Immunological factor development of external genital endometriosis

External genital endometriosis (EGE) is one of the common gynecological diseases of women of reproductive age with a relapsing, progressive course that worsens the quality of life of patients due to pain, emotional imbalance, fear of relapse and possible surgical intervention. Currently, endometriosis is recognized as one of the most common diseases associated with infertility. Thus, among fertile women with preserved childbearing function, the disease is generally diagnosed in approximately 6-7%, while among patients suffering from infertility, its frequency can reach 20-48%.However, the causes that determine reproductive dysfunction in patients with EGE are not well understood. Much attention is currently paid to the role of immunity in the formation of endometriosis. Patients with EGE show changes in both local immunity factors and immunological components of circulating blood.Purpose of the study: the study of factors of innate and adaptive immunity in patients of reproductive age with external genital endometriosis (EGE).The study included 71 patients with various stages of external genital endometriosis, the control group included 24 patients without endometriosis. Determination of the population composition of peripheral blood lymphocytes, the level of monocytes expressing TLR, activation markers, was carried out by laser flow cytometry — Immunotex (France), Caltag (USA), FITC (fluorescein isothiocynate) — labeled CD3, CD4, CD8, CD16, CD19, HLA-DR, CD282, CD284 and PE (phycoerythrin) - labeled with CD25, CD69, CD95, CD107a, CD14.External genital endometriosis is characterized by: at stages I-II of the disease - a violation of the early stages of the innate immune response (an increase in the number of monocytes expressing TLR-4, a violation of the activation and differentiation processes of immunocompetent cells, which is reflected in a decrease in the expression of CD16, CD8, CD16+HLA-DR+, CD16+CD107a+, CD8+CD107a+, at III-IV stages of the disease, there is a decrease in the level of CD16 and activation markers CD69, HLA-DR, CD107a on their surface, which is combined with a decrease in the expression of CD8, CD16, HLADR and CD107a on their surface. CD95+ and CD8+CD95+ were found at various stages of EGE.The results obtained allow us to understand the features of the functioning of innate and adaptive immunity at various stages of external genital endometriosis, and the studied immunological parameters can be used as diagnostic criteria for the formation of various stages of EGE. These data can serve as a theoretical basis for further identification of markers of EGE progression, as well as the mechanisms underlying immune inflammation

Effect of urokinase gene knockout on tissue levels of biogenic amines in mice with melanoma

The research aim was to study the dynamics of biogenic amines in the brain, tumor and intact skin of urokinase (uPA) gene knockout mice on day 21 of the B16/F10 melanoma growth.Material and methods. The study included male and female uPA gene knockout (-uPA, n = 38) and wild type mice (+uPA, n = 61). Melanoma was transplanted subcutaneously. Levels of biogenic amines were studied by ELISA in tissues obtained on day 21 of carcinogenesis.Results and discussion. Intact (-uPA) mice showed an increased total content of biogenic amines: in the skin - due to noradrenaline increase by 4.8 times in males and by 4.9 times in females, histamine - by 3.6 times in males and by 1.6 times (p < 0.05) in females, serotonin - by 3.4 times in males and by 8.3 times in females; in the brain - due to noradrenaline increase by 3.5 times in males and by 3.2 times in females, dopamine by 2.1 times in males and by 2.9 times in females, while histamine content decreased. Melanoma development in (-uPA) mice was characterized by: lower levels of adrenaline with high NA concentrations and an increase in the serotonin metabolism in the brain; higher histamine concentrations in the tumor and higher serotonin levels in the skin; similar to (+uPA) mice levels of adrenaline (males) and noradrenaline in the tumor and higher levels of adrenaline in the tumor and histamine in the skin in (-uPA) females.Conclusions. The uPA gene knockout limits the development of stress at the central regulatory level due to lower levels of A together with increasing serotoninergic mediation in the brain, as well as modulates the immune antitumor response due to higher levels of histamine in the tumor and 5 serotonin in the skin, as a result of lower monoamine oxidase activity, in mice with B16/F10 melanoma

Features of the cytokine profile in adolescents with microvascular complications of type 1 diabetes mellitus

