ObjectiveTo explore the protective effects and possible mechanism of different type of proton pump inhibitors (PPIs) in non-steroid anti-inflammatory drugs (NSAIDs)induced small intestinal injury.MethodsSeventy two SD rats were randomly and equally divided into control group,model group,Omeprazole treated group,Esomeprazole treated group,Rabeprazole treated group and Lansoprazole treated group.Except control group,rats of other groups were gavaged with diclofenac 7.5 mg · kg-1 · d-1,once daily to make NSAIDs related small intestinal injury model.The treated groups were gavaged with Omeprazole 30 mg · kg1· d-1,Esomeprazole 30 mg· kg1 · d-l,Lansoprazole 45 mg · kg1 · d-1 and Rabeprazole 15 mg · kg-1 · d-1 once daily respectively.Continuous administration for five days and then executed,small intestinal tissues were taken and observed for gross and pathological changes.The expression level of nuclear factor erythroid-2-related factor 2(Nrf2) at protein and mRNA level were detected with western blot and real time PCR assay.The qualitative and location of Nrf2 in small intestinal tissue were analyzed by immunohistochemistry staining and the activity of superoxide dismutase (SOD) and malondialdehyde (MDA) in small intestinal tissue were determined with xanthine oxidase method and TBA respectively.ResultsThe successful rate of modeling experiment was 100％.The survival ratio of control group rats,model group,Omeprazole treated group,Esomeprazole treated group,Rabeprazole treated group and Lansoprazole treated group was 12/12,3/12,2/12,1/12,1/12 and 2/12 respectively.The tissue injure scores of Esomeprazole treated group,Rabeprazole treated group and Lansoprazole treated group were significantly lower than that of model group (P＜0.05).The activity of SOD in small intestinal of Rabeprazole treated group was obviously higher than that of model group (P＜ 0.05),while the activity of MDA in Rabeprazole and Esomeprazole treated groups were significantly lower than that of model group (P＜0.05).The results of westen blot indicated that the expression of Nrf2in small intestinal tissue was obviously higher in Rabeprazole treated group than in model group (P＜0.05).The real time PCR results suggested that the expression of Nrf2 at mRNA level in small intestinal tissue of Rabeprazole treated group was obviously higher than those of model and control groups (P＜0.05).Conclusions The protection effects of various PPIs are different in NSAIDs related intestinal injury.There is no obvious protection effect of Omeprazole.Rabeprazole may prevent NSAIDs related intestinal injury by up-regulating expression of Nrf2 and promoting its activation.

ObjectiveTo investigate the effects and molecular mechanism of simvastatin in liver fibrosis model of non-alcoholic fatty liver disease (NAFLD) in vivo and hepatic stellate cell in vitro.MethodsFirstly,the rat liver fibrosis model of NAFLD was established by high-fat diet administration and intervened with simvastatin.The expression of endothelial nitric oxide synthase (eNOS),inducible nitric oxide synthase (iNOS) and Collagen Ⅰ at mRNA and protein level were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.Secondly,quiescent phenotype of LX-2 cell line was induced by promoting adipocyte differentiation medium in vitro,and then the quiescent phenotype of LX-2 cell were treated with transforming growth factor β1(TGF-β1),Nitroso-L-arginine methyl ester (L-NAME) which was NOS inhibitor,simvastatin,TGFβ1 with simvastatin,and L-NAME with simvastatin separately.The changes of eNOS,iNOS,αsmooth muscle actin (α-SMA) and Collagen Ⅰexpressions at mRNA and protein level were determined by RT-PCR and Western blot.ResultsAs modeling time extended,the expressions of eNOS in rat's liver tissue of model group at mRNA and protein level decreased gradually,however the expression of iNOS and Collagen Ⅰ at mRNA and protein level increased gradually,compared with normal control group and the differences were statistically significant (P ＜0.05 and 0.01).By 24weeks,the expressions of eNOS in rat's liver tissue of simvastatin group at mRNA and protein level were increased,the expression of iNOS at mRNA and protein level were decreased and the expression of Collagen Ⅰ at mRNA and protein level were decreased (all P ＜0.05).The expression of eNOS in rat's liver tissue of model group negatively correlated with the expression of Collagen Ⅰ at mRNA and protein level (all P ＜0.01).The expression of iNOS positively correlated with that of Collagen Ⅰ at mRNA and protein level (all P ＜0.01).In LX-2 cell culture,L-NAME inhibited the activation of LX-2,reduced eNOS and iNOS expression and increased α-SMA and CollagenⅠexpression,consistent with the role of TGF-β1.Simvastatin could directly increase the eNOS expression both in quiescent and activated LX-2 cells,decrease iNOS expression,maintain quiescent phenotype and inhibit its activation.ConclusionsSimvastatin ameliorated the genesis and progression of liver fibrosis by increasing eNOS expression in LX-2 cells and reducing iNOS,α-SMA and Collagen Ⅰ expression.

