Myometrial Research Articles

Genomic profile analysis of leiomyomas with bizarre nuclei and fumarate hydratase deficient leiomyomas: Strengths, weaknesses, and limitations of array-CGH interpretation.

A close relationship has been demonstrated between genomic complexity and clinical outcome in uterine smooth muscle tumors. We studied the genomic profiles by array-CGH of 28 fumarate hydratase deficient leiomyomas and 37 leiomyomas with bizarre nuclei (LMBN) from 64 patients. Follow-up was available for 46 patients (from three to 249 months, mean 87.3 months). All patients were alive without evidence of disease. For 51 array-CGH interpretable tumors the mean Genomic Index (GI) was 16.4 (median: 9.8; from 1 to 57.8), significantly lower than the mean GI in LMS (mean GI 51.8, p < 0.001). We described three groups: (1) a group with FH deletion (24/58) with low GI (mean GI: 11 vs. 22,4, p = 0.02), (2) a group with TP53 deletion (17/58) with higher GI (22.4 vs. 11 p = 0.02), and (3) a group without genomic events on FH or TP53 genes (17/58) (mean GI:18.3; from 1 to 57.8). Because none of these tumors recurred and none showed morphological features of LMS we concluded that GI at the cut-off of 10 was not applicable in these subtypes of LM. By integration of all those findings, a GI <10 in LMBN remains a valuable argument for benignity. Conversely, in LMBN a GI >10 or alteration in tumor suppressor genes, should not alone warrant a diagnosis of malignancy. Nine tumors were tested with Nanocind CINSARC® signature and all were classified in low risk of recurrence. We propose, based on our observations, a diagnostic approach of these challenging lesions.

Myometrium infection decreases TREK1 through NHE1 and increases contraction in pregnant mice.

Intrauterine infection during pregnancy can enhance uterine contractions. A two-pore K+ channel TREK1 is crucial for maintaining uterine quiescence and reducing contractility, with its properties regulated by pH changes in cell microenvironment. Meanwhile, the sodium hydrogen exchanger 1 (NHE1) plays a pivotal role in modulating cellular pH homeostasis, and its activation increases smooth muscle tension. By establishing an infected mouse model of Escherichia coli (E. coli) and lipopolysaccharide (LPS), we used Western blotting, real-time quantitative polymerase chain reaction, and immunofluorescence to detect changes of TREK1 and NHE1 expression in the myometrium, and isometric recording measured the uterus contraction. The NHE1 inhibitor cariporide was used to explore the effect of NHE1 on TREK1. Finally, cell contraction assay and siRNA transfection were performed to clarify the relationship between NHE1 and TREK1 in vitro. We found that the uterine contraction was notably enhanced in infected mice with E. coli and LPS administration. Meanwhile, TREK1 expression was reduced, whereas NHE1 expression was upregulated in infected mice. Cariporide alleviated the increased uterine contraction and promoted myometrium TREK1 expression in LPS-injected mice. Furthermore, suppression of NHE1 with siRNA transfection inhibited the contractility of uterine smooth muscle cells and activated the TREK1. Altogether, our findings indicate that infection increases the uterine contraction by downregulating myometrium TREK1 in mice, and the inhibition of TREK1 is attributed to the activation of NHE1.NEW & NOTEWORTHY Present work found that infection during pregnancy will increase myometrium contraction. Infection downregulated NHE1 and followed TREK1 expression and activation decrease in myometrium, resulting in increased myometrium contraction.

Raf kinase inhibitor protein expression in smooth muscle tumours of the uterus: a diagnostic marker for leiomyosarcoma?