Despite advances of modern medical science, the consequences associated with management of complications in type 1 diabetes mellitus (DM1) in children and adolescents represent a serious problem. Common development of microvascular diabetic complications (retinopathy, neuropathy, kidney damage) still remains a sufficient obstacle for achieving high quality of life and social adaptation in the young patients, thus promoting studies of immune mechanisms involved in genesis of microvasculature damage under the conditions of dysmetabolic abnormalities associated with DM1. Our goal was to assess the role of altered cytokine balance in blood serum in development of microangiopathies in adolescents with DM1.140 adolescent patients with type 1 diabetes aged 14-18 years were examined being divided in 2 groups: group I included the patients with glycated hemoglobin (HbA1c) level of > 9.0% (n = 65), and group II which included adolescents with HbA1C level of ≤ 9.0% (n = 75). Each group was divided into subgroups: Ia (n = 50) and IIa (n = 38) included adolescents with diabetic retinopathy, nephropathy or neuropathy, whereas groups Ib (n = 15) and IIb (n = 37) were without microvascular complications. The control group consisted of 36 adolescents with normal body weight, without carbohydrate metabolic disorders, and family history of diabetes mellitus. Determination of TNFα, IL-1β, VCAM-1, fractalkine levels in blood serum was performed by enzyme immunoassay using test systems “RayBiotech” (USA), “BIOSCIENCE” (USA).Development of microangiopathies in adolescents with different glycemic control is associated with increased serum concentration of the factors involved in neoangiogenesis and vascular wall remodeling, i.e., TNFα, IL-1β, VCAM-1, compared with control group (p < 0.05), and a statistically significant decrease in fractalkine level in adolescent patients with either complicated, or uncomplicated DM1. The study allowed us to suggest that occurrence of microvascular complications in adolescents with DM1 is associated with impaired immune response tending for altered cytokine balance towards Th1 type, enhanced intercellular interactions, imbalance of bioregulatory molecules, contributing to development of inflammatory immunoregulatory state. The revealed patterns of laboratory markers, along with assessment of metabolic indices, will enable personalized approaches to early diagnostics of microvascular complications in adolescents with DM1 and prevent their further progression

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

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 genedeficient 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

ВЛИЯНИЕ ХРОНИЧЕСКОЙ НЕЙРОГЕННОЙ БОЛИ НА ДИНАМИКУ ФУНКЦИОНИРОВАНИЯ nO-СИСТЕМЫ В ПРОЦЕССЕ РОСТА МЕЛАНОМЫ B16/F10 У САМЦОВ МЫШЕЙ

Since B16/F10 melanoma demonstrated gender differences in its growth in the presence of chronic neuropathic pain (cnp) and changes in the system of proangiogenic growth factors, the aim of the study was to analyze levels of components of the no-system in male mice during the growth of transplantable B16/F10 melanoma in the presence of cnp.Material and Methods. 66 male mice С57Вl/6 were used in the experiment. A model of subcutaneous growth of B16/F10 melanoma (during 3 weeks) was created in the cnp presence (sciatic nerve ligation). Concentrations of nos-2, nos-3, l-arginine, citrulline, total nitrite, nitrotyrosine and adma were determined by elisa in intact and tumor tissues.Results. A significant increase in levels of no-synthases was revealed in the skin and tumor tissues in the tumor growth with cnp from week 1, as well as a decrease in the level of total nitrite in the skin, multidirectional dynamics of adma and arginine levels, a steadily increased level of citrulline in the skin and tumor in the dynamics of tumor growth with cnp.Conclusions. Male mice with B16 melanoma growing in the presence of cnp demonstrated a more active functioning of the no-system already from week 1, compared to standard tumor growth, which might result in a greater rate of growth of melanoma with cnp. Significantly higher skin and tumor levels of citrulline in males were a distinctive feature, in contrast to melanoma with standard growth, which could be the result of inhibition of arginine synthesis and formation of a tumor auxotrophic for arginine.В связи с выявлением половых различий в росте меланомы B16/F10 на фоне хронической нейрогенной боли (ХрБ) и изменением системы проангиогенных факторов роста цель исследования состояла в изучении уровня компонентов nO-системы у самцов мышей в процессе роста перевивной меланомы B16/F10 на фоне ХрБ.Материал и методы. В эксперименте использовали 66 самцов мышей линии С57Вl/6. Создавали модель подкожного роста меланомы В16/F10 (в течение 3 нед) на фоне состояния ХрБ (перевязка седалищных нервов). В не пораженной опухолью коже и в ткани меланомы определяли концентрации nOs-2, nOs-3, l-аргинина, цитруллина, общего нитрита, нитротирозина и АДМА методом иФА.Результаты. В коже и в ткани опухоли выявлен значительно повышенный уровень nO-синтаз с 1-й нед роста опухоли на фоне ХрБ, сниженный уровень общего нитрита в коже, разнонаправленная динамика уровня АДМА и аргинина, стабильно повышенный уровень цитруллина в коже и в опухоли в динамике роста опухоли на фоне ХрБ.Заключение. У самцов мышей при росте меланомы В16 на фоне ХрБ уже с 1-й нед выявлено более активное функционирование nO-системы, чем при стандартном росте опухоли, что может обусловливать бóльшую скорость роста меланомы при ХрБ. Отличительной особенностью был значительно более высокий уровень цитруллина в коже и опухоли у самцов, в отличие от меланомы при стандартном росте, что может быть результатом ингибирования синтеза аргинина и формирования опухоли, ауксотрофной по аргинину

