Dialysis Method Alters the Expression of MicroRNA‐33a and Its Target Genes ABCA1, ABCG1 in THP‐1 Macrophages

Atherosclerosis and accompanying cardiovascular disease are the first causes of mortality in patients undergoing maintenance hemodialysis. Anti‐atherosclerotic effects of hemodiafiltration (HDF) have been reported. Our study aimed to investigate the effect of serum derived from a healthy group (n = 23), before and after hemodialysis (HD) therapy (n = 23), and before and after HDF therapy (n = 17) on the expression of microRNA‐33a and its target genes adenosine triphosphate‐binding cassette transporter A1,G1 (ABCA1, ABCG1) in THP‐1 macrophages. Meanwhile, blood lipids and high‐sensitivity C‐reactive protein (hs‐CRP) were measured in these groups. Our data showed that the expression of miRNA‐33a was lower (P < 0.05) and ABCA1 and ABCG1 were higher (P < 0.05) in the healthy group than pre‐HD and pre‐HDF. miR‐33a was significantly decreased (P < 0.05) but ABCA1, ABCG1 was significantly increased (P < 0.05) in post‐HDF compared with pre‐HDF, while these parameters in pre‐ and post‐ HD groups did not show any significant change (P > 0.05). High density lipoprotein cholesterol (HDL‐C) was higher and hs‐CRP was lower in the healthy group than pre‐HD and pre‐HDF groups. Moreover, a significant increase of HDL‐C (P < 0.05) and decrease (P < 0.05) of hs‐CRP was shown in post‐HDF compared with pre‐HDF, but HD appeared to have no significant change in these subjects. HDF therapy can downregulate miR‐33a expression, and then result in ABCA1, ABCG1 upregulation and an increase in circulating HDL‐C, leading to a possible anti‐atherosclerosis effect to some extent.     

联合检测抗CCP、RF、抗RA33在类风湿关节炎的临床应用

目的:探讨抗环瓜氨酸肽抗体（抗CCP）、类风湿因子（RF）、抗RA33抗体联合检测在类风湿关节炎（RA）诊断中意义。方法:选取在我院确诊RA患者266例、非RA风湿病患者228例（结缔组织病、强直性脊柱炎、骨关节炎、系统性红斑狼疮、干燥综合征）、健康体检者215名,检测3组抗CCP、RF、抗RA33数值,统计各指标阳性率,并进行统计学分析比较;通过Spearman 相关分析探讨各指标之间的关系;ROC曲线比较各指标的诊断性能;通过平行试验与系列实验探讨3项指标联合检测RA的诊断效能。结果:RA患者各指标数值均显著高于非RA自身免疫疾病患者与健康对照组（P ＜0.01）;Spearman相关分析显示RA患者抗CCP与抗RA33之间不存在关系,RF与抗CCP、RF与抗RA33之间存在显著相关性（P ＜0.01）;ROC曲线分析显示,抗CCP、RF、抗RA33的AUC值分别为0.942、0.922、0.653;3项指标中,阳性率:RF＞抗CCP＞抗RA33,敏感度:RF＞抗CCP＞抗RA33,特异度:抗RA33＞抗CCP＞RF;平行试验敏感度增高（98.50%）,但特异度降低（64.91%）,系列实验敏感度为15.41%,特异度为98.68%。结论:抗CCP、RF、抗RA33联合检测可提高诊断RA的敏感度与特异性,适用于RA的早期诊断,临床上值得推广使用。     

La、hVAP-33、eIF2B γ和HCV IRES特异性小干扰RNA抑制HCV的复制和表达

目的 证实La自身抗原、33kD人类囊相关膜蛋白(hVAP-33)和真核细胞翻译起始因子第3亚单位(eIF2B γ)是HCV在细胞内的协同感染因子,通过抑制Huh7细胞内这些因子的表达可抑制HCV复制和表达. 方法 分别设计合成3条HCV内部核糖体进入位点(IRES)的小干扰RNA(siRNAs),转染Huh7-HCV细胞后筛选出其中沉默效率最高的1条.以HCV假病毒感染Huh7细胞后48 h,分别以上述HCV IRES siRNA和之前试验中已经筛选出的La、hVAP-33和eIF2B γ特异性siRNAs单独或者不同组合转染Huh7-HCV细胞,利用荧光定量PCR方法检测HCV核心基因并计算△△CT值(相对定量法),比较不同siRNAs及组合对目的基因沉默的效率;同时以Western blot观察HCV核心蛋白表达量的差异.结果 La自身抗原与IRES特异性siRNA共转染对HCV表达的抑制效率最高,使HCV核心蛋白基因相对表达量下降了约41％;4种基因特异性siRNAs 对HCV在Huh7细胞中的复制和表达均有不同程度的抑制作用,La、hVAP-33和eIF2Bγ特异性siRNAs分别与IRES siRNA联合均较其单独转染对目的基因的抑制效率高.结论 可以认为La自身抗原、hVAP-33和eIF2Bγ是HCV在宿主细胞内的协同感染因子,通过对Huh7细胞内协同感染因子及HCV IRES基因沉默后可以显著减少HCV的表达.     

