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Typical relationships between the indexes of lipid peroxidation/ antioxidant protection and immune regulation in the patients with chronic atrophic gastritis and stomach cancer 慢性萎缩性胃炎和胃癌患者脂质过氧化/抗氧化保护指标与免疫调节的典型关系
Pub Date : 2020-12-01 DOI: 10.15789/1563-0625-trb-2032
O. Smirnova, V. Tsukanov, A. Sinyakov, O. Moskalenko, N. G. Elmanova, E. S. Ovcharenko
Chronic atrophic gastritis and gastric cancer represent distinct steps of one pathogenic process. The risk of developing cancer of the stomach is directly proportional to the degree of atrophic changes simultaneously detected in antral segment and in the body of the stomach. The role of immune system in transformation of precancerous diseases into cancer is beyond doubt. During development of the malignant disease, the changes in lipid peroxidation systems – antioxidant defense become significant and contribute to the progression of the tumor and the development of metastases. A simultaneous study of lipid peroxidation and antioxidant defense indices along with phagocytic activity will allow us to evaluate relative contribution of these processes to development of chronic atrophic gastritis and gastric cancer. Purpose of the present study was to assess correlations between the lipid peroxidation indices, i.e. antioxidant protection, and chemiluminescent activity of neutrophilic granulocytes and monocytes in chronic atrophic gastritis and gastric cancer. Forty patients with chronic gastritis, 22 patients with chronic atrophic gastritis and 40 patients with gastric cancer were examined. The control group consisted of 50 practically healthy age-matched volunteers. Evaluation of spontaneous and induced production of reactive oxygen species by neutrophils and monocytes was carried out by chemiluminescent analysis. The parameters of lipid peroxidation/ antioxidant protection were determined by spectrophotometric methods. Statistical data processing was carried out using the Statistica v. 8.0 program (StatSoft Inc., USA). The normal distribution of indices was tested using the Kolmogorov–Smirnov method (adjusted by Lillefors). Quantitative indicators, given the normal distribution, were described using the median (Me) and interquartile scatter (Q0.25-Q0.75). To study statistical significance of differences between quantitative characteristics, the Mann–Whitney test was used. To study strength of relationships of these indicators, the Pearson rank correlation coefficient (r) was calculated. The critical significance level (p) when testing statistical hypotheses was taken equal to 0.05. Correlation analysis showed that the weight of positive correlations increases in patients with chronic atrophic gastritis, and it decreases in patients with gastric cancer, the strength of the correlation dependence and new relationships appear between chemiluminescent activity of neutrophils and monocytes in a spontaneous and induced state, and the amounts of malonic dialdehyde, enzyme activities of superoxide dismutase and catalase. In chronic atrophic gastritis and gastric cancer we have established the features of correlation patterns between lipid peroxidation/antioxidant protection indices, and activity of neutrophils and monocytes.
慢性萎缩性胃炎和胃癌是一个不同的致病过程。发生胃癌的风险与同时在胃窦段和胃体检测到的萎缩变化程度成正比。免疫系统在癌前病变转化为癌症中的作用是毋庸置疑的。在恶性疾病的发展过程中,脂质过氧化系统-抗氧化防御的变化变得显著,并有助于肿瘤的进展和转移的发展。同时研究脂质过氧化和抗氧化防御指数以及吞噬活性将使我们能够评估这些过程在慢性萎缩性胃炎和胃癌发展中的相对贡献。本研究旨在探讨慢性萎缩性胃炎和胃癌中脂质过氧化指标(抗氧化保护)与中性粒细胞和单核细胞化学发光活性的相关性。对40例慢性胃炎、22例慢性萎缩性胃炎和40例胃癌患者进行了检查。对照组由50名年龄相当的健康志愿者组成。通过化学发光分析评估中性粒细胞和单核细胞自发和诱导产生的活性氧。采用分光光度法测定脂质过氧化/抗氧化保护参数。统计数据处理使用Statistica v. 8.0程序(StatSoft Inc., USA)。各项指标的正态分布采用Kolmogorov-Smirnov方法进行检验(经Lillefors校正)。定量指标,鉴于正态分布,用中位数(Me)和四分位数散点(Q0.25-Q0.75)来描述。为研究数量特征间差异的统计学意义,采用Mann-Whitney检验。为了研究这些指标之间的关系强度,我们计算了Pearson秩相关系数(r)。检验统计假设时的临界显著性水平(p)为0.05。相关性分析显示,慢性萎缩性胃炎患者的正相关权值增加,胃癌患者的正相关权值降低,自发和诱导状态下中性粒细胞和单核细胞的化学发光活性与丙二醛含量、超氧化物歧化酶和过氧化氢酶活性之间的相关依赖强度增强,出现了新的关系。在慢性萎缩性胃炎和胃癌中,我们建立了脂质过氧化/抗氧化保护指数与中性粒细胞和单核细胞活性的相关模式特征。
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引用次数: 0
Netosis in patients with systemic lupus erythematosus 系统性红斑狼疮患者的Netosis
Pub Date : 2020-12-01 DOI: 10.15789/1563-0625-nip-1978
V. V. Zhelezko, I. Novikova
The article presents the data on assessment of functional features of neutrophils in 34 patients with systemic lupus erythematosus (SLE). Development of neutrophil extracellular traps (NETs) was evaluated in cell cultures incubated in vitro for 30 and 150 minutes (basal levels, NETBAS30 and NETBAS150, respectively), and in the presence of heat-inactivated S. аureus (strain ATCC 25923, 108 CFU/ml) (stimulated levels, NETST30 and NETST150, respectively). NET looks like thin free-lying extracellular fibrillar structures, 2-3 times exceeding the size of unchanged granulocyte. The result was expressed as percentage and relative amount of extracellular traps per 100 counted leukocytes. Phagocytic activity of neutrophils was evaluated as phagocytosis of S. аureus by counting the percentage of neutrophils that engulfed phagocytic index of microbial particles (PI); the average number of phagocytosed objects per neutrophil phagocytic number (PC). ROS-producing activity was determined in the reduction of Nitroblue Tetrazolium tested in spontaneous and stimulated S. аureus variants (NBTBAS and NBTST, respectively). The result was expressed as the percentage of formazan-positive cells per 100 white blood cells. Nitroxide-producing properties were determined using the Crow (1999) method in spontaneous and stimulated samples for the accumulation of the nitrated amino acid tyrosine (3-nitrothyrosine, 3-NTBAS, and 3-NTST, respectively). We revealed a decrease in ROS production, phagocytosis and NO-forming activity of neutrophils associated with increased netosis. Activation of the netosis was observed in cell cultures without stimulation, indicating the in vivo formation of networks in SLE. The NET increase is most pronounced in the patients with lupus nephritis (p < 0.05), and in remission of the disease (p < 0.05). We have revealed a correlation of NET formation parameters with duration and degree of SLE activity (rs = -0.6; p = 0.001, and rs = 0.39; p = 0.02, respectively); autoantibody titers (anti-dsDNA and ANA) (rs = 0.67; р = 0.047 and rs = 0.59; р = 0.034, respectively); prothrombin complex activity (rs = 0.6; p = 0.036), as well and urea and creatinine levels (rs = 0.47; p = 0.037 and rs = 0.39; p = 0.048, respectively). The parameters of NETs can be considered a promising biomarker for verifying the diagnosis of SLE, evaluation of clinical activity, disease severity, and predicting the development of complications.
