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【Objective】 Through bioinformatics methods to analyze the differences in the gene expression profiles of peripheral blood mononuclear cells （PBMCs） between middle-aged and elderly women and normal people， so as to explore the diagnosis and treatment targets of OA. 【Methods】 We downloaded the GSE48556 data set from GEO databases. We utilized the R language to screen out the differentially expressed genes （DEGs） between OA and NC. By gene set enrichment analysis （GSEA）， we obtained the target gene subset. The GO and KEGG pathways of the target gene subset were analyzed by DAVID. We applied STRING and Cytoscape software to construct PPI network. The module analysis was performed by the Mcode and centiscape plug-in， and the key genes were screened out by Cytohubba. 【Results】 By GSEA analysis and P.adjust 0.2， a total of 292 target genes were screened， consisting of 81 upregulated genes and 211 downregulated genes. The GO enrichment analysis of all target genes mainly focused on the biological functions， such as “regulation of NIK/NF-κB”， “monocytes”， “proliferation”， “regulation of apoptosis signaling pathway”， “TNF-mediated signaling pathway”， “regulation of Wnt signaling pathway”， “regulation of MAP kinase activity”， and “regulation of autophagy”. KEGG was mainly enriched in four pathways： cytotoxicity of natural killer cell mediation， TNF signaling pathway， MAPK signaling pathway， and apoptosis. We employed PPI network and related plug-ins to screen out eight core genes highly related to OA inflammation and apoptosis， namely， MAPK1， IL10， PTGS2， IL18， GSK3B， NFKBIA， TNFRSF1A， and EGR1. 【Conclusion】 Bioinformatics analysis revealed that the differences in PBMCs gene expressions between OA and NC were concentrated in the biological events of apoptosis and inflammation， making blood expression profile an effective breakthrough for monitoring OA target markers.
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Objective:To observe the clinical efficacy of modified Erchentang on CXC chemokine ligand receptors (CXCR1/2)and their ligands CXCL8,macrophage inflammatory protein -2(MIP-2) in patients of chronic obstructive pulmonary disease(AECOPD)at acute exacerbation stage,and assess the effect and mechanism of modified Erchentang on anti-inflammatory in patients of AECOPD. Method:This study was a multicenter, randomized single blind, controlled trial. The authors selected 200 cases in conformity to the standards of AECOPD. The AECOPD patients were randomly divided into modified Erchentang group and control group. In addition to the western medicine, modified Erchentang was also given to the modified Erchentang group, and Jizhitangjiang was given to the control group for 14 days. Each group was observed for the alleviation of the symptoms. Euzyme-linked immunosorbent assay (ELISA) was used to determine the levels of CXCL8 and MIP-2 in the patients' plasma of all groups before and after treatment. Western blot were used to detect the levels of CXCR1, CXCR2 and CXCL8 protein in peripheral blood mononuclear cells(PBMCs). Immunocytochemistry (ICC) method was used to detect the expressions of CXCL8, CXCR1 and CXCR2 protein in PBMCs. Result:The level of CXCL8 in plasma, and the expressions of CXCR1, CXCR2 and CXCL8 mRNA and protein in the modified Erchentang group were decreased significantly than those in the control group(PPConclusion:Modified Erchentang has an anti-inflammatory effect on AECOPD. Its mechanism may be related to the down-regulation of the expressions of CXCL8, CXCR1 and CXCR2, the reduction of synthesis and release of CXCL8 and MIP-2, the inhibition of the chemotaxis and activity of inflammatory cells, and the prevention of inflammation progress.
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Objective:To investigate the expression of miR-132 in peripheral blood mononuclear ceils(PBMCs) and plasma of patients with oral lichen planus (OLP) and the relationship of miR-132 with clinical forms of OLP.Methods:40 OLP patients (24 nonerosive OLP,16 erosive OLP) and 40 healthy controls were included.The relative expression of miR132 in peripheral blood mononuclear cells (PBMCs) and plasma was examined by quantitative real-time PC R.Results:The expression of miR-132 in both PBMCs and plasma of OLP patients was significantly higher than those in healthy controls(P ＜ 0.01).miR-132 expression of PBMCs and plasma in erosive OLP group was significantly higher than that in non-erosive OLP group and healthy control(P ＜ 0.01).However,the expression of miR-132 in PBMCs and plasma showed no significant difference between non-erosive OLP group and healthy control (P ＞0.05).Conclusion:miR-132 might be involved in the abnormal immune response of OLP patients,and miR-132 can be utilized as an assistant biomarker for the evaluation of the severity of OLP.
