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 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 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

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

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

Objective To investigate the effect of hepatitis C virus (HCV) genotype on antiviral therapy in patients with human immunodeficiency virus (HIV)/HCV coinfection in Henan province.Methods A total of 129 patients were coinfected with HIV and HCV, among whom, 70 were HCV 1b genotype and 57 HCV 2a genotype.And 131 patients were HIV single infection.Immunological failure rate, virological suppression, CD4+ T lymphocyte counts and liver and renal function after antiretroviral therapy (ART) were compared among the three groups.Flow cytometry was used to count CD4+ T lymphocytes and polymerase chain reaction amplification was used to detect HIV RNA.The liver and renal function were tested by automatic biochemical analysis.Statistical analysis was conducted by χ2 test, analysis of variance and LSD-t method.ResultsImmunological failure rate in HCV 1b group, HCV 2a group and HIV single infection group were 7.14% (5/70), 15.79% (9/57) and 9.92% (13/131), respectively.There was no significant statistical difference among the three groups (χ2=2.59, P>0.05).The CD4+ T lymphocyte counts in three groups were (614±258), (529±245), and (518±243) cells/μL, respectively.The difference was statistically significant (F=3.17, P<0.05).The virus inhibition rates of three groups were 87.0% (HCV 1b), 78.2% (HCV 2a), and 82.3% (HIV single infection).The HIV virus failure rates were 8.6% (HCV 1b), 14.5% (HCV 2a), and 13.1% (HIV single infection).There was no significant difference among three groups (χ2=1.967, P>0.05).The levels of aspartate transaminase, alanine aminotransferase and total bilirubin in HCV 1b group and HCV 2a group were all significantly higher than those in HIV single infection group (F=27.38, 15.22 and 7.33, respectively, all P<0.05), while there was no significant difference between HCV 1b and HCV 2a groups (t=1.27, 0.29 and 1.59, respectively, all P>0.05).Conclusions The main HCV genotypes in patients with HIV/HCV coinfection by blood transmission are HCV 1b and HCV 2a in Henan province.HIV/HCV coinfection does not affect the effect of ART, but could aggravate the liver damage in acquired immune deficiency syndrome patients.

The paper introduces the mutual relationship among the innovation of four dimensions including new medical service concept,new patient interface,new service transmission system and new technology of telemedicine based on the illustration of four-dimensional model of service innovation to the telemedicine service,and makes an analysis by taking the telemedicine service innovation of remote medical center in Henan Province as an example,thus providing reference for the innovative development of telemedicine service.

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.