Estimation of the polymorphism of matrix metal proteinase genes in patients with endometrioid ovarian cysts

Aim. To determine the association of the MMP-9 genetic polymorphism with the risk of developing endometrioid ovarian cysts (ECC). Materials and methods. 55 women aged 19 to 47 were examined. 27 women underwent surgery for ECC (group 1). The control (group 2) consisted of 28 patients without endometriosis and operated because of tubal infertility. Single nucleotide polymorphism was investigated. Genotyping was performed by restriction fragment length polymorphism analysis. Results. We did not find statistically significant differences between the group of patients with ECF and the control group in terms of age 29.0 (25.95; 33.1), 34.5 (29.3; 37.0); p0.05, body mass index 21.2 (19.8; 22.6), 21.95 (20.4; 23.9), p0.05, age of menarche onset 13.0 (12.95; 14.0), 13.0 (12.0; 14.0), p0.05, duration of menstrual bleeding 5.0 (5.0; 5.0), 5.0 (5.0; 6.0), p0.05 for the 1st and 2nd groups, respectively, also dysmenorrhea (2 0.019; p=0.8918), the number of births (2 3.441; p=0.3285) and abortions (2 2.822; p=0.0930) in anamnesis. The frequencies of all studied genotypes of metalloproteinase MMP-9 C (1562) T of the MMP9 gene in the group of patients with ECF and the control group are in HardyWeinberg equilibrium (p=0.99, p=0.43 for 1 and 2 group) which excludes differences in the distribution of genotype frequencies of polymorphic loci of MMP9 genes in the group of patients with ECF and the control group. However, the result could be influenced by the severity of the disease and the size of the study groups. Conclusion. Given the undoubted role of matrix metalloproteinases in the pathogenesis of genital endometriosis, further studies with large samples in various populations are needed.

Therapeutic Mechanism and Clinical Observation of Traditional Chinese Medicine Combined with Interventional Recanalization for Tubal Infertility

To observe the clinical effect of traditional Chinese medicine (TCM) combined with interventional recanalization therapy in the treatment of tubal obstructive infertility, first, different treatment approaches were used on rabbits, and transmission electron microscopy (TEM) indicated that interventional recanalization combined with TCM can significantly ameliorate the pathological condition of the fallopian tube after treatment. Moreover, ELISA disclosed that the treatment could significantly reduce the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) and increase the expression of interleukin-10 (IL-10), which demonstrated that TCM therapy can help against inflammation of the fallopian tubes. PCR array analysis revealed that BMP4, BMPR1A, SMAD2, SMAD3, SMAD4, and KLF10 expressions were upregulated, and SMAD7 expression was downregulated, proving that combined treatment could influence gene expression in the TGF-β family and further regulate the secretion of proteins in SMADs. In addition, a clinical study recorded the fallopian tube patency rate of 165 patients after 12 months. The recanalization rates in the two groups were 81.9% and 53.1%, with the higher rates in the combined medicine enema group. All these findings implied that interventional recanalization combined with TCM preparation has a stronger effect. The mechanism probably involves effects on the expression of genes in the TGF-β/SMAD and BMP/SMAD signaling pathways, with simultaneous regulation of inflammatory factors, thereby improving the ovarian environment and increasing pregnancy rates.

PRETERM BIRTH AND LOW BIRTH WEIGHT IN SINGLETON PREGNANCIES AFTER IN VITRO FERTILIZATION- INFLUENCE OF INFERTILITY AND METHOD OF FERTILIZATION

The aim of our study was to determine the effect of infertility and the method of fertilization on the frequency of preterm birth and low birth weight in singleton pregnancies achieved by in vitro fertilization (IVF). The study was conducted in the period 01.2013 - 12.2017 and included 286 patients with births from singleton IVF pregnancies. Classical IVF received 107 patients (group A1) and ICSI fertilization - 148 (group A2). Cases with tubal infertility were 81 (group B1), and 85 were with male infertility (group B2). In group A1, birth prior to 37 weeks of gestation (w.g.) was found in 33 (30.8%) cases, compared to 26 (17.6%) in group A2 (p=0.029).In group A1, birth weight <2500 g was observed in 27 (25.3%) of cases, and in group A2 - in 17 (11.5%) of cases (p=0.015). In group B1, birth prior to 37 w.g. was established in 26 (32.1%) of the cases, compared to 9 (10.6%) in group B2 (p=0.003). In group B1 there was birth weight <2500 g in 19 (23.5%) of patients, compared to 6 (7.1%) in group B2 (p = 0.009). The average weight of the newborn and average duration of pregnancy was lower in groups with IVF fertilization and tubal infertility compared to ICSI fertilization and male infertility. Tubal infertility and classical IVF fertilization are independent risk factors that increase the frequency of preterm birth and low birth weight in IVF singleton pregnancies.

