1.The role of NF-κB binding element in regulation of NOD2 gene
Liping SUN ; Chaofeng HU ; Jikai ZHANG
Chinese Journal of Microbiology and Immunology 2008;28(7):606-609
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.
2.Klotho: a protein molecule interacting with FPC protein
Liping SUN ; Xinzhou ZHANG ; Chaofeng HU
Chinese Journal of Pathophysiology 2010;26(3):518-523
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.
3.Role of ROS and energy metabolism dysfunction in hepatocyte adipose degeneration in vitro
Chaofeng HU ; Daxing LU ; Liping SUN
Chinese Journal of Pathophysiology 2000;0(07):-
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
4.Klotho:a protein molecule interacting with FPC protein
Liping SUN ; Xinzhou ZHANG ; Chaofeng HU
Chinese Journal of Pathophysiology 1986;0(03):-
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.
5.The spectra and the change of diseases in AIDS patients after antiretroviral treatment in Henan province
Xuan YANG ; Xue ZHANG ; Chaofeng LI ; Yan SUN ; Qingxia ZHAO
Chinese Journal of Primary Medicine and Pharmacy 2015;(14):2118-2121
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.
6.Relationship betweenα-SMA and atrial fibrosis in patients with valvular atrial fibrillation
Junqiang PAN ; Dianxin ZHANG ; Chaofeng SUN ; Jin YANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(2):242-246
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.
7.Role of Huoxue Jiangzhi Recipe in preventing and treating fatty liver in mice
Chaofeng HU ; Liping SUN ; Han ZHOU ; Qinhe YANG ; Daxiang LU
Chinese Journal of Pathophysiology 2015;(6):1087-1092
[ 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.
8.Therapeutic effect of radiofrequency ablation in patients with atrial fibrillation
Tianyu MENG ; Juan GAO ; Jine WU ; Yongrong JIANG ; Chaofeng SUN
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(6):796-800
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.
9.The Model Innovation of Telemedicine Service Based on the Four Dimensional Model
Yunkai ZHAI ; Chaofeng QIAO ; Jie ZHAO ; Dongxu SUN
Journal of Medical Informatics 2017;38(4):1-6
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.
10.Effect of hepatitis C virus genotype on antiviral therapy in patients with human immunodeficiency virus/hepatitis C virus coinfection
Zhaoyun CHEN ; Yan SUN ; Qingxia ZHAO ; Chaofeng LI ; Lin DING
Chinese Journal of Infectious Diseases 2017;35(7):403-406
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.