本文介绍了34例系统性红斑狼疮(SLE)患者中性粒细胞功能特点的评估数据。在体外培养30分钟和150分钟(基础水平,分别为NETBAS30和NETBAS150)的细胞培养物中,以及在热灭活的沙门氏菌(菌株ATCC 25923108CFU/ml)存在下(刺激水平,分别是NETST30和NETST150),评估中性粒细胞外陷阱(NETs)的形成。NET看起来像是薄的游离的细胞外原纤维结构,是不变粒细胞大小的2-3倍。结果表示为每100个计数的白细胞中细胞外陷阱的百分比和相对量。通过计算吞噬微生物颗粒吞噬指数(PI)的中性粒细胞百分比,将中性粒细胞的吞噬细胞活性评估为对S.аureus的吞噬作用;每个中性粒细胞吞噬细胞数量(PC)的被吞噬物体的平均数量。ROS产生活性在硝基蓝四氮唑的还原过程中测定,该还原过程在自发和刺激的硫变体(分别为NBTBAS和NBTST)中进行测试。结果表示为每100个白细胞中甲赞阳性细胞的百分比。使用Crow(1999)方法在自发样品和刺激样品中测定了硝酸化氨基酸酪氨酸(分别为3-硝基甲状腺素、3-NTBAS和3-NTST)的积累产生亚硝酸盐的性质。我们发现中性粒细胞ROS产生、吞噬作用和NO形成活性的降低与网状细胞增多有关。在没有刺激的细胞培养中观察到网状结构的激活,表明SLE体内形成了网状结构。NET的增加在狼疮性肾炎患者中最为明显(p<0.05),在疾病缓解时最为显著(p<0.01)。我们发现NET形成参数与SLE活动的持续时间和程度相关(分别为rs=-0.6;p=0.001和rs=0.39;p=0.02);自身抗体滴度(抗dsDNA和ANA)(分别为rs=0.67;р=0.047和rs=0.59;р=0.034);凝血酶原复合物活性(rs=0.6;p=0.036)以及尿素和肌酐水平(分别为rs=0.47;p=0.037和rs=0.39;p=0.048)。NETs的参数可以被认为是一种很有前途的生物标志物,用于验证SLE的诊断、评估临床活动、疾病严重程度和预测并发症的发展。
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引用次数: 0
Is there a dependence between expression of integrin receptors by peripheral blood immune cells and duration of tuberculous granuloma existence in the patients? 患者外周血免疫细胞整合素受体的表达与结核性肉芽肿的存在时间之间是否存在依赖性?
Pub Date : 2020-12-01 DOI: 10.15789/1563-0625-ita-2088
О. V. Berdyugina
Over recent years, the number of patients with tuberculosis has not decreased in the country and in worldwide. This is due to high resistance of the pathogen and changing mechanisms of bacterial perception by the human immune system thus requiring closer examination of the issue. Cell fusion during the formation of pulmonary tuberculous granuloma involves a large number of adhesive events. Importance of α1β1 integrin has been shown for the granuloma integrity during the chronic phase of infection. It has been proven that pulmonary tuberculous granuloma should be monitored, including with the detection of cells expressing CD11c, since they support the continuous priming of T cells at different stages of infection. The aim of this study was to answer the question, if there is a different expression of integrin receptors by immune cells from the patient’s peripheral blood at different stages of the existence of pulmonary tuberculous granuloma? The study involved 38 people: the first group (control) consisted of 15 practically healthy people; a second group included 11 subjects with pulmonary tuberculous granuloma; the condition was first diagnosed 2 to 10 months before the present study. A third group consisted of 12 patients with pulmonary tuberculous granuloma, with primary diagnosis established 12 to 219 months before this study. All the participants underwent a general clinical blood tests using a 5 Diff Mythic 22 AL analyzer (Cormay, Poland). The adhesion markers CD11b, CD11c were detected with a Coulter Epicx XL instrument (Beckman Coulter, USA). The following peripheral blood cell populations were determined: CD14- CD13lowCD11b+, CD14- CD13lowCD11c+, CD14+CD11b+, CD14+CD11c+, CD45+CD3- CD16+CD56+, CD45+CD3- CD16+CD56+CD11b+. Statistical processing of the results was performed in the Windows 10 operating environment (Microsoft Corp., USA), using Statistica v. 12.5 software (StatSoft, USA). Kruskal–Wallis one-way analysis of variance (pk-w), with differences significant at p < 0.017, as well as the Wald–Wolfowitz test (pw-w) at a significance level of p < 0.05 were used as criteria for assessing differences between the compared groups. In addition, cluster and factor analysis were implemented. When studying the role of β2-integrins, we have found that they play an important role in maintaining the existence of pulmonary tuberculous granuloma. An increase in total number of granulocytes, and CD11b-expressing granulocytes, a decrease in the population of lymphocytes, NK cells and NK cells expressing CD11c proved to be distinctive in cases of pulmonary tuberculous granuloma detected 0.5 years before the study. Characteristic changes observed in the study of peripheral blood in the patients with pulmonary tuberculous granuloma detected 9.5 years before the study were as follows: an increase in the leukocyte population, total monocyte number, as well as CD11band CD11c-expressing monocytes.
