Peripheral Venous Blood Of Patients Research Articles

Effects of primed adipose mesenchymal stem cell‐derived exosomes on immunomodulation in Behcet uveitis

Aims/Purpose: Exosomes of interferon‐gamma (IFN‐ γ) primed (IFN + Exo) and non‐primed (IFN‐Exo) adipose‐derived mesenchymal stem cells (AdMSCs) were studied for their possible significance in the therapy of Behçet Disease (BD) uveitis (BU).Methods: Adipose tissue was used to isolate AdMSCs. Characterization as well as multipotency investigations were carried out. Exosomes of AdMSCs were also identified and characterized. Lymphocytes were isolated from the peripheral venous blood of patients with BD uveitis (n = 15) and healthy controls (n = 15). AdMSCs were preincubated for 48 h with or without IFN‐ γ. Peripheral Blood Mononuclear Cells (PBMC) of BD uveitis patients and healthy controls were cultivated with exosomes separately for 72 h. Flow cytometry was used to assess lymphocyte proliferation, viability, and apoptosis after the culture period. Real‐time polymerase chain reaction (RT‐PCR) was used to assess the expression of interleukin‐10 (IL‐10), interleukin‐17 (IL‐17), transforming growth factor‐ β (TGF‐ β), and interferon‐ γ (IFN‐ γ).Results: In patients with BD uveitis, IFN + Exo inhibited lymphocyte apoptosis. IFN + Exo reduced the viability of BD uveitis T lymphocytes. Both exosomes also raised IL‐10 level as an anti‐inflammatory cytokine and decreased IL‐17, TGF‐ β, IFN‐γ levels as a pro‐inflammatory cytokine in BD uveitis patients' lymphocytes. However, exosomes were ineffective in stimulating T lymphocyte proliferation.Conclusions: Exosomes of AdMSCs stimulated with and without IFN‐ γ decrease the abnormal inflammatory response caused by BD uveitis PBMC. This study's findings have the potential to pave the road for MSC/exosome therapy in BD uveitis.

The Role of Systemic Oxidative Status in Coronary Arterial and Peripheral Venous Blood of Patients with Unstable Angina Pectoris.

(1) Background: We aimed to analyze the oxidative status of patients with unstable angina pectoris (UA), as well as to determine the correlation of these parameters between coronary arterial and peripheral venous blood samples. (2) Methods: The study included 47 human subjects with UA and 45 control subjects. We performed clinical examinations, hemodynamic and coronary angiography measures. Also, in the blood samples, we measured routine laboratory markers and the concentration of pro-oxidants: index of lipid peroxidation (TBARS), superoxide anion radical (O2-), hydrogen peroxide (H2O2) and nitrites (NO2-), while antioxidant parameters were determined from red blood cells: reduced glutathione (GSH), catalase (CAT) and superoxide dismutase (SOD). All parameters were determined spectrophotometrically. (3) Results: Significantly higher values of TBARS and all measured antioxidants SOD, CAT and GSH were observed in the coronary arterial blood of the UA group relative to coronary arterial blood of the control subjects. On the other hand, in the peripheral venous blood samples, a significantly lower GSH value was found in the UA group compared to the control. (4) Conclusions: This study has shown that the majority of changes in all measured redox markers are found in coronary blood, especially related to the activity of antioxidant components. In patients with an unstable form of angina, prooxidants (superoxide anion radical and index of lipid peroxidation) and endogenous antioxidants (catalase, superoxide dismutase and reduced glutathione) are in direct correlation with the course of ischemic disease. Future studies, where participants would be randomized depending on symptom duration, are necessary to confirm these conclusions.

Store-operated calcium entry enhances the polarization and chemotaxis of neutrophils in the peripheral venous blood of patients with bronchial asthma by upregulating ERM protein.

