Search

Background. Core biopsy of the breast is currently considered to be the standard method of obtaining material for morphological and molecular genetic methods. Unfortunately, this method is associated with a number of problems, primarily the risk of complications (bleeding, pneumothorax) and discomfort during manipulation.Aim. To analyze transcriptional signatures of breast tissue samples obtained by fine-needle aspiration biopsy. Materials and methods. Using reverse transcriptase polymerase chain reaction, we studied the mRNA expression level of 60 target genes in 60 samples obtained by fine-needle aspiration biopsy and in 60 corresponding formalin-fixed paraffin-embedded (FFPE) surgical specimens of breast. Samples were obtained from the tumor, adjacent tissue, the so-called tumor bed and formally normal tissue at a distance from the primary lesion.Results. A comparative analysis of transcriptional signatures in samples obtained by fine-needle aspiration biopsy and FFPE specimens (120 samples in total) reveled the strongest correlations between transcriptional signatures in biopsy samples and FFPE specimens of tumors. Very strong correlation in tumor samples was established for one gene (CTSL2); strong for 18 genes (MKI67, MYBL2, NAT1, PTEN, TPX2, PTTG1, UBE2T, CCNB1, ESR1, CCND1, MYC, SCGB2A2, MIA, TRAC, FGFR4, ANLN, GSTM1, PRLR); averages for 28 genes (PGR, AURCA, KRT5, FOXA1, SFRP1, EMSY, EXO1, PAK1, KIF14, ERBB2, MMP11, BCL2, BAG1, TMEM45B, BIRC5, CD274/PDL1, ZNF703, TYMS, CCNE1, TPT1, TMEM45A, BRCA1, BRCA2, ESR2, STS, TNFSF11/RANKL, TNFRSF11B/OPG, TNF); weak for 4 genes (GRB7, EGFR, PGRMC1, CYP19A). The presence of correlations between transcriptional signatures in biopsy samples and FFPE specimens can be established in case of sufficient material corresponding to sample intake control (SIC) ≥5 lg for B2M gene.Conclusion. The ability to conduct molecular genetic research on small samples of breast tissue makes it possible to obtain the material using the most minimally invasive method. And this, in turn, expands the possibilities of “genetic monitoring” of cancer, as well as the possibility of more accurate assessment the risks of malignant tumor development in the settings of benign conditions in women with fibrocystic disease and increased mammographic density.

Background. Atypical ductal hyperplasia is a relatively common breast lesion that increases the risk of breast cancer by 3.5 to 5 times. Genomic rearrangements underlying the development of atypical proliferative lesions and breast cancer lead to gene expression changings.Aim. To determine the mRNA expression profile of neoplasia and oncogenesis driver genes in atypical ductal hyperplasia of the breast.Materials and methods. The real-time reverse transcription polymerase chain reaction was used to explore the expression profile of 46 genes in 107 samples of formalin-fixed paraffin-embedded (FFPE) specimens from atypical ductal hyperplasia, ductal hyperplasia without atypia, ductal carcinoma in situ and normal breast tissue.Results. In atypical ductal hyperplasia, we detected changes in the expression of 22 of 46 studied genes, including ESR1, AR, PRLR, FGFR4, MKI67, CCNB1, KIF14, PAK1, MMP11, GATA3, FOXA1, ZNF703, which were upregulated, and MYC, which was downregulated.Conclusion. The transcriptional signature of atypical ductal hyperplasia was similar to that of ductal carcinoma in situ and breast cancer of luminal subtypes.

