Endometrial Layer Research Articles

Three-dimensional visualization of uterine nerve fiber distribution using fluorescence micro-optical sectioning tomography (fMOST): A pilot study.

This study aimed to observe and quantitatively analyze the morphology and distribution of uterine nerve fibers using fluorescence micro-optical sectioning tomography (fMOST). The goal was to provide an accurate morphological reference for pathological evaluations of uterine nerves. Using fMOST technique, we observed and analyzed the distribution of nerve fibers within the uterus. Our findings revealed a radial dispersion of nerve fibers radiating from myometrium to endometrium. The cervix uteri region exhibited a high density of nerve fibers, displaying terminations in a flower spray pattern. In contrast, nerve fibers in corpus uteri were comparatively sparse. However, we identified a unique "vine-like" pattern of a single nerve fiber extending from myometrium to endometrial layer in areas with concentrated nerves. The fMOST technique is able to effectively elucidate the morphology and distribution of uterine nerve fibers. This method enables three-dimensional visualization of nerves in myometrium and offers a novel approach to observe the pathological changes in uterine nerves.

Evaluation of morphology, apoptosis, and cell proliferation of the uterus in postmenopausal women.

The aim of this study was to evaluate the morphology (atrophy and fibrosis), apopto-sis, and cell proliferation in the uterine wall. The research material came from postmenopausal women who had undergone hysterectomy due to uterine myomas or prolapse of the reproductive organ and were not taking menopausal hormone therapy (MTH). The collected material was divided into three groups. Group I (n=18) con-sisted of uterine sections taken 1 to 5 years after the last menstruation, Group II (n=17) 6 to 10 years after the last menstruation, and Group III (n=15) over 11 years after the last menstruation. To assess morphology and fibrosis, the uterine sections were subjected to hematoxylin and eosin (HE) staining and to Mallory's staining. In addition, we performed a histochemical examination to identify apopto-sis in endometrial and myometrial cells using the TUNEL method. An immunohistochemical analysis of endometrial and myometrial cells was also performed to detect the location of the proliferating cell nuclear antigen (PCNA). Differences in apoptosis were only found in the myometrium between Group I and Group III, and were strongest in Group I myometrial cells, and weakest in Group III. Neither the endome-trium nor the myometrium showed statistically significant differences in the overall percentage of PCNA(+) cells between groups. Morphological changes in the endometrial and myometrial layers of postmenopausal uteri increased with time since the last menstruation.

P-312 Transcriptional profiling of SSEA-1+ endometrial epithelial cells highlights their role in endometrial regeneration, remodelling and homeostasis

Abstract Study question Will the transcriptional profile of SSEA-1+ endometrial epithelial cells (EECs) explain their functional role, which can be explored in an in vitro endometrial organoid model? Summary answer SSEA-1+ EECs demonstrate differentially expressed genes (DEGs) and associated functional pathways expected from a basal stem/progenitor cell (SPC) population, agreeing with the in vitro experiments. What is known already Endometrial SPCs are postulated to reside in the basalis and to be responsible for the full phenotypic and functional restoration of the endometrial functionalis layer. SSEA-1+ EECs assume the postulated basalis SPC niche. Previous studies demonstrated isolated SSEA-1+ EECs to have a higher capacity to generate organoids in 3D matrix and display growth like that observed after endometrial denudation. They have lower steroid hormone expression and higher telomerase activity with longer telomere lengths, all suggesting a SPC phenotype. SSEA-1+ EECs co-express nuclear SRY-box transcription factor 9 (nSOX9) and nuclear b-catenin, suggesting an activated Wnt pathway. Study design, size, duration Endometrial samples were collected from eight pre-menopausal women attending the Liverpool Women’s Hospital for benign gynaecological procedures, aged 30-47 years. These patients were having regular menstrual cycles and had not been on any hormonal therapy for at least three months prior to enrolment. Three further endometrial samples were obtained for 3D EEC organoid culture studies. Participants/materials, setting, methods Magnetic-activated cell sorting was used to isolate SSEA-1 enriched/depleted EECs from freshly harvested endometrial samples. Microarray analysis was performed using Agilent human arrays. DEGs and pathway enrichment analysis were explored using R-studio and Ingenuity Pathway Analysis (IPA). Internal and external validation used RT-qPCR, dual-immunofluorescence, and comparison against publicly available endometrial single cell spatial transcriptomic data. Human EEC organoids models were employed to assess hormonal regulation of SSEA-1 expression using immunohistochemistry. Main results and the role of chance SSEA-1+ EECs have a distinct transcriptional profile, with 1,059 DEGs compared with SSEA-1- EECs. Pathway analysis highlighted their role in endometrial regeneration, adhesion, remodelling, and neovascularisation, enriching for pathways such as ‘extracellular matrix structural constituent’, ‘epithelial to mesenchymal transition’, ‘angiogenesis’, ‘TNFα signalling via NFKB’ and ‘Notch signalling’. IPA’s biological functions demonstrated their involvement in tissue homeostasis, tumour suppression and their more quiescent SPC phenotype, with activation of ‘organismal death’ and inhibition of ‘cell movement/migration’ and ‘cell movement/migration of tumour cell lines’. The activation of canonical pathways such as ‘PTEN signalling’ and ‘endocannabinoid cancer inhibition’ demonstrated their fine equilibrium to drive proliferation during regeneration, whilst providing protection from hyperplastic/carcinogenic transformation. Ten selected DEGs were internally validated using RT-qPCR, demonstrating concordance. Dual-immunofluorescence of gene products of DEGs MMP7, MMP26, C11orf52 and CD47 confirmed co-localisation alongside SSEA-1 in an external biological cohort of endometrial samples. External in silico validation using the Garcia-Alonso et al endometrial single-cell transcriptomics dataset, mapped significantly upregulated DEGs MMP7 and MMP26 to mainly SOX9+ and SOX9+/LGR5+ EECs. EEC organoids exposed to hormones (including oestradiol) demonstrated an induction of proliferation and higher proportion of organoids expressing SSEA-1 (46.8% vs 67%) simulating endometrial glandular regeneration process. Limitations, reasons for caution Freshly isolated/sorted EECs were obtained from women at different menstrual cycle phases. Our organoid model system only contained EECs without the stromal component, thus, was not suitable to assess progestogenic response of EECs that occur in vivo. Wider implications of the findings This study provides novel information on human SSEA-1+ EECs, suggesting their pivotal role in endometrial regeneration. Since endometrial epithelial SPCs make a vital contribution to endometrial repair and pregnancy establishment, characterising of these cells in health will allow the development of targeted novel therapies on aberrant SPCs within gynaecological disease. Trial registration number Not applicable

