Aim of the research. The aim is to study the content of intercellular adhesion molecules (ICAM-1), vascular cell adhesion molecule (VCAM-1) and calprotectin in the blood serum of patients with type 2 diabetes mellitus and various stages of diabetic retinopathy. The aim is also to evaluate the role of these molecules in the pathogenesis of the disease. Materials and methods. Four groups of people were formed: first group (control group) included 21 healthy individuals; second group included 21 patients with prediabetes, third group 21 patients with type 2 diabetes. The fourth group included 63 patients with diabetic retinopathy, and this group was further divided into 3 groups of 21 people each: with non-proliferative stage of DR, with preproliferative stage, with proliferative stage.. The concentrations of ICAM-1, VCAM-1 and calprotectin (MRP8/14) in blood serum were determined using Human Vascular Inflammation Panel 1 multiplex analysis kits from Biolegend (USA). The results were assessed using CytoFlex flow cytometer (USA). The results were calculated using Jamovi version 2.3. Results. In individuals with prediabetes, the content of MRP8/14 was increased by 111,7% (p < 0,001) relative to the control group. In type 2 diabetes without retinopathy, the values of MRP8/14 protein exceed the control group values by 2,7 times (p < 0,001) and those in individuals with prediabetes by 29,2% (p = 0,049). In the group of patients who had non-proliferative stage of DR, the levels of ICAM-1, VCAM-1 and MRP8/14 are higher than control group values in the groups of people with prediabetes and patients with diabetes without complications. During the preproliferative stage, the number of adhesion molecules increases even more; during the proliferative stage, the concentrations of VCAM-1 and calprotectin remain high, and the level of ICAM-1 increases relative to the previous stages. Conclusion. Increasing of MRP8/14 level in diabetes and increasing of ICAM-1 and VCAM-1 concentrations in the initial stage of DR demonstrate the role of these molecules in the initiation of DR in type 2 diabetes. Researching the relationship between these markers and the development of DR can provide additional information to develop strategies for prevention and treatment of DR as well as predicting its complications.

Introduction. Thrombo-inflammatory changes in the alveolar-capillary membrane are significant links in the pathogenesis of respiratory failure in COVID-19. This study investigates whether leukocyte-platelet aggregates contribute to its development and, if so, the mechanisms involved. Aim. To examine the quantitative changes in leukocyteplatelet aggregates in patients with COVID-19 receiving different levels of oxygen support. Materials and methods. The study included 134 COVID-19 patients of varying severity and 20 volunteers examined in the pre-pandemic period. The criterion for dividing the studied patients into groups was the ratio of blood oxygen saturation to its inhaled fraction, investigated by pulse oximetry. Three groups were formed depending on the value of the index: in the first (n=48), the SpO2/FiO2 index was above 450%, in the second (n=55) it ranged from 370 to 449%, and in the third (n=51) the index was up to 369%. The enumeration of blood cells, major leukocyte populations, lymphocyte subpopulations, and plateletleukocyte complexes was performed using the CytoFLEX LX flow cytometer (Beckman Coulter, USA). Results. In the third group, there was a decrease in the number of monocytic aggregates (p<0.001) and their subfractions, as well as the number of lymphocyte rosettes (p=0.015), while the number of lymphocytes (p<0.001) and neutrophil interactions (p=0.05) increased in parallel. In the second group, there was a statistically significant decrease in the total number of monocytic aggregates (p=0.038), and in the third, the number of aggregates with classical monocytes (p=0.012). Conclusion. The number of lymphocyte-platelet and monocyte-platelet aggregates decreased in patients with different types of oxygen support, while neutrophil-platelet aggregates increased and correlated with the SpO2/FiO2 ratio.

AIM:To study the dynamics of cystatin C in patients after procedures on retroperitoneal viscera using laparoscopic and retroperitoneal access. MATERIAL AND METHODS:A prospective study of serum and urine cystostatin C levels in 162 patients with renal cysts using laparoscopic (n=83) and retroperitoneal access (n=79) was carried out. Patients were divided into four groups depending on the duration of the procedure and the gas level. For determination of cystatin C level in serum and urine, flow cytometry was performed using Human Kidney Function Panel 2 Mix and Match Subpanel (for Serum/Plasma Samples). RESULTS:A significant increase in the serum and urine level of cystatin C in groups 3 and 4 was seen on the 1st day after laparoscopic procedures in case of pneumoperitoneum pressure above 12 mm Hg., regardless of its duration. For retroperitoneoscopic procedures, similar changes were found only in group 4 with a gas pressure above 12 mm Hg and duration of the intervention of more than 30 minutes. By day 3, cystatin C levels returned to baseline values in all groups. CONCLUSION:The most significant risk factor for acute kidney injury after laparoscopic and retroperitoneoscopic procedures is the gas pressure. The second most important factor is the duration of the intervention. Pneumoperitoneum has a greater effect on cystatin C levels compared to retropneumoperitoneum.

