[Clinical guidelines «Polycystic Ovary Syndrome¼].

Polycystic ovary syndrome (PCOS) is a polygenic endocrine disorder caused by both genetic and epigenetic factors. Depending on the period of a woman's life, the clinical picture, diagnosis, and treatment tactics of the disease are different. PCOS has a complex of reproductive, metabolic and psychological characteristics. The target audience of these clinical recommendations are obstetrician-gynecologists, endocrinologists, general practitioners, general practitioners. In these clinical guidelines, all information is ranked according to the level of persuasiveness of recommendations and the reliability of evidence, depending on the number and quality of studies on this issue.

[The expression of moesin, p21-activated kinase 4 (PAK 4), matrix metalloproteinases (MMP 2, MMP 9), and CD34 in the eutopic and ectopic endometrium in adenomyosis]. / Ékspressiia moézina, p21-aktivirovannoi kinazy 4 (PAK 4), matriksnykh metalloproteinaz (MMR 2, MMR 9) i CD34v éutopicheskom i éktopicheskom éndometrii pri adenomioze.

OBJECTIVE: To investigate the expression of moesin, p21-activated kinase 4 (PAK 4), matrix metalloproteinases (MMP 2, MMP 9), and CD34 in the eutopic and ectopic endometrium in different forms of adenomyosis. MATERIAL AND METHODS: Fifty uteri removed for diffuse adenomyosis and for adenomyomas were examined in reproductive-aged (n=25) and premenopausal (n=25) women. A comparison group included 20 uteri removed for intramural and subserosal fibroids in reproductive-aged and premenopausal women. The investigators performed histological and immunohistochemical (using antibodies to moesin, PAK 4, MMP 2, MMP 9, and CD34) examinations of the eutopic and ectopic endometrium. RESULTS: Different forms of adenomyosis were characterized by the irregular border of the endometrium and myometrium due to that there were multiple foci of ingrowth of the basal layer of the endometrium through the terminal plate into the myometrium. In both diffuse adenomyosis and adenomyomas, the basal layer of the eutopic and ectopic endometrium differed many (3-8.5) times, showing the higher expression of the enzymes in the epithelial and stromal cells, which affected their invasive activity (moesin, PAK 4, MMP 2 and MMP 9), and the increased number of CD34 cells in its stroma. At the same time, there were no statistically significant differences in their expression in the basal layer of the eutopic and ectopic endometrium in diffuse adenomyosis and adenomyomas. CONCLUSION: The findings favor the theory of the pathogenesis of adenomyosis due to the invasion of the eutopic endometrium into the myometrium.

Diagnostic Value of a Group of MicroRNA Genes Hypermethylated in Ovarian Carcinoma.

The study was designed to determine genes of microRNAs hypermethylated in malignant ovarian tumors and to select new diagnostic and prognostic markers of the disease and effective system of markers. Using methyl-specific PCR and a representative sample of 54 ovarian cancer specimens, we determined 5 microRNA genes (MIR-34b/c, MIR-9-1, MIR-124-3, MIR-129-2, and MIR-107) hypermethylated in the majority of tumor samples in comparison with paired samples of histologically unchanged tissue (48-57% vs. 4-19%, p<0.001). Using ROC-analysis, we selected an effective system of 4 markers for diagnosis of ovarian cancer (MIR-9-1, MIR-124-3, MIR-129-2, and MIR-107) characterized by high sensitivity and specificity (up to 87-94% at AUC=0.92) relative to the conventional norm (54 paired samples of histologically unchanged tissue) and absolute norm (18 ovarian tissue samples from subjects who died from non-tumor diseases). It was also shown that methylation of MIR-129-2, MIR-9-1, and MIR-34b/c genes is significantly (p<0.01) correlated with the clinical stage or the presence of metastases. The results indicate that epigenetic modifications of the studied microRNA genes are involved in the pathogenesis and progression of ovarian cancer and attest to their diagnostic and prognostic potential.

Expression of microRNA Potentially Regulated by AhR and CAR in Malignant Tumors of the Endometrium.

We studied microRNA whose expression can be regulated by carcinogenic compounds. Bioinformatic analysis has detected microRNA potentially regulated by xenosensor receptors AhR (miR-28, miR-30c, miR-30e, miR-139, and miR-153) and CAR (miR-29c, miR-31, miR-185, miR-625, and miR-652). Published data indicate that these microRNAs are oncosuppressors, except miR-31 that can act as an oncogene. The expression of these microRNAs in malignant tumors of the endometrium was studied. The expression of the majority of the studied microRNAs, except miR-652, was 2-3-fold below the normal, which confirms their oncosuppressor function and indicates their involvement in the endometrial carcinogenesis and hence, allows considering them as potential markers of the disease.