P-653 Impact of dydrogesterone use in cycles with low progesterone levels on the day of frozen embryo transfer

Abstract Study question Assessing if Frozen Embryo Transfer (FET) outcomes in hormonal replacement treatment with dydrogesterone (DYD) for low progesterone levels match those with normal progesterone (P4) values Summary answer The live birth rate showed no significant difference between patients with low and normal serum P4 levels in FET, utilizing DYD as rescue therapy What is known already The number of IVF/ICSI cycles and frozen embryo transfers (FET) is increasing. The preparation for FET can occur in natural or artificial cycles, showing no significant differences in clinical pregnancy and live birth rates. Artificial cycles involve exogenous estradiol supplementation, starting after menses, and progesterone (P4) addition post-endometrial proliferation. Numerous studies consistently indicate that diminished P4 levels are correlated with inferior outcomes in hormonal replacement treatment, showing a potential decrease of up to 30% in ongoing pregnancy rates. Despite a suggested rescue strategy, evidence supporting its efficacy using an oral route is insufficient. Study design, size, duration This was a retrospective cohort study, of single embryo transfers (SET) from FET cycles conducted between July 2019 and March 2022, after artificial endometrial preparation with valerate-estradiol and micronized vaginal P4. In cases of low serum P4 (up to 12,5ng/dl) on the planned transfer day, 10 mg of dydrogesterone (DYD) three times daily was added. Solely blastocyst single embryo transfers (SET) were included. The primary endpoint was the live birth rate> 24 weeks. Participants/materials, setting, methods 535 FET-SET cycles were analyzed. Patients with an endometrial layer thinner than 7 mm, endocavitary pathology or serum P4 values< 4ng/dl were excluded. The explanatory variables evaluated were: relevant female characteristics, previous IVF/ICSI cycle and relevant FET cycle parameters (endometrial thickness, serum P4 value on the FET day, days in culture before blastocyst freezing) and use of DYD as rescue therapy. Statistical analysis was performed using IBM®SPSS® statistics v22.0 and R4.2.2 for Windows. Main results and the role of chance Out of the 535 FET cycles analyzed, 136(25.4%) underwent treatment with DYD. There were 337 pregnancies (63%), 207 live births (38.6%) and 130 miscarriages (38.5%). Serum P4 varied from 4.2-32.3 ng/dl, with an average of 14.93 ng/dl (SD 4.52). The best model to explain serum P4 distribution considered a gamma distribution for P4. Female age on the FET day, female ethnicity and female weight seemed to explain P4 variation. A sensitivity analysis on the P4 values was performed. The data were divided into 5 groups according to P4 levels and the rate of live birth per quintile was evaluated. There were no statistically significant differences between the groups. Patients with low P4 which had DYD added had no statistical differences regarding deliveries, 40% vs 36% (p = 0,354). The multivariable model revealed that older females at pick up, a thinner endometrial measure and transferring a day 6 blastocyst were negativey associated with live birth (p < 0,05), but not P4 levels or medication with DYD. A sub-group analyses was performed according to the median value of P4 (9.6 ng/ml) of those to whom DYD was added. No statistically significant differences were identified for biochemical pregnancy 59% vs 60% (p = 1) and live birth rate 34%vs38% (p = 0,653). Limitations, reasons for caution This retrospective study lacks an untreated control group, causing inherent heterogeneity. It’s unclear if there’s an upper limit for P4 levels to restrict DYD supplementation or a minimum level rendering this strategy unsuitable. Monitoring DYD’s fetal/neonatal safety is imperative. Wider implications of the findings The supplementation with oral DYD in hormonal replacement FET cycles is an easy and cheap strategy in patients with for low P4 levels. The duration of DYD administration should be further evaluated. Trial registration number not applicable