Neutrophils can be rapidly recruited and are largely abundant in the airways of patients with asthma. However, whether the polarization and chemotaxis of neutrophils in patients with asthma are abnormal, and the underlying mechanisms, have not been clarified. Pseudopods formation is the initial step of neutrophils' polarization, ezrin, radixin and moesin (ERM) play an important role in the polarization of neutrophils. As an important signaling molecule in cell physiological processes, Ca2+ has been shown to be involved in the polarity changes of neutrophils. This study thus aimed to explore polarization and chemotaxis of neutrophils in patients with asthma and the underlying mechanism. Fresh neutrophils were isolated using standard separation protocols. The polarization and chemotactic activity of neutrophils were observed using Zigmond chamber and Transwell migration assay under linear concentration gradients of N-formyl-methionine-leucine-phenylalanine (fMLP) or interleukin (IL)-8. The distribution of calcium, ERMs and F-actin in neutrophils were observed by confocal laser scanning microscope. The expression of the main components of ERMs (moesin and ezrin) was detected with reverse transcription-polymerase chain reaction (RT-PCR). Compared with those in the healthy control group, the polarization and chemotaxis of neutrophils in the venous blood of patients with asthma were significantly increased, and the expression and distribution of cytoskeletal proteins F-actin and ezrin were abnormal. The expression and function of key components of store-operated calcium entry (SOCE), stromal interaction molecule 1 (STIM1), STIM2, and Orai1 of neutrophils in patients with asthma were significantly increased. The polarization and chemotaxis of neutrophils in the venous blood of patients with asthma are increased. This may be due to the abnormal expression and distribution of ERM and F-actin as a result of abnormal SOCE function.

Comparative characteristics of humoral immunity indicators and their relationship with chelation therapy in thalassemia

Introduction. Data on the increased susceptibility of patients with thalassemia to infections are of great interest to the study of various aspects of the immune status of patients.Aim. The aim of the study was to study the parameters of the humoral immunity and their relationship with chelation therapy in patients with thalassemia.Materials and methods. This study was carried out at the Republican Specialized Scientific and Practical Medical Center for Hematology from 2015 to 2020. The material of the study was the peripheral venous blood of patients with thalassemia. When determining the immunological status of patients with β-thalassemia, we examined 74 schoolchildren, aged 8–12 years. Statistical processing of clinical material was carried out using the statistical package of application programs STATISTICA 10.0.Results and discussion. The results of the study of the content of serum immunoglobulins in the blood of sick children showed a reduced synthesis of IgA (p < 0.001). At the same time, the level of IgM was significantly increased in patients with β-thalassemia, compared with the control (p < 0.001), which indicates the presence of acute inflammation in the body. Also, the results of our studies showed that there is a certain tendency to increase the level of IgG (11.2 ± 0.41 pg/ml in control versus 16.13 ± 0.58 pg/ml), there was a significant increase in the synthesis of this immunoglobulin (p < 0.001), this allows us to state about the autoimmune process. In the group of children with β-thalassemia, the level of C-reactive protein (CRP) was 4 times higher than the values of the control group (3.8 ± 0.1 ng/mg in the control versus 15.33 ± 0.17 ng/mg, p < 0.001). The level of lactoferrin in sick children was reduced by 4.2 times compared with the data of the control group. There is a dysregulation of acute phase protein factors – C-reactive protein and lactoferrin. β-thalassemia is characterized by a sharp increase in the level of pro-inflammatory cytokines – the level of IL-6 is increased by 2.9 times, and the level of IL-18 – by 8.7 times.Conclusion. The relationship between the indicators of the immune status and indicators of hemosiderosis was revealed, there is a significant difference in all the studied indicators, the use of chelation therapy contributes to positive dynamics.

Oxidative stress in patients with pulmonary hypertension

RationalePulmonary arterial hypertension is a devastating cardiovascular disease that is progressive and fatal despite advances in therapy. Animal models have shown that oxidative stress may be pathogenic in pulmonary hypertension. ObjectivesWe hypothesized that oxidative stress, as measured by increased levels of oxidants, including superoxide, may play a role in pulmonary arterial hypertension in humans. We also hypothesized that oxidants are elevated in patients with pulmonary arterial hypertension compared with normal controls and that they correlate with measures of pulmonary arterial hypertension severity. Methods and MeasurementsUsing a novel application of electron paramagnetic resonance spectroscopy to measure free radical oxidants, we initially evaluated whether levels of oxidants from whole blood collected from the pulmonary artery obtained during right heart catheterization would correlate with oxidant levels in peripheral venous blood. We then measured oxidants in peripheral venous blood in patients with pulmonary arterial hypertension and normal controls. Main ResultsResults showed a significant correlation between oxidant levels in blood collected from the pulmonary artery and peripheral vein but no significant elevation of oxidants in pulmonary hypertension patients versus control subjects. Furthermore, there were no correlations between levels of oxidants and severity of pulmonary arterial hypertension, as assessed by symptoms, functional capacity, hemodynamics, and the REVEAL risk score. ConclusionsContrary to previous animal studies and indirect measures of oxidative stress, we were unable to confirm an oxidative environment in patients with pulmonary arterial hypertension, at least when assessed by levels of free radical oxidants, such as superoxide, in whole blood via electron paramagnetic resonance spectroscopy.