[Immunohistochemical predictors of recurrent ovarian endometriomas after laparoscopic surgery]. / Immunogistokhimicheskie prediktory retsidivirovaniia éndometrioidnykh kist iaichnikov posle laparoskopicheskogo operativnogo lecheniia.

OBJECTIVE: To investigate the expression of proliferation and apoptotic factors (Ki-67, Bcl-2), inflammatory factors (NF-kß p65, COX-2), adhesion factors (ß-catenin), estrogen (ER-α) and progesterone receptors (PR-α) in ovarian endometrioma (OE) in patients with recurrent OE by an immunohistochemical assay. SUBJECT AND METHODS: This investigation enrolled 48 reproductive-aged patients with OE. According to the course of the disease during a follow-up period of 1.5 years after surgical treatment, the biomaterial obtained from the examined patients was divided into two groups: 1) an OE capsule from 19 patients with recurrent OE (a study group); 2) an OE capsule from 28 patients without recurrent OE (a comparison group). This investigation used histological and immunohistochemical examinations. The histological analysis of the OE capsule was performed following a standard procedure. Their immunohistochemical analysis was carried out using the Tissue-Tek Quick-Ray kit that allows the preparation of paraffin blocks with a large number of tissue samples (tissue microarrays). Antibodies to Ki-67 (clone 30-9, VENTANA), Bcl-2 (clone 124, VENTANA), NF-kß p65 (clone p65, 'Spring Bioscience Corp.'), COX-2 (clone CX-294, Agilent), ß-catenin (clone 14, VENTANA), ER-α (clone SP1, VENTANA), and PR-α (clone 1E2, VENTANA) were also employed in the investigation. The specimens were prepared according to a standard protocol using a Ventana Ultra immunohistostainer. Positive and negative controls were used to correctly carry out immunohistochemical tests. Statistical analysis was performed using the applied statistical analysis programs Statistica 10.0 and Microsoft Excel. RESULTS: The patients with recurrent OE had a significantly decreased expression of Ki-67 (2.86% vs. 9.69%; р=0.044) in the epithelial component of the OE capsule; a significantly lower expression of NF-kß p65 (2.54 vs. 3.5; р=0.0082) and СОХ-2 (0.231 vs. 1.381; р=0.0025) in the stromal component of the OE capsule, a significantly increased expression of ß-catenin (2.5 vs. 1.59; р=0.017) in the stromal component of the OE capsule; a significantly increased expression of PR-α (188.46 vs. 71.15; р=0.028) in the epithelial component of an OE capsule. The expression of ER-α (stromal component, 266 vs. 256.84; p=0.48; epithelial component, 251.54 vs. 233.85, p=0.82) and Bcl-2 (stromal component, 0.33 vs. 0.25; p=0.85; epithelial component, 0.944 vs. 0.625; p=0.31) in the OE capsule is not statistically significantly different between the study patient groups. CONCLUSION: The immunohistochemical difference in the expression of a number of the markers under study can serve as the basis for a further investigation of these markers as predictors of recurrent OE after surgical treatment. Further investigations of these factors will also be able to examine the molecular mechanisms underlying the pathogenesis of recurrent OE, which will make it possible to affect these mechanisms in order to eliminate the fundamental causes of a recurrence of this disease.

Immunohistochemical Features of O6-Methylguanine-DNA Methyltransferase Expression during Ovarian Endometriosis.

A comparative immunohistochemical study for the expression of O6-methylguanine-DNA methyltransferase (MGMT) was performed in tissues of the eutopic endometrium and ovarian endometriosis. The highest level of MGMT expression in eutopic endometrial tissue was observed in epitheliocyte nuclei during the proliferative phase. In regions of endometriosis the expression of MGMT in epitheliocyte nuclei was shown to increase during stages I and II, but decreased in stages III and IV. The progression of endometriosis was accompanied by a gradual increase of study parameters in the nuclei and cytoplasm of stromal cells. These changes reflect the impairment of DNA reparation, which probably serves as a stage in the development and progression of endometriosis.

[Expansion of secretory cells in the fallopian tubal epithelium in the early stages of the pathogenesis of ovarian serous carcinomas]. / Ékspansiia sekretornykh kletok épiteliia matochnoi truby na rannikh étapakh patogeneza seroznykh kartsinom iaichnika.