Cytoarchitectural differences in reproductive organs of some polycystic ovary-like induced animal models

Polycystic ovary syndrome (PCOS) is the most common gynaecological, endocrine disorder that occurs during reproductive age and is a significant cause of anovulatory infertility. Letrozole is an aromatase inhibitor which negates the action of the aromatase enzyme, which results in the buildup of male hormones (testosterone) in the females, causing hyperandrogenism, which is a hallmark of Polycystic Ovarian Syndrome. Mifepristone (RU486) is a progestin antagonist that acts to arrest the actions of the progesterone hormone, resulting in follicular atresia and anovulation. DHEA is an androgen which was also administered in a bid to cause hyperandrogenism in the rats.This study aimed to evaluate the effects of these hormones on the cytoarchitecture of the ovaries and uterus to assess their various PCOS-like histological features.Animals were grouped mainly into three: Letrozole, Mifepristone and DHEA groups, which were further divided into two subgroups each, administered low and high doses of letrozole orally, Mifepristone and Dehydroepiandosterone (DHEA) subcutaneously. Each of the subgroups also had a comparison control group. Following the completion of administration, the Wistar rats were euthanized, and their ovaries and uterus were collected for histological analysis.Increased proliferation of ovarian follicles was noted in the treated groups compared to control, as well as thickening of the endometrial layer.

The changes in ultrasound parameters during treatment of chronic endometritis depending on the morphological type in women of reproductive age

Objective: to evaluate the dynamics of echographic parameters of structural changes in the endometrium depending on the morphological type of chronic endometritis (CE) before and after etiotropic therapy.Material and methods. A retrospective cohort based on the results of examination of 140 patients of reproductive age with a verified diagnosis of CE before and after complex etiotropic therapy. A pelvic ultrasound was performed in the 1st phase of the menstrual cycle with measurements of the endometrial and uterine volume, followed by a calculation of the percentage endometrial/uterine volume ratio (EV/UCV). The groups of hypoplastic, mixed, and hyperplastic morphotypes were formed according to the values of EV/UCV. The B-mode examination included the assessment of the structure and echogenicity of the uterine cavity mucosa; the line of closure or divergence of the endometrial layers; the external contour of M-echo; the presence of liquid in the uterine cavity, and acoustic artifacts, in particular reverberation, as a sign of gas bubbles.Results. An increase in the thickness and volume of the endometrium, and EV/UCV was obtained after therapy in the hypoplastic morphotype. There were no changes in the thickness or volume of the endometrium in the mixed morphotype. A significant decrease in the thickness and volume of the endometrium, and EV/UCV were found in the hyperplastic morphotype. In all morphotypes, the endometrium remained mostly hyperechoic and heterogeneous, with an abnormal midline.Conclusion. The hypoplastic and mixed morphotypes are characterized by more persistent endometrial changes that are difficult to treat. Pathological signs were eliminated much more often in the hyperplastic morphotype. The results of the study demonstrate the absence of a complete reduction in the analyzed ultrasound criteria of CE after end of treatment for all morphological types.

The regulatory effect of electroacupuncture on endometrial M1-type macrophages in rats with intrauterine adhesions.