Immunohistochemical features of the fallopian tubes’ structure in case of fetuses with a gestational term of 21-28 weeks from mothers with a chronic infection of the lower genital tracts

In the following article we are disclosing immunohistochemical features of the fallopian tubes’ structure in case of fetuses with a gestational term of 21-28 weeks. We have studied the structure of organs of 15 fetuses from mothers with a chronic infection of the lower genital tracts (HILGT) (the group of comparison) relatively to one in case of 15 fetuses from mothers with a physiological pregnancy (the main group). All fetuses had a gestational term of 21-28 weeks and had died intranatally as a result of an acute disorder of uterine-placental circulation. Methods of reearch: organometric, histological, immunohistochemical and statistical. The complex research allowed to reveal a probable decrease of the organometric data as well as indicators of thickness of the main structural components of the organs’ wall in case of fetuses from mothers of the group of comparison relatively to ones in case of fetuses from the main group. By applying histological method we had reveled a disorder in the strucuture of mucous and muscular membranes in the structure of organs of fetuses from mothers with a pathology of pregnancy, which is manifested by decreasing of number of folds, uneven thickness of the mucous membrane as well as disorder in formation of the muscular membrane’ layers. Immunohistochemical method has revealed an increase of apoptotically changed mucosal cells in the fallopian tubes of fetuses from mothers with HILGT. By applying MCAT to endotheline-1 we have revealed an increased glow of endithelial cells in vessels both of arterial and venous types in the strucuture of the organs’ wall in case of fetuses from the group of comparison. During the study on the endocrine activity of organs of fetuses from mothers with complicated pregnancy we have postulated a probable decrease of the cells’ glow’ intensity towards MCAT of progesterone. At the same time, there is also an inflammatory infiltration in the mucous membrane of the fallopian tubes’ wall. The aforementioned changes in the strucuture as well as in the functional activity of fetuses’ organs from mothers’ with HILGT are formed under the influence of chronic hypoxia and endocrine insufficiency of the feto-placental complex, which is taking place in case of this pathology. Histological and immunohistochemical features of the fallopian tubes’ structure in case of fetuses from mothers with complicated pregnancy are indicating a disorder in formation and maturing of the organ as well as they could lead to development of ectopic pregnancy and tubal infertility in the subsequent ontogenesis.

Alterations of Vaginal Microbiota in Women With Infertility and Chlamydia trachomatis Infection

Chlamydia trachomatis (C. trachomatis) is the most common etiological agent of bacterial sexually transmitted infections (STIs) worldwide and causes serious health sequelae such as cervicitis, pelvic inflammatory disease, and even infertility if ascending from the lower to the upper female genital tract. Previous studies have revealed the pivotal role of vaginal microbiota in susceptibility to STIs. However, alterations in the vaginal microbiota in women who are infertile and infected with C. trachomatis remain unknown. This study used metagenomic analysis of sequenced 16S rRNA gene amplicons to examine the vaginal microbial profiles of women with tubal infertility who were C. trachomatis-negative and those who were C. trachomatis-positive pre- and post-antibiotic treatment. Women who were C. trachomatis-negative and deemed healthy were recruited as references of eubiosis and dysbiosis. Women with tubal infertility and C. trachomatis infection presented a unique Lactobacillus iners-dominated vaginal microbiota rather than one dominated by Lactobacillus crispatus and displayed a decrease in Lactobacillus, Bifidobacterium, Enterobacter, Atopobium, and Streptococcus, accompanied by decreased levels of cytokines such as interferon (IFN)-γ and interleukin (IL)-10. This altered vaginal microbiota could be restored with varying degrees after standard treatment for C. trachomatis. This shift could be a predictive vaginal microbiota signature for C. trachomatis infection among females with tubal infertility, while no significant differences in phylum, class, and operational taxonomic unit (OTU) levels were observed between women with tubal infertility who were C. trachomatis-negative and healthy controls. This is the first study to provide data on the association of vaginal microbiota with C. trachomatis infection among women with tubal infertility and highlights unprecedented potential opportunities to predict C. trachomatis infection.

