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Siberian Scientific Medical Journal

2019 year, number 4

1.
MORPHOFUNCTIONAL CHARACTERISTICS OF MITOCHONDRIA AND IMMUNOHISTOCHEMICAL APPROACHES TO THEIR STUDY

Lizaveta Igorevna BON, Nataliya Yevgenyevna MAKSIMOVICH
Grodno State Medical University
Keywords: митохондрии, иммуногистохимические маркеры, энергетика клетки, mitochondria, immunohistochemical markers, cell energy

Abstract >>
Violation of the functions of mitochondria is accompanied by any disease, so further study of the functional characteristics of mitochondria in various pathologies in the clinic and experiment, as well as the search for new diagnostic markers is promising and relevant. The purpose of this review is to summarize and systematize the literature data on morphofunctional characteristics and molecular immunohistochemical markers used to assess the function of mitochondria. The most characteristic feature of these organelles is the presence of a large number of enzymes involved in oxidative phosphorylation and energy supply to the cell. Also, the aim was to allocate so-called mitochondrial diseases associated with genetic, structural, biochemical defects of mitochondria, including those leading to energy deficiency of cells. Mitochondrial diseases are transmitted through the female line, since only the ovum contains mitochondria. Hereditary mitochondrial diseases associated with mutations in the genes encoding the synthesis of mitochondrial proteins - Bart’s syndrome, Kearns - Sayre syndrome, Pearson’s syndrome, and others are known. In addition, mitochondria are involved in the storage and transmission of hereditary information, apoptosis and plastic processes. There are a number of molecular markers, the use of which allows a detailed study of the activity of mitochondria under various experimental effects. The nearly thousand mitochondrial markers are known, but this review encloses the main ones.



2.
LYSOSOME-DEPENDENT CELL DEATH DEFINES SPECIFIC ENDOTHELIAL TOXICITY OF CALCIUM PHOSPHATE BIONS

Daria Kirillovna SHISHKOVA1, Elena Anatol’evna VELIKANOVA1, Rinat Avkhadievich MUKHAMADIYAROV1, Arseniy Evgen’evich YUZHALIN1, Yuliya Aleksandrovna KUDRYAVTSEVA1, Anna Nikolaevna POPOVA2, Dmitriy Mikhaylovich RUSSAKOV3, Anton Gennad’evich KUTIKHIN1
1Research Institute for Complex Issues of Cardiovascular Diseases
2Institute of Coal Сhemistry and Material Science
3Kemerovo State University
Keywords: бионы, фосфат кальция, гидроксиапатит, токсичность, эндотелий, лизосомы, клеточная гибель, атеросклероз, bions, calcium phosphate, hydroxyapatite, toxicity, endothelium, lysosomes, cell death, atherosclerosis

Abstract >>
Aim of the study was to identify the mechanism of specific endothelial toxicity related to calcium phosphate bions (CPB). Material and methods. CPB and magnesium phosphate bions (MPB) were artificially synthesised through supersaturation of culture medium with respective salts and then added to human endothelial cells (EA.hy 926) and murine endothelial cells (2H-11) to study: 1) spatiotemporal aspects of bion internalisation by means of transmission electron microscopy and confocal microscopy; 2) whether blocking of H+-ATPase by lysosomal inhibitor bafilomycin A1 affects endothelial toxicity of bions; 3) expression of caspase-3 and its substrate poly(ADP-ribose) polymerase (PARP-1). Results. CPB were internalized by endothelial cells as early as 1 h upon their addition and were localized in lysosomes; after 4 h, we detected release of calcium ions (Ca2+) from lysosomes to cytosol accompanied by multifold increase in cleaved caspase 3 and its substrate PARP-1. Bafilomycin A1 rescued endothelial cells from death induced by slightly soluble CPB regardless of exposure time and dose; however, freely soluble MPB did not evince endothelial toxicity regardless of bafilomycin A1 addition. Conclusion. Upon internalization by endothelial cells, CPB cause their death due to dissolution in lysosomes and subsequent release of calcium ions into the cytosol, ultimately leading to cleavage of executioner caspases. MPB lack endothelial toxicity because their dissolution does not lead to release of calcium ions. Therefore, specific endothelial toxicity of CPB is defined by lysosome-dependent cell death.



3.
EXPRESSION OF PROTEIN GENES PARTICIPATING IN FIBROPLASTIC PROCESSES IN MICE LUNG DURING THE DEVELOPMENT OF TUBERCULOUS INFLAMMATION

Peter Mikhaylovich KOZHIN1, Anton Vladimirovich CHECHUSHKOV1, Natal’ya Sergeevna ZAYTSEVA1, Marina Valer’yevna KHRAPOVA1, Liliya Aleksandrovna CHERDANTSEVA2,3, Elena Bronislavovna MENSHCHIKOVA1, Aleksandr Vasil’yevich TROITSKY1, Vyacheslav Alekseevich SHKURUPY2,4
1Research Institute for Experimental and Clinical Medicine of Federal Research Center
2Research Institute for Experimental and Clinical Medicine of Federal Research Center
3Novosibirsk Research Institute of Traumatology and Orthopaedics n.a. Ya.L. Tsivyan
4Novosibirsk State Medical University of Minzdrav of Russia
Keywords: туберкулезный гранулематоз, фиброз, окисленный декстран, декстразид, матриксная металлопротеиназа-9, проколлаген типа III, TGF-ОІ, ZEB1, Snai1, tuberculous granulomatosis, fibrosis, oxidized dextran, dextrazide, matrix metalloproteinase 9, type III procollagen, TGF-ОІ, ZEB1, Snai1