Aim: To study the correlations between Cystatin C (Cys-C) level and values of the markers of immune system and inflammation, and clinical manifestations of diabetic retinopathy (DR) in patients with type 2 diabetes mellitus (DM).Material and Methods. 3 groups of patients with type 2 diabetes and different stages of diabetic retinopathy were formed (21 people in each). Group I – with nonproliferative diabetic retinopathy (NPDR), Group II – with pre-proliferative diabetic retinopathy (PPDR), Group III – with proliferative diabetic retinopathy (PDR). The comparison group included patients with type 2 diabetes without vascular complications. Clinical study included: visometry, tonometry, assessing critical flicker fusion frequency, biomicroscopy of the anterior segment of the eye, ophthalmoscopy, biomicroscopy and ultrasound of the retina, crystalline lens, vitreous body, photographic recording of the fundus, optical coherence tomography. The content of Cystatin C (Cys-C), soluble forms of molecules B7.2 (CD86), 4-1BB, CTLA-4, Tim-3, LAG-3, PD-1, PD-L1, Galectin-9, proteins sICAM-1, SAA, NGAL and enzymes (MPO, MMP-2, MMP-9) was examined in the blood serum with the use of multiplex analysis.Results. As DR progresses, the level of Cys-C increases and becomes higher than in individuals with diabetes: with NPDR higher by 94.1% (p < 0.001), with PPDR – higher by 293.6% (p < 0.001). In individuals with PDR, the concentration of Cys-C is maximum. With DR, the amount of PD-1, PD-1L, NGAL, ICAM-1, MMP-9, and MPO increases in the blood serum; as the severity of DR worsens, the levels of ICAM-1, MPO, and MMP-9 increase. Direct correlations were found between the Cys-C values, on the one hand, and the values of some studied indicators, on the other.Conclusions. In type 2 diabetes and DR, the amount of Cys-C in the blood serum increases relative to individuals with diabetes without microangiopathy; in groups with worsening severity of ophthalmopathy, an increase in Cys-C concentration was recorded with a statistically significant difference between the groups. In groups with DR, the level of ICAM-1, MMP-9, and MPO increases with increasing severity. Moderate direct correlations were found between the amount of Cys-C on the one hand and PD-1, PD-L1, as well as the noticeable ones with the values of ICAM-1, NGAL, MMP-9, MPO on the other. A direct noticeable correlation was revealed between the level of Cys-C and the values of the fundus scale.

The aim of the research. To identify changes in tryptophan and cytokine metabolism in synovial joint tissue in rheumatoid arthritis. Research materials and methods. The experiments were performed on 40 WISTAR rats. At the beginning of the study, each animal of the experimental groups received an intraperitoneal injection of collagen type 2 solution (Chondrex, Inc., USA) in Freund's incomplete adjuvant. The content of tryptophan, kynurenine, 3-hydroxykinurinine, L-5-hydrotryptophan by HPLC with fluorimetric and spectrophotometric detection, and the concentration of cytokines: IL1b, IL6, IL10, IL17, IL18, TNFa were determined in the tissues of the synovial membrane of the knee joint. They were determined using Rat Inflammation Panel V02 multiplex assay kits (Biolegend, USA) on a Cytomics FC500 flow cytofluorimeter (Beckman Coulter, USA). Statistical analysis was performed using the Jamovi program version 2.3.Results. The concentration of cytokines: IL1b, IL6, IL10, IL17, IL18, TNFa was found to be high during the acute development of rheumatoid arthritis. With experimental rheumatoid arthritis, the content of tryptophan metabolites along the kynurenine pathway increases and the concentration of metabolites along the serotonin pathway decreases. Direct positive correlations of IL1b, IL6, IL10, IL17, IL18, TNFa with the content of tryptophan metabolites along the kynurenine pathway have been established.Conclusions. Tryptophan metabolism and IL1b, IL6, IL10, IL17, IL18, and TNFa content are altered in rheumotoid arthritis.

