Objective To investigate the role of NF-KB binding element in regulation of NOD2. Methods Promoter region of NOD2 containing the NF-κB binding site was amplified by PCR from human genome DNA and correctly connected to the vector pEGFP-N3 which had been cut out promoter by restriction enzyme to obtain the GFP expression vector driven by human NOD2 gene promoter. The constructed plasmids were transiently transferred into cell line HeLa by LipofectAMINETM2000 and the GFP expression was ob- served by the inversion fluorescence microscope. The NF-κB binding site in the constructed vector pEGFP- N3-NOD2wt was deleted by the QuikChange site-directed mutagenesis kit. The recombinant plasmid mpEG- FP-N3-NOD2 was transiently transferred into cell line HeLa by LipofectAMINETM2000, and the GFP expres- sion was observed by the inversion fluorescence microscope. Results The constructed pEGFP-N3-NOD2wt plasmids and mpEGFP-N3-NOD2 were the same as the design confirmed by restriction digestion and se- quence analysis. The results of the cell transient transfection indicated that different strength of GEP ex- pressed by recombinant plasmids in HeLa cells could be observed. The GFP expression of constructed mpEGFP-N3-NOD2 was lower than that of pEGFP-N3-NOD2wt. Conclusion The GFP expression vector driven by human NOD2 gene promoter which contains the NF-κB binding site, and the site deleted plasmid were successfully constructed. The GFP expression of recombinant plasmid mpEGFP-N3-NOD2, deletion of the NF-KB binding site, was obviously weaken in HeLa. The results indicate that NF-KB binding element may play a positive role in regulation of NOD2 gene, which establishes favourable bases for further study on the mechanism of NOD2 gene expression and regulation.

AIM:To explore the role of reactive oxygen species(ROS)and energy metabolism dysfunction in hepatocyte adipose degeneration induced by alcohol and calf serum(CS).METHODS:The growing L02 cells were treated with different concentrations of alcohol.To screen the proper concentration of alcohol,the proliferation of cells was measured by MTT.Lipid droplets in the cells were observed through oil red staining.Triglyceride(TG)content was detected with analyzed kit.The level of ROS and changes of mitochondrial membrane potential(??m)in cells were tested by flow cytometry.Reversed-phase high performance liquid chromatography(RP-HPLC)was applied to assay the cellular ATP content.RESULTS:Lipid droplets were observed under light microscope in the cells treated with 2% alcohol and 50% CS(A+CS)for 36 h.Compared to control group,the cellular TG and ROS levels in model group markedly increased while ??m and ATP content in cells significantly decreased(P

AIM:To investigate the role of fibrocystin/polycystin (FPC) in autosomal recessive polycystic kidney disease (ARPKD) development by means of screening the protein interaction using yeast two-hybrid approach. METHODS:The constructed pGBKT7-FPC was used as the bait to screen the pre-transformed human fetal kidney cDNA expression library by yeast two-hybrid assay to obtain the host cell protein which interacted with C-terminal region of FPC. The sequence transformation screening experiment was applied to confirm the protein interactions in yeast. RESULTS:After yeast mating and co-transformation screening analysis,Klotho (KL) was selected from the host cells and the interaction between KL and FPC was further confirmed. CONCLUSION:C-terminal region of FPC can interact with KL,which probably provide the approach for further studying the role and biochemistry mechanism of FPC protein in ARPKD.

AIM: To investigate the role of fibrocystin/polycystin (FPC) in autosomal recessive polycystic kidney disease (ARPKD) development by means of screening the protein interaction using yeast two-hybrid approach. METHODS: The constructed pGBKT7-FPC was used as the bait to screen the pre-transformed human fetal kidney cDNA expression library by yeast two-hybrid assay to obtain the host cell protein which interacted with C-terminal region of FPC. The sequence transformation screening experiment was applied to confirm the protein interactions in yeast. RESULTS: After yeast mating and co-transformation screening analysis, Klotho (KL) was selected from the host cells and the interaction between KL and FPC was further confirmed. CONCLUSION: C-terminal region of FPC can interact with KL, which probably provide the approach for further studying the role and biochemistry mechanism of FPC protein in ARPKD.