近年来,该国和全世界的结核病患者人数没有减少。这是由于病原体的高耐药性和人类免疫系统细菌感知机制的变化,因此需要对该问题进行更深入的研究。肺结核性肉芽肿形成过程中的细胞融合涉及大量的粘附事件。在感染的慢性期,α1β1整合素对肉芽肿的完整性具有重要意义。已经证明,应该监测肺结核性肉芽肿,包括检测表达CD11c的细胞,因为它们支持T细胞在不同感染阶段的持续启动。本研究的目的是回答这样一个问题:在肺结核性肉芽肿存在的不同阶段,患者外周血的免疫细胞是否存在整合素受体的不同表达?这项研究涉及38人:第一组(对照组)由15名实际健康的人组成;第二组包括11例肺结核性肉芽肿患者;在本研究前2至10个月首次诊断出这种情况。第三组由12名肺结核性肉芽肿患者组成,在本研究前12至219个月确定了初步诊断。所有参与者都使用5 Diff Mythic 22 AL分析仪(波兰科梅)进行了一般临床血液测试。用Coulter Epicx XL仪器(Beckman Coulter,USA)检测粘附标记CD11b、CD11c。测定以下外周血细胞群:CD14-CD13低CD11b+、CD14-CD13低CD11c+、CD14+CD11b+、CD14+CD11c+,CD45+CD3-CD16+CD56+、CD45+CD3-CD16+CD56+CD11b+。结果的统计处理是在Windows 10操作环境(Microsoft Corp.,USA)中使用Statistica v.12.5软件(StatSoft,USA)进行的。Kruskal–Wallis单向方差分析(pk-w)和Wald–Wolfowitz检验(pw-w)被用作评估比较组之间差异的标准,前者的差异在p<0.017时具有显著性,后者的显著性水平为p<0.05。此外,还进行了聚类分析和因子分析。在研究β2-整合素的作用时,我们发现它们在维持肺结核性肉芽肿的存在中起着重要作用。在研究前0.5年检测到的肺结核性肉芽肿病例中,粒细胞总数和表达CD11b的粒细胞的增加,淋巴细胞、NK细胞和表达CD11c的NK细胞数量的减少被证明是显著的。在研究前9.5年检测到的肺结核性肉芽肿患者的外周血研究中观察到的特征变化如下:白细胞数量、单核细胞总数以及表达CD11b和CD11c的单核细胞增加。
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引用次数: 0
Features of cytokine profile in patients with sarcoidosis 结节病患者的细胞因子谱特征
Pub Date : 2020-12-01 DOI: 10.15789/1563-0625-foc-2064
Lazareva Nm, O. Baranova, I. Kudryavtsev, N. A. Arsentieva, N. Liubimova, T. Ses’, M. Ilkovich, A. Totolian
Sarcoidosis is an inflammatory disease of unknown etiology with damage to the lungs and other organs characterized by development of necrosis-free epithelioid cell granulomas. Granulomatous inflammation characterized by the activation of different immune systems cells, in particular T lymphocytes, and the cytokines production. Our study was aimed at investigating the characteristics of the cytokine profile of blood plasma in patients with sarcoidosis. We studied peripheral blood plasma samples of patients with sarcoidosis (n = 52). The control blood samples were taken from healthy volunteers (n = 22). The level of 46 cytokines (pg/ml) was determined, as follows: IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL- 6, IL-7, IL-9, IL-12 (p40), IL-12 (p70), IL-13, IL-15, IL-17A, IFNα2, IFNγ, TNFα, TNFβ, IL- 1ra, IL-10, EGF, FGF-2, Flt3 Ligand, G-CSF, GM-CSF, PDGF-AA, PDGF-AB / BB, TGFα, VEGF-A, sCD40L, CCL2, CCL3, CCL4, CCL5, CCL7, CCL11, CCL17, CCL20, CCL22, CXCL1, CXCL8, CXCL9, CXCL10, CXCL11, CXCL13, CX3CL1. Significantly higher levels of interleukins and some proinflammatory cytokines were found in the patients with sarcoidosis, i.e., IL-3, 0.70 vs 0.20, p = 0.003; IL-4, 14.37 vs 3.15, p = 0.009; IL-5, 1.06 vs 0.89, p < 0.001; IL-12 (p70), 1.27 vs 0.56, p = 0.028; IL-17A, 1.48 vs 0.43, p < 0.001; IFNα2, 41.79 vs 25.04, p = 0.003; IFNγ, 4.13 vs 1.14, p < 0.001; TNFα, 21.67 vs 6.70, p < 0.001; anti-inflammatory cytokine IL-10, 1.03 vs 0.45, p = 0.019; growth factors: FGF-2, 40.08 vs 30.58, p = 0.008, G-CSF, 24.18 vs 8.21, p = 0.006, and VEGF-A, 42.52 vs 26.76, p = 0.048; chemokines: CCL3, 3.86 vs 1.33, p < 0,001; CCL17, 78.24 vs 26.24, p < 0.001; CCL20, 7.19 vs 5.64, p = 0.021; CCL22, 660.60 vs 405.00, p < 0,001; CXCL9, 4013 vs 1142, p < 0,001; CXCL10, 565.90 vs 196.60, p < 0.001; CXCL11, 230.20 vs 121.10, p = 0.018; CX3CL1, 56.99 vs 5.16, p < 0.001. Peripheral blood chemokine CCL11 levels were significantly lower in patients compared to the group of healthy volunteers: 77.58 vs 124.70, p = 0.022. The features of the cytokine profile in patients with sarcoidosis may indicate their important role in the processes of formation and outcomes of granulomas. These issues require an additional detailed study, comparison with phenotypes, differential course and outcomes of the disease.