Aim: To study the content of tryptophan and the intermediate metabolites of kynurenine and serotonin pathways of its metabolism in the blood plasma of patients with type 2 diabetes mellitus (DM) and non-proliferative diabetic retinopathy as candidates for markers of the early stage of the pathological process.Material and Methods. Three groups of people were analyzed: the first group of 10 patients with type 2 diabetes mellitus and without diabetic retinopathy; the second group of 10 people with type 2 diabetes mellitus and non-proliferative stage of diabetic retinopathy; and the control group of 10 healthy people. The features of the first group were a decrease of macula’s photosensitivity in the fundus and a slight increase in the central thickness of the retina. Patients with diabetic retinopathy tended to have a moderate number of microaneurysms and microhemorrhages, moderately presented intraretinal microvascular anomalies in one quadrant, vein dilatation, clearness of the central retinal vein and its branches. In the macular zone there was an edema with hard exudates in the center and lateral to the fovea centralis. All participants of the study gave blood on an empty stomach in the morning, and after that in we measured the content of tryptophan (TRP), kynurenins ((kynurenine (KYN), 3-hydroxykynurenine (3-HKYN), kynurenic acid (KYNA)) and the level of L-5-hydroxytyrptophan (5HTrp) in blood plasma by HPLC method with fluorimetric and spectrophotometric detection.Results. The groups of people with type 2 diabetes mellitus showed the increasing of TRP level in blood relatively to healthy individuals: by 15.1% (p = 0.032) and 17.9% (p = 0.030) in the first and second groups, respectively. As for the patients of the first group, the content of their KYN was increased by 57.7% (p = 0.012) and KYNA by 33.6% (p = 0.012) relatively to the control and the concentration of 3-НKYN decreased by 18.1% (p = 0.020) relatively to healthy people. As for the patients in the second group, the changes in their level of kynurenines had the same direction, but were more visible. Thus, the concentration of KYN exceeded the same parameters of healthy individuals by 84.5% (p = 0.001) and the parameters of the first group by 18.0% (p = 0.049); the KYNA level increased by 56.6% (p = 0.001) relatively to the control and by 17.3% (p = 0.049) from that of the first group. There was a decrease in the content of 3-HKYN amounted to 18.6% of the control (p = 0.038) and an increase in the concentration of 5HTrp – 193,9% (p < 0.001).

Introduction. The pathogenetic role of biochemical changes in the development of diabetic retinopathy (DR) is undoubted and determines the importance of studies that reveal metabolic disorders of both individual molecules and show the relationships between them. Because of some commonalities between tissue respiration, monosaccharide and tryptophan exchange, and activation of lipid peroxidation (LP), there is an interest in studying the relationship between metabolites of these pathways in retinopathy.Aim. The aim is to study the content of metabolites of the kynurenic pathway and intermediate product of lipid peroxidation in blood, and to reveal the correlation between them and glycated hemoglobin’s (HbA1c) level in retinopathy with type 2 diabetes.Materials and methods. The 1st group (control group) included 21 healthy people; the 2nd group included 21 people with «prediabetes», the 3rd group - 21 patients with type 2 diabetes, the 4th group - 63 people with type 2 diabetes and diabetic retinopathy (DR) of varying severity levels. The content of kynurenines in the blood plasma (kynurenine (KYN), 3-hydroxykynurenine (3-HKYN), kynurenic acid (KYNA)) of all participants, as well as the concentration of malondialdehyde (MDA) was determined by high-performance liquid chromatography using a Shimadzu LC-20 chromatograph (Japan). The level of glycated hemoglobin (HbA1c) was determined using Beckman Coulter AU 480 (USA) biochemical analyzer. The results were calculated using Jamovi program version 2.3.Results. In the group of people with «prediabetes», the values of all studied parameters in the blood plasma were increased in comparison with the control group. In the group of patients with type 2 diabetes, the levels of KYN, 3- НKYN and MDA significantly exceeded those in the group of people with «prediabetes». In the group of people with DR, all indicators remained high, the values of 3-НKYN and KYNA showed a statistical difference compared to the group of patients with diabetes. When conducting a correlation analysis, corellations were identified between the level of HbA1c on the one hand and the values of KYN (r=0.77; p ˂ 0.001), concentrations of 3-НKYN (r=0.80; p ˂ 0.001), KYNA (r= 0.72; p ˂ 0.001) and MDA (r=0.84; p ˂ 0.001) – on the other. There was a correlation between HbA1c level and fundus scale (r=0.82; p ˂ 0.001) and between concentrations (of 3-HKYN) and MDA (r=0.50; p=0.002).Conclusion. An increased level of glycosylated hemoglobin and lipid peroxidation products in the blood during retinopathy against the background of type 2 diabetes, an increase in the concentrations of KYN, 3-HKYN, KYNA, and the presence of correlations between these indicators serve as the basis for proving the relationship of uncontrolled hyperglycemia with changes in cellular respiration and the development of oxidative stress. Hypoxic effects and the accumulation of intermediate metabolic products of the kynurenine pathway contribute to progressive neurovascular damage to the retina, activation of lipid peroxidation processes and inflammation.