AIM: to investigate the frequency of the types of fallopian tubal secretory cell expansion (SCE) in diseases of the reproductive organs and to determine the immunophenotype and biological role of the cells in the early stages of the pathogenesis of high-grade ovarian serous carcinomas (HGOSC). SUBJECTS AND METHODS: The investigation enrolled 287 patients with extraovarian diseases and ovarian serous tumors varying in grade, whose fallopian tubes were morphologically and immunohistochemically examined using p53, Ki-67, PAX2, Bcl-2, beta-catenin, and ALDH1 markers. The material was statistically processed applying the Mann-Whitney test and χ2 test. RESULTS: The rate of secretory cell proliferation (SCP) (more than 10 consecutive secretory cells) and that of secretory cell overgrowth (SCO) (more than 30 consecutive secretory cells) increase with age in all investigated reproductive system diseases. The rate of SCP in the corpus fimbriatum of the patients with HGOSC was 5.9 times higher than that in those with extraovarian disease (p<0.01); when comparing the same patient groups, that of SCO was 3.4 times higher (p<0.05). The immunohistochemical characteristics of the investigated lesions (in scores) were as follows: PAX2 was expressed in the intact epithelium (2.8), in SCP (1.3), in SCO (1.2), in serous tubal intraepithelial carcinoma (STIC) (1.0), and in HGOSC (0.9); Bcl-2 was in the intact epithelium (2.2), in SCP (2.1), STIC (0.9), and in HGOSC (0.6), ß-catenin was in the intact epithelium (0.5), in SCP (2.85), in SCO (2.95), in STIC (0.6), and in HGOSC (0.5); ALDH1 was in the intact epithelium (0.5), in SCP (2.91), in SCO (2.92), in STIC (1.2), and in HGOSC (0.6). There were statistically significant differences with a 95% confidence interval (p<0.05) for: 1) PAX2 between the intact epithelium and pathology (fallopian tube lesions and HGOSC); 2) Bcl-2 between the intact epithelium and SCE (SCP and SCO) and between SCE and HGOSC; 3) beta-catenin between the intact epithelium and SCE (SCP and SCO) and between SCE and HGOSC; 4) ALDH1 between the intact epithelium and SCE, between and SCE and STIC, and between STIC and HGOSC. CONCLUSION: SCE was shown to be an independent intraepithelial lesion. The incidence of this abnormality increased with age and significantly differed in the patients with fallopian tubal lesions in extraovarian diseases from that in those with malignant ovarian serous tumors (by 5.3 times), while these groups showed a three-fold difference in SCO. Thus, SCP may serve as a more sensitive marker for the early stages of the pathogenesis of ovarian serous carcinoma. The studied types of SCE demonstrated multiple molecular events (loss of PAX2 expression and increased Bcl-2, beta-catenin, and ALDH1 expressions), some of which underwent considerable changes, by increasing the severity of a pathological process (loss of ALDH1, and beta-catenin, and bcl-2 expressions). Thus, therapeutic exposure in the early stages of pathogenesis may have a few points of application and just several molecules can serve as independent markers for early pathological changes in the fallopian tubal epithelium.

[The molecular mechanisms and morphological manifestations of leiomyoma reduction induced by selective progesterone receptor modulators]. / Molekuliarnye mekhanizmy i morfologicheskie proiavleniia reduktsii leiomiomy pod vozdeistviem selektivnykh moduliatorov retseptorov progesterona.

AIM: to investigate the molecular mechanisms and morphological substrate of reduced uterine leiomyoma in patients receiving the selective progesterone receptor modulator (SPRM) ulipristal acetate for 3 months, by estimating the immunohistochemical expression of the markers steroid receptor coactivator 1 (SRC-1), nuclear receptor corepressor 1 (NCoR-1), ER, PgR, Ki-67, p16, TGF-ß, and VEGF in tumor tissue. SUBJECTS AND METHODS: The investigation enrolled 75 women with uterine leiomyoma, menorrhagias, and anemia. Group 1 included 40 patients who were treated with ulipristal for 3 months, followed by laparoscopic myomectomy. Group 2 consisted of 35 patients who underwent surgery without previous preparation. The intra- and postoperative parameters and molecular and morphological changes in the myomatous nodules were comparatively analyzed in both groups. RESULTS: After 3 months of therapy initiation, menorrhagia completely ceased, myomatous nodules decreased in size (p<0.05), hemoglobin levels were elevated (p<0.01), and total intraoperative blood loss and operative time decreased in all the patients in Group 1. The morphological substrate of partial leiomyoma reduction was leiomyocyte apoptosis and dystrophy, tumor stroma sclerosis and hyalinosis with diminished Ki-67 expression and elevated p16 in the smooth muscle cells, trophic nodular tissue disorders exhibited by vascular wall sclerosis and lower VEGF and TGF-ß expression, and leiomyocyte hormonal reception dysregulation that made itself evident through the reduced expression of SRC-1 with the unchanged expression of PR and ER and the maintained level of NCoR-1. CONCLUSION: The molecular mechanisms of tumor reduction involved the reduced Ki-67 expression and elevated p16, lower VEGF and TGF-ß, diminished SRC-1 expression with the maintained level of PR, ER, and NCoR-1. Overall, this is suggestive of enhanced apoptosis and reduced leiomyoma proliferation and angiogenesis induced by SPRM and indicative of the expediency of using ulipristal acetate as a preoperative agent for organ-sparing surgery in reproductive-aged patients with uterine myoma, menorrhagias, and anemia.