Research questionWhat is the expression pattern of Raf kinase inhibitory protein (RKIP) in different subtypes of leiomyoma (usual type, cellular, apoplectic or haemorrhagic leiomyoma, leiomyoma with bizarre nuclei and lipoleiomyoma) and leiomyosarcoma specimens, and what is its biological role in leiomyosarcoma cells? DesignLeiomyoma and leiomyosarcoma specimens underwent immunohistochemistry staining. Leiomyosarcoma SK-LMS-1 cell line was RKIP knocked down and RKIP overexpressed, and cell viability, wound healing migration and clonogenicity assays were carried out. ResultsA higher immunohistochemical expression of RKIP was observed in bizarre leiomyomas, than in usual-type leiomyomas. Decreased expression was also found in cellular leiomyoma, with generally absent staining in leiomyosarcomas. Upon RKIP expression manipulation in SK-LMS-1 cell line, no major differences were observed in cell viability and migration capacity over time. RKIP knockout, however, resulted in a significant increase in the cell's ability to form colonies (P = 0.011). ConclusionRKIP distinct expression pattern among leiomyoma histotype and leiomyosarcoma, and its effect on leiomyosarcoma cells on colony formation, encourages further studies of RKIP in uterine smooth muscle disorders.

Application of modified sodium alginate hydrogel for interventional embolization of hemorrhagic diseases

Traditional particulate embolic agents are small in diameter, but can easily embolize the ends of blood vessels, resulting in ischemia and necrosis of normal tissues and organs. The metal spring embolic agent has a larger diameter, but it cannot be degraded and can easily cause permanent damage to blood vessels. Ideally, a bleeding embolism should achieve rapid hemostasis without causing long-term necrosis of organs and tissues. In this study, a modified sodium alginate hydrogel (MSAH) was prepared by mixing an oxidized sodium alginate (OSA) aqueous solution with a carboxymethyl chitosan (CMC) aqueous solution at a ratio of 1:6 in a 38°C bath for 8 min. The feasibility of this modified hydrogel was then tested in an internal iliac artery hemorrhage model using New Zealand rabbits. The MSAH had good adhesion. The hydrogel was injected through a single curved 4F catheter without obvious effects on uterine smooth muscle cell proliferation and apoptosis. The blood flow of the internal iliac artery was restored by long-term degradation of the sodium alginate hydrogel, and no ischemia and necrosis were observed by histopathology. The MSAH prepared using a mixture of OSA and CMC had good adhesion, biocompatibility, and injectability and could be used for target-vessel embolization in an internal iliac artery hemorrhage model. The MSAH can achieve main artery embolization without affecting the peripheral artery blood supply, resulting in both short-term rapid hemostasis and long-term degradation, with no target organ necrosis.

FOXO3a deregulation in uterine smooth muscle tumors

ObjectiveThe present study aimed to investigate FOXO3a deregulation in Uterine Smooth Muscle Tumors (USMT) and its potential association with cancer development and prognosis. MethodsThe authors analyzed gene and protein expression profiles of FOXO3a in 56 uterine Leiomyosarcomas (LMS), 119 leiomyomas (comprising conventional and unusual leiomyomas), and 20 Myometrium (MM) samples. The authors used techniques such as Immunohistochemistry (IHC), FISH/CISH, and qRT-PCR for the present analyses. Additionally, the authors conducted an in-silico analysis to understand the interaction network involving FOXO3a and its correlated genes. ResultsThis investigation revealed distinct expression patterns of the FOXO3a gene and protein, including both normal and phosphorylated forms. Expression levels were notably elevated in LMS, and Unusual Leiomyomas (ULM) compared to conventional Leiomyomas (LM) and Myometrium (MM) samples. This upregulation was significantly associated with metastasis and Overall Survival (OS) in LMS patients. Intriguingly, FOXO3a deregulation did not seem to be influenced by EGF/HER-2 signaling, as there were minimal levels of EGF and VEGF expression detected, and HER-2 and EGFR were negative in the analyzed samples. In the examination of miRNAs, the authors observed upregulation of miR-96-5p and miR-155-5p, which are known negative regulators of FOXO3a, in LMS samples. Conversely, the tumor suppressor miR-let7c-5p was downregulated. ConclusionsIn summary, the outcomes of the present study suggest that the imbalance in FOXO3a within Uterine Smooth Muscle Tumors might arise from both protein phosphorylation and miRNA activity. FOXO3a could emerge as a promising therapeutic target for individuals with Unusual Leiomyomas and Leiomyosarcomas (ULM and LMS), offering novel directions for treatment strategies.