IMMUNE STATUS IN PATIENTS DIAGNOSED WITH COLON CANCER

The article reflects the state of the immune status in people with oncological diseases. The data of laboratory examination of people suffering from oncological neoplasms were used. Peripheral venous blood of patients with oncological pathology of various localization was studied.

Study of the level of tumor markers and mutations in the BRCA1/2 genes in breast cancer patients with metachronous tumors.

e12563 Background: Primary multiple malignant tumors are more common in women with breast cancer (BC). However, the difference in the frequency of mutations in the BRCA1/2 genes in metachronous BC tumors has not been fully studied, and there are also conflicting data on the increase in the level of specific tumor markers in this category of patients. Methods: The survey included 58 patients with BC with II-III clinical stages of the disease, of which metachronous tumors in terms of 6 months or more were found in 26 (44.8%) patients, among them metachronous cancer of the second breast - in 14 (53 .8%), cancer of the body of the uterus - in 7 (26.9%) and ovarian cancer - in 5 (19.2%) patients. In the control group of 32 (55.2%) patients with initially diagnosed BC, with the same stages of the disease. Tumor markers were determined in the peripheral venous blood of patients using reagent kits for ELISA analysis produced in Russia. The study of mutations in the BRCA1/2 genes was performed using sequencing on the Illumina MiSeq platform in tumor samples from waxed histological blocks. Library sequencing was performed using MiSeq Reagent Kit v2. Results: ELISA analysis of the level of tumor markers in patients with BC showed that with the development of metachronous cancer, the studied markers were more common than in the control group. In patients with metachronous tumors, elevated levels of CA-125 occurred in 10 (38.5%) patients, CA-15-3 - in 8 (30.8%), CA-19-9 - in 4 (15.4%), AFP - in 5 (19.2%), CEA - in 6 (23.1%). In the control group, CA-125 was found in 6 (18.8%) patients, CA-15-3 - in 5 (15.6%), CA-19-9 - in 2 (6.3%), AFP - in 1 (3.1%), CEA - in 2 (6.3%). Excess levels significant for diagnosis were observed for CA-125 and CA-15-3 in both groups of patients, but in the group of patients with BC with metachronous tumors, these figures were approximately 2 times higher. In the rest of the patients, the oncomarkers under consideration did not reach values significant for diagnosis. The total incidence of patients with clinically significant germline variants in the BRCA1/2 genes in the control group was 5 (15.6%), of which there were predominantly patients with an increased frequency in the population (5382insC, 4154del4, C61G). The total incidence of BC patients with metachronous tumors with germline and somatic variants in the BRCA1/2 genes was 16 (61.5%), with an increased frequency in the same population, mainly in patients with germline mutations. Conclusions: Dynamic study of the level of tumor markers CA-125 and CA-15-3 and CA-19-9 may be useful in the preventive examination of patients with a history of BC as an early diagnosis of the development of metachronous tumors in this category of patients. The most important diagnostic unfavorable factor in the development of metachronous tumors in the examined BC patients was germline mutations in the BRCA1/2 genes.

The influence of transarterial chemoembolization on serum levels of soluble programed cell death ligand-1 in advanced hepatocellular carcinoma patients.

To investigate the implications of soluble programmed cell death-ligand 1 (sPD-L1) in hepatocellular carcinoma (HCC) patients treated with transarterial chemoembolization (TACE) and to evaluate the potential value of sPD-L1 to guide selection of the optimal time to begin combination therapy with TACE and immune checkpoint inhibitors (ICIs). Thirty-one HCC patients suitable for TACE and 55 healthy volunteers were enrolled in this study. Three milliliters of peripheral venous blood of patients were collected on 1 day before TACE and 3, 7, and 30 days after TACE respectively for assay of sPD-L1 using enzyme-linked immunosorbent assay. The associations of the sPD-L1 level with the clinical features, outcomes, and the fluctuation of sPD-L1 during the treatment were analyzed. The initial sPD-L1 level of patients was significantly higher than that of the control group. And it was significantly associated with BCLC stage, portal venous invasion, tumor size, and number of foci. The sPD-L1 levels of 3 and 7 days after TACE were both significantly higher than the initial level. And that of 30 days after TACE was lower than the initial level, but the difference was not statistically significant. There was no significant difference of sPD-L1 level after embolization with embolic beads of different size. The level of sPD-L1 of CR patients was lower than that of PR, SD patients, but the differences were not significant. The level of sPD-L1 was associated with tumor aggressiveness and outcomes, suggesting its role as a possible predictive biomarker. The increases in sPD-L1 after TACE suggests that combined treatment with TACE and ICIs may be a promising therapeutic strategy in HCC. One week after TACE might be a suitable time to begin the administration of immunotherapy.

