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 investigate molecular typing and drug resistance patterns of 98 Klebsiella pneumoniae (K.pneumoniae) isolated from type 2 diabetes patients complicated with maxillofacial infection,to research the virulence and resistance mechanisms.Methods The study was a prospective study that adopted the method of continuous sampling from fixed location,from March 2010 to October 2012.The maxillofacial surgery patients diagnosed with type 2 diabetes complicated with maxillofacial infection were chosen in 7 hospitals in Zhengzhou as the research object,and a total of 431 pus sample were collected continuously,in which 98 strains K.pneumoniae were isolated and identified.The Kirby-Bauer disk diffusion test was conducted in 98 strains to determine the resistance to 19 antibacterial agents.K.pneumoniae chromosomal DNA were digested by restriction endonuclease Xba Ⅰ and analyzed by pulsed-field gel electrophoresis (PFGE).PFGE patterns of K.pneumoniae strains were analyzed using Fingerprinting software.The relationship between the molecular types and resistance phenotype was observed.The extended spectrum β-1actamase-producing K.pneumoniae were screened out by the double disc synergy test (DDST)Polymerase chain reaction (PCR) was used to detect resistant genotypes,serotype and virulence genes.The purified PCR products of resistant genes were cloned and sequenced.Hypermucoviscosity phenotype of all strains were determined by string test.Results Much severer drug-resistance for K.pneumoniae was identified and the result of extended-spectrum beta-lactamase producing rate was 57.1 ％.Ninety-eight strains were dispatched into 13 groups by PFGE.No dominant bands and specific extended-spectrum beta-lactamase DNA bands were found.The results of PCR showed that among the 56 strains of extended spectrum β-lactamase-producing K.pneumoniae,40 were positive for blaSHV (accounting for 71.4％),28 positive for blaTEM (accounting for 50.5％),21 positive for blaCTX-M (accounting tor 37.5％).The sequencing results were as follows:TEM-1,CTX-M-3 and a variety of SHV.Serotype K1,K2,K3,K5,K20,K54 and K57 and 3 kinds of virulence genes were detected,but not in strong toxicity-based.Hypermucoviscosity positive rate was 31.6％ (31/98).Conclusion Much severer drug resistance of K.pneumoniae in this study was identified and resistant mechanism was complex,in which strong toxicity serotype and virulence geues exist,which need more attention from clinical.

A new wood-degrading fungus Monodictys asperospera(Cooke & Massee) Ellis with a high level of laccase production was chosen to study.This laccase was purified by ammonium sulfate precipitation,DEAE-cellulose and sephacryl S-300.Purification of about 8.1 fold was achieved with an overall yield of 5.7%.Its molecular weight was estimated to be about 77 kD.The optimum temperature and pH of the lac-case activity were 55?C and 6.0,respectively.Kinetic studies of the laccase showed that the Km and the Vmax for using syringaldazine as substrate was 0.163 mmol/L and 0.194 mmol/(L.min),respectively.The carbo-hydrate content was 18.14%.In addition,it was found that laccase activity was significantly inhibited by Cu2+.

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 relation between polymorphism of NOS3 G894T and susceptibility of cancer .Methods Literatures under the inclusion and exclusion criteria about relation between polymorphism of NOS 3 G894T and susceptibility of cancer were collected by computer -based retrieval and manual retrieval .The pools ORs with 95% CI were calculated to assess the association strength between polymorphism of NOS 3 G894T and cancer risk by Meta methods .Sensitivity and publication bias were evaluated .Results 19 literatures with 7679 cases of cancer and 8180 cases as control group were included .The pooled result indica-ted that no significant association between its polymorphism and cancer (ORT versus G=1 .030 ,95% CI=0 .964 -1 .101 ;ORTT versus GG=1 .056 ,95% CI=0 .895-1 .245 ;ORTG versus GG=1 .045 ,95% CI=0 .977-1 .119 ;ORTT/TG versus GG=1 .047 , 95% CI=0 .981-1 .117 ;ORTT versus TG/GG=1 .005 ,95% CI=0 .900-1 .123) .No significant association was observed in sub-group analysis based on ethnicity ,cancer type ,and source of control group .Conclusion There was no relationship between NOS3 G894T polymorphism and cancer susceptibility .

Adenocarcinoma, Clear Cell ; pathology ; Humans ; Kidney Neoplasms ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Thyroid Neoplasms ; secondary

Objective To analyze the mechanism of nasal deformity by reviewing the possible pathogenesis and nasal anatomy and to find the effective and reliable operative methods to correct nasal deformities in unilateral cleft lip.Methods 57 patients (37 males,20 females,and range in age from 12 to 25 years,with mean of 18.6 years) with nasal deformities in unilateral cleft lip were available for this study.The nasal deformities were treated with the following surgical procedures according to the different locations and degree Of nasal deformities.The eompositive techniques included:alveolar bone grafting was taken to correct the collapse the nasaI base;a cortical plate was inserted between the two medial crura of the alar cartilage.According to the nasal contours of non-cleft side,the alar cartilage was resected and suspended to its normal and symmetrical position.The alar cartilaginous ring was reconstruction,which maintained the nostril shape 3 month postoperatively.Results A total of 57 cases were treated by the method above,and were ranged with score by operators and patients.93% of cases were evaluated as satisfaction after operation.The follow-up for 3 to 24 months showed that 52 cases had achieved satisfactory effects,5 cases showed the trend to relapse.Conclusion The cause of nasal deformities in unilateral cleft lip is complex.The study has achieved a significant improvement by synthetical correction of deformities of maxilla,cartilage and soft tissues, and the restoration of nasal-labial muscles.The rigid suspending is more important to maintain the nostril contour and avoid relapse.

Carcinoma, Intraductal, Noninfiltrating ; diagnosis ; pathology ; surgery ; Carcinoma, Pancreatic Ductal ; pathology ; Diagnosis, Differential ; Humans ; Male ; Prostate ; pathology ; Prostatic Intraepithelial Neoplasia ; pathology ; Prostatic Neoplasms ; diagnosis ; pathology ; surgery

Humans ; Meniere Disease

Carcinoma, Transitional Cell ; pathology ; Humans ; Neoplasm Grading ; Urologic Neoplasms ; pathology