Fibroblast Growth Factor 23 is a Potential Prognostic Biomarker in Uterine Sarcoma.

Uterine sarcoma (US) is a highly malignant cancer with poor prognosis and high mortality in women. In this study, we evaluated the expression of human fibroblast growth factor 23 (FGF23) in different US subtypes and the relationship between survival and clinicopathological characteristics. We conducted a comparative analysis of FGF23 gene expression in different pathological types of US. Utilizing a cohort from The Cancer Genome Atlas of 57 patients, a 50-patient microarray dataset (GSE119043) from the Gene Expression Omnibus and a Suining cohort of 44 patients, we analyzed gene expression profiles and corresponding clinicopathological information. Immunohistochemistry was used to examine the expression level of FGF23 in four US subtypes. Survival analysis was used to assess the relationship between FGF23 expression and prognosis in US patients. Compared with uterine normal smooth muscle and uterine leiomyoma, FGF23 expression was significantly upregulated in US and was differentially expressed in four US subtypes. Uterine carcinosarcoma exhibited the highest expression of FGF23 among the subtypes. Survival analysis revealed no correlation between FGF23 expression and either overall survival or progression-free survival in US (P > 0.05). Similar results were obtained from the validation cohorts. Univariate and multivariate analyses showed no significant correlation between FGF23 expression and the US prognosis. Tumor stage, CA125, and tumor recurrence were independent prognostic factors for survival of US patients. FGF23 was highly expressed in US and was promising as a novel potential biomarker for the diagnosis and prognosis of US.

Mefenamic Acid Reduces the Contractility of Isolated Uterine Smooth Muscle of Rat

Objective: Mefenamic acid is a nonsteroidal anti-inflammatory drug widely used for the treatment of pain and inflammation. The drug is indicated in patients with mild to moderate pain and inflammatory diseases, usually toothache, painful menstruation, muscle or joint pains and postpartum pain. There are not enough studies showing the effectiveness of this drug, which is significantly effective in gynecological pains, on the uterine contraction and relaxation mechanism. This study was carried out to investigate the effects of mefenamic acid on the uterine contraction and relaxation mechanism. Materials and Methods: In the study, seven female intact Sprague-Dawley rats in the diestrus period were used. Longitudinal myometrium sections of 1.2 cm length, 2 mm width and 1 mm thickness from animals were suspended in an isolated organ bath containing crebs solution. After the regulation period, mefenamic acid was administered at a dose of 300 μM. Contractile changes were monitored using an isometric transducer. Before and after the application, the area under the curve (AUC), frequency and peak to peak (p-p) values were normalized as % change. Statistical analyzes of the data were performed with the Paired Sample T test in SPSS 22.0 program. Results: Mefenamic acid caused a statistically significant decrease in the p-p, AUC and frequency values of spontaneous uterine contractions at 300 μM dose (p&lt;0.001). Conclusion: Mefenamic acid has an inhibitory effect on uterine contractions. This drug, which is widely used in the clinic for menstrual pain caused by abnormal uterine contractions, especially in young women, may show its analgesic effect by inhibiting uterine contractions.

