Purpose. Purpose: to study the content of pro-inflammatory cytokines and matrix metalloproteinases in the intraocular fluid in patients with advanced stage of primary open-angle glaucoma. Material and methods. 50 patients were examined with a verified diagnosis of the advanced stage of poag. the control group consisted of 30 patients with a diagnosis of uncomplicated cataract. the concentration of the determined biologically active molecules was carried out by the method of flow fluorimetry on a two-beam laser analyzer -Bio-plex 200, Bio-rad, USA. Results. A statistically significant increase in the concentrations of Il-6, 8, 12, 17, mIp-1β matrix metalloproteinase-2 was found in the intraocular fluid of patients with poag. Conclusion. The data obtained indicate the significance of the local inflammatory process, as well as disturbances in the structure of the extracellular matrix in the mechanisms of poag development. Keywords: primary open-angle glaucoma, intraocular fluid, cytokines, matrix metalloproteinases

ЦЕЛЬ: изучить концентрации цитокинов в сыворотке крови у больных сахарным диабетом (СД) 2 типа с атеросклерозом сонных артерий и артерий нижних конечностей. МАТЕРИАЛЫ И МЕТОДЫ: обследовано 65 пациентов с длительностью СД 2 типа не менее года от мо- мента постановки диагноза. Всем пациентам выполнено ультразвуковое дуплексное сканирование сонных артерий и артерий нижних конечностей. На основании данных сканирования, выделяли группы пациентов без атеросклероза (АТ0), с атеросклерозом одного и двух сосудистых бассейнов (АТ1 и АТ2 соответствен- но). В сыворотке крови определяли концентрации 27 цитокинов, вовлеченных в хроническое воспаление, ангиогенез и ремоделирование сосудистой стенки, методом мультиплексного анализа (PlexPro™ Human Cytokine 27-plex, Bio-Rad Laboratories, США). Группу контроля составили 45 лиц без СД и сердечно-сосуди- стых заболеваний. РЕЗУЛЬТАТЫ: средний возраст больных составил 62 [56; 67] года, длительность СД – 10 [6; 16] лет, сред- ний уровень HbA1c – 7,7% [6,8; 9,6]. В группу АТ0 вошли 25 пациентов, в группу АТ1 - 15, в группу АТ2 – 25. Больные группы АТ2 были старше (р=0,0001), имели большую длительность СД (р=0,006) и меньший ИМТ (р=0,0001). У больных СД 2 типа в сравнении с контролем обнаружены более высокие концентрации ин- терлейкинов (IL): IL-1β (p=0,03), IL-2 (p=0,02), IL-8 (p=0,02), IL-9 (p<0,0001), IL-12 (p=0,0002), IL-17A (p<0,0001), основного фактора роста фибробластов (bFGF, p=0,0001), гранулоцитарного колониестимулирующего фактора (G-CSF, p=0,04), гранулоцитарно-макрофагального колониестимулирующего фактора (GM-CSF, p<0,0001), интерферон гамма-индуцибельного белка 10 (IP-10, p=0,03), макрофагального воспалительного белка 1 альфа (MIP-1α, p<0,0001) и RANTES (p=0,0001), выявлены более низкие концентрации IL-15 (р=0,04) и фактора роста эндотелия сосудов (VEGF, p<0,0001). Различий между группами больных СД 2 типа с раз- личной выраженностью/локализацией атеросклероза по большинству параметров не выявлено. В моделях мультиномиального дискриминантного анализа концентрация IL-17A, наряду с возрастом, индексом массы тела, длительностью СД, средней амплитудой колебаний гликемии и скоростью клубочковой фильтрации, являлась предиктором атеросклероза одной и двух локализаций. ВЫВОДЫ: изменения в спектре циркулирующих цитокинов у больных СД 2 типа характеризуются повышением уровня провоспалительных регуляторов, bFGF и снижением уровня VEGF и IL-15. В целом, изменения концентраций цитокинов в сыворотке крови в большей степени ассоциированы с наличием СД, нежели с выраженностью атеросклероза.