Оптимізація діагностики вторинних метаболічних порушень та лікувальної тактики у пацієнтів із анемією злоякісного новоутворення при колоректальному раку

The purpose of the study was to study the content of free fraction of heparin, histamine and serotonin in plasma of peripheral venous blood of patients with malignant neoplasm anemia in colorectal cancer, depending on the severity of anemia. Materials and methods. The material for the study was the blood plasma of 445 patients (228 men and 217 women). Among them, 53 patients (31 women and 22 men) with iron deficiency anemia were examined and included in the first observation group (І) and 392 patients (206 men (52.55 %) and 186 women (47.45 %)) with colorectal cancer whose course of the underlying disease was burdened with malignant neoplasm anemia (ICD-10 code: D63.0) were included in the second observation group (II). Among the patients in the second (II) observation group, there were 222 individuals (119 men and 103 women) with malignant neoplasms of the colon (ICD-10 code: С.18), 29 individuals (16 men and 13 women) with malignant neoplasms of the rectosigmoid junction (ICD-10 code: C.19), 138 individuals (82 men and 56 women) with malignant neoplasms of the rectum (code ICD-10 C.20) and 3 patients (2 men and 1 woman) with malignant neoplasms of the anus (ICD-10 code: C.21). The mean age of the patients was 63.3 ± 1.2 years old. The plasma level of free fraction of heparin of the examined patients was determined using the photocolorimetric method on photoelectric colorimeter 56-M after its preliminary isolation by electrophoretic method according to the appropriate procedure (B. V. Mykhailychenko, S. V. Vydyborets (2000)). The plasma level of free histamine and serotonin of the examined patients was studied using the method of fluorometric analysis on the analyzer “BIAN-130”-“BIAN-100” according to the procedure of B. V. Mykhailychenko, S. V. Vydyborets (1999). Results and discussion. It was found that prior to the initiation of treatment in patients with malignant neoplasm anemia, regardless of the course of colorectal cancer, there was a significant increase in the plasma level of free fraction of heparin, histamine, serotonin (p < 0.001); the ratio of free histamine to free serotonin was also changed in comparison with the values in the control group (p < 0.05), which indicated both an increased release of heparin, histamine and serotonin from the depot, and an impaired inactivation processes of these biologically active substances. Considering all of the above and the quite obvious reasons, namely, the secondary metabolic disorders of serotonin, histamine, heparin which manifested by a significant increase in their plasma level of patients with malignant neoplasm anemia in colorectal cancer, we suggested the need to use a medicinal product in a complex of therapeutic measures which can cause antihypoxic, membrane stabilizing and anti-edema action. Conclusion. Malignant neoplasm anemia in colorectal cancer is accompanied by significant changes in the metabolism of biologically active substances – free fraction of heparin, histamine, serotonin, and the ratio of free histamine to serotonin. It was correctly concluded that in addition to the baseline therapy the administration of arginine glutamate which causes both antihypoxic and membrane-stabilizing action, reliably contributes to the normalization of secondary metabolic disorders of histamine, serotonin and heparin metabolism in malignant neoplasm anemia in patients with colorectal cancer

Результаты исследования содержания 2,3-дифосфоглицерата в эритроцитах пациентов с анемией злокачественного новообразования при раке мочеполовых органов