Https://fz.kiev.ua/index.php?abs=2034

Investigation of the mechanism of neuronal communication underlies the fundamental discoveries that promote health. The studies of oxytocin signaling in neurons from or within different brain areas are directed to explore the role of this neurohormonal modulator in the regulation of synaptic transmission and plasticity, neuronal excitability that contributes to the reproduction, social behavior and learning capacity, anxiety, inflammation and differentiation in the brain. Oxytocin is synthesized in supraoptic and paraventricular nuclei of the hypothalamus and when secreted into the bloodstream from the posterior pituitary, it produces a significant effect on uterine contraction and lactation. At the same time this nonapeptide being released within the limbic system and brain cortex modulates neuronal activity by affecting ion channels on their membranes. The oxytocin receptor is primarily coupled to Gq/11 proteins causing phospholipase C activation, Ca2+ release and store-operated Ca2+ entry. These pathways are central for the regulation of the activity of different types of TRP channels, especially of the canonical subfamily (TRPC). Here we highlight the link between oxytocin signaling, which is particularly well investigated in the myometrium, and receptor-operated TRPC4 and multimodal TRPV4 ion channels that participate in oxytocin-dependent regulation of the uterine smooth muscle contractility under various conditions. Importantly, similarly to oxytocin, these channels have been implicated in neuropathic pain behavior, anxiety, fear and depression. Since similar signal transduction pathways are likely to be functional in neuronal cells, we propose that future studies of oxytocin effects in the CNS should also consider the role of these Ca2+-permeable channels.

A Case Report of a Patient with Uterine Smooth Muscle Tumor of Uncertain Malignant Potential

A Case Report of a Patient with Uterine Smooth Muscle Tumor of Uncertain Malignant Potential

Phytochemical constituents from the roots and lignotubers of Rhoicissus tridentata and their in vitro uterotonic activity

Rhoicissus tridentata is one of the most frequently used plants in preparing Isihlambezo, a herbal drink consumed by many South African women to induce labour and tone the uterus in pregnancy. This study aimed to identify the uteroactive compounds in this plant. Chromatographic purification of the methanol and water extracts from the roots yielded eight compounds, i.e. morin 3-O-α-L-rhamnopyranoside, trans-resveratrol 3-O-β-glucopyranoside, a mixture of asiatic and arjunolic acids, quercetin 3-O-rhamnopyranoside, catechin, β-sitosterol, and linoleic acid. All compounds were evaluated for their uterotonic effects using uterine smooth muscle isolated from stilboestrol-primed Sprague-Dawley rats. The mixture of asiatic and arjunolic acids showed the highest activity with EC50 of 0.02129 μg/mL for amplitude. These results validate the use of R. tridentata in ethnomedicine to facilitate labour in childbirth. Morin 3-O-α-L-rhamnopyranoside and trans-resveratrol 3-O-β-glucopyranoside caused a relaxation of the uterine muscle, which suggests that some compounds in R. tridentata possess opposing activities.

Indicators of uterine contractile activity in highly productive sows using supraphysiological doses of oxytocin

The use of large doses of oxytocin to sows in the early postpartum period is indicated to normalize lactogenesis, increase immune proteins in colostrum and transitional milk, prevent postpartum complications, and increase the viability of the offspring. However, the effect of these doses of myotropic drug on the contractile activity of the uterus is not shown. The aim of the work was to study the contractile function of the uterus of highly productive sows in the early postpartum period and the peculiarities of its reaction to supraphysiological doses of oxytocin. Scientific and production experience was carried out on the basis of a large pig breeding complex. The object of the study was highly productive sows 16 hours after the completion of normal labor. The hormonal drug oxytocin was administered intra-muscularly to animals at a supraphysiological dose of 75 units. Hysterograms of uterine contractions were recorded by internal hysterography before and immediately after administration of the myotropic drug, as well as one hour after injection. On hysterograms, spontaneous contractile activity of the uterus in highly productive sows was characterized by regular, short-term contractions of the optimal amplitude. Activation of the contractile function of the myometrium occurs 5-7 minutes after injection of the drug. After administration of oxytocin, an increase in the amplitude of contraction of uterine smooth muscle cells was noted by 39.17% (p&lt;0.05), while a longer period of contractions was recorded by 31.64%. The frequency of uterine contractions increased 2.0 times (p&lt;0.01). The contract index was significantly 3.66 times (p&lt;0.05) higher than that in relation to the initial level. After 1 hour after oxytocin administration, an increase in the contraction period by 5.42% was noted, with a decrease in amplitude by 37.87% (p&lt;0.01) and the number of contractions by 49.91% (p&lt;0.01), the contractional index decreased by 3.05 times (p&lt;0.01) relative to the previous level. In relation to the initial level, the contract index was 20.0% higher. At the same time, an increase in the duration of contractions by 38.78% and a decrease in their amplitude by 13.53% were noted. Consequently, oxytocin at a dose of 75 ME not violate the contractile ability of the myometrium. The response of the smooth muscle cells of the uterus to the supraphysiological dose of oxytocin is maximally manifested in the first hour after the administration of the myotropic drug.