Abstract >>
The study analyzes the expression of protein genes involved in intracellular signaling pathways associated with a profibrotic response, activation of epithelial-mesenchymal and endothelial-mesenchymal transitions, in modeling tuberculous granulomatosis and pharmaceutical effects. Material and methods. The study was performed on male BALB/c mice aged two months and weighing 18-22 g. Generalized tuberculous granulomatosis was simulated by a single intravenous (retro-orbital) injection of 0.5 mg BCG vaccine in 0.2 ml of isotonic aqueous NaCl solution (SS), after 4 months a part of mice started to receive treatment drugs, after 2 months the animals were sacrificed by decapitation (6 months after BCG vaccine administration) and their lung tissues were collected. Mice were divided into 6 groups, 5 males in each: intact, which were intravenously into the retro-orbital sinus injected with 0.2 ml SS (C); infected with BCG and receiving SS intraperitoneal injections (BS); infected with BCG and received intraperitoneal injections of isonicotinic acid hydrazide solution (INAH) (BI); infected with BCG and received intraperitoneal injections of dextrazide solution (conjugate of 40 kDa oxidized dextran and INAH) (BD); infected with BCG and received intraperitoneal or inhalation injections of solution of molecular-nanosomal pharmaceutical compositions of oxidized dextran (MNPC) (BMP and BMH, respectively). Finally, the expression of mRNA of matrix metalloproteinase 9, type III procollagen, TGF-β, and transcription factors ZEB1 and Snai1 was determined by real-time PCR in mice lung tissue. Results. The modeling of generalized tuberculous granulomatosis was found to be accompanied by the induction of epithelial-mesenchymal transition and the activation of profibrotic processes, which after 6 months was manifested in increase of the type III collagen α1-chain and TGF-β mRNA expression. Administration of traditional (INAH) and original (dextrazide, MNPC) preparations with anti-tuberculosis activity within two months has inhibitory activity of varying severity in relation to various markers of these processes.



4.
BLOOD LIPOPROTEINS AS A PLATFORM FOR TRANSPORT OF HYDROPHILIC AND HYDROPHOBIC COMPOUNDS

Lev Mikhaylovich POLYAKOV, Roman Aleksandrovich KNYAZEV, Aleksandr Vladimirovich RYABCHENKO, Mariya Vladimirovna KOTOVA, Nataliya Viktorovna TRIFONOVA
Institute of Biochemistry of Federal Research Center for Fundamental and Translational Medicine
Keywords: липопротеины очень низкой плотности, липопротеины низкой плотности, липопротеины высокой плотности, цитохалазин B, бензилпенициллин, бензантрацен, бензо(а)пирен, ксенобиотики, транспортные формы, very low density lipoproteins, low density lipoproteins, high density lipoproteins, cytochalasin B, benzylpenicillin, benzanthracene, benzo(a)pyrene, xenobiotics, transport forms

Abstract >>
The paper discusses the transport functions of the main classes of blood plasma lipoproteins (LP) that are not associated with the metabolism of lipids that make up their composition. The aim of the study was to study the ability of various plasma LP fractions (very low (VLDL), low (LDL) and high density (HDL)) to interact with certain hydrophilic and hydrophobic compounds and show the role of LP as transport forms of xenobiotics in the organs and tissues of the body. Material and methods. The studies were performed with tritium-labeled cytochalasin B, benzylpenicillin, benzanthracene, benzo(a)pyrene, ultracentrifugation of human plasma LP fractions, column chromatography; in vivo experiments with intravenous injection of LP complexes with tritium-labeled benzanthracene were conducted. Results. The ability of various classes of LP to form complexes with hydrophilic (cytochalasin B, benzylpenicillin) and hydrophobic (benzanthracene, benzo(a)pyrene) compounds is shown by the method of ultracentrifugation. More than 50 % of the radioactivity of hydrophilic compounds in human blood plasma was represented in the composition of the LDL and HDL fractions, and in the composition of the VLDL fractions it was minimal - 6.3 and 5.1 %, respectively. A significant part of cytochalasin and benzylpenicillin was also present in the protein infranatant - 43.6 and 40.9 %, respectively. The distribution in blood plasma for hydrophobic (benzanthracene, benzo(a)pyrene) compounds was different. More than 80 % of the radioactivity was represented in the composition of the LP fractions. The polar protein infranatant contained 16.1 % of the radioactivity of benzantracene and 13.6 % of benzo(a)pyrene. The features of the lipophilic xenobiotics uptake by organs and tissues were shown in vivo experiments with intravenous injection of complexes of LP with tritium-labeled benzanthracene to rats. The highest specific radioactivity was found in the liver and adrenal glands after the intravenous injection of 3H-benzanthracene in the composition of VLDL and LDL. Twice less uptake of the labeled drug was observed in the testis and kidneys. Radioactivity decreased in the series: lungs, adipose tissue, thymus, heart, and spleen. A feature of the use of HDL as a platform for 3H-benzanthracene is the intense accumulation of lipophilic xenobiotics in steroid-producing organs: the adrenal glands and testis. Conclusion. The results obtained allow us to consider the real possibility of using blood plasma PL as transport platforms for hydrophilic and hydrophobic compounds into the cells of organs and tissues.



5.
ADRENO- AND CHOLINOREACTIVITY OF ERYTHROCYTES AND ESTIMATION OF THEIR CONJUGATION WITH PARAMETERS OF FREE RADICAL BALANCE OF BLOOD IN NON-LINEAR RATS

Evgeniya Vladimirovna KURYANOVA, Andrey Valerievich TRYASUCHEV, David Lvovich TEPLY
The Astrakhan State University
Keywords: адрено- и холинореактивность эритроцитов, свободнорадикальный баланс крови, анаприлин, атропин, нелинейные крысы, adreno- and cholinoreactivity of erythrocytes, free radical balance of blood, anapriline, atropine, non-linear rats

Abstract >>
The values of the β-adrenoreactivity of erythrocytes (β-ARE, 40.6 ± 2.3 rel. units) and the M-cholinoreactivity of erythrocytes (M-ChRE, 7.9 ± 0.3 rel. units), concentration of hemoglobin, the parameters of free radical balance: the level of TBA-reactive products (TBA-RP) and catalase activity in erythrocytes and plasma were determined in male non-linear rats. Sample grouping by clustering largest β-ARE showed that rats with a high β-ARE (>50 rel. units) have a greater concentration of hemoglobin, TBA-RP and CAT in erythrocytes, than rats with an average (30 to 50 rel. units) and low (<30 rel. units) β-ARE. According to the results of the correlation and factor analysis, M-ChRE is predominantly conjugated with erythrocyte (Factor 1), and β-ARE - with plasma characteristics of free radical balance in blood (Factor 3). The introduction of β-adrenergic blocker anapriline (2 mg/kg bw) decreases β-ARE and potentiates the accumulation of TBA-RP in the erythrocytes and plasma, the administration of the blocker of M-cholinergic receptors atropine (1 mg/kg bw) reduces not only M-ChRE, but also β-ARE and promotes the growth of erythrocyte catalase activity. Thus, in the intact state, under the conditions of blockade of adrenergic receptor and cholinergic receptors, dependences have been revealed that make it possible to treat β-ARE and M-ChRE as a link between regulatory influences on erythrocytes and biochemical, including free radical processes in the «erythrocyte - blood plasma».