To observe the effect of electroacupuncture(EA) on endometrial fibrosis and M1-type macrophages in rats with intrauterine adhesions(IUA), so as to explore the possible mechanism of EA in the treatment of IUA. Fifteen female SD rats were randomly divided into blank group, model group and EA group, with 5 rats in each group. The IUA rat model was established by double damage method using mechanical scraping combined with lipopolysaccharide infection. Rats in the EA group were treated with acupuncture at "Guanyuan"(CV4), and EA at bilateral "Zusanli"(ST36) and "Sanyinjiao"(SP6)for 20 minutes each time, once a day, for 3 consecutive cycles of estrus. Five rats in each group were sampled during the estrous period, and the endometrial morphology, endometrial thickness and the number of blood vessels and glands were observed after HE staining. The fibrotic area of the uterus was observed after Masson staining. The positive expressions of Runt-related transcription factor(RUNX1), transforming growth factor-β1(TGF-β1), connective tissue growth factor(CTGF), α-smooth muscle actin(α-SMA), collagen type I(Col-Ⅰ), cluster of differentiation 86(CD86), interleukin-1β(IL-1β), and tumor necrosis factor-α(TNF-α) in endometrial tissue were detected by immunohistochemistry. Western blot was used to detect relative protein expressions of RUNX1, TGF-β1, α-SMA, CD86, and TNF receptor 2 (TNFR2), and real-time fluorescence quantitative PCR was used to detect mRNA expressions of RUNX1, TGF-β1, α-SMA, CD86, and TNF-α in the endometrium. During the estrous phase, the endometrial layer in the model group was damaged, with reduced folds, disordered arrangement of epithelial cells, loose fibrous connective tissue, significant narrowing and adhesions in the uterine cavity, interstitial congestion, edema, and a significant infiltration of inflammatory cells with sparse glands. While uterine tissue structure of the EA group was basically intact, resembling a normal uterus, with more newly formed glands and a small amount of inflammatory cell infiltration. In comparison with the blank group, the endometrial thickness, the number of blood vessels, and the number of glands were significantly decreased(P<0.001) in the model group, while the ratio of uterine fibrosis area, the positive expressions of RUNX1, TGF-β1, CTGF, α-SMA, Col-Ⅰ, CD86, IL-1β, and TNF-α, the protein relative expressions of RUNX1, TGF-β1, α-SMA, CD86 and TNFR2, and the mRNA relative expression levels of RUNX1, TGF-β1, α-SMA, CD86 and TNF-α in the endometrium were significantly increased (P<0.001, P<0.01). Compared to the model group, the endometrial thickness, the number of blood vessels, and the number of glands were significantly increased(P<0.01, P<0.05) in the EA group, while the ratio of uterine fibrosis area, the positive expressions of RUNX1, TGF-β1, CTGF, α-SMA, Col-Ⅰ, CD86, IL-1β and TNF-α in the endometrial tissue, the protein expressions of RUNX1, TGF-β1, α-SMA, CD86 and TNFR2, and the mRNA relative expressions of RUNX1, TGF-β1, α-SMA, CD86 and TNF-α in the endometrium were significantly decreased (P<0.001, P<0.01, P<0.05). EA can improve endometrial fibrosis in IUA rats, which may be related to its function in decreasing the level of endometrial M1-type macrophages and the secretion of related inflammatory factors.

Association of Uterine Tissue Innervation and Peripheral Nerve Density with Adenomyosis Related Pain. A Systematic Review.

Adenomyosis is associated with dysmenorrhea and chronic pelvic pain; however, the triggering mechanisms of painful stimuli and the role of uterine nerve fibers in the manifestation of pain remain poorly understood. The objective of this study was to systematically review the role of uterine nerve fibers' presence and density in the occurrence of pain in patients with adenomyosis. An electronic search was performed using the Embase, PubMed/Medline, and Cochrane databases. We included all studies from inception to November 2023. A total of ten studies that compared uterine biopsies samples of women with and without adenomyosis were included. The biomarker antiprotein gene product 9.5 was decreased or absent in the endometrium of most included women with adenomyosis. None of the included studies observed a difference in neurofilament (NF) staining between the adenomyosis and non-adenomyosis groups. Studies that assessed nerve growth factor (NGF) staining were heterogeneous in design. One study reported no difference in immunohistochemistry staining in any endometrial layer between the adenomyosis and non-adenomyosis groups, while another reported increased staining in the adenomyosis functional endometrial layer, and a third study reported overexpression of NGF, synaptophysin (SYN), and microtubule-associated protein 2 mRNA in focal adenomyosis alone. Preliminary data from poor-quality studies suggest an increase in the uterine density of nerve fibers in patients with adenomyosis. Well-designed studies are essential to assess the cause-and-effect relationship between uterine nerve fibers and pain in patients with adenomyosis.

Effectiveness Of Clittoria Ternatea And Zingibers Officinale Concoction On The Reduction Of Dysmenorrhea In Adolescent Girls

Dysmenorrhea or menstrual pain is abdominal pain originating from uterine abdominal cramps, occurring during menstruation due to sloughing of the endometrial layer. The prevalence of dysmenorrhea varies widely in various countries, ranging from 16-91%; the incidence in Indonesia ranges from 45-95%, while in the Special Region of Yogyakarta, as much as 52%, and in Sleman district, as much as 88.64%. The purpose of the study was to determine the effectiveness of steeping butterfly pea (Clitoria Ternatea) and ginger concoction(Zingibers Officinale) on reducing menstrual pain (dysmenorrhea) in adolescent girls. The research method used was Quasi Experiment Two Group Pretest-Posttest design. The subjects in this study were 90 respondents who experienced menstrual pain and met the criteria. Data analysis using the Wilcoxon test and Mann-Whitney test. The results of this study showed that the average menstrual pain before being given butterfly pea brew was 5.97. After being given Butterfly Pea Brew, the average menstrual pain was 0.51 with a P-Value 0.000 less than 0.05, meaning that there was a difference in menstrual pain felt before and after being given Butterfly Pea Brew. The mean value of menstrual pain before being given a ginger concoction is 6.02. After being given a ginger concoction, it decreased to 0.44 with a P-Value of 0.000 less than 0.05, meaning that there is a difference in menstrual pain felt before and after a ginger concoction. This study concludes that butterfly pea brew and ginger concoction effectively reduce menstrual pain (dysmenorrhea).