Введение. Выяснение механизмов, ответственных за восстановление иммунного ответа при проведении плазмолифтинга у больных, страдающих хроническим пародонтитом, позволит профилактировать и раннее развитие атеросклероза, и ревматоидного артрита, и ряда других патологических процессов, имеющих сходное с пародонтитом основное звено патогенеза. Цель: оценка системных и локальных иммунологических эффектов плазмолифтинга в модели экспериментального пародонтита у крыс. Методика. У самцов крыс Wistar (возраст 18–20 нед, начальная масса тела 200,0±31,5 г.) моделировали экспериментальный пародонтит путем отслойки десны концом копьевидного скальпеля (№ 11) в сочетании с инъекциями 10% раствора этилового спирта с 0,25% раствором новокаином в течении 7 сут. Сформированы 4 группы по 10 особей в каждой: одна контрольная группа и 3 опытные: 1-я опытная группа –животные с экспериментальным пародонтитом, не получающие терапии; 2-я – крысы с экспериментальным пародонтитом, получающие инъекции плазмы. Плазму, в объеме 50 мкл, вводили микрошприцем локально в область альвеолярной кости, на 2 мм ниже десневого края, между правой стороной 1-го и 2-го моляров верхней челюсти (3 инъекции через 3 сут каждая); 3-я группа – животные с индуцированным пародонтитом, леченные пленками фирмы «Диплен» с активными действующими компонентами «Метронидазолом» и «Хлоргексидином» (курс процедур составил 10 сут). Результаты. На 10-е сут индуцированного пародонтита клинически и гистологически регистрировался воспалительный процесс, сопровождающийся резким подъемом уровня цитокинов, более выраженным в гомогенатах тканей. Применение процедуры плазмолифтинга, а также стандартной терапии проявлялось уменьшением фибринозных наложений, санацией пародонтального кармана, формированием вторичного пародонтального прикрепления. Отмечалось ограничение зоны инфильтрации, снижался уровень цитокинов. При этом, терапевтический эффект плазмолифтинга был более выраженным. Заключение. Полученные нами данные свидетельствуют о том, что плазмолифтинг не только снижает местную воспалительную реакцию тканей при экспериментальном пародонтите, но и уменьшает напряженность системного воспалительного процесса. Introduction. Elucidation of the mechanisms that contribute to restoration of the immune response following plasmolifting in patients with chronic periodontitis will allow prevention of early atherosclerosis, rheumatoid arthritis, and other pathological processes that share the main pathogenetic component with periodontitis. Aim: evaluating systemic and local immune effects of plasmolifting in a rat model of periodontitis. Methods. Experimental periodontitis was induced in male Wistar rats aged 18-20 weeks and weighing 200.0±31.5 g at baseline. Periodontitis was modeled by detaching the gums with a spear-headed scalpel tip (sz. 11) in combination with injections of 10% ethanol with 0.25% Novocain for 7 days. Four groups (a control group and 3 experimental groups) of 10 rats each were used: group 1, rats with experimental periodontitis not receiving therapy; group 2, rats with experimental periodontitis injected with plasma (three 50-μl microinjections every third day, locally, into the alveolar bone area, 2 mm below the gingival margin between the right side of the maxillary first and second molars); group 3, rats with experimental periodontitis treated by application of Diplen films with metronidazole and chlorhexidine active ingredients for 10 days. Results. On day 10 of experimental periodontitis, the inflammatory process was clinically and histologically detected. The inflammation was associated with a sharp increase in cytokines that was more pronounced in tissue homogenates. The use of the plasmolifting procedure as well as a standard therapy was manifested by a decrease in fibrin deposits, sanitation of the periodontal pocket, and formation of a secondary periodontal attachment. A restriction of the infiltration zone, and a decrease in cytokines were noted. Also, the therapeutic effect of plasmolifting was more pronounced. Conclusions. Experimental periodontitis is accompanied by the clinical and morphological picture of inflammation and increases in concentrations of both pro- and anti-inflammatory cytokines, that are more pronounced in tissue homogenates. Plasmolifting not only reduces the local inflammatory response of tissues to experimental periodontitis, but also alleviates the intensity of the systemic inflammatory process. However, none of the therapies provided achievement of the intact animal status.