ЦЕЛЬ: изучить концентрации цитокинов в сыворотке крови у больных сахарным диабетом (СД) 2 типа с атеросклерозом сонных артерий и артерий нижних конечностей. МАТЕРИАЛЫ И МЕТОДЫ: обследовано 65 пациентов с длительностью СД 2 типа не менее года от мо- мента постановки диагноза. Всем пациентам выполнено ультразвуковое дуплексное сканирование сонных артерий и артерий нижних конечностей. На основании данных сканирования, выделяли группы пациентов без атеросклероза (АТ0), с атеросклерозом одного и двух сосудистых бассейнов (АТ1 и АТ2 соответствен- но). В сыворотке крови определяли концентрации 27 цитокинов, вовлеченных в хроническое воспаление, ангиогенез и ремоделирование сосудистой стенки, методом мультиплексного анализа (PlexPro™ Human Cytokine 27-plex, Bio-Rad Laboratories, США). Группу контроля составили 45 лиц без СД и сердечно-сосуди- стых заболеваний. РЕЗУЛЬТАТЫ: средний возраст больных составил 62 [56; 67] года, длительность СД – 10 [6; 16] лет, сред- ний уровень HbA1c – 7,7% [6,8; 9,6]. В группу АТ0 вошли 25 пациентов, в группу АТ1 - 15, в группу АТ2 – 25. Больные группы АТ2 были старше (р=0,0001), имели большую длительность СД (р=0,006) и меньший ИМТ (р=0,0001). У больных СД 2 типа в сравнении с контролем обнаружены более высокие концентрации ин- терлейкинов (IL): IL-1β (p=0,03), IL-2 (p=0,02), IL-8 (p=0,02), IL-9 (p<0,0001), IL-12 (p=0,0002), IL-17A (p<0,0001), основного фактора роста фибробластов (bFGF, p=0,0001), гранулоцитарного колониестимулирующего фактора (G-CSF, p=0,04), гранулоцитарно-макрофагального колониестимулирующего фактора (GM-CSF, p<0,0001), интерферон гамма-индуцибельного белка 10 (IP-10, p=0,03), макрофагального воспалительного белка 1 альфа (MIP-1α, p<0,0001) и RANTES (p=0,0001), выявлены более низкие концентрации IL-15 (р=0,04) и фактора роста эндотелия сосудов (VEGF, p<0,0001). Различий между группами больных СД 2 типа с раз- личной выраженностью/локализацией атеросклероза по большинству параметров не выявлено. В моделях мультиномиального дискриминантного анализа концентрация IL-17A, наряду с возрастом, индексом массы тела, длительностью СД, средней амплитудой колебаний гликемии и скоростью клубочковой фильтрации, являлась предиктором атеросклероза одной и двух локализаций. ВЫВОДЫ: изменения в спектре циркулирующих цитокинов у больных СД 2 типа характеризуются повышением уровня провоспалительных регуляторов, bFGF и снижением уровня VEGF и IL-15. В целом, изменения концентраций цитокинов в сыворотке крови в большей степени ассоциированы с наличием СД, нежели с выраженностью атеросклероза.

68 КЛИНИЧЕСКИЕ, МЕТАБОЛИЧЕСКИЕ И ВОСПАЛИТЕЛЬНЫЕ ПАРАМЕТРЫ, АССОЦИИРОВАННЫЕ С МАКРОАНГИОПАТИЕЙ НИЖНИХ КОНЕЧНОСТЕЙ У БОЛЬНЫХ САХАРНЫМ ДИАБЕТОМ 2 ТИПАКоролева Е

Purpose. To study the content of cytokines and matrix metalloproteinases in the intraocular fluid in patients with advanced stage of primary open-angle glaucoma. Material and methods. 50 patients were examined with a verified diagnosis of the advanced stage of POAG. The control group consisted of 30 patients with a diagnosis of uncomplicated cataract. The concentration of the determined biologically active molecules was carried out by the method of flow fluorimetry on a two-beam laser analyzer Bio-Plex 200, Bio-Rad, USA. Results. A statistically significant increase in the concentrations of IL-6, 8, 17, TGF-1, 2, 3, matrix metalloproteinase-2 and tissue inhibitors of matrix metalloproteinases-1, 2, 3, 4 was found in the intraocular fluid of patients with POAG. Conclusion. The data obtained indicate the significance of the local inflammatory process, as well as disturbances in the structure of the extracellular matrix and its remodeling in the mechanisms of POAG development. Keywords: primary open-angle glaucoma, intraocular fluid, cytokines, matrix metalloproteinases, tissue inhibitors of matrix metalloproteinases.