Advances in Clinical Diagnosis and Treatment of Intrauterine Adhesions

Intrauterine adhesion is a kind of high incidence disease caused by many pathogenic factors, which damages the endometrial basal layer and affects the menstruation and fertility of women. In recent years, due to artificial abortion and other types of increased uterine cavity surgery, the incidence of the disease gradually increased. Hysteroscopy is the main method to diagnose intrauterine adhesion. Combining with ultrasound, it can determine the range and degree of intrauterine adhesion and guide the choice of treatment. But it cannot find small changes in the uterine cavity, and for serious adhesions and endometrial injury in patients, hysteroscopy examination results are not good. Accurate assessment of recurrence risk and individualized diagnosis and treatment are important prerequisites to reduce the high recurrence rate and improve the low pregnancy rate. This article reviews the research on clinical diagnosis and treatment of intrauterine adhesions in order to enhance and improve the understanding of the disease and provide a new clinical thinking.

Human umbilical cord-derived mesenchymal stem cells and auto-crosslinked hyaluronic acid gel complex for treatment of intrauterine adhesion.

The purpose of this study was to explore the therapeutic characteristics of mesenchymal stem cells generated from human umbilical cord (hUC-MSCs) when utilized in conjunction with auto-crosslinked hyaluronic acid gel (HA-gel) for the management of intrauterine adhesion (IUA). The goal was to see how this novel therapy could enhance healing and improve outcomes for IUA patients. In this study, models of intrauterine adhesion (IUA) were established in Sprague-Dawley (SD) rats, which were then organized and divided into hUC-MSCs groups. The groups involved: hUC-MSCs/HA-gel group, control group, and HA-gel group. Following treatment, the researchers examined the uterine cavities and performed detailed analyses of the endometrial tissues to determine the effectiveness of the interventions. The results indicated that in comparison with to the control group, both HA-gel, hUC-MSCs, and hUC-MSCs/HA-gel groups showed partial repair of IUA. However, in a more notable fashion transplantation of hUC-MSCs/HA-gel complex demonstrated significant dual repair effects. Significant outcomes were observed in the group treated with hUC-MSCs and HA-gel, they showed thicker endometrial layers, less fibrotic tissue, and a higher number of endometrial glands. This treatment strategy also resulted in a significant improvement in fertility restoration, indicating a profound therapeutic effect. The findings of this study suggest that both HA-gel, hUC-MSCs, and hUC-MSCs/HA-gel complexes have the potential for partial repair of IUA and fertility restoration caused by endometrium mechanical injury. Nonetheless, the transplantation of the hUC-MSCs/HA-gel complex displayed exceptional dual healing effects, combining effective anti-adhesive properties with endometrial regeneration stimuli.

The impact of treatment on endometrium ultrasound findings in reproductive age women with chronic endometritis

Purpose: to evaluate the dynamics of ultrasound parameter of structural changes of endometrium in B-mode before and after therapy in reproductive age women with chronic endometritis.Material and methods. A retrospective cohort study of 158 reproductive age patients (34.5 ± 6.5 years) with a established diagnosis of chronic endometritis was carried out. Ultrasound was performed twice prior and after comprehensive etiotropic therapy in the proliferative phase of the menstrual cycle with measurements of uterus volume, endometrium thickness and volume, with calculation of endometrial/uterine volume percentage ratio. Echogenicity and structure, including the presence of polyps, liquid in the uterine cavity, gas in the endometrial layer or on midline, the line of endometrial layers closure and the contour of endometrial midline were assessed.Results. There was no complete regress of any of pathological signs after treatment, but the frequency of most of them decreased significantly (p &lt; 0.05). At the same time, there was no significant changes in such signs as heterogeneous structure and increased echogenicity of the endometrium (p &gt; 0.05). In addition, a statistically significant decrease of endometrium thickness and volume, as well as endometrial/uterine volume ratio was obtained with a constant uterus volume (p &lt; 0.05).Conclusion. The dynamic decrease in the frequency of revealing of the ultrasound signs in chronic endometritis on the background of comprehensive etiotropic therapy allows to assess a positive therapeutic benefits.

Effects of endometrial versus non-endometrial suturing on isthmocele development; a randomized controlled trial.

ObjectiveIncomplete healing after cesarean section (CS) can result in isthmocele formation. When suturing the uterus, fully folding the wound lips may embed the endometrial layer into the myometrium, leading to isthmocele development. Hence, this study aimed to compare the effects of endometrial and non-endometrial suturing on isthmocele development. Material and methodsThis randomized controlled trial included 274 patients. Women who underwent primary CS were randomly allocated to one of the two study groups: endometrial suturing and non-endometrial suturing. The primary outcome was isthmocele rate at postpartum 6 months. Secondary outcomes were the volume of the isthmocele, thickness of the residual myometrium, menstrual irregularities (intermenstrual spotting), and the relationship between the isthmocele and uterine position. ResultsA total of 159 patients (81 in the endometrial suturing group and 78 in the non-endometrial suturing group) were analyzed. The incidence of isthmocele was significantly lower in the non-endometrial suturing group than in the endometrial suturing group (12 [15.4%] vs. 24 [29.6%] patients; p = 0.032). Menstrual irregularities, such as intermenstrual spotting, were significantly higher in the endometrial suturing group than in the non-endometrial group (p = 0.019). ConclusionUterine closure with non-endometrial suturing was associated with significantly lower isthmocele development and less intermenstrual spotting compared to that with endometrial suturing.

