Objective:To explore the mechanisms by which DNA methylation regulates miR-126 and its host gene EGFL7 in CD4+T cells from patients with systemic lupus erythematosus (SLE).
Methods:We analyzed the expression and the DNA methylation status within promoter region of EGFL7 and miR-126 by real-time qPCR and bisulifte genomic sequencing analysis.
Results:miR-126 and EGFL7 mRNA expression was upregulated in CD4+T cells from SLE compared with that from healthy controls (P<0.01). EGFL7 mRNA level was positively correlated with miR-126 expression in CD4+T cells from SLE (r=0.538, P=0.015). The average methylation level of EGFL7 promoter in CD4+T cells from SLE was lower than that from healthy controls (P<0.05).
Conclusion:hTe upregulation of miR-126 and its host gene EGFL7 expression in CD4+T cells from SLE is associated with the hypomethylation of the EGFL7 promoter.

Objective To estimate the influence of 1,25(OH)2D3 on the proliferative ability of and methylation levels of genomic DNA and proliferation-associated gene promoter in human HaCaT keratinocytes.Methods Some cultured HaCaT cells were treated with 1,25 (OH)2D3 of 10-6,10-7 and 10-8 mol/L for 24 hours,then,methyl thiazolyl tetrazolium (MTT) assay was carried out to evaluate the proliferative activity of cells,and a global DNA methylation quantification kit was used to determine the global DNA methylation level.Real-time PCR was conducted to quantify the mRNA expression of DNA methyl transferases (DNMTs) and methyl-DNA binding domain (MBD) proteins,and methylation-specific PCR (MS-PCR) to evaluate the methylation status of promoter region in the programmed cell death 5 (PDCD5) and tissue inhibitor of metalloproteinase 2 (TIMP2) genes,in HaCaT cells after 24-hour treatment with 1,25 (OH)2D3 of 10-6 mol/L.The HaCaT cells receiving no treatment served as the control.Results Compared with the untreated HaCaT cells,those treated with 1,25(OH)2D3 of 10-6 mol/L showed significantly down-regulated proliferative activity (0.152 ± 0.027 vs.0.290 ± 0.017,P ＜ 0.01),global DNA methylation level (0.187 ± 0.071 vs.0.316 ± 0.049,P ＜ 0.05),DNMT3a and DNMT3b mRNA expression levels (P ＜ 0.01 or 0.05),but markedly upregulated mRNA expression levels of MECP2,MBD2,PDCD5 and TIMP2 (P ＜ 0.01 or 0.05).Moreover,the DNA methylation levels within the promoter region of PDCD5 and TIMP2 genes were significantly lower in HaCaT cells treated with 1,25 (OH)2D3 of 10-6 mol/L than in the control cells (0.38 ± 0.135 vs.0.72 ± 0.121,0.46 ± 0.172 vs.0.68 ± 0.133,both P＜ 0.05).Conclusions 1,25(OH)2D3 may down-regulate the global genomic DNA methylation level of,and modulate the expression of DNA methylationmodifying genes in,HaCaT cells.Furthermore,1,25 (OH)2D3 can decrease the promoter methylation levels but induce the overexpression of PDCD5 and TIMP2 genes,and decelerate the proliferation of HaCaT cells.

Objective To investigate the significance of changes in serum uric acid levels in elderly patients with obstructive sleep apnea hyponea syndrome (OSAHS).Methods The study recruited elderly patients aged 60 years and over undergoing polysomnography (PSG) at the Center for Diagnosis and Treatment of Sleep Disorders of our hospital from January 2012 to December 2014.According to the diagnostic criteria established by the Group of Sleep and Breathing Disorders,Respiratory Diseases Branch,Chinese Medical Association (2011),patients were divided into the simple snoring group,the mild OSAHS group,the moderate OSAHS group and the severe OSAHS group.Levels of serum uric acid,blood glucose,blood lipids and high-sensitive C-reactive protein (hsCRP) were compared between all groups after adjusting for age,gender and underlying diseases.Independent risk factors associated with serum uric acid were analyzed using Spearman' s rank correlation and stepwise multivariate analysis.Results A total of 129 elderly subjects including 100 males and 29 females,with a mean age of (63.6±2.25) years,were enrolled.Of these subjects,23 were grouped as simple snoring,24 as mild OSAHS,39 as moderate OSAHS and 43 as severe OSAHS.Serum uric acid and hs-CRP levels were increased in the severe OSAHS group as compared with the simple snoring group (both P＜0.05).Spearman's rank correlation analysis showed that the apnea-hypopnea index (AHI) and the oxygen desaturation index (ODD were positively correlated with the serum uric acid level (r =0.251 and 0.210,P =0.004 and 0.018,respectively).Stepwise multivariate analysis showed that,with serum uric acid level as the dependent variable,AHI was an independent risk factor for increased serum uric acid levels (β=0.354,P =0.000).Conclusions Serum uric acid levels are elevated in elderly patients with severe OSAHS,and AHI is an independent risk factor for the increased serum uric acid levels.