6.
MORPHOLOGICAL CHARACTERISTICS OF TISSUE MICROREGION OF THE THYROID GLAND AT EXPERIMENTAL HYPOTHYROIDISM

Olga Vladimirovna GORCHAKOVA1, Vladimir Nikolaevich GORCHAKOV2,3, Georgiy Anatolevich DEMCHENKO4, Serik Nauryzbaevich ABDRESHOV4
1Research Institute of Clinical and Experimental Lymphology - Branch of the Institute of Cytology and Genetics SB RAS
2Research Institute of Clinical and Experimental Lymphology - Branch of the Institute of Cytology and Genetics SB RAS
3Novosibirsk State University
4Institute of Human and Animal Physiology of the Scientific Committee of the Ministry of Education and Science of the Republic of Kazakhstan
Keywords: щитовидная железа, тканевой микрорайон, гипотиреоз, thyroid gland, tissue microregion, hypothyroidism

Abstract >>
Research purpose was to study the structures of the tissue microregion of the thyroid gland at an experimental hypothyroidism. Material and methods. The hypothyroidism model was created by mercazolilum intake at a dose of 5 mg/100 g of body mass for a month. The structure of tissue area (follicle, thyrocyte, interfollicular space) with morphometry and calculation of the integral indicators of the thyroid gland functional activity have been investigated by the light-optical and ultrastructural methods. Results and discussion. The morphophysiological structure of the thyroid gland is a tissue microregion. It unites a group of follicles and interfollicular space. The structure of the tissue miroregion have features of structural response to merkazolilum intake. The thyrostatic remedy changes the follicular organization of the thyroid gland to the predominance of small and medium follicles with signs of alteration. Thyrocytes have expansion and deformation of the granular endoplasmic reticulum cisterns, formation of intracellular colloid, reduction in the number of lysosomes and mitochondrions with their disorganization. The ratio of the area of the follicular epithelium, colloid and stroma changes at hypothyroidism. The portion of interstitial increases, there is an accumulation of tissue fluid because of weakening of a lymphatic drainage. Blood and lymphatic microvessels react by reducing the volume density to mercazolilum intake. Conclusion. The merkazolilum thyreostatics effects on morphology and metabolic and transport processes in tissue microregion. It testifies to decrease of thyroid function and the creation of hypothyroidism adequate model.



7.
OBTAINING CHIMERIC VARIANTS HBcAg EXPOSING HIV-1 MPER FRAGMENTS

Andrey Pavlovich RUDOMETOV, Nadezhda Borisovna RUDOMETOVA, Boris Nikolaevich ZAYTSEV, Leonid Rudol’fovich LEBEDEV, Alexander Alekseevich ILYICHEV, Larisa Ivanovna KARPENKO
State Research Center of Virology and Biotechnology В«Vector» of Rospotrebnadzor
Keywords: ВИЧ-1, антигены, рекомбинантный НВсАg, рекомбинантные иммуногены, bNAbs, MPER, HIV-1, antigens, recombinant HBcAg, recombinant immunogens, bNAbs, MPER

Abstract >>
The HIV-1 epidemic is one of the most acute global health problems. For several reasons, an effective vaccine against this infection has not yet been created. Currently, an important direction in the development of a vaccine against HIV / AIDS is the design of immunogens that would be able to induce antibodies that neutralize a high diversity of HIV-1 strains (bNAbs). One approach to creating such immunogens is the construction of chimeric virus-like particles (VLPs) exposing epitopes recognized by bNAbs. The aim of the study was to obtain and characterize chimeric VLPs based on HBcAg, exposing epitopes recognized by bNAbs 2F5 and 4E10. Material and methods. The producing strains of chimeric HBcAg variants were obtained by transforming E. coli BL21 cells with recombinant plasmids carrying the HBcAg genes and containing insertions encoding bNAbs epitopes 2F5 and 4E10. Purification of recombinant proteins was performed using gel filtration on a sepharose CL-6B column. The ability of recombinant HBcAg to form virus-like particles was assessed using electron microscopy. Analysis of the antigenic properties of epitopes in the composition of chimeric variants of HBcAg was performed using immunoblotting. Results. A modified nucleotide sequence of the HBcAg gene was obtained, which included the introduction of unique restriction sites flanking the region of the main antigenic determinant of the core. Based on this genetic construct, three recombinant plasmids encoding chimeric HBcAg variants, including epitopes of bNAbs 2F5 and 4E10, were obtained. Using immunoblotting, it was found that epitopes recognized by bNAbs retain their antigenic properties after insertion into the HBcAg.



8.
EFFECT OF UROKINASE GENE-KNOCKOUT ON GROWTH OF MELANOMA IN EXPERIMENT

Elena Mikhaylovna FRANTSIYANTS, Irina Viktorovna KAPLIEVA, Ekaterina Igorevna SURIKOVA, Irina Valerievna NESKUBINA, Valeria Akhtyamovna BANDOVKINA, Lidiya Konstantinovna TREPITAKI, Nataliya Sergeevna LESOVAYA, Nataliya Dmitrievna CHERYARINA, Yuliya Aleksandrovna POGORELOVA, Lyudmila Anatol’evna NEMASHKALOVA
Rostov Research Institute of Oncology of Minzdrav of Russia
Keywords: нокаут по гену uPA, меланома, хроническая нейрогенная боль, половые особенности, мыши, uPA gene knockout, melanoma, chronic neurogenic pain, gender differences, mice