Absence of Heme Oxygenase-1 Affects Trophoblastic Spheroid Implantation and Provokes Dysregulation of Stress and Angiogenesis Gene Expression in the Uterus.

The enzyme heme oxygenase-1 (HO-1) is pivotal in reproductive processes, particularly in placental and vascular development. This study investigated the role of HO-1 and its byproduct, carbon monoxide (CO), in trophoblastic spheroid implantation. In order to deepen our understanding of the role of HO-1 during implantation, we conducted in vivo experiments on virgin and pregnant mice, aiming to unravel the cellular and molecular mechanisms. Using siRNA, HO-1 was knocked down in JEG-3 and BeWo cells and trophoblastic spheroids were generated with or without CO treatment. Adhesion assays were performed after transferring the spheroids to RL-95 endometrial epithelial cell layers. Additionally, angiogenesis, stress, and toxicity RT2-Profiler™ PCR SuperArray and PCR analyses were performed in uterine murine samples. HO-1 knockdown by siRNA impeded implantation in the 3D culture model, but this effect could be reversed by CO. Uteruses from virgin Hmox1-/- females exhibited altered expression of angiogenesis and stress markers. Furthermore, there was a distinct expression pattern of cytokines and chemokines in uteruses from gestation day 14 in Hmox1-/- females compared to Hmox1+/+ females. This study strongly supports the essential role of HO-1 during implantation. Moreover, CO appears to have the potential to compensate for the lack of HO-1 during the spheroid attachment process. The absence of HO-1 results in dysregulation of angiogenesis and stress-related genes in the uterus, possibly contributing to implantation failure.

International Endometrial Tumor Analysis (IETA) descriptors in the diagnosis of chronic endometritis

Objective: to compare the terms, definitions and measurement methods developed by the IETA group with the ultrasound criteria of chronic endometritis (CE) used in Russia.Material and methods. A retrospective cohort study of 158 reproductive age women with clinical and laboratory diagnosis of CE was carried out. Sonographic examination was performed in the early or middle proliferative phase (cycle day 4–10 days) with the use of Affiniti70 ultrasound system (Philips, the Netherlands) with a multifrequency 3D endocavitary probe. Uterine corpus volume, endometrial thickness and volume were measured, followed by percentage endometrial/uterine volume ratio calculation, the so-called adjusted endometrial volume. Qualitative analysis of grayscale imaging included assessment of endometrial structure and echogenicity; closure or separation of the endometrial layers; contour of endometrial midline; the presence of acoustic artifacts, such as reverberation in the presence of gas or liquid in the uterine cavity, described by a number of authors. Relevant IETA descriptions were searched when assessing all qualitative CE features. In parallel, a qualitative score analysis proposed by the IETA group was carried out.Timely preoperative diagnosis of endometrioic cyst (endometrioma), as well as deep endometriosis remains relevant. The aim of the study was to assess the diagnostic value of ultrasound in patients with endometriomas and assess the combination of them with other foci of external genital endometriosis. The study based on retrospective analysis of a date of 95 patients with ultrasound signs of ovarian endometriomas, who underwent examination in MedicoProfi LLC – Borisov Medical and Diagnostic Clinic (Krasnoyarsk) during the period from January 2019 to October 2023. All of patients underwent surgery , followed by morphological evaluation. In the vast majority of cases, it was possible to detect a combination of endometriomas with one or more foci of deep endometriosis. Superficial peritoneal endometriosis and adhesions were found on surgery in all cases when endometriomas appeared isolated on ultrasound. The results of the study showed: endometriomas combined with deep endometriosis in 96.8% of cases. Thus, ultrasound detection of endometrioma is a very reliable sign of deep endometriosis presence. The “kissing ovaries” symptom in bilateral endometriomas can be considered as an absolutely reliable sign of the uterosacral ligaments endometriosis with specificity of 100% and positive predictive value of 100%. The presence of the “kissing ovaries” sign should be depicted in the conclusion of the ultrasound protocol, since it highly suggestive to obliteration of the pouch of Douglas and involvement of adjacent organs (fallopian tubes, intestines, ureters, etc.) in the endometrioid infiltrates, which is extremely important for the surgery planning, as well as in patients with infertility. There is an obvious need to introduce the extended pelvic ultrasound protocol to the diagnostic algorithm for patients with suspected endometriosis, which will more accurately describe the disease extension.Results. The comparative analysis of endometrium description in CE indicates a similar measurement technique for endometrial and intrauterine lesions thickness, both proposed by the IETA group and used in our country. Most of the IETA descriptors for qualitative ultrasound findings may be used in CE diagnosis. However, there are no some significant ultrasound features for identifying the inflammation, such as marked and partially or completely thickened midline, as well as gas focuses within the endometrium or in the uterine cavity, in IETA description.Conclusion. Terminology standardization allows compare the results and perform multicenter studies followed by meta-analysis for the diagnosis of chronic endometritis, if researchers use the similar descriptors.