Every year, more than 6 million people are diagnosed with and more than 4 million people die from cancer all over the world, which is approximately 10% of the total mortality. The incidence of cancer in Ukraine shows a gradual increase, with the number of newly diagnosed patients 304-308 per 100 thousand people. Bladder cancer ranks second in the world among oncological diseases of the urinary system after prostate cancer. In the later stages of diseases, their course may be associated with malignant neoplasm anemia. The purpose of the study was to study the content of 2,3-diphosphoglycerate in erythrocytes of peripheral venous blood of patients with malignant neoplasm anemia with urothelial bladder cancer and prostate cancer, depending on the severity of anemia; to evaluate the possible diagnostic and prognostic value of the studied indicator. Materials and methods. 96 patients (64 men and 32 women) with urothelial bladder cancer were examined. There were 39 patients (28 men and 11 women) among them, the course of their underlying disease was not accompanied by the presence of anemia (first (I) observation group) and 57 patients (36 men and 21 women), the course of their underlying disease was aggravated by malignant neoplasm anemia (second (II) observation group). Also, 48 men with prostate cancer whose course of the disease was not aggravated by malignant neoplasm anemia (19 men) (third (III) observation group) and 29 men (fourth (IV) observation group) were diagnosed with malignant neoplasm anemia. The age of the patients under the survey was from 22 to 79 years old. All patients were examined after verifying the diagnosis and before starting any treatment. The content of 2,3-diphosphoglycerate in erythrocytes of peripheral venous blood was determined by the method of I. S. Luganova, M. N. Blinov (1975). Results and discussion. The content of 2,3-diphosphoglycerate in peripheral venous erythrocytes of patients with malignant neoplasm anemia due to urothelial bladder cancer and prostate cancer has been studied. It was found that malignant neoplasm anemia in patients with urothelial bladder cancer and prostate cancer is associated with an increase in the studied indicator (p<0.05). The article discusses the possible causes and pathogenetic mechanisms of the identified changes. Conclusion. Patients with urothelial bladder cancer and prostate cancer have an imbalance of energy metabolism that occurs in erythrocytes. This process is manifested by an increase in the amount of 2,3-diphosphoglycerate

Evaluation of circulating CD34+ stem cells in peripheral venous blood in patients with varying degrees of periodontal disease.

Periodontal disease presents a challenge for modern medicine, and research on the use of stem cells as a treatment is currently underway. The study included 45 patients who were given a thorough physical examination. Additionally, an evaluation of their medical history of the disease, degree of progression of periodontal disease, and the level of CRP in the blood was carried out. Patients were divided into 4 groups: 4 patients were in the first group with no periodontal disease, 8 patients in the second group with a moderate level, 20 patients in the third group with an advanced level, and 13 patients in the fourth group were toothless. For each group, the use of stem cells as a treatment of antibody-labeled CD34+ stem cells, lymphocytes, and leukocytes was conducted. A statistically significant positive correlation was observed in CD34+ stem cells in proportion to lymphocytes in the moderate (0.80), in the advanced (0.75), and in the toothless groups (0.70). The ratio of CD34+ stem cells to leukocytes was statistically significant in the toothless group (0.92) and in the advanced group (0.91). A statistically significant increase was noted in the level of CRP in the previously mentioned groups of patients, and the highest concentration of CD34+ stem cells in the advanced group. The highest concentration of CD34+ cells was observed in the group of patients with advanced periodontal disease.

Let7f‑5p attenuates inflammatory injury in invitro pneumonia models by targeting MAPK6.

Pneumonia accounts for ~1.3 million mortalities in children per year worldwide. MicroRNAs are implicated in several diseases, including cancer and pneumonia; however, the role of let7f-5p in pneumonia is not completely understood. In the present study, lipopolysaccharide (LPS) was used to establish an in vitro pneumonia model in A549 and WI-38 cells. The reverse transcription-quantitative PCR (RT-qPCR) and western blotting results demonstrated that let7f-5p expression levels were significantly decreased, whereas MAPK6 expression levels were significantly increased in the peripheral venous blood of patients with pneumonia and in LPS-induced A549 and WI-38 cells compared with healthy volunteers and control cells, respectively. Furthermore, the dual-luciferase reporter assay demonstrated that let7f-5p targeted the 3′-untranslated region of MAPK6. The ELISA and RT-qPCR results demonstrated that let7f-5p mimic ameliorated LPS-induced inflammatory injury in A549 and WI-38 cells, as demonstrated by decreased expression levels of proinflammatory cytokines, including TNF-α and IL-6. In addition, the Cell Counting Kit-8 assay results indicated that let7f-5p mimic ameliorated LPS-induced reductions in cell viability, and the western blotting results demonstrated that let7f-5p mimic reversed LPS-induced activation of the STAT3 signaling pathway. Notably, the aforementioned let7f-5p-mediated effects were reversed by MAPK6 overexpression. Collectively, the results of the present study suggested that let7f-5p inhibited inflammation by targeting MAPK6 in the in vitro pneumonia model, thus let7f-5p may serve as a potential novel therapeutic target for pneumonia.

Association of NM23 polymorphisms and clinicopathological features and recurrence of invasive pituitary adenomas.

