Sleep is the one of the most important vital functions of the body. The study of the primary mechanisms that result in disturbance of the sleep-wake cycle is a priority of modern medical science. It is known that the orexin neurons of the hypothalamus, which send projections to all waking centers, play a leading role in initiating and maintaining this state. Insufficiency of the function of the orexinergic system of the hypothalamus leads to disturbances in the sleep-wake cycle and the development of the severe diseases of narcolepsy and cataplexy. The presented review is devoted to the investigation of possible mechanisms of development of pathological changes in the orexin-containing neurons functions and the diseases caused by them. 

Significant progress in the field of free radical biology and medicine since the middle of the century has been made. Subsequently, the spectrum of diseases and pathological states believed to be associated with free radicals significantly has been expanded. At present, the development of oxidative stress molecularcellular mechanisms investigations is extremely topical for medical science because it involves in a number of pathologies, including social-significance disorders .

The important role of the lipid peroxidation (LPO) system and deficiency of antioxidant defense (AOD) factors in general pathological processes has been proved by authors and their assistants in the laboratories of the Institute of Clinical and Experimental Medicine of Siberian Branch of the Russian Academy of Medical Sciences (Novosibirsk), since 1972. A list of these processes: cold and toxic stress, tumor growth, inflammation, physiologic and pathologic pregnancies (including experimental models). In fact the first clinical investigations were not only in the USSR, but also in the whole world. Several patents proved this.

In the experiments on rodents, the phenomenon of imprinting on the offspring’s LPO and drug metabolism after LPO inducing toxicants action during pregnancy was discovered.

The research series continued at the Scientific Centre for Family Health and Human Reproduction Problems (Irkutsk). Technology for oxidative stress evaluation and the use of antioxidants in the complex therapy of complicated and uncomplicated pregnancy, reproductive disorders in men and women, sleep disorders, sociallysignificant diseases, pediatric practice were created here. The analysis of lipid peroxidation changes during the physiological development of the reproductive function performed. The gender and ethnic dependence of lipid peroxidation was noted. Ethno-specific changes in the LPO-AOD system among the representatives of the Buryat ethnic group at different pathologic conditions (diabetes mellitus type 1, essential arterial hypertension, complicated pregnancy, endocrine infertility, endometriosis, male infertility), were revealed. The LPO-AOD was used to define “relative ethnic norm” for Buryat ethnic group. The “specificity” of features of nonspecific process of free radical oxidation in different pathologic situations was postulated. It is very important for personalized diagnosis, prevention and treatment. 

Antipsychotics are the main drug group in the treatment of disorders of the schizophrenic spectrum. At the same time, this drug group requires long-term use, has a narrow therapeutic breadth and a large spectrum and high severity of undesirable drug reactions, the manifestation of which is largely determined by the patient’s genotype. In this connection, the severity of undesirable drug reactions in drugs of the first and second generation of antipsychotics is different. Atypical antipsychotics, unlike typical antipsychotics, have a significantly lower chance of causing extrapyramidal disorders, though they primarily induce metabolic side effects, consisting of significant weight changes, glucose metabolism dysregulation, and dyslipidemia. These unwanted drug reactions significantly reduce adherence to therapy and can lead to serious complications in the future. At the same time, experience with the use of atypical antipsychotics has shown that the severity of unwanted reactions differs within the group of atypical antipsychotics and, more importantly, even for one drug taken by different patients, is a variable index. The reason for this phenomenon lies is in the polymorphism of the pharmacokinetic and pharmacodynamic factors involved in the metabolism of atypical antipsychotics and the different chemical structure of the drugs. The study of the effect of various combinations of polymorphic variants of the pharmacokinetic and pharmacodynamics factors on the development of antipsychotic-induced metabolic disorders is a complex and high-priority task of personalized medicine. 

The ability to regulate volume in response to changes in intracellular and extracellular osmolarity is one of the fundamental cell functions. Cell volume changes stimulate a variety of intracellular signaling cascades that activate protective and adaptive mechanisms. In general, the regulation of volume occurs due to the transport of osmolytes, which results in the restoration of the intracellular water volume and the normalization of cellular functions. In this review we focused on the physiological significance of regulatory volume decrease (RVD) and regulatory volume increase (RVI) in response to fluctuations of extra- and intracellular osmolarity in the context of epithelial cells. 

According to the current paradigm proposed by Piter Novell [1], carcinogenesis is a process of clonal evolution in which consequent cycles of clonal selection in the adaptive tissue microenvironment give rise to tumors with a variety of genetic and other molecular changes determining the biological behavior of each individual tumor. Selection of clones with different properties provides heterogeneity of cancer cells within one tumor and thus results in a low effectiveness of chemotherapy.

On the average, only 40–60% of cancer patients respond to chemotherapy, and even in the case of complete regression, there is a high probability of tumor recurrence [2, 3]. Increasing the effectiveness of solid tumor therapy and reducing the possibility of recurrence requires not only the use of optimal individual schemes of therapy for each patient, but also the development of combined approaches aimed at both the destruction of tumor cells and antitumor programming of the microenvironment, where immune cells plays a prominent regulatory role. The key cells of the immune system that determine the relationship between tumor cells and the microenvironment, from early stages of tumor growth, including the regulation of neoangiogenesis, and to terminal stage of dissemination of malignant process, are tumor-associated macrophages (TAM) [4– 6]. Identification of the pathways responsible for the tumor-supporting function of macrophages makes it possible to develop therapeutic approaches combining chemotherapy with the macrophage blocking strategy. Inhibition of macrophage infiltration into tumor, their removal with anti-macrophagal agents, and switching off the function of the macrophage colony-stimulating factor can be the macrophage blocking tools. Approaches of simultaneous alteration of cancer stem cells and TAM to abolish chemoresistance and inhibit tumor progression are promising. Strategies for reprogramming of macrophages to switch off towards the antitumor phenotype are developing.

