4 курс / Акушерство и гинекология / Роль_протеина_ALK5_в_профиле_ранних_репродуктивных

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menstruation. The author also evaluated reproductive history (abortions, miscarriages, premature births), the duration of infertility, the number of ART attempts, the type of infertility, the controlled ovarian hyperstimulation protocol, the fertilization method, the total gonadotropin dose, and the serum hCG level 14 days after transfer (IU/L).

Early spontaneous abortion after IVF at 6–8 weeks of gestational age (or 4–6 weeks after embryo transfer) was the criterion for inclusion in the immunohistochemical study of decidual tissue of aborted material for early reproductive losses to detect ALK5 expression (20 patients of the Reproductive Medicine Center). The patients of the Center for Reproductive Medicine underwent anabdominalultrasoundexaminationaftertheembryotransferprocedure3–4weeks after the transfer to verify the presence of an intrauterine pregnancy. If there were no signs of progression within 7 days (hCG growth, fetal heartbeat at ultrasound), a surgical abortion was performed (20 patients). After intravenous anesthesia, vacuum aspiration was performed and the obtained material was sent for examination.

The criterion for inclusion in the control group (20 healthy women) was a planned surgical abortion of a singleton progressive pregnancy that occurred in the natural cycle at their request at 6–8 weeks gestational age without a threat of pathological pregnancy termination.

To obtain reliable data in determining ALK5 expression, the study and comparison groups were selected with regard to their identity according to the following key parameters: age (25–45 years), socioeconomic and marital status, specific features of obstetric and gynecological history, extragenital pathology, etc. The research team obtained women’s informed consent to the use of biological material for scientific purposes.

Exclusion criteria for immunohistochemical studies were the presence of somatic, autoimmune, hereditary genetic, and endocrine diseases, multiple pregnancies, cases of detected congenital anomalies, and women's refusal to use their biological material for scientific purposes.

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were dewaxed, rehydrated, and boiled for antigen extraction using standard procedures. After blocking with 3% (weight/volume) BSA for 1 hour, the sections were incubated with primary antibodies overnight at 4°C. Immunoreactive signals were obtained using the 3,3’-diaminobenzidine (DAB) substrate set.

In the last step, the prognostic and clinical significance of ALK5 expression in the profile of early reproductive loss after ART was evaluated. Differentially expressed genes were determined using a two-tailed t-test (p<0.05) and multiplicity <0.5 and >2, and mapped on a heat map using GenePattern. Gene set enrichment analysis was performed on the KEGG gene sets, and gene sets that met a false discovery rate criterion of <0.25 were selected for further extraction of advanced subsets.

To quantify the results, the micropreparations were photographed in as many non-overlapping fields of view as possible. The pictures were obtained using an x20 lens with a resolution of 1600×1200 pixels, using a Leica microscope and a Leica 425C digital camera. The expression of markers was quantified using the computer program Aperio Image Scope_v9.1.19.1567. The program was calibrated, and after that, the results of immunohistochemical reactions were evaluated based on the indicator “positivity” (in the program interface “positivity”). For each case, the averagepositivityvalueofallimageswascalculatedaccordingtotherulesoffinding the average.

An electronic database of primary data was created using Microsoft Excel 2016 spreadsheets. Statistical data were processed and analyzed using the SPSS Statistics 17.0 software package (Statistical Package for the Social Science).

A descriptive analysis of the results of the study was based on the methods of variance statistics. The study analyzed frequency characteristics (in %) for qualitative parameters. For quantitative parameters, the average levels of indicators were determined, and their variability and statistical significance by means of the arithmetic mean (M) and standard deviation (m) were assessed.

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CHAPTER III. ANALYSIS OF FACTORS ASSOCIATED WITH EARLY REPRODUCTIVE LOSSES IN ASSISTED REPRODUCTIVE TECHNOLOGY

3.1. Etiological risk factors for early reproductive losses in women after assisted reproductive technology based on clinical and statistical analysis

Nowadays, some people who used to struggle with infertility have more therapeutic chances. In recent years, many scientific and technical advances have appeared in the field of infertility treatment. However, the success rates of such methods have shown little progress, despite their use over the past decade. Meanwhile, a significant difference was revealed between a high success rate of ovulation after treatment and a low pregnancy rate. The clinical course of pregnancy in women after ART is the subject of many scientific studies. Pregnancy with high obstetric risk is an understudied issue, and existing publications on this topic do not provide an unequivocal answer regarding the role of various etiological risk factors for early reproductive loss in women after ART. The author studied pregnancy risk factors, including maternal age, BMI, gynecological and somatic diseases, etiology of infertility, response to stimulation, quality and number of embryos replaced, and the type of treatment.

The ages of the women included in the study ranged from 25 to 40 years old (Table 1), with 5% in the study group (SG) and 15% in the control group (CG) aged 25–29 years old. There were 55% of women aged 30 to 34 years old in the SG and 68.33% in the CG. There were 23.3% of women aged 35 to 40 in the SG and 15% in the CG. There were 16.67% of women aged 41 to 45 years old in the SG and 1.67% in the CG.

Table 1

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Table 2 Distribution of women who were included in the retrospective clinical and

statistical analysis by their BMI

Analysis of the data showed that in most of the women included in the study, BMI was normal (18.5–24.9 kg/m2). Every 10th woman was underweight (less than 18.5 kg/m2), and every 3rd woman was overweight (25–29.9 kg/m2). There were cases of firstand second-degree obesity (30.0–34.9 and 35.0–39.9 kg/m2, respectively).

Thus, BMI <20 kg/m2 was recorded in 7.5% of the women who participated in the study, of which 6.67% were in the SG and 8.33% were in the CG. BMI 20– 26 kg/m2 was observed in 54.2% of the studied women, 53.3% of them were in the SG and 55% were in the CG. BMI 26–30 kg/m2 was revealed in 40% of the studied

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The author could not find any statistically significant differences in the direction of professional affiliation and type of labor activity between the SG and CG. It should be noted that the larger share of patients in both groups were engaged in intellectual work (employees, doctors, researchers, and employees of state institutions). There is also a trend towards a larger share of women who were engaged in physical work among SG patients, a statistically significant level of indicators between the main and control groups was not reached.

It was impossible to analyze the presence of bad habits, because this information was not available in the medical records of the studied women.

The analysis of the menstrual cycle of the patients of the study groups showed that the mean age of menarche was 12.7±1.1 years old in the women of the SG as comparedto11.8±1.3yearsintheCG(p>0.05).The menstrualcyclewasestablished immediately in the vast majority of the women included in the study (p>0.05). The study of the functional features of the menstrual cycle did not reveal a direct correlation between the nature of its disorders and the occurrence of early reproductive losses after the use of the therapeutic programs of ART.