Abstract >>
The purpose of the study was to reveal special features of the В16/F10 melanoma growth in urokinase (uPA) gene knockout mice with and without chronic neurogenic pain (CNP). Material and methods. The study included male and female С57ВL/6 mice ( n = 102) and C57BL/6-Plautm1.1BugThisPlauGFDhu/GFDhu mice with uPA gene knockout ( n = 48). Mice of the main subgroups underwent subcutaneous transplantation of В16/F10 melanoma 2 weeks after bilateral ligation of sciatic nerves (CNP model); mice of the same strain with standard melanoma transplantation served as controls. Results and discussion. Survival of uPA gene knockout mice did not differ from that of normal animals - 1.5 times higher in females than in males ( p < 0.05), with melanoma onset in gene-deficient mice a week earlier. The dynamics of tumor growth had pronounced gender differences: in females, the tumor did not grow and its maximal volume prior to death was 1.0 cm3, while tumors in males were characterized by an active growth with two peaks of volume increase (weeks 2 and 4). Melanoma was weakly metastatic - solitary metastases to the lungs (in females) or no metastases, but pulmonary and heart hemorrhages were noted (in males). CNP decreased the survival of uPA gene knockout females, as well as of normal animals, but did not influence the survival of males; primary tumors in gene-deficient mice appeared a few days later than in controls but their growth was more intense, with diminished gender differences. Increased metastasis was manifested by the initiation of metastatic lesions to the lungs and liver in males, with maintained pulmonary hemorrhages, and by increased number of metastatic foci in the lungs together with the appearance of pulmonary hemorrhages in females. Conclusions. The influence of uPA gene knockout on the course of В16/F10 melanoma differs in male and female mice. CNP enhances malignant tumor growth, diminishing gender differences, and activates melanoma metastasis.



9.
GENDER FEATURES OF LAMINA CONTENT IN HUMAN SKIN FIBROBLASTS IN A PROCESS OF CHRONOLOGICAL AGING

Natalya Nikolaevna Golubtsova, Aleksey Vladimirovich Bogdanov, Andrey Germanovich Gunin
Chuvash State University n.a. I.N. Ul’yanov
Keywords: ламин А, ламин В1, ламин В2, фибробласты, кожа, гендерные особенности, хронологическое старение, lamin A, lamin B1, lamin B2, fibroblasts, skin, gender features, chronological aging

Abstract >>
The purpose of the study was to investigate gender peculiarities of the content of fibroblasts immunopositive to lamins A, B1 and B2 in skin samples in a process of chronological aging. Material and methods. 331 skin samples at the period from 20 to 40 weeks of gestation and people from birth to 85 years were examined. The content of positively stained dermal fibroblasts and the expression level of lamins A, B1 and B2 in their nucleus was explored by immunohistochemistry. Results and discussion. The gradual decrease of fibroblasts with positive staining for lamin A and the expression level of lamin A after antenatal period and up to old age depends on the age, but does not have differences, which are related to gender. The lamin B1 positively fibroblasts content is declining reliably from birth to 40 years and then it is increasing as well as the expression level of lamina B1. The indicated changes don’t have reliable variances in gender. The change in the number of lamina B2-positively colored fibroblasts and the expression level of lamina B2 has no statistically valid correlation with gender and age. Thus, there are no gender differences in dynamical age related changes of the content and the expression level of protein-lamina in skin fibroblasts in a process of chronological aging.



10.
ACTIVITY OF FREE RADICAL OXIDATION AND CYCLOOXYGENASE EXPRESSION IN RATS KIDNEYS UNDER EXPERIMENTAL THERAPY OF OXALATE NEPHROLITHIASIS WITH TETRAPEPTIDE Leu-Ile-Lys-Met

Aleksandr Yuryevich ZHARIKOV, Nikolay Nikolaevich YAKUSHEV, Ganna Viktorovna ZHARIKOVA, Olesya Nikolaevna MAZKO, Olesya Gennadyevna MAKAROVA, Valeriy Mikhaylovich BRYUKHANOV, Yuriy Vladimirovich KORENOVSKIY, Olga Vasil’yevna AZAROVA
Altai State Medical University of Minzdrav of Russia
Keywords: мочекаменная болезнь, тетрапептид, окислительный стресс, urolithiasis, tetrapeptide, oxidative stress

Abstract >>
The aim of the study was to investigate the effect of Leu-Ile-Lys-Met tetrapeptide on the activity of free radical oxidation (FRO) and the level of cyclooxygenase-1 (COX-1) and COX-2 expression in rat kidney with experimental oxalate nephrolithiasis. Material and methods. The experiments were carried out on 30 male rats Wistar runoff, divided into 3 groups: the comparison group (10 intact rats), the control group (6 weeks of experimental oxalate nephrolithiasis simulation, n = 10), the experimental group (6 weeks of experimental simulation oxalate nephrolithiasis + oral administration of Leu-Ile-Lys-Met tetrapeptide at a dose of 12 mg/kg from the 3rd to the 6th week, n = 10). The oxalate nephrolithiasis was simulated according to conventional ethylene glycol model. COX-1 and COX-2 concentration in rat urine at baseline and after 6 weeks of experimental nephrolithiasis was determined by enzyme immunoassay using the prostaglandin-endoperoxide synthase 1 and 2 (PTGS 1, 2) kit from «Cloud-Clone Corp.». In rat kidney homogenate, the activity indicators of FRO processes were determined by common methods. Results and discussion. At the background of the Leu-Ile-Lys-Met tetrapeptide, free radical kidney damage was weakened, which was manifested in the complete absence of morphological signs of nephrolithiasis, decrease of thiobarbituric acid-reactive product concentration and overall prooxidant activity as compared to the control group; glutathione peroxidase activity, on the contrary, increased by 1.5 times.



11.
STIMULATION OF REPARATIVE REGENERATION OF SUPPORTING TISSUE UNDER CONDITIONS OF DELAYED APPOSITION OF FRAGMENTS

Svetlana Alexandrovna LEPEKHOVA1, Guanda QIAO1, Oleg Aronovich GOLDBERG2, Nikolay Valer`evich TISHKOV2, Lida QIAO3, Ilya Sergeevich KURGANSKIY1,2, Pavel Olegovich INOZEMTSEV1, Lyudmila Ivanovna FEDOROVA1
1Irkutsk Scientific Center of SB RAS
2Irkutsk Scientific Center of Surgery and Traumatology
3Irkutsk City Clinical Hospital № 1
Keywords: регенерация, ложный сустав, голень, морфометрия костной ткани, биологически активные точки, regeneration, pseudarthrosis, shin, bone morphometry, biologically active points

Abstract >>
The study is devoted to reparative regeneration of supporting tissue in the area of the defect using the methods of acupuncture with stimulation of biological active points when modeling delayed apposition of fragments. Material and methods. The work was performed using laboratory animals of the Chinchilla rabbits, at the age of 6 months. The delayed apposition of shin bone fragments was simulated in all animals. In the dynamics of the study, an assessment of radiological and morphological changes in the area of the bone defect was performed. After the fragments were compared and fixed using animal randomization methods, the animals were divided into two groups - the control and the main group, in which the biologically active points were stimulated. Results and discussion. The evaluation of the supporting tissue regeneration in the defect zone under the influence of proposed biologically active points combination by the method of acupuncture within the conditions of delayed apposition of shin bone fragments has revealed physiological change of regeneration phases with the formation of differentiated bone tissue and bone marrow elements to the 50th day of the study. The 2 fold predominance of bone tissue, 2 fold - the bone marrow substance, a decrease in cartilage tissue, the absence of fibrous connective tissue have been revealed compared with the control group, where the slowdown in the physiological change of regeneration phases and the formation of regenerate supporting tissue in the form of pseudarthrosis have been revealed.