Characteristics of Multipotent Mesenchymal Stromal Cells Isolated from the Endometrium and Endometriosis Lesions of Women with Malformations of the Internal Reproductive Organs.

We isolated and characterized cell cultures from eutopic endometrium and endometriotic lesions of women with malformations of the internal reproductive organs. The cells had fibroblast-like shape and intensively expressed CD90, CD73, CD105, CD44, CD146, and CD117 and were capable of induced adipogenic and osteogenic differentiation in vitro. The obtained cultures exhibited properties of multipotent mesenchymal stromal cells; at the same time, they demonstrated in vitro immunophenotypic differences from cell cultures of eutopic and ectopic endometrium of women without developmental abnormalities, which suggests their functional difference. The cells from eutopic endometrium and from ectopic endometriotic lesions can be used as the model for studying of the etiology and pathogenesis of endometriosis and for testing new drugs for this specific group of patients. Markers CD90 and CD117 were identified as promising molecules for the development of minimally invasive diagnostics of endometriosis based on cell cultures from eutopic endometrium.

[Morphological substrate and pathogenetic mechanisms of pelvic pain syndrome in endometriosis. Part II. Peripheral nerve tissue remodeling in the foci of endometriosis]. / Morfologicheskii substrat i patogeneticheskie mekhanizmy sindroma tazovoi boli pri endometrioze. Chast' II. Remodelirovanie perifericheskoi nervnoi tkani v ochagakh endometrioza.

UNLABELLED: Endometriosis (EM) is morphologically characterized by the development of extrauterine endometrioid heterotopies, the major clinical symptoms of which is chronic pelvic pain, which is a serious problem not only in modern gynecology, but also in public health as a whole. AIM: to investigate neurogenic markers in the foci of EM of various sites and histological structure in women with and without pain syndrome. MATERIAL AND METHODS: The investigation was performed using the operative material (resected segments of the intestine, bladder, rectovaginal septum, and small pelvic peritoneum) obtained from 52 women with an intraoperative and morphologically verified diagnosis of EM and (Group 1) and without (Group 2) pain syndrome. Immunohistochemical examination was made on paraffin-embedded tissue sections in accordance with the standard protocols, by using the antibodies: 1) anti-PGP 9.5 polyclonal rabbit antibodies; 2) mouse anti-human neurofilament (NF) protein monoclonal antibodies (Clone 2F1); 3) mouse anti-nerve growth factor (NGF) monoclonal antibodies; 4) monoclonal mouse anti-human NGF receptor p75 (NGFRp75) antibodies (Dako, Denmark). RESULTS: Our findings demonstrate differences in the expression of PGP 9.5, NFs, NGF, and NGFRp75 in the foci and adjacent tissue in painful and painless EM irrespective of the locations of heterotopies. CONCLUSION: The found molecular features are a manifestation of the remodeling of nerve fibers and nerve endings in the foci of EM and PGP9.5, NGF, and NGFRp75 give rise to nerve fiber neoformation and pain syndrome in EM. At the same time, the immunohistochemical phenotype of EM foci does not depend on their site and reflects the presence or absence of pain syndrome.

Insulin-Like Growth Factors (IGF) and IGF-Binding Proteins (IGFBP) in the Serum of Patients with Ovarian Tumors.

IGF-1, IGF-2, and IGFBP-1,2,3 were assayed in blood serum of patients with malignant ovarian tumors (n=44), borderline ovarian tumors (n=11), and benign ovarian tumors (n=12) as well as in healthy women (n=33). In blood serum of patients with malignant ovarian tumors, the level of IGF-1 was lower and IGFBP-1 was higher than in other groups. In patients with malignant and borderline ovarian tumors, the level of IGFBP-2 was higher than in healthy women and in patients with benign ovarian tumors. There was no correlation between most examined parameters and the clinical and morphological peculiarities of ovarian tumors. The study revealed IGF/IGFBP imbalance in patients with malignant ovarian tumor and showed that IGFBP-2 proved to be a potential diagnostic serological marker w with 90% sensitivity and 90% specificity.

[Nosocomial obstetric infections: Yesterday, today, tomorrow]. / Bol'nichnye akusherskie infektsii: vchera, segodnia, zavtra.

Antibiotic resistance of microorganisms is one of the most acute problems of modern obstetrics. The paper analyzes current antibiotic resistance. It considers the mechanisms of its formation and ways to overcome the resistance.