In the context of the COVID-19 pandemic, scientific interest is growing in studying the impact of the proposed vaccination on women’s reproductive health. As is known, alterations in the state of the immune system and activation of an autoimmune response can lead to reproductive failure in women and potential complications of subsequent pregnancy. Objective: to evaluate the effect of the “Gam-COVID-Vac” on the immune status parameters, the relationship of their changes and the specific immune response to vaccination with the dynamics of the level of autoantibodies in women of reproductive age.The prospective study included 120 women who were vaccinated against COVID-19 with the “Gam-COVIDVac”. The criteria for inclusion in the study were: the age from 18 to 49 years, the absence of COVID-19 in the anamnesis, a negative result of a study on SARS-CoV-2 by PCR and negative results of tests for antibodies of classes G and M to SARS-CoV-2 before vaccination, the absence of pregnancy and serious somatic diseases. The patients were examined twice: immediately before vaccination and 90-100 days after the introduction of the 1st component of the vaccine. The level of IgG antibodies to SARS-CoV-2 after vaccination was assessed using ELISA. Before and after vaccination, the levels of antiphospholipid, anti-nuclear, organ-specific and antihormonal autoantibodies were determined, peripheral blood lymphocytes were immunophenotyped to determine the main subpopulations (CD3, CD4, CD8, CD19, CD5, CD16, CD56), as well as the expression of activation markers of lymphocytes (HLA-DR, CD25, CD147) using monoclonal antibodies.The effectiveness and safety of the combined vector vaccine against COVID-19 were high. Specific IgG antibodies to SARS-CoV-2 were produced in 98.3% of vaccinated women, no serious adverse reactions were observed. After vaccination, there was an increase in the level of some autoantibodies within the reference ranges, only IgM antibodies to phosphatidylethanolamine (PE) and IgG antibodies to DNA increased above the reference values. However, this increase was transient. After vaccination, the following changes in the parameters of the immunogram were observed: an increase in the content of cells with CD3+CD25+, CD19+ phenotype in peripheral blood and a decrease in the content of cells with CD56+CD16+ phenotype within the reference ranges, a decrease in CD147+/CD3+. Weak correlations were noted between these changes in immunogram parameters and the levels of some autoantibodies. The specific antiviral immune response to vaccination did not correlate with the autoimmune response.Vaccination with “Gam-COVID-Vac” is effective and safe and does not lead to disorders in the immune system. The observed increase in the level of autoantibodies to PE and DNA is transient. Changes in the parameters of the immune status within the reference ranges may be due to vaccination and the development of a specific antiviral immune response.

Introduction: The specialists refer unexplained infertility to the so-called diagnosis of exclusion due to the fact that in the process of medical examination of married couples the causes of infertility cannot be established. Material and methods: The clinical characteristics and embryological features of in vitro fertilization programs of couples with unexplained infertility versus the patients with tuboperitoneal infertility were analyzed retrospectively and prospectively. The study group comprised 93 women, who underwent 108 in vitro fertilization programs, and the control group consisted of 45 patients, who underwent 49 in vitro fertilization programs. Results: Significant differences (p<0.05) were found between the groups in anamnestic, clinical, laboratory, and instrumental characteristics. The ovarian stimulation protocols were comparable between the groups of patients. The blastulation rate was considered to be the endpoint in in vitro fertilization programs, and it was significantly lower in the group of women with unexplained infertility (45.53%). In-depth analysis of the embryonic stage of in vitro fertilization programs showed, that low blastulation rate in unexplained infertility was mainly due to the fact that embryos stopped developing about three days after they were cultured. At the same time the morphological assessment showed that the quality of blastocysts was higher in the group of unexplained infertility (66.7%) compared to the group of tuboperitoneal factor of infertility (45.8%). Preimplantation genetic testing for aneuploidy showed similar frequency of detection of euploid embryos (41.7% and 40.0%, respectively). Conclusion: A "clinical portrait" of women with unexplained infertility was described. The low blastulation rate was noted in in vitro fertilization programs for women with unexplained infertility. Given the identified impairments of early embryonic development in unexplained infertility, it is appropriate to recommend the patients to undergo early use of in vitro fertilization with good-quality embryo transfer (>3, AA, AB, BA according to Gardner grading system) on day 5-6 of culture without long-lasting preliminary examination and empirical treatment. [ABSTRACT FROM AUTHOR]