12.
NEW ASPECTS OF THE USE OF PROTEIN TISSUE-SPECIFIC MARKERS IN THE ESTIMATION OF SEVERITY OF COMMUNITY-ACQUIRED PNEUMONIA

Mikhail Ivanovich VOEVODA1,2,3,4, Yuliya Igorevna RAGINO5, Konstantin Yuryevich NIKOLAEV1,4, Olga Sergeevna KHARLAMOVA5, Ekaterina Mikhaylovna STAKHNEVA5, Elena Vladimirovna KASHTANOVA5, Yana Vladimirovna POLONSKAYA5, Dinara Evgenyevna IVANOSHCHUK1,3, Margarita Vitalyevna KRUCHININA1,6, Vladimir Nikolaevich KRUCHININ7
1Research Institute of Internal and Preventive Medicine - Branch of the Institute of Cytology and Genetics of SB RAS
2Federal Research Center for Fundamental and Translational Medicine
3The Federal Research Center Institute of Cytology and Genetics of SB RAS
4Novosibirsk State University
5Research Institute of Internal and Preventive Medicine - Branch of the Institute of Cytology and Genetics of SB RAS
6Novosibirsk State Medical University
7Rzhanov Institute of Semiconductor Physics of SB RAS
Keywords: внебольничная пневмония, тяжелая пневмония, белки легочного сурфактанта А и D, сыворотка крови, парциальный корреляционный анализ, community-acquired pneumonia, severe pneumonia, pulmonary surfactant proteins A and D, blood serum, partial correlation analysis

Abstract >>
The purpose of the study was to assess the associations of proteins of pulmonary surfactants A (SP-A) and D (SP-D) with the severity of community-acquired pneumonia. Material and methods. A survey of 188 patients diagnosed with community-acquired pneumonia (CAP). The average age of the examined patients was 54.3 ± 16.5 years (M ± SD). In 102 patients (54.3 %) severe CAP was verified. All patients underwent clinical, functional, diagnostic and laboratory studies. Results and discussion. It was determined that SP-A and SP-D content was significantly higher in patients with severe CAP compared to patients with a mild course of this disease. The direct association of severe CAP with an elevated SP-AP level, directly associated with an increase in the concentration of C-reactive protein in the blood and erythrocyte sedimentation rate, have been confirmed in early studies. Using partial correlation analysis, it was established that the severe course of CAP is directly related to SP-A ( r = 0.221; p = 0.003) and SP-D content ( r = 0.262; p < 0.001) regardless of age, smoking, and body weight. Thus, direct associations of SP-A and SP-D with a severe course of CAP reflect the high pathogenetic significance of these protective factors in infectious lung damage.



13.
INITIAL RESULTS IN RECURRENT BRAIN GLIOBLASTOMAS MANAGEMENT WITH MAXIMAL SAFE RESECTION FOLLOWED BY INTAOPERATIVE BALLOON ELECTRONIC BRACHYTERAPY

Orkhan Al’zaminogly ABDULLAEV1,2, Aleksey Sergeevich GAYTAN2, Nidal SALIM2, Gleb Sergeevich SERGEEV2, Il’ya Vladimirovich MARMAZEEV2, Evaldas CHESNULIS3, Aleksey Leonidovich KRIVOSHAPKIN1,2
1Novosibirsk State Medical University of Minzdrav of Russia
2European Medical Center
3Klinik Hirslanden, Neurochirurgie Hirslanden
Keywords: глиобластома, резекция глиом, радикальность резекции, лучевая терапия, брахитерапия, интраоперационная лучевая терапия, glioblastoma, glioma resection, radical resection, radiation therapy, brachytherapy, intraoperative radiation therapy

Abstract >>
Brain glioblastomas (GBM) are notorious for their early local recurrence despite of standard combined treatment. Technologies for recurrent GBM management require further development and research. Resection of malignant gliomas must be followed by an adjuvant treatment. Intraoperative balloon electronic brachytherapy (IBEB) has been recently introduced into clinical practice and could be successfully applied to recurrent GBM management. This article presents the initial results of recurrent GBM management with maximal safe resection followed by IBEB. Material and methods. Patients ( n = 11) with recurrent GBM after standard combined treatment were managed with maximal safe microsurgical resection followed by IBEB. The follow-up period after IBEB ranged from 1 to 30 months. Results. The median overall survival for the entire study group of patients included in the statistical analysis ( n = 9) was 27 months (range 17-47 months). In the subgroup of patients ( n = 4) with contrast-enhanced tumor MRI volume after resection ≤ 2.5 cm3, the median local progression free survival (locPFS) was 21.25 months (range 10-30 months). Conclusion. Initial results in recurrent brain GBMs management with resection followed by IBEB seems to be promising, especially in case of contrast-enhanced tumor volume detected on MRI after resection is less than 2.5 cm3 without signs of multifocal tumor growth.