МУЛЬТИПЛЕКСНЫЙ АНАЛИЗ ПАНЕЛИ ЦИРКУЛИРУЮЩИХ ЦИТОКИНОВ У БОЛЬНЫХ САХАРНЫМ ДИАБЕТОМ 2 ТИПА С АЛЬБУМИНУРИЧЕСКОЙ И НЕАЛЬБУМИНУРИЧЕСКОЙ ХРОНИЧЕСКОЙ БОЛЕЗНЬЮ ПОЧЕК1 Климонтов В .В ., 1 Корбут А .И ., 1 Орлов Н .Б

We performed a correlation analysis of the morphometry of mesenteric lymph nodes and cytokines of the thoracic duct lymph in chemically induced breast cancer (BC) and after surgical treatment of BC and neoadjuvant chemotherapy for BC according to the CMF scheme. After surgical treatment, correlations were found between the level of cytokines in the thoracic duct lymph and morphological changes in the mesenteric lymph nodes. The observed correlations can be a result of total resection of the tumor and increased immunomodulatory and antitumor action of cytokines. Comparison of the effect of chemotherapy and surgical treatment revealed correlations between cytokine concentrations in the lymph and structural changes in the lymph nodes, which can be related to weakening of local immunity and antitumor effect of cytokines.

Aim. To study the content of matrix metalloproteinase (MMP)-2, -3, -9 and tissue inhibitors of metalloproteinases (TIMPs) -1, -2, -3, -4 in the aqueous humor of patients with moderate primary open-angle glaucoma (POAG). Materials and methods. The experimental group included 47 patients with verified moderate primary open-angle glaucoma. The control group consisted of 26 patients with uncomplicated cataract. The levels of MMP-2, -3, -9 were determined with Luminex Performance Human MMP Magnetic Panel 3-plex kit (R&D Systems, USA), the concentration of TIMPs-1, -2, -3, - 4 was determined with the Human TIMP Magnetic Luminex Performance Assay 4-plex kit (R&D Systems, USA). The study was carried out using flow-through field fluorometry on a Bio-Plex 200 double-beam laser analyzer (Bio-Rad, USA). Results. The study showed a statistically significant increase in the levels of matrix metalloproteinase-2 and tissue inhibitors of matrix metalloproteinases-1, -2, -3, -4 in the aqueous humor of patients with moderate POAG compared with patients with uncomplicated cataract. Conclusion. The obtained data on high concentrations and imbalance in the levels of matrix metalloproteinases and their tissue inhibitors in the aqueous humor of patients with moderate POAG confirm the role of local inflammation, as well as impairments in the structure of the extracellular matrix and its remodeling in the mechanisms of development of this pathology.