Increased expression of HMGB1 in the implantation phase endometrium is related to recurrent implantation failure

Impaired endometrial receptivity is the main cause of recurrent implantation failure (RIF), however, its underlying mechanism is unclear. In this study, we found that HMGB1 expression was significantly decreased in the implantation phase endometrium in the control group (patients with tubal infertility who successfully achieved conception after the first embryo transfer) (P = 0.006). However, the expression levels of HMGB1 mRNA and protein were significantly upregulated during the implantation phase in endometrial tissues obtained from patients with RIF compared to those in the control group (P = 0.001), consistent with the results of genome-wide expression profiling. Moreover, in vitro assays showed that increased expression of HMGB1 in human endometrial epithelial cells cause marked deficiency in supporting blastocysts and human embryonic JAR cell adhesion, mimicking the process of embryo adhesion. However, overexpression of HMGB1 had no effect on cell proliferation and in-vitro decidualization in a human endometrial stromal cell line (T-HESCs) and in primary human endometrial stromal cells (HESCs). These findings indicate that increased HMGB1 levels suppressed the adhesion capability of epithelial cells, contributing to impaired endometrial receptivity in patients with recurrent implantation failure. This characteristic can be used as a target for detecting and treating recurrent implantation failure in clinical practice.

Endometriosis and Oxidative Stress. Do Reactive Oxygen Species Always Lead to Damages of Gametes and Embryos?

Reactive Oxygen Species (ROS) play a crucial role in the pathogenesis of many reproductive disorders, such as endometriosis on the one hand, but on the other hand they participate in different cellular proliferation processes, too. Endometriosis is an apoptotic endometrial, menstrual cells and lysed erythrocytes-induced inflammatory disease outside the uterine cavity, which activates macrophages leading to ROS production and oxidative stress. However, based on the available literature, the reproductive outcomes are still contradictory. In this study, the demographic, embryological and clinical results of 252 patients suffering from tubal infertility (control), ASRM I-II and III-IV endometriosis were analyzed. Endometriosis was associated with decreased anti-Müllerian hormone level and increased gonadotropin doses during stimulation (p<0.0001). In ASRM III-IV, reduced embryological parameters were observed, which resulted in 13.73% and 15.21% decrements in the implantation rates, 19.96% and 23.89% in the clinical pregnancy rates of patients suffering from ASRM III-IV endometriosis in comparison with control or ASRM I-II, respectively. In addition, miscarriage rates were 19.04%, 29.03% and 38.46% in control, ASRM I-II and ASRM III-IV, respectively. In our study, the supposed altered oxido-reduction environment of gametes and embryos obviously exerted negative effects on the embryological and clinical parameters, but these effects could not be observed in case of mild endometriosis with low level of stress.

P–601 Anovulatory patients with PCOS have lower euploidy rates compared to those with hypothalamic amenorrhea and to normo-ovulatory patients