Thus, an extremely wide range of regulatory and effector activity and high functional plasticity of macrophages promise the development of macrophage-targeted therapeutic agents to modulate relationships between tumor and microenvironment to prevent the tumor progression. 

This review and discussion article focuses on the physiology and pathology of connective tissue system and its interaction with mononuclear phagocyte system (MPS) in initiation and development of fibrosis. The study presents the perspective on fibrotic conditions as hypertrophied nonspecific body reactions aimed at maintaining homeostasis and initiated by pathologic (in terms of scale and duration of manifestations) activation of MPS cells resulting from their endocytosis of biocompatible factors of different origin accompanied by nonspecific activation of expression, synthesis, and secretion of lysosomal enzymes and growth factors by MPS cells. This perspective is supported by the authors’ own findings as well as evidence published in studies of other authors. Authors’ findings support and illustrate the above conclusions using data on the modulation of functional status of macrophages and fibroblasts as well as data on prevention and correction of fibrotic processes in different experimental pathologic conditions. 

This review presents current classification, functions of main groups of mitogen-activated protein kinases (MAPK) and summarizes data on the ways of their activation and functioning, giving particular emphasis to p38 MAPK. The authors consider the influence on these signaling cascades as a promising direction for activation of connective tissue growth. This article summarizes international practices on the activation and blocking of intracellular cascades and also the authors’ own experience in this field. In particular, the article shows that p38 MAP-kinase stimulation while JNK inactivation causes accelerated formation of connective tissue in the area of postoperative surgical scar. The authors prove the opportunity to manage connective tissue growth influencing MAPK cascades – prolonged blockade of p38 МАРК reduces scar width and collagen fiber density in the area of postoperative scars and decreases intensity of adhesions in the abdominal cavity in abdominal trauma.

Therefore, considering the importance and flexibility of MAP-kinase mechanisms of cell growth regulation and differentiation, studying the use of these mechanisms in biological processes (such as inflammation, apoptosis, regeneration) and the development the methods of management of these processes show promise. Using stimulators and inhibitors of MAP-kinase mechanisms is a promising new direction in treatment of the diseases with pathogeny related to the disorder of cellular differentiation, proliferation, excessive cytokine production and regulation of connective tissue growth.

This study proposes a theory of immune regulation of regeneration of damaged tissues. An assessment of the role of macrophages, lymphocytes, mast cells, platelets, and endothelial cells in the reconstruction of the structure of the functional element of the defective organ. Discusses the mechanisms of interaction of immune cells during regeneration. Presents the main factors determining the differentiation of stem cells. Apoptosis is considered as one of the components of the recovery process. 

Purpose. Glucose metabolism is tightly regulated in the brain. Aberrant glucose metabolism is an important feature of neurodegenerative diseases, as inAlzheimer’s disease. The transport of glucose to the cell membrane is realized through the activity of insulin-regulated aminopeptidase (IRAP) which controls transfer of glucose transporter to the plasma membrane. IRAP is considered as one of the key markers of insulin resistance in Alzheimer’s disease. However, the question of the mechanism of the action of the IRAP remains open. The aim of the study was to study the effect of IRAP expression on cells of the neuronal and glial lineage, glucose transporter (GLUT4) expression in the brain amygdala on emotional memory in animals with experimental Alzheimer’s disease.

Materials and methods. The study was performed with two experimental models of Alzheimer’s disease in mice. The experimental group was mice of the CD1 line, males aged 4 months (Alzheimer’s disease model with the intra-hippocampal administration of beta-amyloid 1-42 (1 µl) bilaterally in the CA1 area). The control group was mice of the CD1 line, males aged 4 months (sham-operated animals with the intrahippocampal administration of Phosphate buffered salin (1 µl) bilaterally in the CA1). The genetic model of Alzheimer’s disease is the B6SLJ-Tg line mice (APPSwFlLon, PSEN1*M146L*L286V) 6799Vas, males aged 4 months. The control group consisted of C57BL/6xSJL mice, males aged 4 months. Evaluation of emotional memory was carried out using “Fear conditioning” protocol. Expression of molecule-markers of insulin-resistance in the amygdala was studied by immunohistochemistry followed by confocal microscopy.

Results. Aberrant associative learning and emotional memory was revealed in animals with an experimental model of Alzheimer’s disease. A decrease (p ≤ 0,05) of IRAP expression on cells of neuronal and glial nature, associated with GLUT4 down-regulation was detected in amygdala of brain in animals with experimental Alzheimer’s disease.

Conclusion. Decreased number of IRAP-immunopositive neuronal and astroglial cells, as well as IRAP+/ GLUT4+ in cells of amygdala in animals with an experimental model of Alzheimer’s disease, indicates the development of insulin resistance in amygdala of brain, which was in correlation with the hippocampus in performing cognitive functions and memorizing associated with emotionally colored events. 

The purpose of the study is to evaluate the cellular and molecular parameters of bone remodeling in the blood as potential markers of undifferentiated forms of connective tissue dysplasia

Materials and methods. The structural and functional status of cellular elements of in vitro culturing of mononuclear leukocytes of peripheral blood in adolescent athletes connected with phenotypic manifestations of undifferentiated connective tissue dysplasia (UCTD) were investigated. 25 pupils of sport schools from 10–14 years old (main disciplines: figure skating, gymnastics, athletics) were examined with the help of express analysis. The average age of the examined adolescents was (12,0 ± 1,7) years. Clinical examination of adolescents allowed ranking the UCTD signs on a scale of 4–11,5 points.