Objective To study the effects of Genistein - magnetic - nanoparticles at different concentrations on the growth and ex-pression of focal adhesion kinase (FAK) in human hepatocellular carcinoma cell line HcpG2. Methods Activities of HepG2 cells treated by Genistein - magnetic - nanoparticles were examined by MTT assay. The expression of FAK mRNA and protein was respectively detected by immunohistochemistry staining and reversed transcriptase polymerase chain reaction (RT - PCR) . Results Genistein - magnetic -nanoparticles at a concentration of 10 -80mg/L inhibited the proliferation of HepG2 cells, with obvious concentration - dapendent and time - dependent effect relationships (P < 0.05). After treatment with various concentrations of Genistein - magnetic - nanoparticles for 48h, the relative level of FAKmRNA of Genistein - magnetic - nanoparticles and control groups was 1.242 ± 0.031,1.213 ± 0.443,0.834 ± 0.044,0.652 ± 0.039,0.446 ± 0.041, and 1.443 ± 0.053 (F = 21.97 ,P < 0.05), respectively. The expression of FAK protein in the cells was decreased, which showed an obvious a concentration - effect relationship. Conclusion Genistein - magnetic - nanoparticles can reduce the mRNA and protein expressions of FAK and inhibit the proliferation of HepG2 cells.

Objective To study the effects of Genistein-magnetic-nanoparticles at different concentrations on the growth and expression of focal adhesion kinase(FAK) in human hepatocellular carcinoma cell line HepG2.Methods Activities of HepG2 cells treated by Genistein-magnetic-nanoparticles were examined by MTT assay.The expression of FAK mRNA and protein was respectively detected by immunohistochemistry staining and reversed transcriptase polymerase chain reaction(RT-PCR).Results Genistein-magnetic-nanoparticles at a concentration of 10-80mg/L inhibited the proliferation of HepG2 cells,with obvious concentration-dapendent and time-dependent effect relationships(P

Objective To investigate the familial incidence of multiple primary carcinoma in Chinese hereditary nonpolyposis colorectal cancer patients (HNPCC) in Northeast China.Methods By family line investigation,multiple primary carcinoma (MPC) spectrum' s characteristics of 509 patients in 85 families registered in strict conformity with the HNPCC Amsterdam criteria Ⅱ were analyzed retrospectively.Results Of the 85 HNPCC families,multiple primary carcinoma developed in 55 patients in 25 families,among them 45 patients had metachronous carcinoma in 17 families,16 patients had synchronous carcinoma occurred in 12 families,6 patients with both synchronous carcinoma and metachronous carcinoma in 4 families.Conclusions Multiple primary carcinoma developed in significantly high incidence in Northeast China in Chinese hereditary nonpolyposis colorectal cancer patients,the most common MPC are colorectal cancer and endometrial cancer.