14.
COMPARATIVE ANALYSIS OF MARKERS EFFICIENCY OF VISCERAL OBESITY DETECTION IN MEN WITH METABOLIC SYNDROME

Denis Andreevich DEEV, Boris Borisovich PINKHASOV, Vera Georgievna SELYATITSKAYA, Tatyana Remonovna MATSIEVSKAYA, Iraida Georgievna MOKHOVA
Federal Research Center for Fundamental and Translational Medicine
Keywords: мужчины, висцеральное ожирение, метаболический синдром, окружность талии, отношение окружности талии к окружности бедер, толщина эпикардиального жира, men, visceral obesity, metabolic syndrome, waist circumference, waist to hip circumference ratio, epicardial fat thickness

Abstract >>
The purpose of the study was to carry out comparative and quantitative assessment of surrogate markers of visceral obesity and their connection with the severity of clinical, biochemical and hormonal disorders in men with the metabolic syndrome and obesity. Material and methods. 136 men, aged from 23 to 68 years, who were patients of therapeutic clinic were observed. Clinical, anthropometric, biochemical and hormonal screening was taken. Transthoracic echocardiography was performed in all patients. Criteria of international experts’ consensus in the fields of cardiology and endocrinology (2009) were used to identify metabolic syndrome. Results and discussion. It was shown that among the all surrogate markers of visceral obesity (waist circumference; waist circumference measured in the lying position; waist circumference measured without the thickness of the subcutaneous tissue; the ratio of waist circumference to the circumference of the hips; sagittal abdominal diameter; thickness of epicardial adipose tissue) the largest number of statistically significant correlations with the clinical-biochemical, hormonal and adipokine characteristics of men was found for the waist circumference, measured in the lying position, and the waist circumference, measured without the thickness of the subcutaneous tissue. Intermediate position among the estimated markers was taken by a waist circumference measured in a standing position. Fewer number of correlation connections were revealed for thickness of the epicardial fat, sagittal abdominal diameter and the ratio of waist circumference to the circumference of the hips. Conclusion. The received results demonstrate objectivity of the selection of the surrogate markers of the severity of visceral obesity reviewed in the study, among which the waist circumference measured in the lying position had the greatest efficiency.



15.
MENINGIOMAS OF THE CEREBELLAR TENTORIUM: THE RESULTS OF SURGICAL TREATMENT

Vyacheslav Vladimirovich STUPAK1, Ivan Sergeevich KOPYLOV1, Ivan Viktorovich PENDYURIN1, Sergey Borisovich TSVETOVSKIY1, Dmitriy Andreevich DOLZHENKO2, Aleksandr Vasilyevich NOVOKSHONOV3, Timur Mazambekovich SHOGUNBEKOV4
1Novosibirsk Research Institute of Traumatology and Orthopedics n.a. Ya.L. Tsivyan of Minzdrav of Russia
2Regional Clinical Hospital
3Regional Clinical Center of the Miners Health Protection
4State Novosibirsk Regional Clinical Hospital
Keywords: нейрохирургия, менингиома, намет мозжечка, задняя черепная ямка, опухоль головного мозга, neurosurgery, meningioma, tentorium, posterior cranial fossa, brain tumor

Abstract >>
Aim of the study was to evaluate the surgeries of tentorial meningiomas in Neurosurgical Department No.1 of the Novosibirsk Institute of Traumatology and Orthopedics. Material and methods. Study includes 64 patients with tentorial meningiomas of different location, treated surgically from 2008 to 2017. Among them 55 (85.9 %) were female patients and 9 were male (14.1 %) with average age of 62.9 years in females and 56.7 years in males. For diagnostic purposes, the patients were performed brain magnetic resonance imaging and/or computed tomography with or without contrast agents. In this series, 83 % of the tumors were of large or giant (with diameter of more than 4 cm). In 63 cases the tumors were typical meningiomas and only 1 case was a malignant anaplastic meningioma. Results. All patients were treated surgically in one or two steps. Overall, there were 70 standard microsurgical resections. Electromyography with Neuropack-2 («NIHON KOHDEN Corp.», Japan) was performed to control brain stem function during the surgery, with short latency auditory evoked potentials (SLAEPs) received. When sinus walls were affected, the tumor matrix was processed with 20W neodymium laser in coherent mode and ablation. In 63 cases, gross-total resection was achieved: Simpson I (76.6 %, n = 49) and Simpson II (21.8 %, n = 14). One patient (1.6 %) was treated with subtotal resection because of complicated surgical approach and massive transverse sinus and sinus confluence bleeding. Post-operative mortality was 3.1 % (2 cases). SLAEP data did not demonstrate any significant irritation or damage to the brain stem with performed surgical approaches. There were no infections of the surgical wound, meninges, brain or CSF leaks after the surgery. Discussion. Tentorial meningiomas are rare brain tumors treated with gross-total tumor resection when possible. Performed surgeries resulted in mild functional decrease in quality of life, but Karnofsky scale for each patient was returning to its original state in 2 weeks. Complications, including neurological disability, presented in 19 % ( n = 12) of the patients, and post-operative mortality did not exceed the common literature data. Most of the focal neurological disabilities have disappeared in 2 weeks after the surgery.



16.
RADIOSURGICAL TREATMENT OF BENIGN SMALL EYELID TUMORS

Svetlana Vagovna SAAKYAN, Marina Panaetovna KHARLAMPIDI, Elena Borisovna MYAKOSHINA, Robert Alexandrovich TATSKOV, Georgiy Alexandrovich GUSEV
Helmholtz Moscow Research Institute of Eye Diseases
Keywords: доброкачественные опухоли век, радиохирургия, benign tumors of the eyelids, radiosurgery

Abstract >>
Aim of the study - to evaluate the long-term results of the radiosurgical method using microsurgical techniques for the treatment of small benign eyelid tumors. Material and methods. In Ocular Oncology Centre of Helmholtz Moscow Research Institute of Eye Diseases for the period from 2012 to 2018 year 248 patients were examined and treated (108 children aged 12 ± 1.2 years and 140 adults aged 46 ± 8.1 years) with benign eyelid tumors: pigmented (nevus), vascular (capillary hemangiomas), epithelial (follicular keratosis, keratopapilloma). The Surgitron EMC radiosurgery device was used as surgical aids. All patients used microsurgical technique. The longitudinal size of the tumors was 3.4 ± 0.5 mm, transverse - 6 ± 0.8 mm. All patients after surgical treatment underwent histopathological study. The observation period is on average 6 ± 1.4 years. Results. In 133 (53.6 %) of 248 patients, nevi were detected, in 76 (30.7 %) - keratopapilloma and inverted follicular keratosis, in 39 (15.7 %) - capillary hemangioma. Various modifications of radiosurgical treatment were used depending on the location of the tumor, its size, the degree of activity using different modes and powers from 1 to 4 Wt. Intraoperative complications were not noted. The early postoperative period was accompanied by the presence of signs of mild inflammation, which was stopped by local therapy. The formation of a tender scar was observed on average after 7 ± 2 days. Two cases (0.8 %) of recurrence of keratopapilloma were noted 2 years after the first operation. Tumors were removed by radio excision. Conclusion. Benign eyelid tumors, especially of melanocytic origin, are subject to obligatory dynamic observation, and if minimal signs of their progression are detected, their surgical removal is indicated. Radiosurgical treatment using microsurgical techniques is the method of choice in the treatment of small benign eyelid tumors and depends on the size of the tumor, its localization and the nature of the tumor process. The use of the radiosurgical method with obligatory microsurgical technique for treating benign tumors of small sizes with the observance of the rules of radicalness and antiblasticity allows maintaining visual functions with simultaneous surgical treatment, providing normal anatomical and topographic correlations of the anterior segment of the eye and high quality of life of the patient.