结节病是一种病因不明的炎症性疾病,对肺部和其他器官造成损害,其特征是形成无坏死的上皮样细胞肉芽肿。以激活不同免疫系统细胞,特别是T淋巴细胞和产生细胞因子为特征的肉芽肿性炎症。我们的研究旨在研究结节病患者血浆中细胞因子谱的特征。我们研究了结节病患者(n=52)的外周血血浆样本。对照血样取自健康志愿者(n=22)。测定46种细胞因子的水平(pg/ml),如下:IL-1α、IL-1β、IL-2、IL-3、IL-4、IL-5、IL-6、IL-7、IL-9、IL-12(p40)、IL-12、p70、IL-13、IL-15、IL-17A、IFNα2、IFNγ、TNFα、TNFβ、IL-11a、IL-10、EGF、FGF-2、Flt3配体、G-CSF、GM-CSF、PDGF-AA、PDGF-AB/BB、TGFα、VEGF-A、sCD40L、CCL2、CCL3、CCL4、CCL5、CCL7。CCL11、CCL17、CCL20、CCL22、CXCL1、CXCL8、CXCL9、CXCL10、CXCL11,CXCL13、CX3CL1。结节病患者的白细胞介素和一些促炎细胞因子水平显著较高,即IL-3,0.70比0.20,p=0.003;IL-4,14.37 vs 3.15,p=0.009;IL-5,1.06 vs 0.89,p<0.001;IL-12(p70),1.27 vs 0.56,p=0.028;IL-17A,1.48 vs 0.43,p<0.001;IFNα2,41.79 vs 25.04,p=0.003;IFNγ,4.13 vs 1.14,p<0.001;TNFα,21.67 vs 6.70,p<0.001;抗炎细胞因子IL-10,1.03 vs 0.45,p=0.019;生长因子:FGF-2,40.08 vs 30.58,p=0.008;G-CSF,24.18 vs 8.21,p=0.006;VEGF-A,42.52 vs 26.76,p=0.048;趋化因子:CCL3,3.86 vs 1.33,p<0001;CCL17,78.24 vs 26.24,p<0.001;CCL20,7.19对5.64,p=0.021;CCL22660.60对405.00,p<0001;CXCL9,4013对1142,p<0001;CXCL10,565.90 vs 196.60,p<0.001;CXCL11230.20对121.10,p=0.018;CX3CL1,56.99 vs 5.16,p<0.001。与健康志愿者组相比,患者的外周血趋化因子CCL11水平显著降低:77.58比124.70,p=0.022。结节病患者细胞因子谱的特征可能表明它们在肉芽肿的形成过程和结果中的重要作用。这些问题需要额外的详细研究,与表型、不同病程和疾病结果进行比较。
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引用次数: 2
Role of IL13 genetic polymorphism in the development of bronchial asthma in children IL13基因多态性在儿童支气管哮喘发病中的作用
Pub Date : 2020-12-01 DOI: 10.15789/1563-0625-roi-1986
S. Tereschenko, M. Smolnikova, E. Kasparov, E. Shakhtshneider, M. A. Malinchik, O. Konopleva, S. Smirnova
Bronchial asthma is a multifactorial disease, with both environmental factors and genetic predisposal affecting its development. A number of gene associations have been obtained between polymorphisms of cytokine genes produced by different types of immune cells and asthma development. Interleukin-13 is involved in allergic inflammation, increased bronchial hypersensitivity, regulation of eosinophil levels and IgE production by B cells, thus making it promising for studying IL13 gene polymorphisms in bronchial asthma coupled to development of the disease. The aim of this study was to investigate possible association between asthma and IL13 rs1800925 polymorphism in the children of Caucasian origin in Eastern Siberia. Four groups of patients with asthma were examined (mean age 12.8±1.2 years): with a controlled (n = 95) and uncontrolled course (n = 107), with severe (n = 71) and moderate severity (n = 131) diseases. The control group consisted of healthy individuals: children (n = 33) and adults (n = 102). DNA was isolated with sorbent method; genotyping was carried out using RT-PCR using specific oligonucleotide primers and fluorescent TaqMan probes. The allele and genotype frequencies were compared by the χ-square test using an online calculator. The odds ratio (OR) with a 95% confidence interval (CI) was performed to link genetic markers with pathological phenotypes. The CT IL13 rs1800925 genotype was shown to be associated with moderate asthma and cases of uncontrollable clinical course, whereas the TT genotype was associated with severe asthma. Thus, rs1800925 polymorphism of IL13 gene (the T* variant is known to be associated with increased IL-13 expression) may be associated with bronchial asthma in children. Our data are consistent with results of other authors. E.g., Liu Z. et al. revealed an association between rs1800925 IL13 and the risk of developing asthma in children, with CT and TT genotypes being more common in the patient group. Radhakrishnan A. et al., was studied rs1800925 IL13 in adult population of Malaysia and found that the T* allele frequency in the group of patients significantly exceeds the frequency of this allele in the control group. Thus, the results of our study showed that IL13 rs1800925 polymorphism is associated with bronchial asthma in children, especially, with level of its control and severity of the disease.
支气管哮喘是一种多因素疾病,环境因素和遗传易感性都影响其发展。不同类型的免疫细胞产生的细胞因子基因多态性与哮喘发展之间存在许多基因关联。白细胞介素-13参与变应性炎症、支气管超敏反应增加、嗜酸性粒细胞水平调节和B细胞产生IgE,因此研究il -13基因多态性与支气管哮喘的发展有希望。本研究的目的是调查东西伯利亚高加索裔儿童哮喘与IL13 rs1800925多态性之间的可能关联。四组哮喘患者(平均年龄12.8±1.2岁):控制病程(n = 95)和未控制病程(n = 107),严重病程(n = 71)和中度病程(n = 131)。对照组由健康个体组成:儿童(n = 33)和成人(n = 102)。用吸附法分离DNA;利用特异性寡核苷酸引物和荧光TaqMan探针进行RT-PCR分型。等位基因和基因型频率的比较采用χ 2检验,使用在线计算器。采用95%置信区间(CI)的比值比(OR)将遗传标记与病理表型联系起来。CT IL13 rs1800925基因型与中度哮喘和不可控临床病程相关,而TT基因型与重度哮喘相关。因此,IL-13基因rs1800925多态性(已知与IL-13表达升高相关的T*变异)可能与儿童支气管哮喘有关。我们的数据与其他作者的结果一致。例如,Liu Z.等人揭示了rs1800925 IL13与儿童哮喘发生风险之间的关联,CT和TT基因型在患者组中更为常见。Radhakrishnan A.等对马来西亚成年人群rs1800925 IL13进行了研究,发现患者组T*等位基因频率明显超过对照组该等位基因频率。因此,我们的研究结果表明,IL13 rs1800925多态性与儿童支气管哮喘有关,特别是与其控制水平和疾病严重程度有关。
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引用次数: 0
Search for genetic markers of predisposition to psoriasis and psoriatic arthritis 寻找易患银屑病和银屑病关节炎的遗传标记
Pub Date : 2020-12-01 DOI: 10.15789/1563-0625-sfg-2050
M. Smolnikova, A. A. Barilo, M. A. Malinchik, S. Smirnova
Psoriasis (PS) and psoriatic arthritis (PsA) are interrelated diseases that occur in approximately 30% of patients and are characterized by the presence of a systemic inflammatory reaction that occurs as a result of a violation of the functional state of the immune system. With the advent of new technologies, several new pro-inflammatory cytokines, such as IL-23, IL-31, and IL-33, which play an important role in the pathogenesis of the psoriatic process, have been discovered and characterized. It was determined that single nucleotide polymorphisms (SNPs) in the promoter regions of the IL23, IL31 and IL33 genes play an important role in controlling the expression of relevant cytokines involved in the immunopathogenesis of psoriatic disease. The purpose of the study: to analyze the distribution of genotypes and allelic variants of polymorphisms of the IL23A (rs2066808), IL23R (rs2201841), IL31 (rs7977932) and IL33 (rs7044343), in order to search for genetic