Objective To evaluate the efficacy of eucalyptol-limonene-pinene enteric capsule on stable chronic obstructive pulmonary disease (COPD) in the elderly.Methods A total of 120 patients with stable COPD were randomized into ELP group (n=63) with conventional treatment combined ELP and control group (n=57) with conventional treatment without ELP.Primary outcome was the number of exacerbations per year, and secondly outcomes were lung function parameters and St.George's Respiratory Questionnaire (SGRQ).Results After 1 year of treatment, numbers of exacerbations per patient per year were lower in ELP group than in control group (0.70 times/y vs.1.21 times/y, Z=-3.887, P=0.000).The proportion of exacerbation-free patients were 25.4％(16/63) in ELP group and 5.3％ (3/57) in control group (x2 =9.103, P=0.003).The differences in the forced vital capacity (FVC), forced expiratory volume in one second (FEV1), and FEV1/ FVC ratio (FEV1％) between 5 different time points measurements were statistically significant in both groups (ELP group: F=15.429, 36.389, 33.278, respectively, all P=0.000;control group: F=54.238, 94.213 and 83.774, respectively, all P=0.000).Difference in FEV1/FVC ratio between 5 different time point was statistically significant in control group(F=2.766, P=0.043), but not in ELP group(F=0.861, P=0.451).After treatment, SGRQ score was decreased in both group, and difference in symptom score was statistically significant between the two groups (t=2.109, P =0.037).The adverse reactions were found in 3 cases in ELP group, and in 2 cases in control group with no statistically significant difference between the two groups [3 (4.8％) vs.2 (3.5％), x2 =0.013, P=0.732].Conclusions Long-term oral administration of eucalyptol-limonene-pinene enteric capsule can significantly decrease exacerbation frequency, improve quality of life, delay the deterioration in lung function, and have good safety in elderly patients with stable COPD.

OBJECTIVE: To investigate the effect of total glucosides of peony (TGP) on expression and DNA methylation status of ITGAL gene (CD11a) in CD4(+) T cells from patients with systemic lupus erythematosus (SLE). METHODS: CD4(+) T cells were isolated by positive selection using CD4 beads. CD4(+) T cells were treated by TGP at 0, 62.5, 312.5 and 1562.5 mg/L for 48 h. The MTT method was used to assess cell viability; mRNA expression level was measured by realtime-PCR; protein level of CD11a was measured by flow cytometric analysis; DNA methylation status was assayed by bisulfite sequencing. RESULTS: No significant change in cell viability was found in CD4(+) T cells among the different concentration groups (P>0.05). Compared with control, the mRNA and protein levels of ITGAL were down-regulated significantly in SLE CD4(+) T cells treated with TGP (1562.5 mg/L) (P< 0.01). Furthermore, the extent of DNA methylation of ITGAL promoter was increased in TGP (1562.5 mg/L) treated CD4(+) T cells compared with control group (P<0.01). CONCLUSION TGP can repress CD11a gene expression through enhancing DNA methylation of ITGAL promoter in CD4(+) T cells from patients with SLE. This observation represents a preliminary step in understanding the mechanism of TGP in SLE therapy.
CD11a Antigen ; genetics ; metabolism ; CD4-Positive T-Lymphocytes ; immunology ; metabolism ; DNA Methylation ; drug effects ; Down-Regulation ; drug effects ; Glucosides ; pharmacology ; Humans ; Lupus Erythematosus, Systemic ; genetics ; immunology ; Paeonia ; chemistry ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; genetics ; metabolism

OBJECTIVE: To construct a special luciferase reporter to detect DNA methylation regulatory activity in FCER1G gene promoter regulatory element. METHODS: We constructed special full and mock methylated FCER1G gene promoter regulatory luciferase reporters by patch-methylation, and detected DNA methylation regulatory activity by comparing the luciferase activity of full-methylated luciferase reporters with mock-methylated reporters. RESULTS: We successfully constructed the full and mock methylated FCER1G gene promoter regulatory luciferase reporters. The ratio of luciferase activity between the full methylated and the mock methylated was (0.36±0.07):1 (P<0.001). CONCLUSION FCER1G promoter activity is methylation-sensitive and is regulated by DNA methylation.
Base Sequence ; DNA Methylation ; Gene Expression Regulation ; Genes, Reporter ; Humans ; Luciferases ; genetics ; Molecular Sequence Data ; Promoter Regions, Genetic ; genetics ; Receptors, IgE ; genetics ; metabolism

Transient receptor potential cation channel 6（TRPC6）gene is mainly expressed in renal podiocytes.Its variation can lead to steroid-resistant nephrotic syndrome（SRNS），and the specific pathogenesis is not clear.These children have poor response to hormones and immunosuppressants，with lack of specific therapeutic drugs，and poor prognosis.In recent years，it has been found that some drugs can slow down disease progression by inhibiting the expression of TRPC6 gene or its downstream signaling pathway，and the discovery of TRPC6 protein-specific blockers may be the hope of treating such children.This review focuses on the pathogenesis of SRNS induced by TRPC6 gene variation in children and the research progress of drug therapy.