Pituitary adenomas (PAs) are intracranial tumors, deriving from anterior pituitary cells. Previously, expression of non-metastasis-23 (NM23) gene has been shown to correlate with the progression of PAs. In this study, we aim to determine whether there is association between specific NM23 polymorphisms and invasive pituitary adenoma (IPA). Genotypes of rs2302254 and rs16949649 of NM23 were identified in the peripheral venous blood of patients by PCR-RLFP. Next, the correlation between specific genotypes of rs2302254 and rs16949649 and risk of IPA was investigated. Finally, the correlations between NM23 polymorphisms and tumor size, Ki67 LI and recurrence of IPA were analyzed with 3 to 24months follow-up for the enrolled patients. We observed that the TT genotype at rs16949649 correlated closely with a high risk of IPA, while CC and CT genotypes reduced the risk of IPA. CC genotype at rs2302254 increased the risk of IPA, while CT and TT genotypes reduced the risk of IPA. Trs16949649Crs2302254 haplotype of NM23 was found to be a high-risk haplotype for IPA. TT genotype at rs16949649 and CC genotype at rs2302254 were associated with higher rates of tumors larger than 20mm, Ki67 LI and tumor recurrence. Taken together, the present study provides evidence that NM23 polymorphisms are closely associated with the incidence and recurrence of IPA. Specifically, TT genotype at rs16949649 and CC genotype at rs2302254 are risk factors of IPA. NM23 polymorphisms could therefore be used as a reference for clinical diagnosis and prognosis of IPA.

Decreased ALCAM expression and promoter hypermethylation is associated with preeclampsia.

Preeclampsia (PE) is a major obstetrical complication that results in maternal and fetal morbidity and mortality. Aberrant epigenetic modifications are widely involved in the pathogenesis of PE. Previously, the activated leukocyte cell adhesion molecule (ALCAM) was reported to be required for blastocyst implantation but has not been described in the context of pathological pregnancy. This study explored the expression of ALCAM and its methylation levels in the placentas and peripheral venous blood of patients with PE from a Chinese Han population. The mRNA and protein expression levels of ALCAM were downregulated in the PE placentas compared with the control placentas (P < 0.05). The methylation rate of the ALCAM gene promoter was considerably elevated in the placentas (P = 0.003, odds ratio (OR) = 0.264, 95% confidence interval (95% CI) [0.108-0.647], cases n = 47, controls n = 53) and peripheral blood (P = 0.007, OR = 0.455, 95% CI [0.256-0.806], cases n = 100, controls n = 100) of the PE patients compared with those of the normotensive women, suggesting a negative relationship between ALCAM methylation and gene transcription. Moreover, the transcriptional expression of ALCAM was dramatically increased by demethylating treatment in trophoblastic cells. ALCAM is expected to be involved in the pathogenesis of PE through methylation regulation.

ESTIMATION OF LEVEL OF SUBPOPULATION CD14&lt;sup&gt;+&lt;/sup&gt;/CD16&lt;sup&gt;+&lt;/sup&gt; MONOCYTES IN PATIENTS WITH JUVENILE DEPRESSIONS

At present, at least two phenotypically different subpopulations of blood monocytes have been identified: "classical" monocytes with the phenotype CD14 ++ /CD16 - and "nonclassical" (pro-inflammatory) with the phenotype CD14 + /CD16 + . It has been established that monocytes CD14 + /CD16 + participate in latent or so-called systemic non-infectious inflammation in the human body along with blood neutrophils and microglial cells of the brain, which are the analogue of monocyte/macrophage system in the brain. Monocyte function disorders are considered as one of the links in the neurodegenerative process underlying the neuroinflammatory hypothesis of the development of schizophrenia. In this connection, the aim of this research was to study in patients with juvenile depression the amount of pro-inflammatory monocytes in terms of the expression level of CD14 + /CD16 + receptors as well as the activity of leukocyte elastase and a 1-proteinase of elastase inhibitor. It was also interesting to determine the mechanisms of their interaction in the pathogenesis of immunosuppression, in particular, to identify a possible connection of pro-inflammatory monocytes with the activity of leukocyte elastase and a 1-proteinase of elastase inhibitor in patients with juvenile depression at the early stage of disease. 27 male patients aged 17-23 years were examined. The disease diagnostics was carried out in accordance with the criteria of ICD-10 according to which all patients have depression with the syndrome of attenuated psychotic symptoms ( Attenuated Psychotic Syndrome) in the structure of psychopathological disorders. 12 mentally healthy age- and gender-matched persons were examined as controls. Monocytes were obtained by the conventional method of adhesion to the plastic surface from mononuclear cells isolated from peripheral venous blood of patients and healthy controls by centrifugation in a ficoll-urographin density gradient (ρ = 1.077). In the blood of patients and healthy controls, the level of monocytes with the phenotype CD14 + /CD16 + , the enzymatic activity of leukocyte elastase and the functional activity of α1-proteinase inhibitor were studied. The clinical and laboratory study showed an increase of more than twofold in the level of the pro-inflammatory subpopulation of CD14 + /CD16 + monocytes in patients compared with its level in healthy individuals. The increase was accompanied by an elevation of leukocyte elastase and a 1-proteinase inhibitor activity. A positive correlation was found between the increase of surface receptors CD14 + /CD16 + expression on monocytes and elevation of the leukocyte elastase activity. These results confirm participation of CD14 + /CD16 + monocytes in the development of latent or non-infectious immune inflammation, and determine the mechanisms of possible interaction between monocytes and neutrophils in the development of immune inflammation in patients with juvenile depressions with the Attenuated Psychotic Syndrome.