markers of predisposition to psoriasis and psoriatic arthritis. Materials and methods. The genotyping of the patients was conducted: psoriasis (PS, n = 77), median age 31.0 years (27.0-43.0), psoriatic arthritis (PsA, n = 99), median age 49.0 years (39.0-56.0) and practically healthy residents of Krasnoyarsk (n = 103), a median age of 32.0 years (24.0-38.0). DNA was isolated from whole venous blood using a standard sorbent kit. Genotyping of single nucleotide polymorphisms IL23A (rs2066808), IL23R (rs2201841), IL31 (rs7977932), IL33 (rs7044343) was carried out using real-time PCR using specific oligonucleotide primers and fluorescentlylabeled probes. Results and discussion. The frequencies of allelic variants of the studied cytokine genes in the control group obtained during the study correspond to their distribution in Caucasoid populations – the alleles IL23A * T, IL23R * T, IL31 * C, IL33 * C prevail. When comparing the distribution frequency of allelic variants of the IL23A, IL23R, IL31, IL33 genes, we did not obtain statistically significant differences between patients and the control group. Conclusions. Despite the fact that when comparing the distribution frequency of allelic variants of the IL23A, IL23R, IL31, IL33 genes, we did not obtain statistically significant differences between the patients and the control group, there are results worthy of attention. So, in patients with PS, the frequency of the C * IL23A allelic variant (rs2066808) is lower than in the population sample, which may indicate its specific role in relation to the development of the disease. All this dictates the need to continue research with the assessment of other SNPs and increase the sample of patients in search of potential genetic markers of psoriatic disease.
牛皮癣(PS)和银屑病关节炎(PsA)是相互关联的疾病,大约30%的患者发生,其特征是由于免疫系统功能状态的破坏而出现全身性炎症反应。随着新技术的出现,一些新的促炎细胞因子如IL-23、IL-31、IL-33等在银屑病发病过程中起重要作用的细胞因子被发现并被表征。结果表明,在银屑病的免疫发病过程中,il - 23、il - 31和il - 33基因启动子区域的单核苷酸多态性(snp)在控制相关细胞因子的表达中起重要作用。本研究目的:分析IL23A (rs2066808)、IL23R (rs2201841)、IL31 (rs7977932)和IL33 (rs7044343)基因型和多态性等位变异的分布,寻找银屑病和银屑病关节炎易感性的遗传标记。材料和方法。对患者进行基因分型:银屑病(PS, n = 77),中位年龄31.0岁(27.0 ~ 43.0岁);银屑病关节炎(PsA, n = 99),中位年龄49.0岁(39.0 ~ 56.0岁);克拉斯诺亚尔斯克市实际健康居民(n = 103),中位年龄32.0岁(24.0 ~ 38.0)。采用标准吸附试剂盒从全静脉血中分离DNA。采用特异性寡核苷酸引物和荧光标记探针,采用实时荧光定量PCR技术对IL23A (rs2066808)、IL23R (rs2201841)、IL31 (rs7977932)、IL33 (rs7044343)的单核苷酸多态性进行基因分型。结果和讨论。研究中获得的对照组细胞因子基因等位变异频率与它们在高加索人群中的分布一致,等位基因以IL23A * T、IL23R * T、IL31 * C、IL33 * C为主。在比较IL23A、IL23R、IL31、IL33基因等位变异的分布频率时,患者与对照组间差异无统计学意义。结论。虽然在比较IL23A、IL23R、IL31、IL33基因等位变异的分布频率时,我们并没有得到患者与对照组之间有统计学意义的差异,但也有值得关注的结果。因此,在PS患者中,C * IL23A等位基因变异(rs2066808)的频率低于人群样本,这可能表明其在疾病发展中的特定作用。所有这些都表明,需要继续研究其他snp的评估,并增加患者样本,以寻找银屑病的潜在遗传标记。
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引用次数: 0
Effect of IL4-589C>T, FCGR2A-166His>Arg, DEFB1-20G>A, DEFB1-52G>A gene polymorphisms on TNFα, IL-1β, IL-4, and IL-10 contents in the patients with primary hip osteoarthrosis IL4-589C>T、FCGR2A-166His>Arg、DEFB1-20G>A、DEFB1-52G>A基因多态性对原发性髋关节骨关节病患者TNFα、IL-1β、IL-4、IL-10含量的影响
Pub Date : 2020-12-01 DOI: 10.15789/1563-0625-eoi-2034
A. Miromanov, T. Zabello, N. Miromanova
Our objective was to study the effects of IL4-589C>T, FCGR2A-166His>Arg, DEFB1-20G>A, DEFB1-52G>A gene polymorphisms upon content of TNFα, IL-1β, IL-4, and IL-10 in primary osteoarthrosis of the hip joints. We performed a survey of 100 patients of Russian ethnicity (average age 61.3±8.5 years) with primary coxarthrosis at the stage III-IV who lived in the Trans-Baikal region. The control group (n = 100), were local residents, comparable by age (60±8.3 years), gender, habitation place and nationality. The exclusion criteria were as follows: close relationship; other types of osteoarthritis (post-traumatic, rheumatoid, metabolic, etc.); dysplastic syndromes and phenotypes; acute and chronic inflammatory diseases at the exacerbation stage; diabetes mellitus; osteoporosis; vascular diseases; obesity; malignant neoplasia; alcohol abuse. Along with clinical examination, the following laboratory methods were applied: immunological techniques, i.e., determination of TNFα, IL-1β, IL-4, IL-10; genetic testing using polymerase chain reaction, e.g., a point mutation of the IL4 gene at the 589(C>T) position, FCGR2A at 166(His>Arg) site, DEFB1 at the 20(G>A) and 52(G>A) positions. DNA from the peripheral blood of patients was used for the molecular genetic analysis. Radiographic examination was also carried out. The data were statistically processed using STATISTICA 6.1 software package (StatSoft, USA), Microsoft Office Excel 2019 for Windows 10. The differences were considered statistically significant at p ≤ 0.05. Results. The -589T/T genotype of IL4-589C>T gene polymorphism indirectly contributes to higher content of TNFα and IL-1β for primary osteoarthritis of the hip joints. The patients with -166Arg/Arg genotype have a 1.3-fold increase of certain cytokine concentrations, e.g., TNFα and IL-1β, as compared with -166His/Arg genotype, and, conversely, lower content of IL-4 and IL-10 (1.3- fold) in comparison with -166His/His genotype. The patients with -20A/A genotype showed higher levels of TNFα and IL-1β, respectively, 1.2 and 1.3 times, compared with -20G/G genotype, and 1.3 times versus the -20G/A genotype. Conclusions: 1. The presence of -589T/T genotype of the IL4-589C>T gene polymorphism and the -20A/A genotype of the DEFB1-20G>A gene polymorphism contributes to a high content of TNFα and IL-1β in the blood serum, and the carriage of -166His/His FCGR2A-166His>Arg gene polymorphism is associated with both higher level of TNFα, IL-1β, and a low concentration of IL-4, IL-10. 2. Complex carriers of FCGR2A166HisArg x DEFB152AA x DEFB120AA x IL4589TT genotypes in the patients with primary coxarthrosis increases the contents of TNFα, IL-1β cytokines by 1.5 and 1.7 times, respectively.