Инсулиноподобные факторы роста и их белки-переносчики в почках крыс при экспериментальном диабете, злокачественном росте и их сочетании

Persistent hyperglycemia resulting from diabetes mellitus causes microvascular lesions and long-term diabetic complications, such as nephropathy.The aim of the study was to analyze the levels of insulin-like growth factors (IGFs), their carrier proteins (IGFBP), and markers of kidney tissue damage (IL-18, L-FABP, cystatin C, NGAL, and KIM-1) in male rats with diabetes mellitus, tumor growth, and their combination.Materials and methods. The study included white outbred male rats (n = 32) weighing 180–220 g. The animals were divided into four groups (n = 8 each): group 1 – intact animals; controls (2) – animals with diabetes mellitus; controls (3) – animals with Guerin carcinoma; experimental group (4) – animals with Guerin carcinoma against the background of diabetes mellitus. Levels of IGF-1, IGF-2, IGFBP-1, IGFBP-2 and markers of acute kidney injury (IL-18, L-FABP, cystatin С, NGAL, and KIM-1) were determined in the kidney homogenates using enzyme-linked immunosorbent assay.Results. Increased levels of acute kidney injury markers were found in the kidneys of male rats with diabetes mellitus alone and in combination with Guerin carcinoma. In the animals with diabetes mellitus, the levels of IGF-1, IGFBP-1, and IGFBP-2 were decreased on average by 1.3 times, and the level of IGF-2 was increased by 2.1 times compared with the values in the intact male rats. The elevation of IGF-2 / IGF-1 on average by 2.8 times indicated increasing hypoglycemia in the kidney tissue of the animals with diabetes mellitus and in the experimental group with diabetes mellitus and Guerin carcinoma. In the kidney tissues of the rats with Guerin carcinoma, IGF-1 and IGF-2 were elevated on average by 1.5 times, and IGFBP-2 was decreased by 1.7 times. In the animals with malignant tumors growing against the background of diabetes mellitus, IGF-2 and IGFBP-1 were increased by 2.3 and 1.7 times, respectively, and the levels of IGF-1 and IGFBP-2 were similar to those in the intact animals.Conclusion. The study demonstrated abnormalities in the metabolic profile of the kidneys in male rats with experimental diabetes mellitus, Guerin carcinoma, and their combination.Устойчивая гипергликемия в результате сахарного диабета вызывает повреждение микрососудов и долгосрочные диабетические осложнения, такие как нефропатия.Целью настоящего исследования явилось изучение уровня инсулиноподобных факторов роста (IGF), их белков-переносчиков (IGFBP) и маркеров повреждения (IL-18, L-FABP, цистатина С, NGAL, КИМ-1) в ткани почек самцов крыс при сахарном диабете, опухолевом росте и их сочетании.Материалы и методы. В исследование включены самцы белых беспородных крыс (n = 32) массой 180– 220 г, разделены на четыре группы по 8 особей в каждой. Группа 1–интактные животные, контрольная группа (2) –животные с сахарным диабетом, контрольная группа (3) – животные с карциномой Герена, основная группа (4) – животные с карциномой Герена на фоне сахарного диабета. В гомогенатах почек методом иммуноферментного анализа определяли IGF-1, IGF-2, IGFBP-1, IGFBP-2 и маркеры острого повреждения почек: IL-18, L-FABP, цистатин С, NGAL, KIM-1.Результаты. При сахарном диабете в самостоятельном варианте и сочетанном с ростом карциномы Герена у самцов крыс в почках установлено повышение уровня маркеров острого повреждения почек. При развитии сахарного диабета уровень IGF-1, IGFBP-1 и IGFBP-2 был снижен в среднем в 1,3 раза, а уровень IGF-2 повышен в 2,1 раза относительно показателя у интактных самцов. Повышение IGF-2/IGF-1в среднем в 2,8 раза свидетельствовало о нарастании гипогликемии ткани почек животных при сахарном диабете и в группе с сахарным диабетом и опухолью Герена. При опухоли Герена в ткани почек самцов уровень IGF-1 и IGF-2 был повышен в среднем в 1,5 раза, а уровень IGFBP-2 снижен в 1,7 раза. При сочетанном развитии злокачественной опухоли на фоне сахарного диабета содержание IGF-2 и IGFBP-1 было повышено в 2,3 и 1,7 раза соответственно, а IGF-1 и IGFBP-2 не отличались от показателей у интактных животных.Заключение. Обнаружены нарушения метаболического состояния ткани почек самцов при развитии сахарного диабета, опухоли Герена и их сочетания