Results. A comparison of questionnaire survey results and an evaluation of bone remodeling distant markers allowed the revelation of 2 groups in the distribution of adolescent athletes: those with minimal signs of UCTD (scores lesser than 7 points – 10 pupils), and those with expressed UCTD phenotype (scores are equal or more than 7 points –15 pupils). Significant statistical decrease in the content of collagen type I degradation products (CrossLaps) (by 80%) and ionized calcium (by 5%) has been determined in the peripheral blood of adolescent athletes with expressed UCTD phenotype. In conditions of short-term 72-h cultivation of mononuclear leukocytes in the presence of a 3D matrix imitating the properties of the mineral substance of the regenerating bone tissue, morphofunctional features of cellular reaction in adolescent athletes with clinical manifestations of UCTD, as well the heterogeneity of the cell population associated with the appearance of cells with an osteoblast-like phenotype in the blood have been revealed. The results of investigation propose the use of distant cellular and molecular parameters of bone remodeling to screen the mechanisms and dynamics of dysplastic disorders in adolescent athletes. 

The study was aimed at comparative assessment of cardioprotective properties of various necroptosis inhibitors in the isolated perfused rat heart subjected to global ischemia-reperfusion.

Materials and Methods. The study was performed on 38 male Wistar rats weighting 250–300 g. The following necroptosis inhibitors were tested: necrostatin-1 (Nec-1), necrostatin-5 (Nec-5), necrostatin-1s (Nec1s), and necrosulfonamide (NSA). All tested substances were administered intraperitoneally (i.p.) 1 hour prior to heart perfusion. Control animals were treated either with the vehicle (dimethyl sulfoxide, DMSO) or with 0,9% sodium chloride solution (Controls). The dose of necroptosis inhibitors was calculated on the basis of effective concentration (EC50) data. One hour after i.p. injection, the animals were anesthetized, the hearts were rapidly excised, the aorta was cannulated and retrogradely perfused according to Langendorff. After stabilization, the perfusion was stopped for 35 minutes, which was followed by 2 hours of reperfusion. Prior to stabilization, fluid-filled polyethylene balloon was placed into the left ventricle for left ventricular pressure registration. Coronary flow was measured at baseline and during reperfusion by means of perfusate collection. The volume of necrotic myocardium was expressed as a percentage of triphenyltetrazolium chloride-negative tissue relative to the entire heart volume.

Results. The volume of myocardial necrosis and functional heart parameters were not different between Controls and DMSO group. All tested necroptosis inhibitors demonstrated infarct-limiting effect. However, there were no differences between the groups. The volume of necrotic myocardium was (50,5 ± 7,82)%, (29,9 ± 3,42)%, (27,7 ± 3,42)%, (30,6 ± 3,82)%, and (34,7 ± 5,82)% in DMSO, Nec-1, Nec-5, Nec-1s, and NSA groups, respectively (p < 0,01 vs. DMSO group).

Nec-1s and NSA were shown to improve functional recovery of the heart after ischemia. In particular, left ventricular developed pressure and coronary flow rate were higher in Nec-1s and NSA groups (p < 0,01 compared with Controls and DMSO), while end-diastolic pressure was lower in Nec-1s and NSA groups vs. Controls and DMSO (p < 0,01).

Conclusions. It has been shown that Nec-1, Nec-5, Nec-1s, and NSA administration prior to global ischemiareperfusion results in comparable infarct size limitation. In addition to infarct size limitation, Nec-1s and NSA are able to improve postischemic left ventricular function. This fact, along with low toxicity and optimal EC50, makes Nec-1s and NSA perspective candidates for preclinical and clinical development as cardioprotective agents. 

Rationale. According to modern understanding, oxidative stress plays an essential role in the underlying mechanisms for the emergence of type 1 and type 2 diabetes and development of complications in these conditions. The sources of reactive oxygen species in diabetes are protein glycosylation, mitochondrial respiratory chain, membrane-bound NADPH oxidase, and other enzymes. An important enzymatic source of the superoxide anion radical and Н2О2 is xanthine oxidoreductase. Under physiological conditions, it exists mainly in the xanthine dehydrogenase form and may be reversibly or irreversibly converted to xanthine oxidase, following oxidative modification with formation of disulfide bonds in the protein molecule. In rodents, xanthine oxidoreductase in adipose tissue is expressed at a higher rate, as opposed to in other tissues.

The aim of the study was to establish the role of xanthine oxidase in development of oxidative stress in rat adipocytes in experimental type 1 diabetes.

Materials and methods. The study was conducted in male Wistar rats with experimental diabetes induced by two different diabetogenic agents – alloxan and streptozotocin. Serum concentration of insulin in the control and experimental rats was determined by the immunoradiometric assay. The levels of glucose and uric acid were measured by the enzymatic methods. The concentration of lipid hydroperoxides, primary products of lipid peroxidation, was detected by the FOX-2 method. The activity of xanthine oxidase in isolated adipocytes of epididymal adipose tissue and the expression of xanthine dehydrogenase mRNA were determined by fluorometry.

Results. The rats from the experimental group developed hyperglycemia-induced oxidative stress. The rise in the lipid hydroperoxide concentration in adipocytes was observed against the backdrop of the increased xanthine oxidase activity. The boost in the enzymatic activity under oxidative stress in adipocytes of the experimental rats was accompanied by the increase in the proportion of the xanthine oxidase activity in the total xanthine dehydrogenase plus oxidase activity. It resulted in the elevated xanthine oxidase/xanthine dehydrogenase activity ratio. Oxidative stress in rat adipocytes with experimental type 1 diabetes caused oxidative post-translational modification of the enzyme and its conversion from the xanthine dehydrogenase form to the xanthine oxidase one. It resulted in the subsequent increase in reactive oxygen species production. The inhibitor of xanthine oxidase reduced the level of lipid hydroperoxides in rat adipocytes. Thus, xanthine oxidase may be a potential target to protect from oxidative stress in rat adipose tissue in type 1 diabetes.