17.
THE CHARACTERISTICS OF CYTOKINE STATUS IN PATIENTS WITH DEEP PHLEGMON OF THE NECK AND SECONDARY ACUTE MEDIASTINITIS UNDER DIFFERENT METHODS OF TREATMENT

Mariya Gennadievna PUSTOVETOVA1, Yuriy Vladimirovich CHIKINEV2,3, Evgeniy Aleksandrovich DROBYAZGIN2,3, Tatiana Sergeevna KUTEPOVA3
1Federal Centre for Cardiovascular Surgery n.a. S.G. Sukhanov
2Novosibirsk State Medical University of Minzdrav of Russia
3State Novosibirsk Regional Clinical Hospital
Keywords: глубокая флегмона шеи, медиастинит, лаваж озонированным раствором, про- и противовоспалительные цитокины, deep neck phlegmon, mediastinitis, lavage with ozonated solution, pro- and anti-inflammatory cytokines

Abstract >>
The aim of the study was to improve the results of treatment of patients with deep neck phlegmon and acute secondary mediastinitis. Material and methods. 81 case histories of patients with deep neck phlegmon and 40 case histories of patients with secondary acute mediastinitis who underwent examination and treatment in the regional clinical hospital, Novosibirsk, for the period from 2007 to 2017 were analyzed. The results of treatment were evaluated by the dynamics of changes in the serum concentration of pro- and anti-inflammatory cytokines TNF-α, IL-1ß, IL-6, IL-8, IL-4, IL-1β/IL-4. Results and discussion. When applying the method of lavage of deep neck and mediastinum cellular spaces with ozonated 0,9 % sodium chloride solution, there is a 1,2-3-fold faster normalization of cytokine status compared to standard lavage, which indicates a more pronounced decrease in the intensity of the acute inflammatory process in the postoperative period.



18.
FEATURES OF HEART RATE regulation IN ADOLESCENTS WITH ESSENTIAL HYPERTENSION

Alina Mikhaylovna VALEEVA1, Oleg Viktorovich SOROKIN2, Zhanna Valerievna NEFEDOVA3, Victoria Gennad’evna KAYNARA4, Lyudmila Mikhaylovna PANASENKO3, Lyudmila Petrovna DERYAGINA3, Nina Petrovna KAREVA3
1Novosibirsk State University
2National Ayurvedic Medical Association
3Novosibirsk State Medical University
4City Children’s Clinical Hospital № 1
Keywords: подростки, эссенциальная гипертензия, кардиоинтервалография, вариабельность сердечного ритма, adolescents, essential hypertension, cardiointervalography, heart rate variability

Abstract >>
Cardiointervalography is an important method of analyzing the regulation of heart rate variability, which allows evaluating the balance of the entire spectrum of humoral-metabolic and vegetative effects, both in condition of physiological rest and in various functional tests. It is noted that in hypertensive patients, in most cases, there is a chronic predominance of ergotrophic metabolic reactions that reflect the system tension in the regulatory mechanisms. The aim of our study was to conduct a comparative analysis of the features of the mechanisms of heart rate regulation in adolescents with confirmed essential hypertension and their healthy age mates. Material and methods. The study involved 50 adolescents (12-16 years of age) - 26 boys and 24 girls with labile essential hypertension (EHT) and stage 1 HT on the background of the standard medical correction of blood pressure (enalapril 0,58 mg/kg per day). Each participant of the study and control group underwent recording of at least 512 cardio cycles in the state of physiological rest in a sitting position, as well as registration of blood pressure. Results and discussion. We have found that for adolescents with EHT a significantly lower (by 15 %) average duration of the heart cycle is typical in combination with a decrease in the variation range, which indicates some rigidity of the heart rate regulatory mechanisms. The relative decrease in the proportion of high-frequency spectrum components in comparison with the low-frequency and very low-frequency ranges in adolescents with hypertension also indicated a tendency to predominance of sympathicotonic effects with the predominant involvement of the structures of the diencephalic region and the brain stem. At the same time blood pressure inversely correlated with the index of the variation range of cardiointervals, reflecting the lower adaptive potential of the heart rate regulation mechanisms. Conclusion. We have found regular changes in the regulation of heart rate in adolescents with hypertension, that can be taken as markers of its progressive development and used by clinicians to develop more personalized approaches to medication treatment.



19.
LIPID INFILTRATION OF HEPATOCYTES IN THE MORPHOGENESIS OF CHRONIC HEPATITIS C

David L’vovich NEPOMNYASHCHIKH1, Olga Alexeevna POSTNIKOVA1, Ekaterina Eduardovna ABRAMOVA1, Olga Igorevna DOROVSKAYA2
1Novosibirsk State Medical University of Minzdrav of Russia
2State Novosibirsk Regional Clinical Hospital
Keywords: HCV-инфекция, липидосодержащие гепатоциты, генотипирование, HCV infection, lipid-containing hepatocytes, genotyping

Abstract >>
The aim of the study was to investigate structurally functional features of lipidic infiltration of hepatocytes in a morphogenesis of chronic hepatitis C. Material and methods. A clinical and morphological study of 199 patients with HCV infection markers (139 men and 60 women aged from 20 till 65 years) was conducted. In all cases, a comprehensive study including analysis of blood biochemical parameters, serological markers of HCV replication in blood and liver, viremia level, number of the infected hepatocytes and HCV genotypes was performed. Diagnostic complex included clinical, biochemical, immunoserological methods. Results and discussion. The results of the analysis of structural and functional features of lipid-containing hepatocytes and indicators of replication of virus particles of chronic hepatitis C are provided. For its reproduction HCV enters into complex relationships with the metabolic apparatus of the cell, in which lipid metabolism and related structures - lipid droplets play an important role. The only structural marker that correlates with indicators of virus replication is subcytolemmal small-vesicular lipid infiltration, which can reflect a virus-induced increase in lipid metabolism in the host cell for the effective production of infectious active viral particles.