ObjectiveTo explore the role of fatty acid on endoplasmic reticulum (ER) stress and to detect the influence of tauroursodeoxycholic acid (TUDCA) on the expression of ER related genes in steatosis hepatocytes by establishing different models of hepatic steatosis.MethodsHealthy adult hepatocyte cell line HL-7702 was taken as research object.The model of hepatic steatosis was established with palmitic acid alone or mixed fatty acids.The cell viability was measured and calculated through CCK-8 positive cell proliferation assay.The cell morphology and steatosis was observed.The intracellular triglyceride content was tested with the triglyceride determination kit.With TUDCA intervention,the relative expression of Glucose-regulated protein 78 (GRP78) and CCAAT/Enhancer binding protein homologous protein (CHOP) at mRNA level was determined by real-time PCR.ResultsThe viability of hepatocyte was influenced once the concentration of mixed fatty acid reached 0.5 mmol/L or palmitic acid was 0.125 mmol/L.The effect of palmitic acid alone was stronger than that of mixed fatty acid on hepatocyte injury.The content of intra-hepatocyte triglyceride gradually increased in mixed fatty acid group,which only significantly increased after treated for 12 hours in palmitic acid alone group and then there was no significant change.There was no significant difference in relative expression of GRP78 and CHOP at mRNA level in various concentrations and treated time of mixed fatty acid group.After treated with palmitic acid at 0.5 mmol/L,intra-hepatocyte relative expression of CHOP at mRNA level increased obviously,however there was no effect on GRP78mRNA expression.After treated with palmitic acid at 1.0 mmol/L,both intra-hepatocyte GRP78 and CHOP mRNA relative expression increased.There was no significant difference in GRP78 and CHOP mRNA relative expression before and after ER stress in TUDCA intervented low dose palmitic acid group.There was significant difference in CHOP mRNA relative expression before and after ER stress in TUDCA intervented high dose palmitic acid group (at 12hrs:8.6400 to 5.1032; at 24hrs:13.7948to 6.4928,P=0.042 and 0.017),while no significant difference in GRP78 mRNA expression.ConclusionAt same fatty acid concentration,the larger propotion palmitic acid has,the more severe injury hepatocyte has.The regulation of palmitic acid on intra-hepatic ER stress is a time and dose dependent manner.TUDCA may improve palmitic acid induced ER stress to some degree.

ObjectiveTo explore the therapeutic effects and value of endoscopic variceal ligation and tissue glue injection therapy in esophageal and gastric fundal varices.Methods184 patients with severe esophageal varices underwent endoscopic variceal ligation treatment,and 32 cases of those accompanied with gastric fundal varices were treated with tissue glue injection therapy.All patients were followed-up for 6-months to observe the therapeutic effects and complication of endoscopic variceal ligation and tissue glue injection therapy.ResultsThe effective rate of endoscopic variceal ligation in severe esophageal varices was 71.74 ％ ( 132/184 ),the rate of acute hemostasis was 95.00％(57/60)and the rate of complication was 2.17 ％ (4/184).The effective rate of tissue glue injection in gastric fundal varices was 100％ (32/32) and the rate of complication was 21.88％ (7/32) (7 cases with refractory ulcers in injection site,2 of refractory ulcers cases with bleeding).There was no perforation and severe infection complications.ConclusionEndoscopic variceal ligation and tissue glue injection therapy have good therapeutic effects in esophageal and gastric fundal varices.