我们的目的是研究IL4-589C >t、FCGR2A-166His >arg、DEFB1-20G >a、DEFB1-52G >a基因多态性对原发性髋关节骨关节病患者TNFα、IL-1β、IL-4和IL-10含量的影响。我们对100名居住在贝加尔地区的俄罗斯族(平均年龄61.3±8.5岁)III-IV期原发性关节关节病患者进行了调查。对照组(n = 100)为当地居民,年龄(60±8.3岁)、性别、居住地、民族具有可比性。排除标准为:关系密切;其他类型的骨关节炎(创伤后、类风湿、代谢性等);发育不良综合征和表型;急性加重期急慢性炎性疾病;糖尿病;骨质疏松症;血管疾病;肥胖;恶性肿瘤;酒精滥用。结合临床检查,采用以下实验室方法:免疫学技术,即检测TNFα、IL-1β、IL-4、IL-10;利用聚合酶链反应进行基因检测,例如,il - 4基因在589(C>T)位点、FCGR2A在166(His>Arg)位点、DEFB1在20(G> a)和52(G> a)位点发生点突变。从患者外周血中提取DNA用于分子遗传学分析。同时进行x线检查。使用STATISTICA 6.1软件包(StatSoft, USA)、Microsoft Office Excel 2019 for Windows 10对数据进行统计处理。p≤0.05认为差异有统计学意义。结果。IL4-589C >t基因多态性的-589T/T基因型间接导致原发性髋关节骨关节炎中TNFα和IL-1β含量升高。与- 166his /Arg基因型患者相比,- 166arg /Arg基因型患者某些细胞因子如TNFα和IL-1β浓度增加1.3倍,相反,IL-4和IL-10含量较- 166his /His基因型低(1.3倍)。-20A/A基因型患者的TNFα和IL-1β水平分别比-20G/G基因型高1.2倍和1.3倍,比-20G/A基因型高1.3倍。结论:1。-589T/T基因型的IL4-589C>T基因多态性和-20A/A基因型的DEFB1-20G>A基因多态性的存在导致血清中TNFα和IL-1β含量高,-166His/His FCGR2A-166His>Arg基因多态性的携带与TNFα、IL-1β水平高和IL-4、IL-10浓度低有关。2. 原发性关节关节病患者中携带FCGR2A166HisArg x DEFB152AA x DEFB120AA x IL4589TT基因型的复合携带者,其TNFα、IL-1β细胞因子含量分别增加1.5倍和1.7倍。
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引用次数: 0
Using CAST-test to investigate human specific hypersensitivity to the anthrax pathogen 采用cast试验研究人对炭疽病原体的特异性超敏反应
Pub Date : 2020-12-01 DOI: 10.15789/1563-0625-uct-2058
D. G. Ponomarenko, E. Rakitina, M. Kostyuchenko, O. Logvinenko, A. Ryazanova, L. Aksenova, N. P. Buravtseva, I. Tyumentseva, S. A. Kurcheva, A. Kulichenko
We present the results of applying functional cytometric test of antigen-stimulated activation basophils to assess specific immunological reactivity in the people with anthrax, and immunized with anthrax vaccine. As a criterion for antigen-specific basophil activation, we measured expression of the CD63 membrane receptor, which reflects the process of anaphylactic basophil degranulation. To determine spontaneous and antigen-induced activation of basophils (CCR3+CD63+), a FlowCAST reagent kit (Buhlmann laboratories AG, Switzerland) was used. Anthraxin, an experimental anthrax allergen (a hydrolysate the Bacillus anthracis STI-1 strain), manufactured by the Stavropol Anti-Plague Institute, was used as a specific antigen. As based on clinical and experimental data, a threshold value of > 10% of anthraxin-activated (CCR3+CD63+) basophils was accepted for the in vitro immunodiagnostic CAST test, as a laboratory criterion for the subjects exhibiting specific immune response, i.e., IgE-mediated sensitization. It was shown that, in anthrax patients within one week after onset of the disease (3-7 days), a positive CAST result was obtained in 92.3% cases; the levels of specific basophil activation with anthraxin averaged 37.9% (12.01 ÷ 78.9%). Immunological examination of individuals three weeks (21 days) after vaccination against anthrax revealed CAST-positivity in all the vaccinated persons. Intensity of anthraxin-induced basophil activation the vaccinated subjects was ranged from 10.87 to 30.03%, averaging 17.86%. The overall values of spontaneous and specific activation ranged within 12.39 ÷ 41.46%. The study opens prospectives for implementation of basophil antigenic activation test in the Flow CAST format in diagnostics of anthrax and to identify specific immune rearrangements after vaccination in humans, as an index of actual vaccination rates. Usage of CAST test with anthraxin makes it possible to identify anthrax patients at the early stages (2-4 days after onset of the disease) including, among patients with an increased CCR3+CD63+ background values, evaluation of immunological efficiency in the cohorts at risk for vaccination. At the same time, it was found that a significant decrease in diagnostic sensitivity of CAST test could be observed in the patients immune to anthrax pathogen who received intensive antibacterial and pathogenetic therapy at the early stages of infection, including glucocorticosteroids (anti-inflammatory drugs) and desensitizing agents that inhibit the degree of hypersensitivity development and its expression.