Epithelial ovarian cancer (OC) ranks first in the number of deaths among diseases of the female reproductive organs. Identification of OC at early stages is highly beneficial for the treatment but is highly challenging due to the asymptomatic or low-symptom disease development. In this study, lipid extracts of venous blood samples from 41 female volunteers, including 28 therapy-naive patients with histologically verified high-grade serous ovarian cancer at different stages (5 patients with I-II stages; 23 patients with III-IV stages) and 13 apparently healthy women of reproductive age, were profiled by high-performance liquid chromatography mass spectrometry (HPLC-MS). Based on MS signals of 128 differential lipid species with statistically significant level variation between the OC patients and control group, an OPLS-DA model was developed for the recognition of OC with 100% sensitivity and specificity R2 = 0.87 and Q2 = 0.80. The second OPLS-DA model was developed for the differentiation between I-II OC stages and control group with R2 = 0.97 and Q2 = 0.86 based on the signal levels of 108 differential lipid species. The third OPLS-DA model was developed for the differentiation between I-II OC stages and III-IV stages based on the signal levels of 99 differential lipid species. Various lipid classes (diglycerides, triglycerides, phosphatidylchlorines, ethanolamines, sphingomyelins, ceramides, phosphatidylcholines and phosphoinositols) in blood plasma samples display distinctly characteristic profiles in I-II OC, which indicates the possibility of their use as marker oncolipids in diagnostic molecular panels of early OC stages. Our results suggest that lipid profiling by HPLC-MS can improve identification of early-stage OC and thus increase the efficiency of treatment.

Aim. To study long-term results of surgical treatment of patients with familial adenomatous polyposis (FAP) with the cell reconstruction of the rectal mucosa.Materials and methods. 57 FAP patients were subjected to treatment, which involved colproctectomy, the preservation of the lower rectal ampulla, mucosectomy and the reconstruction of the mucosa by cell transplantation. Endoscopic monitoring was carried out, with the endoscopic observation covering the period of 19–120 months (median — 44.3 months). Morphological and immunohistochemical studies were conducted. The long-term functional results of treatment (anorectal manometry (profilometry)) were studied. The patients were surveyed using the SF-36 questionnaire to monitor the quality of their life.Results. Our results show that the use of cell transplantation leads to the reconstruction of the rectal mucosa over a fairly short time: in 44/57 (77.2 %) patients, the endoscopic picture corresponded to the unchanged rectal mucosa 4 weeks after the surgery. In 13/57 (22.8 %) patients, a complete mucosal reconstruction was achieved 8–12 weeks after the surgery. The absence of polyp growth in the preserved part of the rectum was observed. Late complications developed only in 5 (9.4 %) patients. Good functional results (acceptable frequency of defecation, lack of signs of anal incontinence and nocturnal defecation) were observed in 48/53 (90.6 %) patients. The quality of life was at a fairly high level in 90.6 % of patients.Conclusion. The proposed method of FAP treatment allowed the immediate and long-term treatment results to be improved significantly.

Aim. Comparative evaluation of species identification of microorganisms by MALDI-TOF mass-spectrometry and automatic biochemical analyzer VITEK2 Compact30. Materials and methods. Species identification of18 400 isolates of microorganisms (staphylococci, streptococci, enterococci, enterobacteria, nonfermenting gram-negative bacteria, lactobacilli, anaerobes, yeast fungi, neisseriae), isolated from vagina of pregnant and non-pregnant women and from newborns, was carried out. Identification of the isolated microorganisms was carried out by automatic bac-teriologic analyzer VITEK2 Compact30 (BioMerieux, France) and MALDI-TOF-MS analysis method on AutoflexIII (Bruker Daltonics, Germany) mass-spectrometer. Results. Comparative identification of 2005 isolates of microorganisms was carried out. Sequencing of ribosomal RNA was used as a reference method. Authenticity of species identification my MALDI-TOF-MS analysis method was: for staphylococci (95.8%), enterococci (97.5%), enterobacteria (98.4%), nonfermenting gram-negative bacteria (93.6%), P-hemolytic staphylococci (93.8%), lactobacilli (92.8%), yeast fungi (99.9%). Conclusion. Introduction of MALDI-TOF-MS analysis technology into practical work of microbiological laboratories exceeds previously used methods of microbiological testing in terms of speed, cost and authenticity of identification of a wide spectrum of microorganisms.

