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 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.

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 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.

Aim To investigate the the effect of ginsenosides of stem and leaf(GSL) on mouse fatty liver and its mechanism.Methods Mice were randomly divided into normal control group(NC,fed with normal diet) and model group(fed with high-fat diet).Twelve weeks later,when the establishment of experimental model of fatty liver was confirmed,the model mice were subdivided into 4 subgroups: HF group(fed continuously with high-fat deit),NT group(fed with normal deit),GSL1 group(fed with normal deit and treated intragastrically with GSL 50 mg?kg-1?d-1),GSL2 group(fed with normal deit and treated intragastrically with GSL 100 mg?kg-1?d-1).Mice in NC,HF and HT groups were given distilled water by gastric perfusion.Two weeks later,all mice were killed,and blood was collected for measuring serum TC,TG,HDL-C,LDL-C,ALT contents,liver tissue for determining hepatic TC,TG,MDA SOD levels.In addition,liver index and pathology were also observed.Expression of PPAR? and CYP2E1 mRNA in liver was examined by RT-PCR.Results In GSL2 group,GSL significantly reduced liver index,serum lipid,hepatic lipid and MDA contents,and elevated SOD activity.Moreover,GSL obviously improved pathological change,increased PPAR? mRNA and suppressed CYP2E1 mRNA levels in the liver.In NT group,hepatic lipid and MDA contents remained high and SOD activity is low,although liver histology somewhat improved.In GSL1 group the results were similar to those in NT group.Conclusions These results suggest that GSL might be effective in the treatment of hepatic adipose infiltration of mice.It may be associated with increasing PPAR? mRNA,then decreasing serum lipid and hepatic lipid;and inhibiting CYP2E1 mRNA expression in the liver,thus suppressing lipid peroxidation.

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

[ ABSTRACT] AIM:To explore the role of Huoxue Jiangzhi Recipe in preventing and treating fatty liver in mice and its underlying mechanisms.METHODS:Healthy Kunming mice were fed with high-fat diet and treated intragastrically with different doses of Huoxue Jiangzhi Recipe ( compound of ginseng, panax notoginseng and rhizoma gastrodiae, named as GST) for 2 weeks.The levels of blood lipids and triglyceride ( TG) in hepatic tissues were measured.Meanwhile, liver in-dex and hepatic pathology were observed.The optimized dosage of Huoxue Jiangzhi Recipe was determined by the experi-ments.The mice were divided into normal control group ( NC group, fed with normal diet) and model group ( fed with high-fat diet) .The model mice were subdivided into 3 subgroups 12 weeks later:HF group ( fed continuously with high-fat di-et) , ND group ( fed with normal diet) , GSL group ( fed with normal diet and treated intragastrically with GSL) .The mice in NC, HF and ND groups were given distilled water by gastric perfusion.Two weeks later, all mice were killed, and blood was collected for measuring serum total cholesterol (TC),TG,high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol ( LDL-C) contents, hepatic TC, TG, malondialdehyde ( MDA ) levels and superoxide dismutase ( SOD) activity were detected.Moreover, liver index and hepatic pathology were also observed.The mRNA expression of peroxisome proliferator-activated receptor alpha (PPARα) and cytochrome-P450 2E1 (CYP2E1) in the liver was examined by RT-PCR.RESULTS:GST significantly decreased serum lipid, hepatic lipid and MDA levels and elevated SOD activi-
ty.Furthermore, GST markedly reduced liver index, improved hepatic adipose infiltration, increased PPARαmRNA ex-pression and inhibited CYP2E1 mRNA expression.CONCLUSION:GST is effective in the treatment of fatty liver in mice by up-regulating PPARα, thus reducing serum and hepatic TG levels, down-regulating CYP2E1 and inhibiting lipid peroxi-dation.