Conclusions. Oxidative stress in adipose tissue of the rats with alloxan- and streptozotocin-induced diabetes is determined, to a certain extent, by the increased expression of xanthine dehydrogenase mRNA as well as by post-translational oxidative modification of the enzyme activity from dehydrogenase to oxidase. Allopurinol, the inhibitor of xanthine oxidase, decreases the alloxan-induced elevated level of lipid hydroperoxides in rat serum and isolated adipocytes. It indicates a crucial role of xanthine oxidase in development of oxidative stress in adipocytes in experimental type 1 diabetes. 

The paper presents the results of investigation of immunotoxic action of interferon α-2b, immobilized in polyethylene glycol by means of electron-beam synthesis technology.

Materials and methods. In this study we used following methods: a preliminary assessment of immunotoxicity after single administration in wide range of doses, a study of the effect of the drug on the weight and cellularity of central and peripheral immune organs, an assessment of the phagocytic activity of peritoneal macrophages, a study of the effect of the drug on the phagocytic activity of neutrophils, an estimation of the level of serum hemagglutinins to sheep erythrocytes, an evaluation of the effect of the drug on the delayed-type hypersensitivity reaction induced by trinitrobenzenesulfonic acid.

Results. We have shown that the course application of the drug based on immobilized interferon α-2b in tenfold and hundredfold therapeutic doses has no immunotoxic action. At the same time, single administration of the drug at various doses and the therapeutic course application in tenfold dose leads to the stimulation of humoral immune responses. The course application of this drug in tenfold and hundredfold therapeutic dose enhances the phagocytic activity of peritoneal macrophages and peripheral blood neutrophils of experimental animals. 

The study aimed at evaluating the predisposition for development of different variants of SLE based on an inflammatory response in intact mice.

Materials and methods. Low-dose systemic inflammatory response was modeled by the administration of lipopolysaccharide (LPS) of E. coli (strain 111: B4) by intact B6D2F1 recipients in the dose of 10 ng/mouse or 1 μg / mouse. We evaluated the number of leukocytes, neutrophils, and lymphocytes in the blood, the index of neutrophils / lymphocytes (ratio N/L), and the level of free DNA (cf DNA cell free) in dynamics. The first day after LPS injection we induced SLE model: the female B6D2F1 mice were injected 60–70 x 106 spleen cells of the DBA / 2 parent mice twice with interval five days. Three months after the induction of the model, the level of protein in the urine was measured twice: the mice with proteinuria 3 mg/ml and more were assigned to the group SLEnephritis+, and mice with less than 3 mg/ml of protein in the urine were assigned to the group SLEnephritis–.

Results. It was established that administration of LPS does not change the frequency of nephritis in mice. The retrospective analysis of the number of leukocytes, neutrophils, and lymphocytes does not allow the prediction of the development of nephritis. We observed a significant increase in the frequency and the absolute value of the N/L index at week 8 relative to 4 weeks four weeks before the appearance of proteinuria in the group of mice SLEnephritis+ in contrast to mice SLEnephritis– when LPS was administered at a dose of 10 ng/mouse. Mice SLEnephritis+ and SLEnephritis– show different kinetics of the inflammatory response by an increase in the N/L index every subsequent hour relative to the previous one when LPS was administered at a dose of 1 μg/mouse. The dynamics of the cfDNA level is similar to the kinetics of an index N/L in the group SLEnephritis+ and the SLEnephritis–. The data obtained indicate the possibility of using of the index N/L and cfDNA level as a parameter of inflammatory response in mice and prediction of the susceptibility the development of nephritis. 

The purpose of the study was to analyze the expression of proteins-regulators of the cell cycle (p53 and p21) and receptors for endothelial (VEGFR) and epidermal (EGFR) growth factors depending on the expression of galectin-1 in tumor cells in gastric cancer.

Materials and methods. 52 patients with diagnosis of gastric cancer were examined. We used the tissue samples of malignant neoplasms of the stomach, which were obtained during surgical intervention. In tumor tissue the expression of galectin-1, p53 and p21 proteins, and receptors of VEGF and EGF were evaluated by immunohistochemistry. The obtained results were analyzed by statistical methods.

Results. It is shown that in patients with gastric cancer accompanied by overexpression of galectin-1, high expression of mutant protein p53 and EGFR in tumor tissue was recorded significantly more frequently. Low expression of VEGFR was observed in all patients with gastric cancer regardless of the level of expression of galectin-1 by tumor cells. Association of galectin-1 expression with expression of protein p53 and EGF receptor by stomach tumor cells was set.

Conclusion. The relationship between overexpression of galectin-1 and overexpression of mutant p53 protein (not showing antioncogenic properties) and receptor for epidermal growth factor (EGFR) by stomach tumor cells is a negative prognostic criterion of the neoplastic process in gastric cancer. 

Diabetic retinopathy (DR) is one of the leading causes of avoidable blindness worldwide. Chronic low-grade inflammation and abnormal angiogenesis, which is considered to be principal components of DR pathogenesis, is accompanied by imbalance in cytokine production. It was shown that single-nucleotide polymorphisms (SNPs) in the promoters of cytokine genes affect the cytokine production in normal and pathological conditions. Accordingly, we aimed to assess the frequencies of combinations of SNPs from promoters of cytokine genes in type 2 diabetic subjects with and without DR.

Materials and methods: We enrolled 201 caucasian subjects with type 2 diabetes, including 90 ones with non-proliferative or preproliferative retinopathy and 111 subjects without DR. Eight SNPs, located in the gene promoters of TNFA (rs1800630, rs1800629, rs361525), IL1B (rs1143627), IL4 (rs2243250), IL6 (rs1800795) and IL10 (r1800896, rs1800872), were investigated. In groups of patients with and without DR, the frequencies of alleles, genotypes and their combinations, odds ratios and specificity were calculated. The networks between genes and quartiles of glycated hemoglobin A1c (HbA1c) were visualized with Cytoscape.