Иммунофенотипическая характеристика моноцитов периферической крови пациентов с болезнью Альцгеймера и синдромом мягкого когнитивного снижения

Aim : to study the features of subpopulation composition of peripheral blood monocytes in patients with Alzheimer's disease (AD) and with amnesic mild cognitive impairment syndrome (aMCI). Material and methods : 20 AD patients aged 73.8 ± 6.8 years, and 10 patients with aMCI aged 61.8 ± 9.6 years enrolled in the study were examined. 16 mentally and somatically healthy age- and gender-matched persons were examined as controls. The peripheral venous blood of patients and controls was the material for the study. The relative number of monocytes was determined with the help of FC-500 Flow Суtofluorometer [Beckman Coulter, USA] using the same settings for all samples. Results : the study revealed a significant decrease of the relative number of classical subpopulations (p &lt; 0.01) and an increase of the proinflammatory monocytes level (p &lt; 0.01) in the group of AD patients compared with the control group. The severity of changes of the monocytes subpopulation composition depended on the dementia severity. The sharpest change of the monocytes composition compared to controls was typical for patients with moderate dementia (p &lt; 0.001). An increase of the subpopulation level of proinflammatory monocytes was also revealed in patients with aMCI (p &lt; 0.05). Conclusion : the study results have revealed the redistribution of monocyte subpopulations in AD and aMCI patients: the decrease in the classical monocytes subpopulation on the background of the increase of the monocytes subpopulation with proinflammatory activity.

THE STRUCTURAL FEATURES OF VASCULAR ENDOTHELIUM IN ACUTE CEREBRAL ISCHEMIA

The aim of the research was to study the number and structural properties of desquamated endothelial cells (DECs) in the peripheral blood in carotid ischemic stroke (CIS) and carotid transient ischemic attacks (TIAs) and its connection with the marker of endothelial dysfunction - endothelin-1. We examined 35 patients with the first CIS, on days 1st and 10th, and also 34 patients with symptomatic carotid TIAs, on days 1st and 10th of the observation. Middle age of the examined patients with a CIS was 63,7±1,0. Middle age of the examined patients with the ТIАs was 54,7±1,0. 25 practically healthy persons were examined as a group of control. Neurologic deficit was assessed with the National Institutes of Health Stroke Scale (NIHSS). DECs were estimated by CD34 immunobead capture in the peripheral venous blood of patients and persons of control group. We studied the level of endothelin-1 in the peripheral venous blood of patients and persons of control group using the enzyme immunoassay using the Biomedica (Austria) during the first 24 hrs and on day 10. Statistical processing of the obtained results was carried out using statistical analysis package Statistica. In this case, the mean value, the standard error and the correlation analysis were determined. Samples were compared using the Student's criterion (t) and the correlation coefficient (r). During an examination of 35 patients in the acute period of CIS and 34 patients with carotid TIAs using the immunocytochemical method the number of DECs was studied in venous blood. The quantitative analysis of vascular endothelium in acute cerebral ischemias showed its statistically unreliable differences in CIS and TIAs. A conclusion is drawn about the general mechanisms of endothelial dysfunction in CIS and TIAs. The number of DECs significantly correlates with the terms of disease. Regress of this indicator is noted in patients by the end of follow-up in both observation groups. During the first 24 hrs in patients with CIS and TIAs density of DECs of blood directly correlates with the level of endothelin-1 blood. The endothelin-1 level tends to decrease by the 10th day of observation and the correlation force with the DECs level is correspondingly reduced.