Цель исследования - анализ корреляции морфометрии брыжеечных лимфатических узлов и концентрации цитокинов в лимфе грудного протока при химиотерапии рака молочной железы, хирургическом лечении и последующей химиотерапии. Методика. Рак молочной железы индуцировали введением N-метил-N-нитрозомочевины 5 раз с интервалом 7 сут подкожно в область 2-й молочной железы справа. Курс химиотерапии проходил по схеме CMF. Корреляцию между концентрациями 24 цитокинов лимфы и числом клеток структурных зон лимфатических узлов оценивали по коэффициенту ранговой корреляции Спирмена. Результаты. После химиотерапии РМЖ, по сравнению с РМЖ без лечения, морфологические преобразования в лимфатических узлах свидетельствуют о снижении активности местного иммунного ответа. Исследование корреляции концентрации цитокинов в лимфе со структурными изменениями в лимфатических узлах выявило зависимости направленные на повышение иммуномодулирующего и противоопухолевого действия цитокинов. После оперативного лечения РМЖ и последующей химиотерапии, по сравнению только с химиотерапией РМЖ, выявлены положительные связи иммунобластов с цитокином GRO/KC в герминативных центрах, цитокина IL-6 - с митотически делящимися клетками в герминативных центрах и мозговых тяжах, IL-5 - с иммунобластами в мозговых тяжах, хемокина MIP-1a - со зрелыми плазматическими клетками в мозговых синусах. Увеличено количество иммунобластов, средних и малых лимфоцитов в герминативных центрах, возросло количество малых лимфоцитов, незрелых и зрелых плазматических клеток в мозговых синусах. Увеличены площади мозговых тяжей и паракортикальной зоны. Выявлена корреляция: цитокина IL-1α с малыми лимфоцитами, IL-6 с иммунобластами, IL-7 и IL-18 - со средними лимфоцитами, GRO/KC - с иммунобластами, IL-17 - с макрофагами в Т-зависимой зоне; IL-7 и IL-18 - с иммунобластами, IL-12 - с макрофагами, MIP-1a и MCP-1 со зрелыми плазматическими клетками в мозговых синусах. Заключение. После оперативного лечения РМЖ c последующей химиотерапией, по сравнению только с химиотерапией РМЖ, выявлены взаимозависимости концентрации цитокинов в лимфе грудного протока с морфологическими изменениями в брыжеечных лимфатических узлах, которые могут указывать на повышение активности местного звена иммунного ответа. The aim of this study was to analyze correlations of the morphometry of mesenteric lymph nodes with cytokine concentrations in thoracic duct lymph in chemotherapy and surgical treatment with subsequent chemotherapy of breast cancer. Methods. Breast cancer was induced by subcutaneous injection of N-methyl-N-nitrosourea 5 times with 7-day intervals, into the region of the 2nd breast on the right. The course of chemotherapy was performed according to the CMF scheme. Correlations between concentrations of 24 cytokines of the lymph and cells of lymph node structural regions were estimated by the Spearman rank correlation coefficient. Results. After chemotherapy for breast cancer compared to untreated breast cancer, morphological transformations in lymph nodes indicated decreased activity of the local immune response. Analysis of correlations between lymph concentrations of cytokines and structural changes in lymph nodes identified relationships aimed at increasing the immunomodulatory and antitumor effects of cytokines. After surgical treatment of breast cancer and subsequent chemotherapy compared to chemotherapy alone, positive correlations were found for immunoblasts with cytokine GRO/KC in germinative centers, for cytokine IL-6 with mitotically dividing cells in germinative centers and medullary cords, for IL-5 with immunoblasts in medullary cords, and for chemokine MIP-1a with mature plasma cells in medullary sinuses. Numbers of immunoblasts and medium and small lymphocytes were increased in germinative centers whereas numbers of small lymphocytes and immature and mature plasma cells were increased in medullary sinuses. Areas of medullary cords and the paracortical zone were increased. Correlations were found for cytokine IL-1α with small lymphocytes, for IL-6 with immunoblasts, for IL-7 and IL-18 with medium lymphocytes, for GRO/KC with immunoblasts, for IL-17 with macrophages in the T-dependent zone, for IL-7 and IL-18 with immunoblasts, for IL-12 with macrophages, and for MIP-1a and MCP-1 with mature plasma cells in medullary sinuses. Conclusion. After surgical treatment of breast cancer and subsequent chemotherapy compared to chemotherapy alone, cytokine concentrations in lymph of the thoracic duct were observed to correlate with morphological changes in mesenteric lymph nodes, which may indicate increased activity of the local immune response.

Aim of the study was to investigate the content of matrix metalloproteinases 2, 3, 9, in the intraocular fluid of patients with a developed stage of primary open-angle glaucoma. Material and methods. The study included 47 patients with advanced stage of primary open-angle glaucoma. The diagnosis is based on ophthalmological examination, including visual acuity, binocular ophthalmoscopy, spheroperimetry, echoophthalmography, optical coherence tomography, intraocular pressure measurement. Exclusion criteria were acute and exacerbation of chronic inflammatory diseases of the organ of vision, diabetic retinopathy, neovascular glaucoma, uveitis, hemophthalmia, autoimmune and tumor processes of any localization, as well as acute and exacerbations of chronic inflammatory diseases of any organs and systems. Patients who took for normalization of intraocular pressure drugs containing analogues of prostaglandins, that could activate the inflammatory process, were excluded from the study. 26 patients with a diagnosis of uncomplicated cataract made up a comparison group. All patients underwent determination of matrix metalloproteinases (MMP) 2, 3, 9 in the intraocular fluid. Results. Concentration of MMP-2 but not MMP-3 or MMP-9 is statistically significantly higher in the intraocular fluid of patients with a developed stage of primary open-angle glaucoma compared to individuals with uncomplicated cataracts. This may indicate importance of MMP-2 in the development of a local aseptic inflammatory process, impaired extracellular matrix and «remodeling» of the tissue structures of the organ of vision, leading to disruption of the outflow of intraocular fluid, which is an important factor in the pathogenesis of primary open-angle glaucoma.