Results. We revealed 36 combinations of SNPs with different frequencies in the groups of patients with and without DR. In the combinations associated with DR with a specificity >99% homozygous GG genotype in position -174 of IL6 (rs1800795), CT genotype in position -31 of IL1B (rs1143627), СС genotype in position -592 of IL10 (rs1800872), CT genotype in position -590 of IL4 (rs2243250) were the most frequently detected variants. High levels of HbA1c (>9.8%) were associated with combinations of CA genotype in position -863 TNFA (rs1800630) with homozygous variants GG in positions -238 and -308 TNFA (rs1800629, rs361525).

Conclusion. Genetic combinations, associated with DR in type 2 diabetic subjects, include homozygous variants in polymorphic positions of the gene promoters of IL6 (rs1800795), IL1B (rs1143627) and TNFA (rs1800630, rs1800629, rs361525). The combinations of the variants of cytokine genes with a high level of HbA1c can be an important mechanism for realizing of individual genetic predisposition to the development of DR. 

Objective. To analyze the relationship between allelic variants of angiogenesis factor genes and development of genital endometriosis (GE), its clinical evidence and treatment efficacy.

Material and methods. 417 women with laparoscopically verified GE (main group) and 112 women without GE (control group) were examined. Among 358 women in both groups (251 with GE and 107 without GEregions of polymorphic angiogenesis factor genes G-1154A VEGF, 405C VEGF, T-604C KDR and G-735A Ang2 were identified by RFLP-analysis.

Results. It has been established that carriage of separate polymorphic angiogenesis factor genes VEGF G-1154A (allele А and genotypes GA, АА) and G-405C (allele С and genotype GC), KDR T-604C (allele С and genotypes ТС and СС), Ang2 G-735A (allele А and genotypes СА and GА) and their combinations predispose to GE development in women of reproductive age. Carriage of genotype GA of polymorphism G-1154A of VEGF gene predisposes to the development of dysmenorrhea during GE. Pelvic pains and infertility during GE as well as efficacy of hormone treatment for these problems are not associated with carriage of separate polymorphisms of angiogenesis factor genes G-405C and G-1154A of VEGF gene, T-604C of KDR gene, G-735A of Ang2 gene. The most potent combination of polymorphisms of angiogenesis factor genes which predisposes to the development of GE is VEGF1154GА/KDRTС/Ang2АA, while VEGF1154АА/ KDRСС/Ang2GG combination predisposes to the development to endometriosis-associated infertility, and VEGF1154GG/KDRТC/Ang2AA combination predisposes to efficient treatment of infertility during GE.

Conclusion. Polymorphic variants of angiogenesis factor genes G-1154A and 405C VEGF, T-604C KDR, G-735A Ang2 (separately or in their combinations) predispose to GE development. They are associated with its clinical evidence and treatment efficacy. For the formation of risk groups of GE development for primary prevention of GE and effective treatment of endometriosis-associated infertility it is advisable to identify combinations VEGF1154GА/KDRTС/Ang2АA, VEGF1154АА/KDRСС/Ang2GG, and VEGF1154GG/KDRТC/ Ang2AA of angiogenesis factors. 

Purpose of the study. Comparative study of the phenotypic and functional properties of dendritic cells (DC) in groups of patients with rheumatoid arthritis (RA) receiving disease-modifying drugs, or biological drugs and (or) pulse therapy with high doses of glucocorticoids.

Materials and methods. The study included 39 patients with RA and 20 age-appropriate and semihealthy donors. Nineteen patients were treated with standard disease-modifying drugs (BMP) in the form of monotherapy or in combination (group PA1) at the time of the examination, 20 – biological preparations or pulse-therapy with glucocorticoids (group PA2). In the latter case, the examination was carried out for 2–7 days after the last injection of methylprednisolone.

Results. In this study, the properties of DC generated from monocytes under the action of IFN-α (IFN-DK) for RA are described for the first time. It has been established that the general feature of DC in the PA1 and PA2 groups are signs of immaturity of the DC, manifested by increased CD14 expression and a decrease in the proportion of mature (CD14-CD83 +) DC. Despite the differences in DC in the PA1 and PA2 groups, both cell types retain in vitro sensitivity to dexamethasone, the treatment of which leads to a significant inhibition of TNF-α production and a decrease in allostimulant activity of the DC. Thus, IFN-DK in RA patients receiving medical therapy is characterized by the presence of tolerogenic properties, which are most pronounced when used in a program of treatment of biological agents or pulse therapy with corticosteroids. 

The aim of the study was to analyze the influence of glucocorticoid (GC) dexamethasone (Dex) on changes in CD4+ T-cells expressing the surface molecule of activation (CD25, CD71, HLA-DR and CD95) and their ability to produce proinflammatory mediators in cultures of TCR-stimulated CD3+CD45RO+ T-lymphocytes obtained from healthy donors and patients with rheumatoid arthritis in vitro.

Materials and methods. The study included 50 patients and 20 healthy donors. T-cell cultures (CD3+ CD45RO+) were obtained from mononuclear leukocytes of immunomagnetic separation (MACS® technology). As an activator of T-lymphocytes, antibiotic particles with biotinylated antibodies against CD2+, CD3+, CD28+, which simulate the process of costimulation of T cells by antigen-presenting cells, were used. The following concentrations of dexamethasone (2, 8, 16, 32, 64 mg) were used in the experiment. The change in the immunophenotype of T-lymphocytes was analyzed by flow cytofluoometry. The secretion of CD3+CD45RO+ T-cells of proinflammatory cytokines IL-2, IFNγ, TNFα, IL-17 and IL-21 was evaluated by enzyme-linked immunosorbent assay.