Beneficial effect of refined red palm oil on lipid peroxidation and monocyte tissue factor in HCV-related liver disease: a randomizer controller study

A large amount of endotoxin can be detected in the peripheral venous blood of patients with liver cirrhosis, contributing to the pathogenesis of hepatotoxicity because of its role in oxidative stress. The present study aimed to test the effect of the supplementation with red palm oil (RPO), which is a natural oil obtained from oil palm fruit (Elaeis guineensis) rich in natural fat-soluble tocopherols, tocotrienols and carotenoids, on lipid peroxidation and endotoxemia with plasma endotoxin-inactivating capacity, proinflammatory cytokines profile, and monocyte tissue factor in patients with chronic liver disease. The study group consisted of sixty patients (34 males and 26 females; mean age 62 years, range 54-75) with Child A/B, genotype 1 HCV-related cirrhosis without a history of ethanol consumption, randomly enrolled into an 8-week oral daily treatment with either vitamin E or RPO. All patients had undergone an upper gastrointestinal endoscopy 8 months before, and 13 out of them showed esophageal varices. Both treatments significantly decreased erythrocyte malondialdehyde and urinary isoprostane output, only RPO significantly affected macrophage-colony stimulating factor and monocyte tissue factor. Liver ultrasound imaging did not show any change. RPO beneficially modulates oxidative stress and, not least, downregulates macrophage/monocyte inflammatory parameters. RPO can be safely advised as a valuable nutritional implementation tool in the management of chronic liver diseases.

A preliminary study on the relationship between circulating tumor cells count and clinical features in patients with non-small cell lung cancer.

To evaluate the clinical value of circulating tumor cells (CTC) count in peripheral venous blood of patients with non-small cell lung carcinoma (NSCLC). A total of 50 NSCLC patients who were diagnosed in Wuxi No. 2 People's Hospital from January 2013 to December 2013 were selected as the NSCLC group, 35 patients with lung benign tumor as the benign group, and 28 healthy subjects as the normal control group. Venous blood samples (3 mL) were collected in all subjects for counting the CTC, and a result of ≥8.7 was judged to be positive. The relationships between the positive rate of CTC and the age, sex, pathological type, and clinical stage of NSCLC were analyzed. CTC count was significantly higher in NSCLC group than in benign group and normal control group. In NSCLC patients, CTC count was not significantly correlated with sex, age, or the pathological type (P>0.05) but was closely related to clinical stage (P<0.01). Among NSCLC patients, CTC count significantly increased along with tumor progression. CTC count shows certain correlation with the clinical features of NSCLC and thus can, to certain extent, reflect the status of the disease.

Cytokine inhibition in chronic fatigue syndrome patients: study protocol for a randomized controlled trial.

BackgroundChronic fatigue syndrome (CFS) is a medically unexplained syndrome for which no somatic or pharmacological treatment has been proven effective. Dysfunction of the cytokine network has been suspected to play a role in the pathophysiology of CFS. The disturbances of the cytokine network detected in CFS patients are highly variable, in part due to the lack of adequate controls in many studies. Furthermore, all studies have been performed on peripheral venous blood of patients. As cytokines mainly act in tissues, for example, the brain, the information that can be derived from peripheral blood cells is limited. The information regarding the possible role of cytokines in the pathophysiology could come from intervention studies in which the activities of relevant cytokines are reduced, for example, reducing interleukin-1, interleukin-6 or tumor necrosis factor. In this study, the clinical usefulness of anakinra, an IL-1 antagonist, will be assessed in patients with CFS.Methods/DesignA randomized placebo-controlled, double-blind trial will be conducted. Fifty adult female patients meeting the Centers for Disease Control (CDC) criteria for CFS and without psychiatric co-morbidity will be included. After inclusion, patients will be randomized between treatment with anakinra (recombinant human interleukin-1 receptor antagonist) or placebo. Each group will be treated for 4 weeks. Outcome measures will be assessed at baseline, after 4 weeks of intervention, and 6 months after baseline assessment. The primary outcome measure will be fatigue severity at 4 weeks, measured with the validated Checklist of Individual Strength (CIS). Secondary outcome measures are functional impairment, physical and social functioning, psychological distress, pain severity, presence of accompanying symptoms, and cytokine and cortisol concentrations.DiscussionThis is the first randomized placebo-controlled trial that will evaluate the effect of interference with IL-1 on the experience of fatigue in patients with CFS. The results of this study may expand treatment options for patients with CFS, for whom graded exercise therapy and cognitive behavioral therapy are the only evidence-based interventions that exist at this moment.Trial registrationClinicaltrials.gov: NCT02108210. Clinicaltrials.gov registration date: 8 April 2014. EudraCT: 2013-005466-19