Purpose. To study the presence of carotenoids in the tissues of prenatal human eye in the course of its development. Material and methods. The eyes were obtained from abortive human fetuses of 12-28 weeks of pregnancy. The fetuses were brought to the Kulakov Research Center of Obstetrics, Gynecology, and Perinatology, the Russian Academy of Medical Sciences, from licensed institutions of the Ministry of Public Health, working under the Russian Federation legislation on citizen health protection, and in accordance with the approved list of medical indications. The ages of the fetuses were determined by an obstetrician. Totally, 49 samples of eye tissues from human fetuses at different pregnancy ages were used. The content of carotenoids and their oxidized forms was measured by high-performance liquid chromatography (HPLC) technique. Results. The presence of lutein and/or its oxidized forms was estimated by HPLC in the tissues of prenatal human eyes (cornea, sclera, optic nerve, retinal pigment epithelium with choroid, ciliary body with iris, vitreous body). Conclusion. The finding of lutein or oxidized forms of lutein in tissues of the human eye in the course of prenatal development is of essential interests for understanding physiology of the prenatal and postnatal eye.

The autoimmune reactions against sperm, which are accompanied by the elaboration of antisperm antibodies (AsAb), are one of the causes of male infertility.Objective: to specify the role of different factors (other than obstruction) in the etiology of male immune infertility.Subjects and methods. Clinical and laboratory studies were made in 536 males from infertile couples (their age 18–45 years); a control group comprised fertile men whose wives were 8–16 weeks pregnant (n = 82). Their sperms were examined in accordance with the WHO recommendations. AsAbs in the sperm were determined by a MAR test. Oxidative stress was estimated using luminol-dependent chemiluminescence. Chromosome damage was identified from DNA fragmentation by chromatin dispersion in an inert agarose gel, by making a microscopic examination of halo formation after acid DNA denaturation and nuclear protein lysis. The blood levels of interferons (IFN) and their natural and in vitro induced production were determined by the Campbell method. Reproductive tract infections were diagnosed by polymerase chain reaction.Results. There is a high significant correlation between the quantity of AsAb, on the one hand, and previous orchitis, as well as subclinical testicular injury, on the other hand. There was no correlation between varicocele and AsAb, but in the former, the risk of immune infertility and orchitis increases after injury; varicocelectomy promotes a reduction in AsAb with the higher degree of varicocele and a less marked autoimmune process. AsAbs are observed during Chlamydia infection with the increased production of IFN-γ. Cryptorchidism and orchiopexy, parotitis, epididymitis, herniotomy, chronic bacterial prostatitis, and other potential risk factors for decreased male fertility had no significant impacts on an odds ratio for developing immune infertility. In the majority (41 %) of cases, immune infertility seems to be idiopathic, but it is accompanied by the hyperproduction of reactive oxygen species and DNA damage (fragmentation).Conclusion. Male immune infertility is multietiological: blood-testis barrier damage, cross reactions with microorganisms, and immunopathological reactions. Autoimmune reactions against sperm can be considered to be both a cause and a sequence of the bad semen quality caused by genetic and different endo- and exogenous factors.

Aim of investigation. To study pathogenic mechanisms and morphological signs of rectal mucosa reparation after rectal mucosectomy and application of cellbased biotechnologies at surgical treatment of familial adenomatosis coli (FAC).Material and methods. The new method of FAC surgical treatment which provides preservation of part of the mucosa-deprived rectum is presented. For reconstruction of mucosa of the rectum cell-based allotransplantation was applied. Morphological mechanisms of mucosal reparation were investigated in 34 FAC patients (main and control groups).Results. The reparation of mucosa in demucosated rectum at application of cell-based transplantation develops in shorter terms in comparison to control group. It is carried out by largeintestinal, smallintestinal types, by squamous cell differentiation and by combination of all types. At reparation cells with signs of stem cells take part.Conclusions. Original method can be applied for treatment of FAC as it delays polyps and rectal cancer development for many years.

AIM: To compare the efficiency of using different interventional procedures for a barren marriage in case of secretory azoospermia (SAS)/MATERIAL AND METHODS: The trial enrolled 110 patients aged 25 to 54 years with SAS. The levels of sexual hormones were determined; ovarian punch biopsy was performed/RESULTS: Decreased volume of the ovaries was noted in 85.3% of the patients; diffuse or focal changes in their parenchyma were seen in 65.7%. There was an increase of follicle-stimulating hormone levels and a reduction in testosterone ones. Hyperestrogenemia was identified in 26.5% of the patients. The level of inhibin B was lower in all the patients. The efficiency of standard testicular sperm extraction for cryopreservation was achieved in 28.7% of the patients and that of a microsurgical technique was in 56.7%. The wives of 37 out of the 57 patients included in an assisted reproductive technology program were more than 12 weeks pregnant after 95 intracytoplasmic sperm injection cycles/CONCLUSION: The microsurgical technique is most effective in achieving the successful treatment of patients with azoospermia in order to prevent male infertility.