Steady-state Approximations for Hodgkin-Huxley Cell Models: Towards Multi-scale Models of Uterine Smooth Muscle

The Tong, et al. 2011 publication presents a comprehensive mathematical model of electro- mechanical behaviour in a uterine smooth muscle cell. It incorporates ion channel currents into a classic Hodgkin-Huxley membrane voltage formulation as well as Ca2+ concentration dynamics with pumps and exchangers. Mechanical force generation by way of the Hai-Murphy smooth muscle cell model is computed based on the Ca2+ concentrations. The ‘full’ Tong, 2011 model (FTM) includes nineteen ordinary differential equations for the ion channel activation/inactivation variables out of a total of 105 equations overall. An effort at reducing the FTM into a smaller number of equations for computational efficiency – the ‘reduced’ Tong model (RTM) – aims at reproducing the overall behaviour of the FTM without excessive detail. We present here the CellML implementation of the RTM for simulating the FTM within the context of the reduced model’s primary publication as derived from the Tong, et al. 2011 model.

Steady-state Approximations for Hodgkin-Huxley Cell Models: Towards Multi-scale Models of Uterine Smooth Muscle

The Tong, et al. 2011 publication presents a comprehensive mathematical model of electro- mechanical behaviour in a uterine smooth muscle cell. It incorporates ion channel currents into a classic Hodgkin-Huxley membrane voltage formulation as well as Ca2+ concentration dynamics with pumps and exchangers. Mechanical force generation by way of the Hai-Murphy smooth muscle cell model is computed based on the Ca2+ concentrations. The ‘full’ Tong, 2011 model (FTM) includes nineteen ordinary differential equations for the ion channel activation/inactivation variables out of a total of 105 equations overall. An effort at reducing the FTM into a smaller number of equations for computational efficiency – the ‘reduced’ Tong model (RTM) – aims at reproducing the overall behaviour of the FTM without excessive detail. We present here the CellML implementation of the RTM for simulating the FTM within the context of the reduced model’s primary publication as derived from the Tong, et al. 2011 model.

Uterine smooth muscle tumors: a multicenter, retrospective, comparative study of clinical and ultrasound features

ObjectiveTo evaluate a wide range of clinical and ultrasound characteristics of different uterine smooth muscle tumors to identify features capable of discriminating between these types.MethodsThis was a retrospective, multicenter study...

Practical Effect of Cervical Softening Decoction Combined with Delivery Ball Approach in Promoting Vaginal Delivery in Primiparous Women