ObjectiveTo investigate the relation between aplysia Ras homolog Ⅰ(ARH Ⅰ )gene and NF-κB,and explore the possible mechanism in gastric cancer pathogenesis and development.Methods Human embryonic kidney cell line 293T cells were co-transfected with pre-constructed pIRES2-EGFP-ARH Ⅰplasmid and NF-κB luciferase reporterplasmid. The activity of NF-κB transcription was determined according to luciferase reaction.The expression of ARH Ⅰ in MKN-28 cells was interfered by siRNA. The phosphorylation level of NF-κB and the changes of NF-κB expression in nuclear was detected by Western blot.ResultsThe activity of NF-κB transcription was significantly down-regulated by ARHⅠoverexpressing in 293T cells.In MKN-28 cell line,the phosphorylation level of NF-κB and its expression in nuclear at protein level was significantly upregulated.ConclusionsThe activity of NF-κB transcription can be down-regulated by ARH Ⅰ,and which may reduce the phosphorylation level of NF-κB,and restrict it from entering the nucleus,and thus play a role in tumor suppression.

Objective To investigate the health-related quality of life (HRQoL) and its influencing factors in patients with inflammatory bowel disease (IBD).MethodsThe IBD patients underwent cross-sectional survey through Chinese version of inflammatory bowel disease questionnaire (C-IBDQ),the HRQoL of IBD patients were evaluated and analyzed with correlation analysis and multiple-linear regression analysis.Results A total of 100 IBD patients finished the questionnaire (UC 80,CD 17,ID 3).The overall score of HRQoL in IBD patients were (175.9±37.9),score of each item decreased,especially in the aspect of emotional function. There was no significantly difference in total scores of HRQoL between UC patients and DC patients (P＞0.05).The HRQoL total scores and each item score of patients in active period were significantly lower than those in remission period (149.4 ± 35.2,195.1 ± 26.7 ; P＜0.05).The HRQoL scores of patients with the duration less than one year were significantly lower than those over one year (P＜0.05).The HRQoL scores of severe patients were significantly lower than those of mild patients (P＜0.05).The HRQoL scores of patients whose medical cost mainly paid by themselves were lower than those paid by medical insurance,there were significant differences in social function and economic burden between these two groups.Multiple linear regressions analysis indicated that the influential factors of HRQoL were disease severity and activity,duration,and the payment.ConclusionsThe HRQoL of IBD patients are impaired in varying degrees.The influential factors of HRQoL were disease severity and activity,duration,and the payment.The HRQoL of IBD patients is obviously affected by economic burden.

Objective To establish Helicobacter pylori (Hp) infection induced chronic obstructive pulmonary disease (COPD) rat model,and to explore the role of Hp in the pathogenesis of COPD.Methods40 Wistar rats were randomly divided into double modeling group (Hp infection,smoked and intratracheal instillation of lipopolysaccharide),COPD group (smoked and intratracheal instillation of lipopolysaccharide),Hp infected group and control group.The lung function,cytokines level in serum and bronchial alveolar lavage fluid (BALF),Hp related genes expression in bronchial and lung tissue were detected.And Hp in bronchial and lung tissue was isolated and cultured.Results The lung tissue of both COPD group and double modeling group accorded with COPD pathological characteristics,and the latter was more apparent.The lung function of COPD group and double modeling group decreased more significantly than that of control group and Hp infected group (all P＜0.05),and which was more obvious in double modeling group than that of COPD group (P＜0.05).Along with the Hp colonization density increased,Ri and Re value of double modeling group increased (r=0.785 and 0.905),the value of Gdyn,PEF and FEV0.3/FVC decreased (r=-0.975,-0.959and -0.976).Compared with control group,IL-6,IL-8 and TNF-a cytokines levels in serum and bronchoalveolar lavage fluid of other groups increased significantly (all P＜0.05),and within the groups,double modeling group increased most significantly (all P＜0.05).Hp UreC gene was only amplified in part of bronchi and lung tissue of double modeling group,no Hp and suspicious bacteria colonies were isolated and cultured.ConclusionsHp not directly colonized in bronchi and lung tissue,which aggravated inflammation through increasing the serum and bronchoalveolar cytokines level of COPD rat model.Which caused the deterioration in lung function of COPD group.