Цель: Изучить эффективность золедроновой кислоты в терапии постменопаузального остеопороза (ПМО) с учетом полиморфизма гена фарнезилдифосфат-синтетазы (FDPS). Методы исследования: клинический, определение биохимических маркеров костного метаболизма (БМКР) - электрохемилюминесцентным методом («Хоффманн-Ла Рош ЛТД», Швейцария). Измерение МПК проводили с помощью двухэнергетической рентгеновской абсорбциометрии позвонков поясничной области (L1-L4) и шейки бедренной кости (Neck left, Neck total left). Молекулярно-генетический (определение однонуклеотидного полиморфизма гена FDPS (rs2297480) - методом «примыкающих проб» с анализом кривых плавления (ЗАО «НПФ ДНК-Технология, Россия)). Результаты. Обследовано 225 женщин с постменопаузальным остеопорозом. Средний возраст составил 59 (5466) лет. Средняя продолжительность периода постменопаузы 7 (2-13) лет. ИМТ=27,2(23,6-29,05) кг/м2. Распределение аллелей (АА:АС:СС) составило 55,1(n=144):39,5(n=103):5, 4(n=14). Базальные уровни биохимических маркеров костного ремоделирования (БМКР) соответствовали периоду постменопаузы и статистически значимо не различались в зависимости от генотипа FDPS. Средние показатели МПК в поясничном отделе позвоночника и/или шейке бедра соответствовали остеопорозу. Всем пациенткам проводили терапию золедроновой кислотой (zol) в дозе 5 мг в виде внутривенной инфузии 1 раз в 12 месяцев в течение 2 лет. Дополнительно все пациенты получали кальций 1000 мг/сут и витамин Д3 800МЕ/сут. Реакция БМКР на инфузию zol различалась в зависимости от генотипа FDPS. Через 6 месяцев после 1-ой инфузии достоверно более выраженное снижение B-Crosslaps 92 (91-93)% отмечено у пациентов с С/С генотипом (p=0,056), по сравнению с 82% у носителей аллеля А (p=0.07). Через 9 месяцев после инфузии B-Crosslaps у носителей аллеля А повышались до 75%, остеокальцина - 44,4% от исходного. Тогда как у носителей генотипа C/C как B-Crosslaps - (87%), так и osteocalcine -(53,6%) оставались по-прежнему низкими (р=0,056). К 12 месяцу после инфузии уровни B-Crosslaps и osteocalcin у носителей аллеля A повышались и соответствовали премено-паузальным значениям, тогда как у носителей С/С генотипа оставались в пределах ниже нижней границы нормы для пременопаузы. У пациентов с СС генотипом на фоне выраженного снижения БМКР после 2ой инфузии золедроновой кислоты произошло снижение МПК на 5,6%. Выводы. Различная реакция на терапию золедроновой кислотой у пациенток с ПМО связана с полиморфизмом гена FDPS (rs2297480). Для пациентов с генотипом С/С характерно «гиперторможение» костного обмена в ответ на терапию золедроновой кислотой, что возможно, было причиной отрицательной динамики МПК в поясничном отделе позвоночника на втором году терапии золедроновой кислотой.

Aim of investigation. To develop reconstructed mucosa of the rectum for improvement results of surgical treatment of familial adenomatosis polyposis coli (FAP).Material and methods. Overall 27 patients with FAP underwent surgical treatment including colectomy, demucosation of rectum and allogenic transplantation of fetal intestinal epithelium cells and mesenchyma of various origin to develop reconstructed mucosa. In 5 patients the ileorectal anastomosis, in 22 – small-intestinal reservoirs were created.Results. During period from 4 to 12 wks development of rectal mucosa was observed in all patients. At follow-up from 1 year to 5 years no proliferation of polyps was revealed in 26 (96,3%) patients.Conclusions. Cell transplantation can successfully be applied for development of reconstructed rectal mucosa. Keeping part of rectum after colectomy and formation of smallintestinal reservoir allows to keep natural intestinal passage with good functional results.