In Traditional Chinese Medicine, Cervical Softening Decoction is derived from a modified “Wan Bing Hui Chun” formulation and consists of natural herbal ingredients such as Angelica sinensis, Ligusticum chuanxiong, Leonurus heterophyllus, and Cortex Daphnes. The decoction regulates Qi, promotes blood circulation, and facilitates fetus descent. Genkwanin is an active compound in L. heterophyllus and previous studies identified this compound in ethanolic extracts from rat uterine smooth muscle cell membrane fractions. To further understand the pharmacological activities of Cervical Softening Decoction, we examined genkwanin effects on isolated rat uterine contractions and protein expression in rat tissue with postpartum hemorrhage complicated by multiple organ dysfunction syndrome (MODS). We also explored the practical effects of Cervical Softening Decoction combined with a birthing ball approach on promoting natural delivery in primiparous women. First, an isolated rat uterine smooth muscle contraction model was generated and three groups established: oxytocin group (0.002 U/mL), conventional genkwanin dose group (3 μg/mL), and high genkwanin dose group (6.0 μg/mL). Changes in uterine smooth muscle contraction and relaxation amplitudes were recorded before and after administration. Second, 50 rats were used to establish the following groups: (1) control group, (2) postpartum hemorrhage with MODS model group (MODS group), and (3) a postpartum hemorrhage with MODS treated with genkwanin group (genkwanin group). Western blotting was used to detect and compare tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 protein expression levels in lung tissue from groups. Finally, 84 primiparous women admitted to our hospital between January 2020 and December 2021 were selected and divided into two groups based on their obstetric interventions. The control group (n = 42) received routine obstetric intervention, while the observation group (n = 42) received Cervical Softening Decoction and delivery ball interventions. Delivery conditions were compared across groups. In basic studies, genkwanin doses significantly increased contraction and relaxation values in isolated rat uterine smooth muscle (P &lt;0.05). Moreover, the genkwanin contractile effects at conventional and high doses were comparable with oxytocin. TNF-α and IL-6 protein expression in MODS group lung tissue was significantly higher when compared with the control group, while expression in the genkwanin group was significantly reduced when compared with the MODS group (P &lt;0.05). In clinical studies, the observation group showed significantly higher natural delivery rates and Labor Agentry Scale (LAS) scores when compared with the control group. In contrast, cesarean section rates, first and second stage labor duration, total labor duration, and Facial Pain Scale (FPS) scores were significantly lower in the observation group when compared with the control group (P &lt;0.05). In basic studies, genkwanin elicited significant contraction effects on isolated rat uterine smooth muscle and effectively inhibited inflammatory responses in rats with postpartum hemorrhage complicated by MODS. In clinical studies, combined Cervical Softening Decoction and delivery ball generated promising practical effects in promoting vaginal delivery in primiparous women. The intervention significantly increased the probability of a natural delivery, shortened labor duration, relieved maternal pain, and enhanced maternal control during delivery.

Diagnosis and treatment of intravenous leiomyomatosis

Intravenous leiomyomatosis is a rare type of tumor that is histologically benign but biologically invasive. It originates from the smooth muscle of the uterine or the uterine vein. It can grow through the uterus and extend into the pelvic cavity, or grow along the veins without invading the wall of the venous vessel itself. The tumors are estrogen-dependent and can metastasize through the bloodstream. Thus, in addition to continuous growth, some tumors exhibit isolated growths in the venous system and heart chambers or show disseminated growth in the lungs, although distant metastasis to other regions usually do not occur. Currently, there is limited research on this disease, the majority of which are case reports, surgical experience summaries, and differentiation from ordinary gynecological myomas in terms of pathogenesis and radiological diagnostic experience. There are two main theories on the origin of the disease: uterine smooth muscle and smooth muscle of the uterine veins. Some studies have verified the role of estrogen, progesterone receptor-related pathways, and angiogenesis in the development of the disease. The clinical symptoms of this disease are varied, depending on the affected area. In the early stages, when the tumor only affects the pelvic cavity, patients show mild symptoms resulting from pelvic organ compression. When it progresses to the inferior vena cava and heart, patients show more complex symptoms resulting from venous return obstruction, cardiac obstruction, and hemodynamics appearing. Different institutions have proposed different disease staging and classification strategies for different clinical purposes. Some are based on the affected area of the lesion; others are based on the size of the tumor. Although surgery remains the main treatment for this disease, the specific surgical approach, adjuvant drug therapy, and prognosis still need further exploration.