20.
MORPHOFUNCTIONAL FEATURES OF VENTRICULAR ARRHYTHMIAS IN PATIENTS WITH POSTINFARCTION CARDIOSCLEROSIS AND ARTERIAL HYPERTENSION, DEPENDING ON THE AFFECTED CORONARY REGION

Alina Mikhaylovna VALEEVA1, Natalya Viktorovna SHLYAKHTINA1,2, Olga Sergeevna VOLODICHEVA3
1Novosibirsk State Universuty
2Meshalkin National Medical Research Center of Minzdrav of Russia
3City Clinical Hospital of Emergency Medical Care N 2
Keywords: желудочковые нарушения ритма, артериальная гипертензия, постинфарктный кардиосклероз, электрокардиография, эхокардиография, коронароангиография, внезапная сердечная смерть, ventricular rhythm disorders, arterial hypertension, post-infarction cardiosclerosis, electrocardiography, echocardiography, coronary angiography, sudden cardiac death, ventricular rhythm disorders, arterial hypertension, post-infarction cardiosclerosis, electrocardiography, echocardiography, coronary angiography, sudden cardiac death

Abstract >>
Acute and chronic perfusion disorders, the presence of hypertension are among the main risk factors for the development of electrical instability of the myocardium, in particular ventricular arrhythmias (VA), the progressive course of which increases the risk of sudden cardiac death. Changes in a number of electro- and echocardiographic indicators are recommended to be considered as predictors of the development of life-threatening arrhythmias. Purpose of the study was to study VA features, myocardial remodeling processed and lipid metabolism in patients with arterial hypertension (AH) and myocardial infarction, depending on the affected coronary region. Material and methods. The study involved 50 patients 40-80 years old with post-infarction cardiosclerosis, AH, ventricular arrhythmias episodes, of which 25 people are patients with atherosclerotic lesions of left coronary artery (LCA), 25 people are with the lesions of right coronary artery (RCA). Exclusion criteria: dilated cardiomyopathy, decompensated valvular defects, arrhythmogenic dysplasia, idiopathic ventricular tachycardia, myocarditis. Research methods: echocardiography, coronary angiography, Holter monitor (24-Hour ECG monitoring), biochemical data. Results and discussion. The tendency to the presence of more pronounced pathological changes of heart rate turbulence due to an increase of turbulence slope, dominance of the sympathetic division of the autonomic nervous system against the background of a significantly larger number of paired ventricular extrasystoles have been revealed in the group of patients with atherosclerotic lesions of LCA. That reflects organic and functional changes in the myocardium. The absence of significant changes in the duration and dispersion of the QT interval of the compared groups confirmed the low informativeness of these criteria in the prediction of the VA. The significantly lower value of interventricular septum thickness, left ventricular myocardial mass, and larger size of the right ventricle (0.15 ms) have been revealed along with the above mentioned changes in the group due to the slightly different course of the AH, myocardial remodeling processes. However, more significant changes in the lipid profile, in particular an increase in the level of total cholesterol and blood triglycerides have been registered in the group of patients with PCA. Conclusion. The more pronounced pathological shift of heart rate turbulence due to the increase of turbulence slope up to 4.4 ms/RR on the background of more paired ventricular extrasystoles has been determined in the defeat of the LCA. The significantly lower value of interventricular septum thickness, left ventricular myocardial mass, and larger size of the right ventricular have been observed in the group of patients with atherosclerotic lesions of the LCA in comparison with group of patients with lesions of the PCA.



21.
INCIDENCE OF TEENAGE YOUNG MEN BY THE RESULTS OF MEDICAL EXAMINATION AT INITIAL MILITARY REGISTRATION

Anatoliy Ivanovich BABENKO1, Evgeniy Gennadyevich SHVEDOV2, Sergey Mukhtarovich BEKMURZOV3, Evgeniy Anatolyevich BABENKO1
1Research Institute for Complex Problems of Hygiene and Occupational Diseases
2Military Registration and Enlistment Office of the Novosibirsk Region
3425 Military Hospital of Minoborony Russia
Keywords: юноши-подростки, медицинское освидетельствование, распространенность патологии, teenage young men, medical examination, prevalence of pathology

Abstract >>
Teenagers’ health, the level and structure of incidence reflect totality of the influence of the factors and their living conditions, upbringing and training, a way of life, medical care. The results of medical examination of young men at initial military registration to a sufficient extent give an idea of the prevalence of pathology among this contingent and in many respects characterize the quality of labour and defendable potential. Material and methods. The results of medical examination of teenagers by the military physician board of the Novosibirsk Regional Military Registration and Enlistment Office from 2000 to 2014 with the total number of the examined subjects 252.2 thousand were used. The dynamic and comparative analyses of the indices for three time periods along with the assessment of the prevalence of the diseases by the disease classes, separate nosological groups and also the distribution of young men by the medical classification were carried out. Results. It was established that on average 62.7 % of the examined subjects had this or that pathology. At the same time, 22.5 % of all the diseases were revealed for the first time. It was shown that 87.7-88.7 % of all the pathology accounted for 8 basic classes of diseases. Among the diseases limiting the medical classification the main ones were mental and behavioral disorders and also the diseases of the musculoskeletal system and connective tissue. The temporary unfitness was determined by endocrine, nutritional and metabolic diseases, and the complete one was defined by mental disorders, diseases of the nervous system, endocrine pathology, diseases of the spine and their aftereffects, disorders of refraction and accommodation of the eyes, visual impairment, bronchial asthma, mental retardation, flat foot and other foot deformations, degenerative diseases of the nervous system, which accounted for 63.0 % of all. Conclusion. The study showed the feasibility of using the results of the medical examination to assess the health of teenage young men and the inclusion of these indices in social and hygienic monitoring.