EFFECTIVENESS OF COMPLEX THERAPY OF HYPERPROLACTINEMIA IN WOMEN OF THE SUMY REGION, ASSOCIATED WITH STRESS AGAINST THE BACKGROUND OF ARMED AGGRESSION

Introduction. Ukrainians have been living under martial law for the last two years. As a result of a constant stress, physical and especially mental health is affected. Сhronic stress can cause functional hyperprolactinemia, which is characterized by an increase of the level of prolactin in the blood. Elevated serum prolactin is a level that is &gt; 25 ng/ml or 530 mIU/L, which affects the functioning of various levels of the reproductive system. The purpose of the study: to study the effectiveness of complex therapy of hyperprolactinemia in women of the Sumy region, which is associated with chronic stress against the background of armed aggression. Materials and methods. The study was conducted at the Municipal Non-Profit Enterprise "Clinical Maternity Hospital of the Blessed Virgin Mary" and the Municipal Non-Profit Enterprise "Central City Clinical Hospital". A comprehensive clinical and laboratory examination was carried out of 81 women of the Sumy region who had reproductive plans, aged from 22 to 39 years old, who were divided into the following clinical groups: I (main) group, which included 31 women – was prescribed anti-stress therapy, drugs with Vitex agnus castus and cognitive behavioral therapy with a psychotherapist. II (comparison) group – 28 women who received anti-stress therapy and drugs with Vitex agnus castus and III (control) group – healthy women (n = 22). Exclusion criteria were the age of the patients less than 22 and more than 39 years, pituitary tumor confirmed (using magnetic resonance imaging) at the time of the study, taking medications that could affect the results of the studied hormones. Research results. The analysis of the obtained prolactin level data indicated that in the majority of women from the study groups, the prolactin level was elevated within the range of 24–35 ng/ml. Before treatment, the average concentration of prolactin in patients of group I was 36.74±2.72 ng/ml (р&lt;0,05), after 3 months, a decrease in concentration was noted to 20.54±1.74 ng/ml (p&lt;0,05). In the comparison group, the average concentration of prolactin before treatment was 31.64±2.48 ng/ml (р&lt;0,05), while after 3 months, it equaled 27.04±2.02 ng/ml (p&lt;0,05). Also, in patients of the I group, the level of estradiol, progesterone and a thickness of the endometrium were increased. Conclusion. The scheme of a differentiated approach to the treatment of Sumy region women with hyperprolactinemia against the background of stress, which we have outlined, contributes to the normalization of prolactin, estrogen, progesterone levels, and the growth of the functional endometrial layer.

Role of Bacterial and Viral Infections and Co-Infections in Miscarriages

Abstract Aim To investigate the potential role of the following bacterial/viral panel (Chlamydia trachomatis, Ureaplasma urealyticum/parvum, Mycomplasma hominis/genitalium, Gardnerella vaginalis, HSV1/2, EBV, CMV, VZV, HHV6, HHV7, HHV8) as causative factors for miscarriages in women by testing endometrial biopsies. Anaerobic and aerobic microorganisms causing dysbiosis and endometrial bacterial colonization by unbalanced growth were additionally tested. Materials and methods In total, 65 patients with a history of early and late miscarriages were analyzed. DNA extractions, real-time qPCR, agarose gel-electrophoresis were applied. Comparative analysis of the current with previously obtained data on the described panel in menstrual tissue samples was performed. Results In 64,6% of all tested endometrial biopsies bacterial and/or viral pathogens were detected. In 49,23% of all tested samples we found bacterial, while in 15,3% – viral pathogens. These results are similar to our previous data on menstrual tissue samples of infertile women – 61,1% infected, as 48,8% had bacterial and 22,2% had viral pathogens. Gardnerella vaginalis and Ureaplasma parvum were detected in 31,25% and 3,12% of all bacterial infected endometrial biopsies, significantly lower in comparison to the estimated rate of 69,31% and 61,36% on menstrual tissue. Anaerobic and aerobic dysbiosis were detected in 53,33% and 27% of the bacterial infected endometrial samples. In 13,33% a dysbiosis with a mixed etiology was found, while in 7% a dysbiotic condition with a totally absent findings of targeted bacteria and Lactobacillus was observed. EBV, CMV, HHV6 and HHV7 were detected in 30%, 30%, 20% and 20% of the positive for viral factors endometrial biopsies and in 40%, 7,5%, 10% and 42,5% in menstrual tissue samples. In the current study 62,5% bacterial co-infection and 12,5% bacterial/viral co-infection variants were found. Infections with the rest of the target pathogens were not detected in the endometrial biopsies. In contrast to the endometrial biopsy results, Mycomplasma hominis, Ureaplasma urealyticum and HSV2 were detected in our previous research on menstrual tissue samples. Conclusions Our research suggests a possible dysbiosis as a consequence of bacterial/viral endometrial colonization, associated with miscarriages. We prove that menstrual tissue, containing parts of the functional endometrial layer, is a reliable and accurate noninvasive sample for infectious screening of the upper genital tract.