ADAM33基因单核苷酸多态性及单体型与慢性阻塞性肺疾病的关联性研究

目的 探讨中国华北地区汉族人群ADAM33基因S1、S2位点单核苷酸多态性及单体型与慢性阻塞性肺疾病(COPD)及肺功能的关联性.方法 应用DNA直接测序的方法,对90例COPD患者和90名健康对照者的ADAM33基因S1、S2位点基因型进行检测;应用SHEsis在线软件构建单体型并进行单体型关联分析.结果 ①病例组和对照组中S1位点基因型及等位基因频率分布比较差异无统计学意义(P＞0.05),S2位点基因型及等位基因频率分布比较差异有统计学意义(P ＜0.05).②Logistic回归分析表明:ADAM33基因S1位点不同基因型COPD发生的相对危险度比较差异无统计学意义(P＞0.05);S2位点不同基因型COPD发生的相对危险度比较,差异有统计学意义(P＜0.05),其中G/G+C/G基因型的OR值为2.364(95％CI 1.251～4.466).③COPD病例组S2位点基因型与肺功能相关临床指标的关系显示:3种基因型FEV1％预计值比较差异无统计学意义而FEV1/FVC比较差异有统计学意义,其中G/G基因型与C/C、C/G基因型相比FEV1/FVC下降更明显.④SHEsis在线软件对S1、S2位点进行单体型分析结果显示,单体型CG在COPD组和对照组中比较差异有统计学意义(P＜0.05).结论 在中国华北地区汉族人群中,ADAM33基因与COPD的易感性有关,但与疾病的严重程度无关.     

The possible role of interleukin-33 as a new player in the pathogenesis of contrast-induced nephropathy in diabetic rats

Introduction: Patients with diabetic kidney disease (DKD) are more prone to contrast-induced nephropathy (CN). Apoptosis and autophagy were found to be essential in the pathogenesis of DKD. Interleukin-33 (IL-33) is a cytokine, but its role in DKD and CN is unknown. As IL-33 is modulated by apoptosis, we aimed to determine the relationship between IL-33 apoptosis and autophagy in DKD with CN. Materials and methods: Thirty male Sprague–Dawley rats were enrolled and randomly allocated into three groups. The first group was comprised of healthy rats (HRs), whereas the other two groups were made up of diabetic rats (DRs) and diabetic rats with CN (DRs?+?CN). All groups except the HRs received 50?mg/kg/day of streptozotocin (STZ). The DRs?+?CN group was induced by administering 1.5?mg/kg of intravenous radiocontrast dye on the 35th day. Results: We observed increased IL-33 in the kidney tissue following induction of CN in the DRs. The DRs showed moderate immunopositivity, and the DRs?+?CN showed severe immunopositivity for caspase-3, cleaved caspase-3, caspase-8, caspase-9, LC3B, and Beclin-1 in tubular cells and glomeruli. The DRs also showed moderate immunopositivity in tubular cells, and the DRs?+?CN group showed severe immunopositivity for IL-33 in tubular cells. Increased caspase-3 was found in both glomeruli and tubuli; however, we could not demonstrate IL-33 in glomeruli. This could be secondary to inactivation of IL-33 via increased caspase-3 activity. Conclusion: The release of IL-33 from necrotic cells might induce autophagy, which can further balance the effects of increased apoptosis secondary to CN in DKD.     

Allergen-Dependent Differences in ILC2s Frequencies in Patients With Allergic Rhinitis

PurposeGroup 2 innate lymphoid cells (ILC2s) are a novel population of lineage-negative cells that induce innate type 2 responses by producing the critical Th2-type cytokines IL-5 and IL-13 in response to IL-25 and IL-33 stimulation. ILC2s accumulation in the peripheral blood of patients with allergic rhinitis (AR) is controversial; the precise role of ILC2s in the immunopathogenesis of AR is still not clear. We investigated the role of ILC2s in phenotypic AR sensitized to distinct allergens.MethodsFlow cytometric analysis of the peripheral blood of 7 healthy controls (HCs), 9 patients monosensitized to house dust mite (HDM), and 8 patients monosensitized to mugwort was performed to quantify ILC2s frequency. Peripheral blood mononuclear cells (PBMCs) were isolated from HDM-AR and mugwort-AR patients, and Lineage^- and Lineage⁺ cells were separated using a fluorescence-activated cell sorter (FACS). IL-5 and IL-13 levels in the supernatants of PBMCs, and Lineage^- and Lineage⁺ cells stimulated with IL-25 and/or IL-33 combined with IL-2 in vitro were assessed using the Milliplex magnetic bead kit.ResultsThe percentage of ILC2s was significantly elevated in HDM-AR patients compared to mugwort-AR patients and HCs, while no significant difference was found between mugwort-AR patients and HCs. IL-33±IL-25 plus IL-2 induced a significantly greater release of IL-5 and IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. IL-25 plus IL-2 also induced a significantly greater release of IL-13 in the PBMCs of HDM-AR patients compared to PBMCs of mugwort-AR patients. Stimulation with IL-33 and/or IL-25 combined with IL-2 also induced a significantly greater IL-5 and IL-13 release from Lineage^- cells compared to Lineage⁺ cells.ConclusionsAR patients sensitized to HDM or mugwort allergen have distinct phenotypic and functional profiles in ILC2s frequencies. ILC2s mediate major type 2 immunity in the development of HDM-AR and may be a potential therapeutic target.