Objective To investigate the expression of α-smooth muscle actin (α-SMA)in patients with valvular atrial fibrillation and study the relationship betweenα-SMA and atrial fibrosis in patients with valvular atrial fibrillation (AF).Methods For this study we enrolled 84 consecutive patients with rheumatic heart disease who were to receive cardiac surgery.The patients were divided into AF group (AF,n=39)and sinus rhythm group (SR, n=45).Their clinical data including baseline demographics,routine laboratory test and echocardiographics were collected before surgery.The right atrial tissue (0 .3-0 .5 cm3 )was disserted during the surgery.Right atrial fibrosis was observed by Masson staining.The mRNA expression ofα-SMA in atrial tissue were determined by Real-time quantitative PCR.Western blot was used to measure the protein expression ofα-SMA in atrial tissue.Results The two groups did not significantly differ in sex ratio,age,blood pressure,blood biochemical indicators or other aspects of medical history (P>0.05).However,left and right atrium diameters in AF group were significantly larger than those in SR group (P<0 .05 ).Masson staining suggested that collagen volume fraction and collagen content were significantly higher in AF group than in SR group (P<0 .05 ).The mRNA and protein expressions ofα-SMA in right atrial tissue were obviously higher in AF group than in SR group (coefficients P<0 .05 ).The mRNA and protein expressions ofα-SMA from right atrial tissue in the 84 patients were positively correlated with collagen content (coefficients of 0.587 and 0.607;P=0.029,0.014,respectively).Conclusion There is significant atrial fibrosis in patients with valvular atrial fibrillation,which is closely related to up-regulated expression ofα-SMA gene.

Objective To explore the change and the spectra of opportunistic infections in AIDS patients after antiretroviral treatment in He′nan province.Methods Inpatients of post treatment with disease in accordance with the criteria were selected from the Sixth People′s Hospital of Zhengzhou.A retrospective study was carried out, including the content such as gender,age,infection route,treatment time,the date of complication,and CD4+T lympho-cyte counts when the patient with complications.Disease were divided into four group:opportunistic infections,adverse drug reactions,tumor,cardio-cerebrovascular disease and diabetes mellitus.Statistic case number and incidence of every kinds of the disease in the different stages of HAART.Results (1)1 001 cases of patients were involved in the result analysis.People of 41-60 years old,the peasants and peasant workers,accounted for 57.4%and 78.9%.The infection route was mainly through blood transmission and blood collection, accounting for 83.45%.Most of the patients come from Zhumadian, Xuchang, Zhoukou, Nanyang, which were high -incidence area in He′nan.( 2 ) In 1 001 patients of post treatment, the rate of opportunistic infections was 58.4%, while adverse drug reactions was 30.5%.Bacterial pneumonia,extrapulmonary tuberculosis,oral cavity fungal infection and tuberculosis were common in opportunistic infections of patients of post treatment.The main adverse reactions were allergic dermatitis,liver injury and myelosuppression.The liver cancer case was the highest ( 8/29 ) .( 3 ) 1 001 cases were divided into 6 groups according to different count of treatment time,during 3 months of treatment,the incidence rate of opportunistic infec-tions and adverse drug reactions was the highest,accounted for 75.2% and 41.6%.After 5 years of treatment,the incidence rate of cardio-cerebrovascular disease and diabetes mellitus was 6.39%.With the prolonging of HAART time,incidence of opportunistic infections and adverse drug reactions remarkably reduced,but the occurence rate car-dio-cerebrovascular disease and diabetes mellitus increased.Conclusion In AIDS patients after HAART in He′nan at present,opportunistic infections and adverse drug reactions are main diseases and used to occur during three months after HAART.Attention will be focused on tumor, cardio -cerebrovascular disease and diabetes mellitus in AIDS patients of long-term survival.