Study question How do euploidy rates differ in anovulatory women with polycystic ovarian syndrome (PCOS) and hypothalamic hypogonadism (HH) compared to normo-ovulatory women undergoing IVF/ICSI? Summary answer Patients with PCOS have a significantly lower euploidy rate compared to patients with HH and patients with tubal factor infertility. What is known already Previous studies have demonstrated similar blastocyst conversion rates in women with PCOS and tubal factor infertility. Reported aneuploidy rates in preimplantation genetic testing cycles are similar in women with PCOS and tubal infertility. There are no data on blastocyst conversion or aneuploidy rates in women with HH. While PCOS and HH are different physiologic processes, patients with these disorders are reported together to SART and to the CDC National ART Surveillance System under the diagnosis of “ovulatory dysfunction”. Study design, size, duration: Retrospective cohort study of all autologous IVF and ICSI cycles for patients with oligo-anovulation (PCOS, n = 552 and HH, n = 48) and normo-ovulation (tubal factor infertility, n = 423) from 1/1/2012 to 6/30/2019. A total of 1023 cycles from 720 patients were analyzed. Participants/materials, setting, methods Cycle outcomes, including number of oocytes, mature oocytes, blastocysts and euploid blastocysts were assessed for each diagnosis. Adjusted relative risks (aRR) and 95% confidence intervals (CI) were calculated adjusting for age, BMI, AMH, and stimulation protocol. Poisson regression was used for counts and with an offset for ratios. Patients contributing multiple cycles were accounted for using general estimating equations. Main results and the role of chance PCOS patients were given a lower starting dose of gonadotropins and received less total gonadotropins compared to patients with tubal factor infertility or HH, but had similar stimulation durations as tubal-factor patients. Patients with HH received higher total doses of gonadotropins and had longer stimulation durations. PCOS patients had significantly more oocytes retrieved and a higher number of blastocysts than patients with tubal factor infertility (18.9 vs. 13.6 aRR 1.16 95% CI: 1.05–1.28 and 6.6 vs. 3.7 aRR 1.32 95% CI 1.10–1.57, respectively). Patients with HH had a similar number of oocytes retrieved and number of blastocysts compared to tubal factor patients. The blastocyst conversion rate was higher for PCOS than tubal (59.4% vs. 49.7%), but not significantly different (aRR 1.04 95% CI: 0.94–1.15). Blastocyst conversion and euploidy rates were similar for HH and tubal factor patients (51.9% vs. 49.7% and 39.1% vs. 44.9%, respectively, aRR 1.01 95% CI: 0.81–1.26 and aRR 1.05 95% CI: 0.85–1.31, respectively). In the adjusted model, patients with PCOS had a significantly lower euploidy rate than patients with tubal infertility (aRR 0.75 95% CI: 0.58–0.96). Patients with HH also had a significantly higher euploidy rate compared to women with PCOS (aRR 1.41 95% CI: 1.05–1.89). Limitations, reasons for caution This study is limited by its retrospective nature and the small sample size of women with hypothalamic hypogonadism. Additionally, these data represent outcomes from a single academic center, so generalizability of our findings may be limited. Wider implications of the findings: Cycle outcomes differ for ovulatory dysfunction patients with PCOS as compared to those with HH. HH patients require higher total doses of gonadotropins and longer stimulations to achieve similar cycle outcomes as normo-ovulatory patients. While PCOS patients have more embryos, the percent of euploid blastocysts is lower. Trial registration number Not applicable

Prevention of peritoneal pelvic adhesion formation in women of reproductive age

The objective: a prevention of the peritoneal pelvic adhesions formation in women of reproductive age in the postoperative periodMaterials and methods. 132 women with tubal infertility were examined: І group – main one (n=68) and ІІ group – comparison group (n=64). In the postoperative period, the patients of the main group received treatment and prevention measures according to the developed approach (intraoperative administration of anti-adhesion gel 50 ml and 1 suppository of the tiotriazolin a day rectally from the 1st day after surgery for 14 days), and the comparison group – according to the traditional approach. 30 healthy women were examined as a control group.The level of fibrinogen, fibrinogen /fibrin degradation products (FDP), protein-bound hydroxyproline and free hydroxyproline were studied in blood. Pain syndrome was assessed on a 10-point scale.Results. An increase in fibrinogen and FDP levels in the plasma of all examined women were found. But in the main group the concentration of FDP from 4 hours after intervention was significantly higher (5.31±0.24 mg/l) compared to the comparison group (4.53±0.17 mg/l). The level of protein-bound hydroxyproline in the blood serum of the examined women demonstrated its significant increase in the comparison group 12 hours after surgery, with a maximum after 24 hours and maintaining the concentration in 1.9 times higher after 3 days (p<0.05). The level of free hydroxyproline in the main group was significantly (in 1.4–1.6 times) higher than in the comparison group.The increase of the fibrinolytic capacity (p<0.05) in the peritoneal fluid was found during postoperative period. The period of hyperemia in II group lasted 2 times longer than in I group. Also, II group predominates in the number of patients with hyperthermia for more than 3 days – 10 (15.6 %), compared with I group – 4 patients (5.8 %). The duration of stay in the hospital dominated in II group with a standard treatment regimen in the postoperative period – 3.1 %, compared with I group – 1.9 %.The patients after operation in I group evaluated postoperative pain on average by 1.8 points (p=0.00), in II group – at 3.6 points (p=0.06). Synthetic opioid analgesics and nonsteroidal anti-inflammatory drugs were administered intramuscularly for analgesia. The patients in II group required more injections per patient.Conclusions. The proposed approach is effective, pathogenetically reasonable and can be used as a pathogenetic prevention of adhesions formation in women of reproductive age.