我们介绍了应用抗原刺激的活化嗜碱性粒细胞的功能性细胞术测试来评估炭疽患者和炭疽疫苗免疫患者的特异性免疫反应性的结果。作为抗原特异性嗜碱性粒细胞活化的标准,我们测量了CD63膜受体的表达,它反映了过敏性嗜碱性颗粒的脱颗粒过程。为了确定嗜碱性粒细胞(CCR3+CD63+)的自发和抗原诱导的活化,使用FlowCAST试剂盒(Buhlmann laboratories AG,Switzerland)。使用由Stavropol抗瘟疫研究所生产的实验性炭疽过敏原炭疽菌素(炭疽杆菌STI-1菌株的水解产物)作为特异性抗原。根据临床和实验数据,在体外免疫诊断CAST试验中,蒽环素活化(CCR3+CD63+)嗜碱性粒细胞的阈值>10%被接受,作为表现出特异性免疫反应(即IgE介导的致敏)的受试者的实验室标准。结果表明,在炭疽病患者发病后一周内(3-7天),92.3%的病例CAST结果呈阳性;炭疽毒素对嗜碱性粒细胞的特异性激活水平平均为37.9%(12.01÷78.9%)。接种炭疽疫苗三周(21天)后对个体进行的免疫检查显示,所有接种者的CAST阳性。蒽环素诱导接种受试者嗜碱性粒细胞活化的强度在10.87%至30.03%之间,平均17.86%。自发和特异性激活的总体值在12.39÷41.46%之间。该研究为在炭疽诊断中采用Flow CAST格式进行嗜碱性细胞抗原激活测试以及确定人类接种疫苗后的特异性免疫重排作为实际疫苗接种率的指标开辟了前景。使用含蒽环素的CAST测试可以在早期阶段(发病后2-4天)识别炭疽患者,包括在CCR3+CD63+背景值增加的患者中,评估有接种风险的人群的免疫效率。同时发现,对炭疽病原体免疫的患者,在感染早期接受强化抗菌和病原学治疗,CAST试验的诊断灵敏度显著降低,包括糖皮质激素(抗炎药)和抑制超敏反应发展程度及其表达的脱敏剂。
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引用次数: 0
Proven and less studied hematopoietic and vasoactive growth factors in retinal capillary hemangioma 已证实的和较少研究的造血和血管活性生长因子在视网膜毛细血管瘤
Pub Date : 2020-12-01 DOI: 10.15789/1563-0625-pal-2002
V. Neroev, N. Balatskaya, A. Novikova, M. V. Ryabina, P. A. Ilyukhin
Pathogenesis of retinal capillary hemangioma has not been sufficiently studied at the present time. Therefore, the study of cytokine levels in biological fluids seems to be very relevant in order to increase knowledge about the mechanisms of the disease development and searching for targeted therapies. The content of hematopoietic and vasoactive growth factors in blood serum, lacrimal fluid, and vitreous body was studied in patients with retinal capillary hemangioma. A total of 26 patients with retinal angiomatosis were examined. The samples of blood serum (n = 23) and lacrimal fluid (n = 10) from practically healthy people aged 22 to 46 (27.4±1.4 years) were used as a control. To perform comparative assessment of cytokine concentrations in the vitreous body of patients with retinal capillary hemangioma, were used samples of the vitreous body from 6 patients (average age 33±4.7 years; from 21 to 49 years) with rhegmatogenous retinal detachment. To measure the cytokine concentrations, we applied multiplex analysis technique using the xMAP platform with LuminexxPONENT 3.1 program and ProcartaPlex sets (eBioscience, Austria). A detailed characteristic of vasoactive factors in capillary retinal hemangioma was obtained as a result of this work. Some disorders in chemokine regulation were identified. There was a significant increase in serum concentrations of three vasoactive factors, i.e., PDGF-BB, HGF, and PIGF-1, with a decrease in chemokines (MCP-1, MIP-1α, and MIP-1β). The frequencies of PIGF-1 and MIP-1α detection also significantly differed from the control group. SCF was significantly more often determined in patients with retinal angiomatosis only at the systemic level. Correlations between PDGF-BB and PIGF-1, as well as PIGF-1 and MIP-1β were shown. A significant increase in VEGF-A, HGF, VEGF-D, as well as MCP-1 concentrations was shown in the lacrimal fluid. The inversion of PDGF-BB concentrations in serum and lacrimal fluid was noted. Analysis of intraocular cytokine levels revealed a significant increase in VEGF-A and HGF concentrations, with marked decrease in MIP-1α and MIP-1β. PDGF-BB in 100% of cases was determined only in vitreous body of patients with retinal angiomatosis. With respect to the revealed characteristic shifts of HGF/SF intraocular production in retinal capillary hemangioma, it seems relevant to search ways for its inhibition, thus providing potential basis for a new therapeutic strategy in treatment of retinal angiomatosis.