ObjectiveTo obtain stable animal models and observe Helicobacter hepaticas (Hh)colonization and pathological claracteristics,through infecting different mice strains with Hh.Methods SPF-class male BABL/c Cr,SCID/Cr and C57BL/6 Cr mice were inoculated 0.2 ml Hh standard strain ATCC51450 bacterial suspension (1 × 108CUF/ml),inoculated for 3 times with 48 hours intervals,the control group was fed with the same volume of PBS.Mice were executed at 4 weeks,8weeks and 16 weeks since last Hh inoculation,and mice esophagus,stomach,jejunum,ileum,cecum,colon,liver and pancreas tissue were taken for histopathology examination,Micro-aerobic bacteria isolation,culture and identification and Hh specific 16S rRNA gene amplification.Results The colonization rates of Hh in cecum after inoculated in BALB/c Cr mice and SCID/Cr mice at 4 weeks,8 weeks and 16 weeks were all 8/8,colonization rates in colon at 4 weeks,8 weeks and 16 weeks were 4/8,5/8,5/8 and 3/8,6/8,5/8 respectively,colonization rates in ileum and jejunum at 16 weeks were 1/8,colonization rates in liver at 8 weeks and 16 weeks were 2/8,3/8 and 2/8,2/8respectively.The colonization rates of Hh in cecum after inoculated in C57BL/6 Cr mice at 4 weeks,8weeks and 16 weeks were 1/8,2/8 and 2/8 respectively,colonization rates in colon at 8 weeks and 16weeks were 1/8,2/8 respectively.Compared with C57BL/6 Cr mice,the inflammatory changes in liver,cecum and colon were more significant in Hh infected BALB/c Cr and SCID/Cr mice (P＜0.01),and histological scores gradually increased as infection time extended (P＜0.05,P＜ 0.01 ).The histological scores were significantly higher in those with colon and liver Hh bacterial colonization than those without Hh bacterial colonization (P＜0.05).The histopathological score of cecal tissue was positively correlated with the density of Hh colonization.ConclusionDifferent mice strains are with different susceptibility to Hh,and better Hh infection model can be obtained in Hh inoculated BALB/c Cr and SCID/Cr mice.

ObjectiveTo explore the mechanism of clopidogrel in human gastric epithelial cell line (GES-1) injury.MethodsSet up GES-1 cells monolayer culture model.Then the GES-1 cells were divided into negative control group,U0126 intervented group,clopidogrel intervented group and combined intervented group (U0t26 treated firstly then clopidogrel intervented).The cell proliferation and apoptosis in each group was examined by methyl thiazolyl tetrazolium (MTT) assay and Flow cytometry.TheexpressionofphosphorylatedERK1/2ineachgroupwasdetectedby immunocytochemistry method,and the expression quantity of phosphorylated ERK1/2 in each group was measured by western blot.ResultsThe result of MTT assay showed that compared with negative control group,the proliferation of GES-1 cells was inhibited in U0126 group,clopidogrel group and combined intervented group,and the inhibition percentage was 21.8％ ±2.7％,46.3％ ± 3.4％ and 82.9 ％ ± 0.8 ％ respectively ( F=615.556,P =0.000 ).The result of immunocytochemistry indicated that the expression of p-ERK in U0126 group,Clopidogrel group and combined intervented group decreased compared with negative control group,which was 10.80±1.64,7.20± 1.64,4.40±0.89and 1.40±0.55 respecitively (F=49.426,P=0.000).The result of western blot and immunocytochemistry was of the same trend.Conclusion In GES-1 cell model,clopidogrel may injureGES-1 cells through MAPK/EPK signal transduction pathway.

ObjectiveTo investigate the effects of protein phosphatase 2A (PP2A) inhibitors on the viability of pancreatic cancer cell line PANC-1 and its mechanism.MethodsPANC-1 cells were treated with PP2A inhibitors Cantharidin or Okadiac acid.The activity degree of NF-κB pathway was tested by Western blot.NF-κB pathway was blocked from all sectors by PP2Acα plamid transfection,NF-κB inhibition of protein kinase α (IKKα) and NF-κB inhibitor α (IκBα) dominant negative mutant and p65 interfering plasmid.Cell viability was determined by MTT.ResultsPP2A inhibitors could induce phosphorylation of IKKα,further phosphorylation of IκBα and degradation and followed by the release of p65 into nucleus.When PP2Acα,IKKα dominant negative mutant and IκBα dominant negative mutant were overexpressed,or p65 was interfered,the inhibition rate of Cantharidin on cell viability decreased (31.85±13.37) ％,(23.48±8.98)％,(22.63±5.81)％ and (20.88±3.24)％respectively,and the inhibition rate of Okadiac acid on cell viability decreased (40.17 ± 11.65)％,(27.34±14.28)％,(24.85±3.39)％ and (27.08±3.81)％ respectively.ConclusionsPP2Ainhibitors play a role in preventing pancreatic cancer through PP2Acα/IKKα/IκBα/p65 pathway.