Results. The general suppressor effect of Dex on CD3+CD45RO+ T-cell cultures mediated by a decrease in the number of CD4 + T cells expressing activation molecules (CD25) and proliferation (CD71), as well as inhibition of the production of inflammatory mediators: IFNγ, IL-2 and TNFα. It is shown that against the background of TCR activation Dex increases the number of CD4+CD95+HLA-DR+ cells in CD3+CD45RO+ cultures obtained from RA patients and does not change their content in the control. The correlations between the number of proinflammatory factors (IL-17, IL-21 and TNFα) in CD4+CD45RO+CD95+HLA-DR+ T cells in supernatants of cell cultures in RA patients indicate the presence of a pro-inflammatory potential of this population of T cells. We assume that the resistance of CD4+CD45RO+CD95+HLA-DR+ T cells in RA patients to the suppressor effect of GC generally leads to the preservation and enhancement of the functionality of autoreactive cells in the pathogenesis of RA. 

The aim of this research was to investigate the regenerative potential of stem and progenitor cells derived from the pancreas and testes in metabolic disorders.

Materials and methods. The experiments were performing on C57Bl/6 mice. Metabolic disorders (MD) were modeling by streptozotocin and fat diet. Morphological methods were used to assess morphopathological changes in the pancreas and testicular tissue and fertility. We investigated the insulin expression in the islets of Langerhans, CD16 in testes by immunohistochemical methods. We evaluated the blood lipids, level of glucose, inflammatory mediators, testosterone and a glucose-dependent insulinotropic polypeptide levels in biological samples by biochemical methods and ELISA. We used cytometric methods to study the surface markers of stem and progenitor cells, culture methods and transplantation test to investigate the regenerative potential of stem and progenitor cells.

Results. After streptozotocin injection and fatty diet we observed the metabolic imbalance of lipids, testosterone, glucose and insulin resistance in C57BL/6 male mice. The inflammation, type 2 diabetes, asthenoand oligozoospermia were developed and the fertility index was decreased after the metabolic disorders. We observed the increase of oligopotent β-cell precursors (CD45- TER119- CD133+CD49flow) and precursors of hemangiogenesis (CD45- TER119- cKit-1+Flk-1+) count in the pancreas, spermatogonial stem cells (CD117- CD90+ and CD117+CD90+) and precursors of hemangiogenesis (CD45- TER119- cKit-1+Flk-1+) in the testes in MD. Stem and progenitor cells had a high clonal activity and self-renewal capacity, the ability to differentiate into mature cells in vitro, the effective engrafment in injured tissue.

The aim of this investigation is the comparative study of changes in the surface phenotype of lymphocyte in patients with different forms of atopic diseases and latent sensitization.

Materials and methods. The study was conducted on peripheral blood lymphocytes from 22 latent sensitization patients, 30 pollinosis patients, 44 atopic bronchial asthma patients and 36 atopic dermatitis patients. The control group consisted of 26 healthy people.

The results of our studies demonstrate that atopic diseases are different not only in clinical manifestations, but also in mechanisms of disturbances in the functions of the immune system. Comparative study of surface markers of lymphocytes in patients with different forms of atopic diseases revealed a significant increase of surface changes of lymphocyte phenotype according to the severity of the clinical manifestations of the disease. All patients with studied forms of atopy had an increase in content of B-lymphocytes expressing CD72 marker in the peripheral blood and of lymphocytes expressing early activation antigens CD23, CD25, CD71 and adhesion receptor CD54. The developments of pollinosis, atopic bronchial asthma and atopic dermatitis are accompanied by an additional increase levels of lymphocytes expressing the late activation marker HLADR, the CD38+ precursors of plasma cells, and of lymphocytes carrying surface immunoglobulins in peripheral blood. In the blood of patients with atopic disease during exacerbation with evident clinical symptoms revealed a significant increase in all the studied populations and subpopulations of B-lymphocytes. Patients with latent sensitization had increasing blood lymphocytes expressing the CD95 receptor of Fas inducing apoptosis and low content of cells expressing its ligand CD178. Content of CD95+ lymphocytes in peripheral blood at atopic bronchial asthma and atopic dermatitis patients is reduced, and CD178+ lymphocytes increased, reflecting infringement of Fas-dependent apoptosis in severe atopic diseases.

The goal is to study the effects of pro-inflammatory agents on the expression of NLRP3 and the structural integrity of cerebral microvessel endothelial cells in vitro.

Materials and methods. The study was conducted on a culture of cerebral endothelial cells. The cells were isolated from Wistar rat brains. Polyinosinic-polycytidylic acid (RolyI: C) – 20 µg / ml was added to cerebral microvessel endothelial cells. In another series of experiments, cells incubated with cerebrospinal fluid (CSF) obtained from patients with enteroviral meningitis (100 µl) were used as a comparison group. CSF was standardized for protein concentration by the Lowry method. The protein concentration was 1 µg / ml. As a control group, cerebral endothelial cells have been cultured in a standard medium. We used the culture insert for 12-well plates. After 24 and 72 h of culturing, we measured transendothelial electrical resistance (TEER) in the endothelial layer. Expression of NLRP3 inflammasomes was assessed with immunocytochemistry 24 hrs from the beginning of cell culture. We used double indirect immunoenzymatic staining method according to the manufacturer’s protocol. Primary antibodies to NLRP3 (Abcam, USA, ab51952) in 1: 100 dilution, secondary antibodies labeled with Alexa Fluor 488 (Abcam, USA, ab150117) in dilution 1: 200 have been applied. Visualization was performed by confocal laser microscopy microscope Olympus FV10i (Olympus, Japan).

Results. TEER decreased in PolyI:C and CSF-treated cells after 24 hr from the beginning of incubation. After 72 hours, TEER was significantly lower in these groups compare to the control one. NLRP3 expression was maximal in the cells treated with CSF. After incubation of the cells with PolyI:C, NLRP3 expression in cerebral endothelial cells was elevated compare to the control group, but did not reached the level seen in CSF-treated cells.