A new affine inhibitor of sodium pump thiacalix[4]arene С-1193 increases the intracellular concentration of Ca ions and modifies myometrium contractility

The methods of enzymatic and kinetic analysis were used to demonstrate that thiacalix[4]arene-bis-hydroxymethylphosphonic acid С-1193 had the inhibitory effect (І0.5 = 42.1 ± 0.6 nM) on Na+,K+-ATPase activity in the plasma membrane of myometrium cells with no effect on the relative activity of other ATPases localized in this subcellular structure. The method of confocal microscopy and Са2+-sensitive fluorescent probe fluo-4 were used to demonstrate that thiacalix[4]arene С-1193 increased the intracellular concentration of Ca ions in the immobilized uterine myocytes. The tenzometric studies proved that С-1193 (10 and 100 μМ) increased the isometric phasic contractions, induced via the paths of both electromechanical (depolarization with high-potassium solution) and pharmacomechanical (application of uterotonic hormone oxytocin, neurotransmitter acetylcholine or selective agonist of muscarinic acetylcholine receptors cevimeline) coupling. Application of thiacalix[4]arene С-1193 as a selective and effective inhibitor of Nа+,K+-ATPase may be useful both for studyng the regulation of ion homeostasis in smooth muscle cells and creation of new uterotonics based on the calixarene core. Keywords: contractility mechanokinetics, myometrium, Na+‚K+-АТРase, thiacalix[4]arene

Hemostatic Effect of Intrauterine Instillation of Oxytocin in Hysteroscopic Myomectomy: A Randomized Double Blinded Control Trial

Introduction: Uterine myomas are benign, smooth muscle tumors of the uterus that are affected by estrogen and progesterone. Most fibroids are asymptomatic and require no intervention or further investigations. Fibroids arise from the uterine smooth muscles then migrate to a place of lower resistance becoming subserosal or submucous myomas or remains intra-myometrial. Aim of the Work: The main aim of this study was to investigate whether the intrauterine instillation of oxytocin into the distention medium during hysteroscopic myomectomy will reduce blood loss and improve surgical visibility. Patients and Methods: This Randomized controlled double blinded clinical trial was conducted at Early Cancer Detection Unit (ECDU) at Ain Shams University hospitals. This study included 60 women who were scheduled to undergo a hysteroscopic myomectomy were divided into two equal groups: Group A (Oxytocin group): 30 women underwent a hysteroscopic myomectomy with the use of 10 IU of oxytocin for every 1000 ml of the distending medium. Group B (Placebo group): 30 women underwent hysteroscopic myomectomy with the use of a sterile bacteriostatic water ampule for every 1000 ml of the distending medium. Results: Groups were comparable in demographic data (in terms of age) and surgical data sin term of (myoma size and number and duration of surgery) and intraoperative fluids and there was no statistically significant difference between groups. There were significant difference in Hgb and Hct drop in each group but control group showed more mean difference than oxytocin group. Our results showed that groups were comparable in post-operative need of blood transfusion and 2nd surgery and there was statistically significant difference between groups in post-operative Hgb and HCT. Groups were comparable in surgeon evaluation in term of visual fluid clarity and there was statistically significant difference between groups in surgeon rating bleeding. Our results showed that groups were comparable in complications in term of (uterine and medical) and there was no statistically significant difference between groups regarding complications. Conclusion: Complications during diagnostic hysteroscopy are common. Intrauterine instillation of oxytocin into the distention medium during hysteroscopic myomectomy reduce blood loss and improve surgical visibility. More studies with varied dosages and methods are needed to validate these findings.

Effects of polydimethylsiloxane membrane surface treatments on human uterine smooth muscle cell strain response