Antifouling and Injectable Granular Hydrogel for the Prevention of Postoperative Intrauterine Adhesion.

Postoperative intrauterine adhesion (IUA), caused by endometrial basal layer injury, is one of the main causes of female infertility. The excessive deposition of fibrin as well as fibroblast is considered the root cause of IUA. However, few clinical strategies are effective in preventing extracellular matrix (ECM) deposition at endometrial wounds that include protein and cell deposits. Herein, the injectable granular poly(N-(2-hydroxyethyl) acrylamide) (PHEAA) hydrogel (granular PHEAA gel), which presents excellent antifouling properties and remarkably prevents protein and cell adhesions, is used to prevent postoperative IUA. The granular PHEAA gel with a jammed network structure exhibits outstanding injectability and superior stability. Compared with the IUA group, the granular PHEAA gel can promote regeneration of the endometrium while reducing the area of endometrial fibrosis. Immunohistochemical staining experiments indicate that the granular PHEAA gel can improve the proliferation of the endometrium, promote vascularization, and enhance anti-inflammatory effect in IUA rats. And the granular PHEAA gel can effectively slow down the fibrosis of uterine tissue. Importantly, the number of embryos is significantly increased after injecting granular PHEAA gel, inferring that there is an obvious reproductive function recovery of injured endometrium.

Treatment strategies for intrauterine adhesion: focus on the exosomes and hydrogels.

Intrauterine adhesion (IUA), also referred to as Asherman Syndrome (AS), results from uterine trauma in both pregnant and nonpregnant women. The IUA damages the endometrial bottom layer, causing partial or complete occlusion of the uterine cavity. This leads to irregular menstruation, infertility, or repeated abortions. Transcervical adhesion electroreception (TCRA) is frequently used to treat IUA, which greatly lowers the prevalence of adhesions and increases pregnancy rates. Although surgery aims to disentangle the adhesive tissue, it can exacerbate the development of IUA when the degree of adhesion is severer. Therefore, it is critical to develop innovative therapeutic approaches for the prevention of IUA. Endometrial fibrosis is the essence of IUA, and studies have found that the use of different types of mesenchymal stem cells (MSCs) can reduce the risk of endometrial fibrosis and increase the possibility of pregnancy. Recent research has suggested that exosomes derived from MSCs can overcome the limitations of MSCs, such as immunogenicity and tumorigenicity risks, thereby providing new directions for IUA treatment. Moreover, the hydrogel drug delivery system can significantly ameliorate the recurrence rate of adhesions and the intrauterine pregnancy rate of patients, and its potential mechanism in the treatment of IUA has also been studied. It has been shown that the combination of two or more therapeutic schemes has broader application prospects; therefore, this article reviews the pathophysiology of IUA and current treatment strategies, focusing on exosomes combined with hydrogels in the treatment of IUA. Although the use of exosomes and hydrogels has certain challenges in treating IUA, they still provide new promising directions in this field.

CRITICAL ANALYSIS OF RAJASWALA PARICHARYA IN RELATION TO MEN-STRUAL DISORDERS A CROSS-SECTIONAL SURVEY STUDY

In women, menstruation is a common phenomenon that periodically repeats. During this phase, there is shedding of the endometrial layer which flows out of the body through the vagina along with blood. Due to these physio-logical changes and hormonal fluctuations, normal activities become stressful and demand measures to over-come the stress. Rajaswala Paricharya explained in Ayurveda can be considered as one such measure which is conceptualized to overcome this stress and maintain the health of women. This proposes various indications and contraindications of foods and activities during this phase. Since this Paricharya helps to overcome the most vul-nerable phase of the Menstrual cycle it might conceptually have positive long-term effects on the menstrual and gynecological health. This relation is studied through a cross-sectional survey study with a sample size of 100 women belonging to Shivamogga City, Karnataka. The 24 rules explained under Rajaswala Paricharya across different classics were considered and the survey was conducted to collect the data relating to their following of Rajaswala Paricharya along with data on menstrual and gynecological disorders. We find a marginal difference in the occurrence of gynecological disorders among women who have followed more than 80% of the mentioned rules. 44.44% of these women have any of the gynecological disorders where as 55.56% have no gynecological disorders. But in consideration of symptoms during menstruation, the women following a higher number of rules have a lesser number of menstrual symptoms. Women following more than 65% of the mentioned rules have an average of 2-3 symptoms, whereas women following less than 65% of Rajaswala Paricharya have an average of 4 menstrual symptoms. This represents the significance of following Rajaswala Paricharya.