Conclusion. PolyI:C and CSF obtained from the patients with viral meningitis induce disruption of cerebral microvessels endothelial layer integrity with the corresponding rise in NLRP3 expression in the cells, thereby suggesting mechanism of blood-brain barrier impairment in neuroinfection. 

Purpose of work. To perform a genome-wide association study of loss of heterozygosity (LOH) with monoresistance genes expression during neoadjuvant chemotherapy (NAC) in breast cancer.

Materials and methods. The study involved 68 patients with breast cancer. The tumour stages were IIAIIIB. RNA was extracted from tissue specimens (before and after NAC) using RNeasy Plus mini Kit (Qiagen, Germany). Expression profiling of the RRM1, ERCC1, TOP1, TOP2a, TUBB3, TYMS, BRCA1 genes was carried out using quantitative real-time PCR (qPCR). DNA was extracted from 68 biopsy specimens of tumour tissues using QIAamp DNA mini Kit (Qiagen, Germany). LOH status was detected using microarray analysis using high density DNA-chip manufactured by Affymetrix CytoScanTM HD Array company.

Results. As a result of the study of loss of heterozygosity was evaluated in 13815 genes. The frequency of LOH varied from 0% to 63%. The highest incidence of heterozygosity loss events is characteristic for genes of 16, 17 and the X-chromosome. Our study established that the phenomenon of loss of heterozygosity in monoresistance genes (BRCA1, ERCC1, RRM1, TOP1, TOP2A, TUBB3 and TYMS), is not associated with their level of expression in the tumor. A statistical association has been found between LOH and level of expression of the studied genes in 54 genes. Among them it is necessary to note genes encoding miRNA and «zinc fingers» involved in the regulation of transcription of many genes, transmembrane drug transporters and ion channels, genes of the MAP kinase signaling pathway, and others.

Conclusion. The results of this study allow for the more exact determination of the expression picture of monoresistance genes in the tumor and the indication of new candidate genes which are involved in the regulation of the expression of these genes. Evaluation of the loss of heterozygosity in tumor tissue can be used as an additional criterion for personalizing chemotherapy. 

Aim. The changes of blood cytokine profile in patients with ischemic heart disease (IHD) with different development rates and formation of systemic inflammatory response (SIR) after coronary artery bypass grafting by using cardiopulmonary bypass (CPB) are analyzed in this article.

Materials and methods. The patients with slowly progressive of IHD (20 patients) and rapidly progressive of IHD (20 patients) were examined. The concentration of interleukine (IL) 1β, IL-1ra, IL-4, IL-6, IL-8 and tumor necrosis factor (TNF) α in blood plasma were evaluated by ELISA at patients with IHD before surgery and at 6 and 24 h after surgery.

The results of the study showed that concentration of IL-1β, IL-6, IL-8, TNF-α and IL-1ra in blood plasma increases in patients with IHD of both groups before surgery. The concentration of IL-4 in the blood saved in the normal range before the operation in the case of slow disease progression, but maximum increase in content of proinflammatory (TNF-α, IL-6) and anti-inflammatory (IL-4, IL-1ra) cytokines in the blood and the IL-1ra/IL-1β ratio was detected in a rapidly developing of IHD. It was noticed that after coronary artery bypass grafting in patients with long history case of IHD the content of IL-1β, IL-8, TNF-α, IL-1ra, IL-4 increased with a normalization of the IL-6 concentration in the blood; in patients with a short period of IHD increase of IL-1 concentration and high content of IL-6 are combined with remaining unchanged level of IL1β, IL-8, IL-4 and negative dynamics of the TNF-α concentration in the blood. Thus, the operation in the СРВ in the case of IHD with prolonged course induces the formation of SIR, typical for acute inflammation, and coordinated anti-inflammatory response, and in the case of short period of coronary disease progress this operation causes SIR, characteristic for chronic inflammation, and uncoordinated anti-inflammatory response. 

The issue of sustainability is relevant for all types of businesses and organizations. Long-term development has always been and remains one of the most difficult tasks faced by organizations. The implementation the provisions of international standards ISO series 9000 has proven to be effective. The ISO standards are concentrated on the global experience for sustainable success of organizations. The standards incorporated all the rational that has been accumulated in this field of knowledge and practice. These standards not only eliminate technical barriers in collaboration and have established standardized approaches, but also serve as a valuable source of international experience and ready management solutions. They became a practical guide for the creation of management systems for sustainable development in organizations of different spheres of activity.

Problem and purpose. The article presents the author’s approach to the problem of sustainable development health of the organization. The purpose of this article is to examine the approaches to management for sustainable success of organizations and to describe a model of sustainable development applied in research healthcare institutions providing high-tech medical care.

Methodology. The study used general scientific methods of empirical and theoretical knowledge, general logical methods and techniques and methods of system analysis, comparison, analogy, generalization, the materials research for the development of medical organizations.

The main results of our work are to first develop the technique of complex estimation of activity of the scientific-research institutions of health and deploy key elements of the management system that allows the level of maturity of the management system of the institution to be set in order to identify its strengths and weaknesses, and to identify areas for improvements and innovation, and to set priorities for determining the sequence of action when implementing process improvement. Factors required for use in the management of the organization are critical to sustainable success.

Conclusion. The authors observed that the implementation of the quality management system, continuous its maintenance for 15 years, and its transformation in the management system of sustainable development has improved all key performance indicators of the Novosibirsk NRITO. Positive results are confirmed by the results of self-evaluation management system for key elements, i.e. the determination of the level of maturity and tangible improvement at this level during the period of implementation of the system of sustainable development from the second to level 4 (advanced control system). 

Introduction. One of the main national postulates, including domestic health care, is the thesis that the cadres decide everything. At the same time, in recent years, insufficient attention has been paid to human resources for health in scientific research.