Objective To assess the therapeutic effect and safety of radiofrequency catheter ablation of atrial fibrillation.Methods We analyzed the clinical data of 47 patients with atrial fibrillation who underwent radiofrequency catheter ablation between March 2013 and January 2008,in the First Affiliated Hospital of Xi'an Jiaotong University.In the average 32 months’follow-up,Holter monitering and echocardiography were reviewed for the left atrial diameter.Results The immediate success rate of catheter ablation for atrial fibrillation was 82.6%.The long-term success rate was 65%,the rate of paroxysmal atrial fibrillation was 69.7%,and the rate of longstanding persistent atrial fibrillation was 42.9%.After ablation,the left atrial diameter were markedly reduced compared with that before treatment [(36.3 ± 4.3 )mm vs .(38.1 ± 5.9 )mm)(P < 0.001 ).The patients with recurrent atrial fibrillation were older than those without recurrence,their left atrial diameter was bigger,and the prevalence rate of hypertension was higher (P <0.05).The average atrial fibrillation load was 14.9% after ablation compared with 46.1% before (P <0.05).Conclusion Radiofrequency catheter ablation is an effective and safe treatment of atrial fibrillation,especially for paroxysmal atrial fibrillation.The left atrial diameter was significantly decreased after radiofrequency catheter ablation compared with that before the ablation.

Objective:To investigate the effect of pioglitazone on blood pressure,blood glucose and beta cell function and the mecha- nism of oxidative stress in hypertensive patients with impaired glucose tolerance(IGT). Methods:One hundred and sixty patients were divided into two groups:pioglitazone group and control group.The levels of fasting plasma glucose(FPC),2 hours postprandial glucose(2hPG),blood pressure,Homa I3,Homa IR,blood malondiahtehyde (MDA)and superoxide dismutase before and after pioglitazone treatment were compared. Results:After pioglitazone treatment for 8 weeks,FPG,2hPG,FInS,2hPInS,GHbAlc,mean systolic and diastolic blood pressure,Homa IR and blood MDA were significantly decreased(P

Objective:To investigate the expression of fibrillin-1 (FBN-1) in the atrium tissue of the patients with rheumatic heart valve disease complicated with atrial fibrillation(AF), and to explore its relationship with atrial fibrosis in the patients with valvular atrial fibrillation.Methods:Eighty-four consecutive patients with rheumatic heart valve disease underwent cardiac surgery were enrolled in this study.The patients were divided into AF group(n=39) and sinus rhythm group(SR group, n=45).The clinical data of patients were collected before operation.The right atrium tissue (0.3-0.5 mm3) was disserted during operation.The degrees of right atrial fibrosis of the patients in two groups were observed by Masson staining.Western blotting method was used to measure the protein expressions of FBN-1 in atrium tissue of the patients in two groups.Results:There were no significant differences in the gender ratio, age, blood pressure, blood biochemical indicators and other aspects of medical history between two groups(P>0.05);the diameters of left and right atrium of the patients in AF group were significantly larger than those in SR group(P<0.05).The Masson staining results showed that there was obvious fibrosis in AF group, and the collagen volume fraction and collagen level in AF group were significantly higher than those in SR group (P<0.05).The expression level of FBN-1 in right atrium tissue in AF group was obviously higher than that in SR group(P<0.05).The expression level of FBN-1 protein in right atrium tissue of the patients with valvular atrial fibrillation was positively correlated with the collagen level(r=0.544,P=0.021).Conclusion:There is obvious atrium fibrosis in the patients with valvular atrial fibrillation;it is closely related to the up-regulation of the expression of FBN-1 gene.

Objective To investigate the clinical effects of Alprostadil combined with Shuxuetong treatment on patients with diabetic peripheral neuropathy,and its influence on malondialdehyde(MDA),superoxide dismutase(SOD) and total antioxidative capability(TAOC).Methods Totally 160 patients with DPN were divided into two groups: treatment group(100 cases) and control group(60 cases).The levels of blood SOD activity,serum MDA and serum TAOC were determined before and after treatment.Results The symptoms in both groups were significantly improved,but the total effective rate in treatment group was significantly better than that in control group(80.0% vs.41.7%,P