视网膜毛细血管瘤的发病机制目前尚未得到充分的研究。因此,研究生物体液中的细胞因子水平对于增加对疾病发展机制的认识和寻找靶向治疗方法似乎非常重要。研究了视网膜毛细血管瘤患者血清、泪液和玻璃体中造血因子和血管活性生长因子的含量。本文对26例视网膜血管瘤病患者进行了检查。选取22 ~ 46岁(27.4±1.4岁)实际健康人群的血清(n = 23)和泪液(n = 10)作为对照。为了比较评价视网膜毛细血管瘤患者玻璃体中细胞因子的浓度,采用6例患者(平均年龄33±4.7岁;年龄从21岁到49岁),伴有孔源性视网膜脱离。为了测量细胞因子浓度,我们采用多重分析技术,使用xMAP平台和LuminexxPONENT 3.1程序和ProcartaPlex套件(eBioscience, Austria)。血管活性因子在毛细血管视网膜血管瘤中的详细特征是这项工作的结果。发现了趋化因子调节中的一些紊乱。血清中三种血管活性因子(PDGF-BB、HGF和PIGF-1)浓度显著升高,趋化因子(MCP-1、MIP-1α和MIP-1β)浓度降低。PIGF-1和MIP-1α的检测频率也与对照组有显著差异。在视网膜血管瘤患者中,SCF通常只在全身水平上被确定。结果显示PDGF-BB与PIGF-1、PIGF-1与MIP-1β之间存在相关性。泪液中VEGF-A、HGF、VEGF-D以及MCP-1浓度显著升高。注意到血清和泪液中PDGF-BB浓度的反转。眼内细胞因子水平分析显示VEGF-A和HGF浓度显著升高,MIP-1α和MIP-1β浓度显著降低。100%的病例中PDGF-BB仅在视网膜血管瘤患者的玻璃体中检测。针对视网膜毛细血管瘤中HGF/SF在眼内生成的特征性变化,寻找抑制HGF/SF的途径,为视网膜血管瘤病治疗提供新的治疗策略提供潜在依据。
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引用次数: 0
Immunophenotype and metabolism are linked in peripheral blood neutrophils from patients with kidney cancer 免疫表型和代谢与肾癌患者外周血中性粒细胞有关
Pub Date : 2020-11-27 DOI: 10.15789/1563-0625-iam-2037
A. Savchenko, A. Borisov, I. Kudryavtsev, I. Gvozdev, A. Moshev
The aim of the present study was to analyze the relationships between expression of activation and adhesion receptors on peripheral blood neutrophils, and intracellular activity of some neutrophil enzymes in patients with kidney cancer (KC). Patients and methods: the KC patients (n = 72) (T3N0M0, clear-cell type) were examined prior to surgical treatment at the Krasnoyarsk Regional Oncology Center. The diagnosis was verified histologically for all KC patients. The phenotype of blood neutrophils was studied using flow cytometry. The surface receptor expression levels of the neutrophils were evaluated by mean fluorescence intensity. NAD and NADP-dependent dehydrogenases activities in purified peripheral blood neutrophils were measured by bioluminescent method. Results: we have found that the phenotypic alterations in circulating KC patients’ neutrophils appeared along with inhibition of main intracellular metabolic processes and were closely linked with them. The features of the phenotypic imbalance in the neutrophils from KC patients were associated with a decrease in blood cells expressing adhesive (CD11b and CD62L) and functional (CD64 and HLA-DR) receptors. Moreover, the patient’s neutrophils expressed CD11b, CD16 and HLA-DR on their cell surface more intensively, than neutrophilic leukocytes from control group. These phenotypic changes in KC patients’ blood neutrophils occurred in parallel with pronounced decrease in immature cells numbers. The metabolic changes of neutrophil cytoplasmic compartment in KC patients were determined by a decrease in Glu6PDH activity (a key and initializing enzyme of the pentose phosphate cycle) and NADH-LDH (anaerobic glycolysis). Mitochondrial metabolism in neutrophils of KC patients was characterized by multidirectional changes in the activity of NAD- and NADP-dependent glutamate dehydrogenases (decreased activity of NAD-dependent and increased activity of NADP-dependent) and a decrease in NADH-MDH activity. The established features in mitochondrial enzymes activities suggest some disturbances of NAD-dependent processes that could lead to down-regulation of aerobic energy processes. We guess that the decreased activity of plastic and energy processes in blood neutrophils of KC patients could affect the receptor expression levels. By means of correlation analysis, we have found that the relationships in KC patients were determined by negative effects of NADHGDH and NADH-LDH activities upon expression of activation and adhesion receptors in blood neutrophils. Of these enzymes, only glutathione reductase activity in neutrophils from KC patients was positively linked with the CD23 and HLA-DR expression. Thus, an increase in activity of energy processes (e.g., coupling the tricarboxylic acid cycle to amino acid metabolism) in blood neutrophils from the patients with kidney cancer could stimulate expression levels of activation and adhesion receptors and potentially increase antitumor activity of neutrophils.
本研究的目的是分析肾癌(KC)患者外周血中性粒细胞活化和粘附受体的表达与细胞内某些中性粒细胞酶活性的关系。患者和方法:手术前在克拉斯诺亚尔斯克地区肿瘤中心检查KC患者(n = 72) (T3N0M0,透明细胞型)。所有KC患者的诊断经组织学证实。用流式细胞术研究血液中性粒细胞的表型。用平均荧光强度评价中性粒细胞表面受体表达水平。用生物发光法测定纯化后的外周血中性粒细胞中NAD和nadp依赖性脱氢酶的活性。结果:我们发现循环KC患者中性粒细胞的表型改变伴随着细胞内主要代谢过程的抑制而出现,并与之密切相关。KC患者中性粒细胞表型失衡的特征与表达黏附(CD11b和CD62L)和功能性(CD64和HLA-DR)受体的血细胞减少有关。此外,患者的中性粒细胞在其细胞表面表达CD11b, CD16和HLA-DR比对照组的中性粒细胞更密集。KC患者血液中性粒细胞的这些表型变化与未成熟细胞数量的显著减少同时发生。KC患者中性粒细胞胞质室的代谢变化是通过葡萄糖6pdh(戊糖磷酸循环的关键初始化酶)和NADH-LDH(厌氧糖酵解)活性的降低来确定的。KC患者中性粒细胞线粒体代谢的特点是NAD依赖性谷氨酸脱氢酶和nadp依赖性谷氨酸脱氢酶活性的多向变化(nadp依赖性活性降低,nadp依赖性活性升高)和NADH-MDH活性降低。线粒体酶活性的既定特征表明,nad依赖性过程的一些干扰可能导致有氧能量过程的下调。我们推测KC患者血液中性粒细胞可塑性和能量过程活性的降低可能影响受体的表达水平。通过相关性分析,我们发现KC患者的相关性是由NADHGDH和NADH-LDH活性对血液中性粒细胞活化受体和粘附受体表达的负面影响决定的。在这些酶中,只有KC患者中性粒细胞中的谷胱甘肽还原酶活性与CD23和HLA-DR表达呈正相关。因此,肾癌患者血液中性粒细胞能量过程活动的增加(例如,三羧酸循环与氨基酸代谢的耦合)可以刺激活化和粘附受体的表达水平,并可能增加中性粒细胞的抗肿瘤活性。
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Medical immunology (London, England)
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