometrial cancer samples defined by immunohistochemistry and MMR genomic status. DesignWe applied the Idylla MSI test to 126 early-stage endometrial cancer cases with MMR testing by immunohistochemistry and genomic characterization (methylation in MLH1 and sequence alterations in MLH1, PMS2, MSH2 and MSH6). Individual markers and overall specific performance indicators were explored. ResultsThe Idylla platform achieved a higher global concordance rate with MMR genomic status than with immunohistochemistry (75 % and 66 %, respectively). Sensitivity and specificity are also higher (75 % vs 66 % and 96 % vs 90 %, respectively). Clustering analysis split the patients into 2 well-differentiated clusters, the pMMR and the dMMR group, represented by MLH1/PMS2 loss and the MLH1 methylated promoter. Overall, immunohistochemistry and MMR genomic status identified more dMMR cases than did the Idylla test, although correlations were improved with a modified Idylla test cut-off. ConclusionsPerformance of the Idylla test was better correlated with MMR genomic status than MMR immunohistochemistry status, which improved with a modified test cut-off. Further studies are needed to confirm the cut-off accuracy.

Single-cell transcriptomic analysis of immune cell dynamics in the healthy human endometrium

The microenvironment of the endometrial immune system is crucial to the success of placental implantation and healthy pregnancy. However, the functionalities of immune cells across various stages of the reproductive cycle have yet to be fully comprehended. To address this, we conducted advanced bioinformatic analysis on 230,049 high-quality single-cell transcriptomes from healthy endometrial samples obtained during the proliferative, secretory, early pregnancy, and late pregnancy stages. Our investigation has unveiled that proliferative natural killer (NK) cells, a potential source of endometrial NK cells, exhibit the most robust proliferative and differentiation potential during non-pregnant stages. We have also identified similar differentiation trajectories of NK cells originating from proliferative NK cells across four stages. Notably, during early pregnancy, NK cells demonstrate the highest oxidative phosphorylation metabolism activity, and, in conjunction with macrophages and T cells, exhibit the strongest type II interferon response. With spatial transcriptome data, we have discerned that the most robust immune-non-immune interactions are associated with the promotion and inhibition of cell proliferation, differentiation and migration during four stages. Furthermore, we have compiled lists of stage-specific risk genes implicated in reproductive diseases, which hold promise as potential disease biomarkers. Our study provides insights into the dynamics of the endometrial immune microenvironment during different reproductive cycle stages, thus serving as a reference for detecting pathological changes during pregnancy.

Efficacy of colchicine and melatonin in the treatment of rat endometriosis model: An animal study

ObjectiveThis study investigates the therapeutic effects of colchicine and melatonin on endometriotic implants in an experimentally created endometriosis model in rats. Study designForty-four adult female Wistar albino rats weighing between 260 and 300 g, 8 weeks old, were selected for the study. The unilateral uterine horn of rats with a bicornuate uterus was excised for 1 cm, washed with sterile saline, incised longitudinally, and the endometrium was exposed. A 0.5*0.5 cm endometrial tissue sample taken with a scalpel was implanted with suturing (4/0 Vicryl) to the abdominal wall. Forty-four rats were divided into four groups. Group 1 was randomized as the endometriosis group (control), Group 2 as endometriosis + colchicine treatment, Group 3 as endometriosis + melatonin treatment, and Group 4 as the endometriosis + melatonin + colchicine treatment group. The colchicine (Sigma Chemical Co., St Louis, Missouri) group was administered orally at a dose of 0.1 mg/kg, and the Melatonin group orally at a dose of melatonin (20 mg/kg per day). Treatment continued daily for 30 days. ResultsIn the post-treatment focal diameter measurements, the endometrial focal diameter in the colchicine and colchicine + melatonin group was significantly lower than the control group (p=0.026). Bcl-2 levels of the colchicine group were lower than the control group and the melatonin group (p=0.021). ConclusionColchicine and melatonin reduce adhesion to the peritoneal surface in ectopic endometrial cells. It also acts by increasing apoptosis and decreasing cell survival.

