1.DNA methylation status of miR-126 and its host gene EGFL7 in CD4+T cells from patients with systemic lupus erythematosus
Yunsheng LIANG ; Sha ZHAO ; Gongping LIANG ; Ming ZHAO ; Qianjin LU
Journal of Central South University(Medical Sciences) 2013;38(8):793-797
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.
2.Molecular typing, drug resistance and virulence analysis of Klebsiella pneumoniae isolated from Type 2 diabetes mellitus complicated with maxillofacial infection
Yingying LI ; Liang MING ; Hongchun LIU ; Yunkai KANG ; Zhao MING
Chinese Journal of Laboratory Medicine 2014;37(2):136-139
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.
3.Purification and Characterization of Laccase from Monodictys asperospera (Cooke & Massee) Ellis
Yi-Ning WANG ; Guo-Zhu ZHAO ; Yue-Ming ZHAO ; Xiao-Liang DI ; Xiang-Ming XIE ;
Microbiology 2008;0(11):-
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+.
4.Correlation between frustration tolerance and personality of college students and its promotion strategies
Ming GUO ; Baojia CHEN ; Yao WANG ; Liang ZHAO
Chinese Journal of Medical Education Research 2013;(5):527-530
Objective To investigate the correlation between the frustration tolerance and personality of medical college students and to discuss its promoting strategies.Methods Totally 460 college students were selected randomly.Questionnaire of frustration tolerance and scale of Eysenck personality inventory(EPQ) were used and the measurement data were compared with correlation analysis and regression analysis of SPSS 13.0.Results Correlation was significant between frustration tolerance (total score and 6 inducing factors) and personality (EPQ factors) of college students ; personality (EPQ factors)of students could predict their frustration tolerance and corresponding t value was -4.85,P < 0.01 (psychoticism),6.93,P < 0.01 (extraversion),-11.15,P < 0.01 (neuroticism).Conclusions Personality of college students are greatly influenced by their frustration tolerance.Targeted anti-frustration ability training should be conducted according to the different personality characteristics.
7.Correction of nasal deformities in unilateral cleft lip
Bin YANG ; Bin XIONG ; Ming ZHAO ; Liang ZHANG
Chinese Journal of Medical Aesthetics and Cosmetology 2010;16(4):233-236
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.
9.Role of endothelial cell injury in the pathogenesis of diabetic nephropathy
Peng-ming ZHAO ; Jian-qin WANG ; Yao-jun LIANG
Chinese Journal of Diabetes 2016;24(7):669-672
[Summary] Diabetic nephropathy is one of the major chronic microvascular complications of diabetes ,which is the leading cause of end‐stage renal disease ,as well as the main cause of death in diabetic patients. Glomerular endothelial cell is an important component of the glomerular filtration barrier ,which is directly related to the materials of circulation ,and it can be easily damaged by glucose ,lipid and inflammatory factors. Under the hyperglycemia ,the PKC pathway ,the polyol pathway and oxidative stress were activated ,producing an excess of advanced glycation end products and reactive oxygen species ,which damage the endothelial nitric oxide synthase ,reduce the generation of nitric oxide ,while produce a large number of Ang Ⅱ. Ang Ⅱ damage the endothelial cell. In addition ,there are crosstalk between glomerular endothelial cells and endothelial cells ,which also cause endothelial cell injury. Here ,we reviewed the role of endothelial cell injury in the pathogenesis of diabetic nephropathy.
10.Effect 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
Yanling JIANG ; Ming ZHAO ; Gongping LIANG ; Litao WANG ; Yuwen SU
Chinese Journal of Dermatology 2013;46(12):885-888
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.