In the United States, 1 in 10 infants are born preterm. The majority of neonatal deaths and nearly a third of infant deaths are linked to preterm birth. Preterm birth is initiated when the quiescent state of the uterus ends prematurely, leading to contractions and parturition beginning as early as 32 weeks, though the origins are not well understood. To enable research and discovery of therapeutics with potential to better address preterm birth, the capability to study isolated cell processes of pregnant uterine tissue in vitro is needed. Our development of an in vitro model of the myometrium utilizing human uterine smooth muscle cells (uSMCs) responsible for contractions provides a methodology to examine cellular mechanisms of late-stage pregnancy potentially involved in preterm birth. We discuss culture of uSMCs on a flexible polydimethylsiloxane (PDMS) substrate functionalized with cationic poly-l-lysine (PLL), followed by extracellular matrix (ECM) protein coating. Previous work exploring uSMC behavior on PDMS substrates have utilized collagen-I coatings, however, we demonstrated the first exploration of human uSMC response to strain on fibronectin-coated flexible membranes, importantly reflecting the significant increase of fibronectin content found in the myometrial ECM during late-stage pregnancy. Using the model we developed, we conducted proof-of-concept studies to investigate the impact of substrate strain on uSMC cell morphology and gene expression. It was found that PLL and varied ECM protein coatings (collagen I, collagen III, and fibronectin) altered cell nuclei morphology and density on PDMS substrates. Additionally, varied strain rates applied to uSMC substrates significantly impacted uSMC gene expression of IL-6, a cytokine associated with instances of preterm labor. These results suggest that both surface and mechanical properties of in vitro systems impact primary human uSMC phenotype and offer uSMC culture methodologies that can be utilized to further the understanding of cellular pathways involved in the uterus under mechanical load.

FRI393 Mapping Endometrial Cell-type-specific Disease Signatures A nd Endometrial Organoids In Polycystic Ovary Syndrome

Abstract Disclosure: G. Eriksson: None. C. Li: None. S. Risal: None. H. Pui: None. S. Torstensson: None. A. Linden Hirschberg: None. S. Petropoulos: None. Q. Deng: None. E. Stener-Victorin: None. Women with polycystic ovary syndrome (PCOS) suffer from reduced fertility linked to implantation failure and miscarriage, as well as endometrial cancer, all associated with endometrial dysfunction. We hypothesize that cell-type-specific endometrial dysfunction in insulin-resistant and hyperandrogenic women with PCOS contributes to the observed endometrial dysfunction and that treatment aimed at improving insulin sensitivity and decreasing androgen excess has the potential to reverse identified alterations.To uncover cell-type-specific disease signatures and molecular pathways for PCOS-specific endometrial dysfunction, we first extracted single-nuclei (sn) from frozen endometrial biopsies taken in the proliferative phase (day 7-10) from controls (n=5) and women with PCOS (n=10) at baseline and in PCOS after 16 weeks of metformin (n=7) or lifestyle management (n=3) for snRNA-sequencing. The 10x Genomics protocol allowed us to sequence ≈10,000 nuclei/sample and ≈20,000 reads/nuclei. A total of 248,694 nuclei were captured and 6 major cell types were identified. The 3 largest cell clusters were i) stromal cells (124,055 nuclei), ii) epithelial cells (105,095 nuclei) and iii) immune cells (13,596), whilst iv) uterine smooth muscle cells, v) endothelial and vi) lymphoid cell clusters consisted of &amp;lt;3000 nuclei. Subsetting the epithelial cells revealed the functional luminal, glandular and ciliated cell types as well as proliferative cells. In the immune cell cluster, both myeloid and lymphoid lineage cells were identified, of which uterine NK-cells (uNK) and macrophages (uM) were the largest populations. Several differentially expressed genes (DEGs) of the epithelial subtypes, uNKs, uMs and stromal cells were identified in women with PCOS compared with controls. DEGs in cell types between PCOS cases and controls were enriched in pathways related to cilium organization in the ciliated epithelium, extracellular matrix structure in stromal cells, and cysteine-type endopeptidase activity in uM type 1. Of note, both 16-week of metformin treatment or lifestyle management restored multiple DEGs in each subtype. Second, to validate the identified molecular target, we have established 3D endometrial organoids (EOs) from fresh endometrial biopsies of control and PCOS women. After several passages, EOs were cryopreserved to create a biobank for future functional analyses of the molecular targets. The EOs can be re-established after thawing and immunofluorescence staining shows that the EOs consist of an intact proliferative basolateral epithelial membrane. This rigorous mapping of the human PCOS endometrium improves our understanding of cellular complexity and specific cell types. It provides new mechanistic insights into disease-specific endometrial dysfunction(s), and several of the identified dysfunctions can be reversed by current first-line interventions. Presentation: Friday, June 16, 2023