Materials and methods. An analysis of the data provided by the health authorities of the regions of the Siberian Federal District (SFO) for the preparation of materials for the meeting of the Coordination Council on Health of the Interregional Association “Siberian Agreement”.

Results. In general, according to the SFD, the index of the population’s supply of physical persons to physicians is 36,9 per 10 thousand of population, which is somewhat lower than the average Russian indicator (37,2). In general, according to the SFD, the indicator of the population’s availability of doctors for the occupied positions is 56,6 per 10 thousand of the population. In most regions, the part-time ratio exceeds the Russian average (1,4).

In general, in the SFD, the provision of the population with doctors in outpatient care is only 17,2% higher than in inpatient care. The staffing level is quite high (90,4%), but to a large extent it was achieved due to a high level of part-time (1,5). The staffing of doctors in outpatient care is 4,7% lower than in inpatient care. At the same time, the higher staffing of physicians in inpatient care is largely due to part-time work (a factor of 1,7 and 1,4 respectively).

Staffing by district doctors is quite high and amounts to 93,7% with a very low level of intercourse (1,1). The staffing of the specialists in the SFO is also quite high (89,8%), while it was largely achieved due to the high level of part-time (1,5). The staffing of doctors in outpatient care is 4,7% lower than in inpatient care. At the same time, the higher staffing of physicians in inpatient care is largely due to part-time work (a factor of 1,7 and 1,4 respectively).

According to the SFD, the provision of the population with average medical personnel is 124,7 per 10 thousand of the population. The provision of the population with medium-level medical personnel in outpatient and inpatient care is almost the same (56,7 and 56,9 per 10 thousand of the population, respectively). The staffing level of the average medical personnel is quite high (93,7%), to a certain extent, it was achieved through intercourse (1,3). At the same time, the average medical staff in outpatient care is 3,6% lower than in a stationary clinic with practically the same level of part-time (1,2 and 1,3 respectively)

The staffing of nurses in the SFO is quite high (94,4%), to a certain extent, it has also been achieved through part-time work (1,3). At the same time, the average medical staff in outpatient-polyclinic care is 3,6% lower than in the stationary one with practically the same level of part-time (1,2 and 1,3 respectively). In general, according to the SFO, there are 2,5 employed nurses for one doctor of clinical specialties. In inpatient care, the number of nurses per doctor is 3,6, and in the outpatient clinic – 1,9.

Discussion. It seems more expedient to talk not about the absolute shortage of personnel, but about the relative, which, to a large extent, is caused by disproportions in the placement and use of personnel.

Conclusion. By regions, the greatest number of deviations from the mean values for the SFO was revealed by the following parameters: qualification of doctors; Provision by medium-sized medical personnel; Availability of doctors, including medical specialists and staffing of doctors. In staffing in all regions of the SFO, priority is given to inpatient care over outpatient and outpatient care. The adequately high staffing is largely due to the high level of part-time, especially medical specialists. Practically in all regions of the SFD there is not enough high level of qualification of doctors and nurses. In all regions of the SFO, there is a significant shortage of nurses with a sufficiently high level of provision with average medical personnel in general. 

Introduction. An important role in health care management was taken by the service of managing the volume of medical care for this population group and the progressive development of the opportunities of the district doctor and GP for most of the population which requires only general medical assistance while maintaining all the possibilities for the consultant to get this patient’s advice.

Materials and methods. An expert assessment of the organization’s problems, prospects for development and the main directions for optimizing the outpatient care were conducted among the heads of health authorities, state outpatient clinics and the oblast’s top specialists (111 experts).

Results. An expert assessment of the problems and perspectives of the organization of out-patient and polyclinic care in the territorial polyclinics obtained the following results. At present, the health of the population is monitored by: citizens themselves – 23,1%; District doctors – 7,9%, medical specialists – 1,2%, no one follows – 67,8%. At the same time, according to experts, citizens themselves should monitor citizens’ health – 81,3% and district doctors – 18,7%. With a deterioration in health status or an exacerbation of the disease, patients most often turn to: the district doctor – 84,1%; The average medical worker working on the site – 2,7% and the specialist doctor – 13,2%. At the same time, according to experts, patients with worsening health or exacerbation of the disease most often need to contact the local doctor – 88,3% and the average medical worker working on the site – 11,7%.

The share of unreasonable visits is: a district doctor – 28,1%, house calls – 56,4%, doctors-specialists – 37,3%. Thus, according to experts’ estimates, the share of unjustified visits of district doctors is 28,1%, of the doctor’s call at home – 56,4%, and of specialist doctors – 37,3%, which indicates a low level of organization of work of polyclinics. The proportion of visits by patients to district doctors that could be resolved: on pre-hospital admission, is 24,7%, by telephone consultation, is 24,5%. Thus, the share of visits to district doctors, which could be resolved at a pre-hospital reception and by telephone consultation, is almost half (49,2%).

The proportion of visits by patients to medical specialists that could be resolved by telephone consultation: between the district doctor and medical specialists is 11,2%, the number of patients by telephone by medical specialists is 12,4%. Thus, the share of visits by specialist doctors, which could be resolved by telephone consultation between the district doctor and specialist doctors and patient consultations by telephone by specialist doctors, is almost a quarter of all visits (23,6%).

Discussion. The data obtained are in some ways consistent with the results of a number of authors who are optimizing outpatient care in terms of the unreasonableness of visits, the possibility of transferring part of the functions of primary general medical reception to specially trained middle medical personnel, using telephone consultations between patients and between doctors, etc.

Conclusion. The results of the expert assessment of the organization of outpatient care helped to identify some approaches to optimizing its delivery in the territorial polyclinics. This is optimization of the work of district doctors; Transfer of a certain part of the functions performed by district doctors to district nurses and paramedics; The development of a system for counseling patients and district doctors by medical specialists of polyclinics and other medical organizations. 

Subject Index of the 16th Volume.

Authors Index of the 16th Volume.