We performed a correlation analysis of the morphometric parameters of mesenteric lymph nodes and cytokine content in the lymph of thoracic duct in rats with chemically induced breast cancer. The study showed that activity of the local immune response in the lymph nodes in breast cancer is aimed at antitumor protection. In breast cancer, the area of the paracortical zone remained at the level of the intact group, while the area of lymphoid nodules with germinative centers and the area of medullary substance increased; the number of macrophages in the thymus-dependent zone and zone responsible for humoral immunity also increased. The following positive correlations were revealed: in germinative centers and medullary substance, number of mitotic cells correlated with cytokine IL-5 content and the number of medium lymphocytes correlated with the content of chemokine MIP-1α; in the germinative centers, the number of immunoblasts correlated with the level of cytokine GRO/KC, in the paracortical zone, the number of macrophages correlated with the level of chemokine MCP-1, the number of reticular cells correlated with IL-6 and M-CSF content; in medullary substance, the number of small lymphocytes and mature cells plasma cells (their content was reduced) correlated with the level of chemokine GRO/KC, which can be caused by their migration from the lymph node.

We studied the levels of serum and lymph cytokines involved in the pathogenesis of BC. BC was induced by injection of N-methyl-N-nitrosourea to Wistar rats. The animals underwent surgery, or received polychemotherapy (cyclophosphamide, methotrexate, and 5-fluorouracil), or surgical treatment was combined with polychemotherapy; in a special group, Panagen (fragmented DNA) was added to polychemotherapy. Cytokine concentration in the lymph was measured using Bio-Plex Pro Rat Cytokoness 24-Plex Assay test system (Bio-Rad). In rats with BC, the content of most studied cytokines (IL-1β, IL-2, IL-4, IL-6, IL-7, IL-12, IL-13, MIP-1α, MIP-3α, RANTES, TNFα, and MCP-1) in the lymph and blood was significantly higher, while the content of IL-10 and GRO/KC was lower than in intact animals. Surgical resection of the tumor led to a significant decrease in the content of both pro- and anti-inflammatory cytokines in the lymph. Polychemotherapy led to a significant decrease in the content of IL-1β, IL-4, IL-6, IL-7, MIP-1α, MIP-3α, and RANTES in the serum and lymph. Comparison of the cytokine content in the serum and lymph of operated animals after polychemotherapy with and without Panagen showed that the content of most cytokines (IL-5, IL-6, IL-7, IL-10, IL-13, IL-17A , IL-18, GRO/KC, IFNγ, and MIP-3α) was higher after Panagen administration.

The aim of the study was to fulfill correlation analysis of morphometry of the mesenteric lymph nodes and the concentration of cytokines in the lymph of the thoracic duct in breast cancer induced by intramammary administration of N-methyl-N-nitrosourea, chemotherapy according to the CMF scheme (cyclophosphamide, methotrexate, 5-fluorouracil). The results of the study. At breast cancer revealed positive correlation: in the germinative centers and medullary cords of cytokine IL-5 with mitotically dividing cells, chemokines MIP-1α with average lymphocytes, in the germinative centers of immunoblasts with cytokine GRO/KC, in the paracortical zone chemokine MCP-1 with macrophages, reticular cells with IL-6 and M-CSF, in the medullary sinuses chemokine GRO/KC with small lymphocytes and mature plasma cells (number which decreases). All this may indicate the activity of the local immune response in the lymph nodes aimed on the antitumor protection. After chemotherapy of breast cancer, compared with breast cancer without treatment, revealed positive relationship, which may indicate increased immunomodulatory and antitumor actions of cytokines: correlation of interferon IFNγ with small lymphocytes (number which increased) and macrophages in the germinative centers and mitotically dividing cells in the medullary cords, correlation in the germinative centers of immunoblasts with MIP-1α and increased of number small lymphocytes in T-dependent zone lymph nodes, correlation in medullary cords of interleukin IL-17 with mature plasma cells (number which increased) , correlation of interleukin IL-18 with mature plasma cells in medullary sinuses. Conclusion. Study of the correlation of the concentration of cytokines in the lymph of the thoracic duct with structural changes in the mesenteric lymph nodes revealed dependencies aimed at increasing the immunomodulating and antitumor effects of cytokines.