In open prospective study we determine the efficacy of zoledronic and ibandronic acids on bone metabolism markers and bone mineral density in women with postmenopausal osteoporosis. 225 women with postmenopausal osteoporosis treated with zoledronic acid in dose 5 mg, intravenously, once a year and 64 women treated ibandronic acid were followed up. The investigators assessed clinical and medical history data, evaluated the gynecological and somatic statuses, and made laboratory and instrumental studies: clinical, biochemical blood tests, general urinalysis, determination of serum bone metabolism markers (B-Crosslaps, osteocalcin), osteodensitometry and electrocardiography, therapist ’s examination if needed. Therapy with zoledronic acid 5 mg once a year and ibandronic acid 150 mg once a month was followed by a significant reduction in resorption markers (B-Crosslaps) just following one month. Formation markers decreased to a lesser degree, which was indicative of preserved positive bone remodeling balance during the therapy. Determination of biochemical markers of bone remodeling permits assessment of an individual patient response to antiresorbtive therapy just in the first month after infusion. Zoledronic acid or ibandronic acid therapy exerts a rapid and potent effect on bone metabolism, mainly by inhibiting the resorption processes.

Дидрогестерон – высокоселективный прогестаген, использующийся при широком спектре гинекологических заболеваний. Из всех стероидных рецепторов дидрогестерон значительно взаимодействует только с прогестерон-рецепторами. Результаты проведенного нами ранее исследования моделей структур комплекса прогестин–рецептор позволяют предположить, что ключевым моментом высокой селективности и специфичности дидрогестерона является уникальное для этой молекулы позиционирование метильной группы С19. Некоторые другие препараты прогестинов взаимодействуют и с другими типами рецепторов и как следствие приводят к нежелательным побочным эффектам. Побочными эффектами применения некоторых прогестинов являются вирилизация, являющаяся результатом активации андрогенных рецепторов, а также нежелательная активация эстрогеновых рецепторов. В настоящей работе мы приводим результаты полногеномного исследования транскрипционных эффектов активации прогестероновых, андрогенных и эстрогеновых рецепторов. Результаты показывают, что эти три типа рецепторов активируют существенно разные группы генов. Путем анализа функционального связывания установлены гены, предположительно влияющие на вирилизацию или феминизацию плода. Результаты исследования подчеркивают, что использование разновидностей прогестинов, побочно активирующих эстрогеновые или андрогеновые рецепторы, нежелательно при беременности и вмешивается в гормональные процессы развития плода. И, наоборот, применение во время беременности высокоселективных прогестинов (например, дидрогестерона) является безопасным для правильного развития плода как женского, так и мужского пола.

The paper considers the late sequels of preeclampsia that is one of the main causes of poor perinatal outcomes. The idea that this gestational complication shows no noticeable impact on the female health status in the late postpartum periods has long been prevalent. At the same time, recent studies suggest that this cohort of women will have a high cardiovascular risk in future life, which requires the formulation of their management strategy. Epidemiological data are given on late female morbidity and mortality due to coronary heart disease, stroke, and renal failure.

Пациентки, страдающие привычным невынашиванием беременности, составляют группу риска по реализации нарушений имплантации и плацентации в I триместре беременности. Подобные нарушения чаще всего обусловлены иммунными взаимодействиями между эндометрием и клетками развивающегося трофобласта [1–3]. Женщины, которым проводится экстракорпоральное оплодотворение (ЭКО), как правило, имеют гормональные проблемы, страдают хроническими воспалительными заболеваниями и как следствие – спаечными процессами в области малого таза, нередко перенесли одно или несколько оперативных вмешательств на матке, яичниках и трубах. При этом трудности с наступлением беременности, решаемые благодаря использованию вспомогательных репродуктивных технологий, сочетаются с иммунологическими проблемами, что в ряде случаев ведет к повторным ранним репродуктивным потерям. У пациенток с повторными потерями плода формируется так называемый порочный круг, когда иммунопатологический процесс активируется с самых ранних этапов развития беременности