Динамика маркеров острого повреждения почек при использовании эпидуральной блокады во время резекции в условиях тепловой ишемии

Objective: to investigate the time course of changes in the early biomarkers of acute kidney injury in patients with clinically localized cancer during partial nephrectomy, as electively indicated, under thermal ischemia with prior epidural block.Materials and methods. To analyze the nephroprotective effect of an epidural block in kidney resection with warm ischemia, markers of acute kidney injury (cystatin C, interleukin 18, NGAL, L-FABP and KIM-1) were studied by ELISA in the blood and urine of 35 patients with local cancer with an epidural block (main group) and 37 patients with local cancer without an epidural block (control group) before surgery and 40 min after its beginning and on days 1 and 3 of the postoperative period. All patients were divided into 2 groups by the levels of cystatin C in the blood serum: 1000 ng/ml and lower, and over 1000 ng/ml.Results. Epidural block during the perioperative period in kidney resection with warm ischemia for patients with local cancer had an obvious nephroprotective effect allowing maintaining the initial renal functional parameters, in contrast to the standard disease management.Цель исследования – изучение динамики ранних биомаркеров острого повреждения почек у больных клинически локализованным раком при резекции почки по элективным показаниям в условиях тепловой ишемии с предварительным проведением эпидуральной блокады.Материалы и методы. Для изучения нефропротективного влияния эпидуральной блокады при резекции почки и тепловой ишемии в крови и моче 35 больных локализованным раком с проведением эпидуральной блокады (основная группа) и 37 пациентов с локализованным раком без проведения эпидуральной блокады (контрольная группа) до вмешательства и через 40 мин после начала операции, а также на 1-е и 3-и сутки послеоперационного периода методом иммуноферментного анализа были исследованы 5 маркеров острого повреждения почек: цистатин С, интерлейкин 18, NGAL, L-FABP и КIМ-1. По фоновому уровню цистатина С в сыворотке крови все больные были разделены на 2 группы: с уровнем показателя ≤1000 нг/мл и >1000 нг/мл.Результаты. Применение эпидуральной блокады в периоперационном периоде у больных локализованным раком при резекции почки в условиях тепловой ишемии оказывает нефропротективный эффект, позволяя сохранить функциональные показатели почек на исходном уровне в отличие от стандартного ведения больных