Objective Clinical manifestation of the inflammatory process in its relation to biochemical markers (total cysteine [Cys], cysteine-glycine [CysGly], glutathione [GSH], glutamate-cysteine [Glu-Cys], homocysteine [Hcy], the ratio of reduced to oxidized glutathione [GSH/GSSG], the ratio of reduced to oxidized cysteine [CySH/CySS], malondialdehyde-oxidized low-density lipoproteins [MDA-oxLDL]) has been studied in patients with coronavirus disease 2019 (COVID-19). Material and methods 48 patients with mild to severe COVID-19 and 20 healthy volunteers were included in our research. The participants were divided into 4 experimental groups according to inflammation intensity estimated based on the serum levels of interleukin 6 (IL-6). Results All 4 groups showed the prevalence of male patients and elevated serum levels of IL-6 (by 54.6%). There was no comorbidity in patients with mild COVID-19 (nasopharyngitis symptoms) and in healthy control subjects. 50% of patients with lung damage had accompanying diseases. Alterations of aminoethyl metabolism were detected in COVID-19 patients: as reflected by the decreased levels of Cys, CysGly, and Glu-Cys and the increased levels of GSH as compared to the control group. Conclusion Elevation of IL-6 over 7.5 pg/mL was associated with decreased GSH/GSSG and CySH/CySS ratios indicating enhanced oxidative stress and was followed by protein oxidation, specifically MDA-oxLDL.

The role of chemokines involved in the processes of neuroinflammation is not well understood in schizophrenia, especially in patients with the first episode of the disease. The aim of the study: to study the content of pro-inflammatory chemokines in the blood of patients with the first episode of paranoid schizophrenia. We examined 18 patients with a diagnosis of F20.09, the control group – 35 people. Determination of the content of 13 chemokines in blood serum was carried out by flow fluorometry. Blood sampling was carried out before the start of therapy. In patients with the first episode of schizophrenia before the appointment of therapy, an increase in the blood level of CCL4 (MIP-1β) by 1.1 times, CXCL9 (MIG) – by 1.4 times, CCL11 (Eotaxin), CXCL5 (ENA-78), CXCL10 (IP-10) – 1.5 times, CXCL1 (GRO-α) – 1.6 times, СCL20 (MIP-3α) – 2.1 times, CXCL8 (IL-8) – 21.0 times. The results obtained indicate the importance of neuroimmune inflammation in the manifestation of schizophrenia with the involvement of chemokines in this process.

Aim. To investigate formation of neutrophil extracellular traps (NETs) and their impact on fibrinolysis in patients with colon cancer.Materials and methods. The study was performed in two groups. The experimental group consisted of patients with stage 2–3 non-metastatic colon cancer (n = 17, average age – 67 years). The control group included healthy volunteers matched by sex and age (n = 30, average age – 68 years). An experimental model was created from the whole blood. It included platelet-poor plasma and an isolated culture of neutrophils, previously induced to NETosis by adding 100 nmol PMA. The samples were incubated for 4 hours, then the test tubes were centrifuged to pellet cells and their remnants, and the plasma was transferred for subsequent examination. The plasma incubated with intact neutrophils was used as a control. The levels of interleukin-8 (IL-8) and P-selectin glycoprotein ligand-1 (PSGL-1) were used to determine the degree of cell activation. NETosis was confirmed by enzyme-linked immunosorbent assay (ELISA) and fluorescent microscopy. Fibrinolysis was assessed using the thrombodynamics test. The results were compared with the levels of fibrinolytic system components measured by flow cytometry.Results. In the control group, NETosis induction contributed to pronounced neutrophil activation that was accompanied by an increase in the IL-8, PSGL-1, and plasminogen levels, a decrease in PAI-1, and enhancement of fibrinolysis, compared with the intact samples. Higher levels of IL-8, PSGL-1, plasminogen, and PAI-1 and intensified fibrinolysis were detected in the intact samples. However, PMA-induced NETosis did not result in an increase in the degree of activation and significant changes in the given parameters.Conclusion. NETosis promotes both formation and lysis of fibrin clots. However, in cancer patients, suicidal NETosis does not contribute to fibrinolysis due to intracellular protease depletion, which may be one of the mechanisms causing hypercoagulation and insufficient fibrinolysis in cancer.