Utility of culture and molecular methods using AllplexTM Bacterial Vaginosis Plus Assay (SeegeneⓇ) as a tool for endometriosis, infertility and recurrent pregnancy loss diagnosis

Endometriosis, infertility, or recurrent pregnancy loss (RPL) are entities characterised by a decrease in Lactobacillus spp. and an increase in bacterial vaginosis-associated bacteria, (BVAV) according with 16S rRNA sequencing studies. However, the use of nucleic acid amplification tests (NAAT) as a tool for diagnosis algorithms is unknown. Seventy-four patients were included, with a median age of 36.5 years old (IQR: 34-39) including infertility (n=31), endometriosis (n=25), or RPL (n=18), for culturing and NAAT using the Allplex™ Bacterial Vaginosis Plus (ABVP) assay (SeegeneⓇ) with endometrial samples. The objective was determining the utility of ABVP assay for diagnosing the entities. Forty-six microorganisms were isolated from 31 out of 74 patients (41.9 %). Twenty-five endometrial samples (33.8 %) were positive for some targets included in the ABVP-assay, with median Ct value ∼37 (IQR: 31.3-37.1) and Qt value 1.43 Log10copies/reaction (IQR:1.1-2.6). For Lactobacillus species, sensitivity and specificity were 80 % and 84 %, respectively. Gardnerella vaginalis, 63.6 % and 95.7 %. No significant increase in BVAV was detected in any of the gynaecological entities. The ABVP and culture based algorithm did not show utility as a tool for endometriosis, infertility, or RPL diagnosis.

Analyzing 10-year time trends for Hysterectomy and Oophorectomy: Focus on Endometrial sampling and risk factors for Endometrial Cancer

Analyzing 10-year time trends for Hysterectomy and Oophorectomy: Focus on Endometrial sampling and risk factors for Endometrial Cancer

An altered uterine microbiota with endometrial hyperplasia

BackgroundEndometrial hyperplasia (EH) is a precursor to endometrial cancer, and the role of the microbiome in its development is unclear.ResultsThe present study investigated the uterine microbiome in patients with benign uterine conditions and endometrial hyperplasia. A significant structural shift in the uterine microbiome of patients with endometrial hyperplasia compared to those with benign conditions was found. Delftia, Serratia and Stenotrophomonas were significantly enriched in endometrial hyperplasia samples and associated with the presence of endometrial hyperplasia.ConclusionsThe novel finding suggested that increased abundance of Delftia, Serratia and Stenotrophomonas is associated with the presence of endometrial hyperplasia. Further investigation is needed to determine the value of these microbes as biomarkers for endometrial hyperplasia.

Revolutionizing chronic endometritis diagnosis: real-time polymerase chain reaction unveils microbial pathogens in Indian women with abnormal bleeding and reproductive challenges

BackgroundThis study aimed to assess the utility of real-time-polymerase chain reaction (PCR) for diagnosing chronic endometritis (CE) by targeting 11 prevalent pathogens and to compare the outcomes with conventional culture-based diagnosis. Study DesignA retrospective analysis was conducted on 500 patients with clinical conditions such as abnormal bleeding, in vitro fertilization failure, recurrent implantation failure, recurrent miscarriage, and recurrent pregnancy loss. The prevalence of 11 key pathogens associated with CE was evaluated in endometrial biopsy samples. ResultsIn our study, PCR identified 318 cases (63.6%) positive for at least one of the 11 investigated pathogens, while culture-based methods detected 115 cases (23%). Predominant pathogens detected by PCR included Enterococcus faecalis (E. faecalis) (19%), Escherichia coli (E. coli) (6.8%), Staphylococcus aureus (S. aureus) (9%), Mycoplasma hominis (5%), Mycoplasma genitalium (6.2%), Streptococcus agalactiae (S. agalactiae) (4.2%), Ureaplasma urealyticum (4%), nontuberculous Mycobacterium (5.2%), Mycobacterium tuberculosis (1.2%), Neisseria gonorrhoeae (0.6%), and Chlamydia trachomatis (2.4%). Standard culture methods identified E. faecalis (10.8%), S. aureus (6.2%), E. coli (3.8%), and S. agalactiae (2.2%). ConclusionThe DICE panel proves itself as a swift, precise, and cost-effective diagnostic tool for detecting both culturable and nonculturable endometrial pathogens in CE. Demonstrating superiority, the Molecular method outshines microbial culture, ensuring accurate and sensitive detection of CE-associated pathogens, harmonizing seamlessly with histology and hysteroscopy findings.