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The ELISPOT assay is increasingly used for assessing cellular immune responses in clinical trials of HIV-1 or cancer vaccines. However, to date, data from clinical trials do not consistently show that immune responses are correlated with clinical endpoints. This is due in part to the lack of assay standardization and validation across laboratories and therefore, a quality control panel is required to establish competency and comparability amongst different laboratories. In this study peripheral blood mononuclear cells (PBMCs) from healthy individuals were screened and frozen in liquid nitrogen. The recovery and viability of the PBMCs and the frequencies of interferon (IFN)-γ-secreting cells after CEF peptide pool stimulation were detected after various intervals in seven different laboratories. The recovery and viability did not differ significantly after different intervals. Although the frequencies of IFN (interferon)-γ-secreting cells among thawed PBMCs (peripheral blood mononuclear cells) fluctuated after CEF peptide pool stimulation at different intervals, they were not significantly decreased compared with those among fresh PBMCs. However, the viabilities, recoveries and frequencies of IFN-γ-secreting cells differed significantly among the seven laboratories. Our results indicate that cryopreserved PBMCs could be used as a quality control panel for ELISPOT. However, the procedures for ELISPOT need to be standardized amongst different laboratories.
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Objective To evaluate the methylation status at CpG site -55 in the interferon-gamma (IFN-7) gene promoter and its effect on IFN-7 expression in chronic hepatitis B. Method The authors recruited 30 patients with UBeAg-positive chronic hepatitis B (CHB), 30 HBeAg-negative CHB patients, and 30 healthy blood donors. Pyrosequeneing was used to determine the methylation status at CpG site -55 in the IFN-γ gene promoter following bisulfite treatment of DNA in peripheral blood mononuclear cells (PBMCs). The expression of IFN-γ was analyzed by real-time RT-PCR and ELISA. HBV DNA in PBMCs was detected by nested PCR. Results The methylation level at CpG site -55 in the IFN-γ gene promoter was significantly increased, resulting in subsequent down-regulation of the expression of this cytoldne in CHB. The methylation level at CpG site -55 was significantly higher in HBeAg-positive patients than in HBeAg-negative ones (P<0.01) and was also significantly higher in PBMCs from HBV DNA-positive patients than from HBV DNA-negative ones (P<0.01) ; the methylation level at CpG site -55 was positively correlated with the amount of HBV DNA in serum (P<0.01). Oonclusion IFN-γ gene expression appears to be regulated by methylation of the IFN-γ gene promoter in CHB; the methylation level at CpG site -55 is associated with HBV infection.
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Objective: To evaluate the methylation status at CpG site -55 in the interferon-gamma (IFN-γ) gene promoter and its effect on IFN-γ expression in chronic hepatitis B. Method: The authors recruited 30 patients with HBeAg-positive chronic hepatitis B (CHB), 30 HBeAg-negative CHB patients, and 30 healthy blood donors. Pyrosequencing was used to determine the methylation status at CpG site -55 in the IFN-γ gene promoter following bisulfite treatment of DNA in peripheral blood, mononuclear cells (PBMCs). The expression of IFN-γ was analyzed by real-time RT-PCR and ELISA. HBV DNA in PBMCs was detected by nested PCR. Results: The methylation level at CpG site -55 in the IFN-γ gene promoter was significantly increased, resulting in subsequent down-regulation of the expression of this cytokine in CHB. The methylation level at CpG site -55 was significantly higher in HBeAg-positive patients than in HBeAg-negative ones (P < 0.01) and was also significantly higher in PBMCs from HBV DNA-positive patients than from HBV DNA-negative ones (P < 0.01); the methylation level at CpG site -55 was positively correlated with the amount of HBV DNA in serum (P < 0.01). Conclusion: IFN-γ gene expression appears to be regulated by methylation of the IFN-γ gene promoter in CHB; the methylation level at CpG site -55 is associated with HBV infection.
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Objective To explore the expression of interleukin 13(IL-13) mRNA in peripheral blood mononuclear cells (PBMCs) and its association with serum C-reaction protein(CRP) in patients with ulcerative colitis(UC). Methods IL-13 mRNA expression in PBMCs was detected by RT-PCR in patients with UC(21 cases of active UC and 10 cases of quiescent UC)and 20 healthy subjects. At the same time, the serum level of CRP was detected by ELISA. Results The expression level of IL-13 mRNA in PBMCs of patients with active UC was significantly lower than that in PBMCs of patients with quiescent UC and healthy subjects (6.45?1.23 vs 14.72?2.12 or 15.17?2.38,P0.05). The expression level of IL-13 mRNA in PBMCs of active UC patients was negative correlation with serum level of CRP (r=-0.589,P