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Significant changes in cellular and humoral immunity are observed in new coronavirus infection (COVID-19). The cytokine storm develops in cases of severe clinical course, being accompanied by significantly increased levels of pro-inflammatory cytokines, often associated with suppression of immune response. At the same time, the prediction of the immune status is an urgent task, thus allowing timely correction of current therapy. The aim of our research was to evaluate predictive capability for the immune system changes on the 6th day of COVID-19 disease when using standard therapy, or with addition of immunocorrector thymalin to the treatment regimen. A retrospective study was conducted in 87 patients with severe COVID-19. All patients were divided into 2 groups, i.e., (1) controls who received basic treatment; (2) basic treatment supplied with thymalin (10 mg, intramuscular injections daily for 5 days). Assessment of severity and clinical course of COVID-19, and basic treatment regimen for the patients corresponded to current version of the interim Guidelines from the Ministry of Health of the Russian Federation Prevention, diagnosis and treatment of a new coronavirus infection COVID-19. Laboratory studies included complete blood counts, immunogram parameters with the calculation of the ratio of certain types of leukocytes were performed on the 1st and 6th days of observation. Statistical evaluation was made using scripts of the specialized statistical analysis language R (http://cran.r-project.org) version 4.1.3. The blood parameters were evaluated using the binary classification method. The changes in parameters of cellular immunity were classified by means of ROC-analysis. We have found that the basic therapy of severely ill COVID-19 patients was not followed by recovery of immune status on the 6th day from the start of treatment. At marginal level, one can only suggest a probable prediction of increase in WBC and MON counts, a decrease in CD4+, NK and CD8+perNK, as well as the CD4+/CD8+ ratio. Addition of thymalin to the basic therapy is largely accompanied by the normalization of immunogram parameters. At the same time, it is possible to predict, with excellent rating, an increased number of T-LIM, including CD4+ and B-LIM, and, with good rating, an increase in the total numbers of LIM, as well as CD8+, HLA-CD3+DR+ and NK cells. The data obtained in severe cases of COVID-19 make it possible to predict changes in immune status, and, hence, the course of the disease, at a high degree of probability.