Purpose. To study the content of cytokines and growth factors in the intraocular fluid of patients with developed stage of primary open-angle glaucoma (POAG).Materials and methods. 56 patients with a verified diagnosis developed stage of primary open-angle glaucomawere examined. The control group consisted of 30 patients with a diagnosis of uncomplicated cataract. A concentration of 17 cytokinesand 3 isoforms of the transforming growth factor (TGF) β was determined using a Bio-Plex Pro™ Human Cytokine 17-plex Assay and Bio-Plex Pro™ and TGFβ 3-plex Assay kit by flow-through fluorometry on a Bio-Plex 200, Bio-Rad double beam laser analyzer, USA.Results. Astatistically significant increase was shown in the concentrations of cytokines and growth factors (interleukins (IL) 4, 6, 7, 8, 12, 17, TGFβ 1, 2, 3, macrophage inflammatory protein 1 β) in the intraocular fluid of patients with developed stage of the primary open-angle glaucoma in respect to data obtained from the study of the intraocular fluid of the persons with uncomplicated cataract, as well as a statistically significant decrease in the concentrations IL-2, IL-10, granulocyte-macrophage colony-stimulating factor.Conclusion. In the pathogenesis of primary open-angle glaucoma, the activity of the local chronic inflammatory process is determined. This is confirmed by statistically significant changes in the studied cytokines and growth factors. Increase in the concentrations of the studied representatives of the superfamily of transforming growth factors-beta, which have anti-inflammatory activity, the ability to stimulate proliferation, cell growth, synthesis of extracellular matrix proteins, etc., attests to their importance in the mechanisms of primary open-angle glaucoma development. Increase concentrations of IL-7 in the intraocular fluid of patients with primary open-angle glaucoma allows us to assume participation in the pathogenesis of the primary open-angle glaucoma of this autocrine mediator of activation of the growth of lymphatic structures.

We studied the relationships between body composition parameters and plasma levels of pancreatic, gut, and adipose tissue hormones regulating energy balance and glucose metabolism in diabetic db/db mice (BKS.Cg-Dock7 m+/+ Lepr db /J). The body composition parameters in mice aged 8, 12, and 16 weeks were assessed by magnetic resonance imaging. The concentrations of insulin, glucagon, ghrelin, glucagon-like peptide-1, glucose-dependent immunotropic peptide, leptin, resistin, and plasminogen activator-1 were measured by multiplex analysis at the age of 8 and 16 weeks. In comparison with non-diabetic control ( db/+ ), db/db mice demonstrated high fat mass and reduced lean body mass and water content. In 8- and 16-week-old db/db mice, the levels of leptin ( p <0.001), insulin ( p <0.01), and glucagon-like peptide-1 ( p <0.05) were elevated and the concentration of ghrelin ( p <0.05) was reduced. The body weight and fat mass positively correlated with the levels of leptin, insulin, plasminogen activator-1, and glucagon-like peptide-1 and negatively correlated with ghrelin concentration. The results provide further details for characteristics of db/db mice, a widely used model of obesity and type 2 diabetes mellitus.

AIM:Ghrelin, a peptide hormone mostly produced by stomach, plays an important role in regulation of feeding behavior, energy balance and glucose homeostasis.THE AIM:to determine the relationships between fasting serum levels of ghrelin, body composition, adipose tissue endocrine function and glucose variability (GV) in type 2 diabetic subjects with and without obesity.MATERIALS AND METHODS:We observed 124 individuals with type 2 diabetes, including 42 non - obese subjects and 82 patients with obesity. Thirty non - obese healthy subjects were acted as control. The concentrations of ghrelin, leptin, resistin, and visfatin in the fasting serum were determined by Multiplex analysis. Body composition was assessed with DEXA. The 24-hour and nocturnal GV parameters were derived from continuous glucose monitoring.RESULTS AND DISCUSSION:Ghrelin levels in patients with diabetes were decreased significantly as compared to control (p.