Введение. Внутрисосудистое формирование нейтрофильными лейкоцитами внеклеточных ДНК-сетей (англ. neutrophil extracellular traps, NETs) является одной из причин развития иммунотромбоза – тромбообразования, ассоциированного с реализацией иммунных механизмов резистентности. NETs выступают как интегральное звено, объединяющее в единый процесс функционирование систем иммунитета и гемостаза, что приобретает особое значение в зонах роста злокачественных новообразований. Цель исследования: уточнение патогенетической роли литического нетоза в процессе коагуляционного гемостаза, а также выявление изменения локальной концентрации основных цитокинов в условиях формирования NETs in vitro. Материалы и методы. Измеряли плазменную концентрацию цитокинов: интерлейкинов (англ. interleukin, IL) – IL-1β, IL-2, IL-4, IL-6, IL-10, свободного активного трансформирующего фактора роста-β1 (англ. transforming growth factor, TGF-β1), фактора некроза опухоли-альфа (англ. tumor necrosis factor-alpha, TNF-α), интерферона-γ (англ. interferon-γ, IFN-γ), хемокинов (CXCL8, CCL2, CXCL10), факторов I, II, III, IX, XI и XIII свертывания крови, а также методом тромбодинамики оценивали показатели коагуляции до и после экспериментального моделирования литического нетоза in vitro. Культуру нейтрофильных лейкоцитов и плазму получали из цельной венозной крови 30 больных с впервые выявленным раком толстого кишечника II–III стадии без метастазов. Группу сравнения составили 40 добровольцев, сопоставимых по полу и возрасту, не имевших злокачественного процесса. Уровень цитокинов и факторов свертывания крови в образцах плазмы измеряли методом проточной цитометрии с использованием панелей мультиплексного анализа. Результаты. В обеих группах выявлено ускорение коагуляции на 5,5% и удлинение времени задержки роста фибринового сгустка без изменения концентраций I, II, III, IX и XIII факторов свертывания крови. Значимо увеличилось плазменное содержание CXCL8, IL-2 и свободного активного ТGF-β1 в образцах, где нейтрофилы погибли путем литического нетоза, в сравнении с исходным плазменным уровнем указанных молекул, а также в сравнении с образцами, содержавшими покоящиеся клетки. Заключение. Процесс литического нетоза нейтрофилов сопровождается ускорением I–III этапов коагуляции плазмы по внешнему пути и сопровождается значительным локальным повышением концентрации CXCL8 и в меньшей степени IL-2 и ТGF-β1, что потенциально может явиться одним из механизмов развития и поддержания хронической гиперкоагуляции и хронического воспаления, а также реализации проопухолевых свойств NETs. Background. Intravascular neutrophil extracellular traps (NETs) releasing represents one of the mechanisms of immunothrombosis – thrombi formation associated with immune reactions. NETs act as integrating link between immunity and hemostasis, thus acquiring particular importance in the sites of malignancy. Objectives: to reveal whether the suicide NETosis can modify clotting parameters and the plasma level of essential immunoregulatory factors in vitro. Patients/Methods. The plasma concentration of interleukins (IL) – IL-1β, IL-2, IL-4, IL-6, IL-10, free active transforming growth factor (TGF-β1), tumor necrosis factor-alpha (TNF-α), interferon-γ (IFN-γ), chemokines CXCL8, CCL2, CXCL10 and I, II, III, IX, XI, XIII coagulation factors were measured before and after of experimental modeling of suicide NETosis in vitro. Coagulation process was assessed by thrombodynamics method. Cell culture of neutrophils and plasma were obtained from whole venous blood of 30 patients with newly diagnosed stage II–III colon cancer without metastases. The comparison group consisted of 40 healthy, age and gender matched volunteers. Plasma level of cytokines and clotting factors were measured by flow cytometry using multiplex assay panels. Results. Increasing of clotting rate by 5.5% and prolongation of lag-time were detected in both groups. There were no variations of coagulation factors level. The significant increase of plasma level of CXCL8, IL2 and free active TGF-β1 were detected in both groups in samples with netotic cells, when compared with their baseline concentration and with samples, contained resting cells. Conclusions. Suicide NETosis is accompanied by acceleration of initial stage of fi brin clot formation through external way and by strong increase in CXCL8 concentration and less increase in IL-2 and TGF-β1 that can be considered as one of the ways how NETs contribute to hypercoagulation state and chronic inflammation development and also implement their tumorgenicity.

Comorbidity is a common condition in medical practice. The presence of comorbid conditions in a patient makes a great contribution to the course and prognosis of the underlying disease, as well as the development of complications [12]. At the same time, diseases occurring with broncho-obstructive syndrome and arterial hypertension syndrome are widespread throughout the world [13, 14]. The aim of this study was to assess the degree of endothelial dysfunction associated with impaired nitric oxide synthesis, based on the state of the NO/NOS/ADMA system and genetic polymorphisms of the nitric oxide synthetase gene in patients with bronchial asthma combined with essential hypertension. Materials and methods Three groups of patients, 96 people each, were formed, corresponding in age and gender structure with a diagnosis of bronchial asthma (BA group), essential hypertension (EH group) and their joint course (BA+EH group). Serum concentrations of asymmetric dimethylarginine, symmetric dimethylarginine, genetic polymorphisms of nitric oxide synthase (synthases NOS3 786T/C and NOS3 894G/T) were determined. To determine the role of other factors in increasing the concentration of ADMA and SDMA, multiple linear regression analysis was performed. Results: according to the results of the study, for a group of patients with a combination of bronchial asthma and hypertension, there are increased levels of ADMA and SDMA, the T allele NOS3 786C/T is more common in hypertension, in multiple linear regression significant factors in increasing ADMA are the presence of hypertension, bronchial asthma, body mass index and glomerular filtration rate, for SDMA - the presence of hypertension, asthma and glomerular filtration rate. Conclusions: The comorbidity of bronchial asthma and arterial hypertension is associated with an increased serum concentration of ADMA and SDMA. The presence of the TT genotype of the NOS3 786C/T polymorphism is associated with an increased incidence of arterial hypertension in patients with asthma.