Aim To build a simple experiment model of rat colon injury induced by acetic acid in vitro and to observe the effects of melatonin on this model.Methods On the basis of the establishment of rat colon injury model with acetic acid in vitro,we observed the colon mucosal damage caused by different concentrations of acetic acid(0,0.1%,0.2% and 0.4%) for different time(5,10,15,20,25 and 30 min),determined the content of lactic dehydrogenase(LDH),malondiadehyde(MDA) and mucus in medium,and examined the histological changes of colon mucosa by Alacian Blue method.On the basis of the establishment of this model,the experiment was designed into normal group,model group(acetic acid of 0.4%,time for 30 min) and melatonin treatment group(the final concentration 0.1,1.0、10 mmol?L~(-1)),and the indicators described above were detected to investigate the protective effects of melatonin.Results The LDH content of medium elevated gradually and the colon mucosal epithelial cells were injured by acetic acid in dose-and time-dependent manner,the mucosal edema,colon-wall depth and epithelium lose were observed at the same time,the MDA content of medium enhanced and mucus reduced correspondingly,but no significant change of the mucin expression in mucosal epithelial cells were observed.Pretreatment with melatonin reduced the release of LDH and MDA while promoted the secretion of mucus.Conclusion Colon mucosa of rats was injured by acetic acid in dose-and time-dependent manner in vitro.Acetic acid can impair the mucosal-mucus barrier by oxidating injured cell memberane.Melatonin can strengthen the barrier function of colon mucosa by its anti-oxidant action,attenuate the direct damage on colon mucosa of acetic acid.

Aim To establish the method of detecting the Arylamine N-acetyltransferase-1(NAT1) genotype and its distribution of polymorphism,and analyze the correlation between the genotype and the phenotype in a Chinese Hans population.Methods The peripheral blood samples from 140 Han people were collected and analyzed for NAT1 genotypes by multiple PCR and PCR-RFLP method.The NAT1 enzyme kinetics in leukocytes in 32 persons with different genotypes from 140 Han people,were determinated for NAT1 phenotype by HPLC,the values of intrinsic clearance(Clint) and V_(max) and Michaelis constant(K_m) of NAT1 were calculated for evaluation of para-aminobenozic acid as a specific substrate.Results The NAT1 genotype of Chinese Hans population was distinguished accurately by multiple PCR and PCR-RFLP methods,and was not interfered by the interaction of several restriction endonucleases.The allelic frequencies of NAT1~*3,NAT1~*4,NAT1~*10 and NAT1~*11 from 140 Han people,were 0.082,0.496,0.40 and 0.022,respectively.The frequency of NAT1~*4 allele was significantly lower than that of Caucasian populations,but higher than that of Southeast Asia and African.Frequencies of NAT1~*3 and NAT1~*11 allele were comparable with those of many Asian populations,but the frequency of NAT1~*10 allele was higher than that of in Europe and America populations.Compared with the activity of wild genotype NAT1 ~*4/~*4,activities of the homozygote or heterozygote NAT1~*10 genotypes which include the NAT1 ~*4/~*10,the NAT1 ~*10/~*10 and the NAT1 ~*10/~*3 were higher significantly(P

4 time percentage of 24 h were observed among the three different genotype groups after a single dose or repeated doses of rabeprazole. And the ratio of these pH dates between day 1 and day 8 ranged from 85% to 110%.Conclusions In Chinese healthy Han people, CYP 2C19 genetic polymorphism has not any significant influence on acid inhibitory efficacy and the metabolism of rabeprazole. Rebeprazole could rapidly achieved maximum acid inhibitory efficacy in the subjects with different CYP2 C19 genotype status.

Objective To investigate the expressions of Smad3 and Smad7 in patients with ulcerative colitis(UC)and their relation with clinicopathology.Methods The expressions of Smad3 and Smad7 were measured by immunohistochemistry with SABC method in 60 UC specimens and 16 normal colonic tissues.The association of expressions of Smad3 and Smad7 proteins with clinical staging,lesion extent and pathologic grading were retrospectively analyzed.Results The expression of Smad3 was significantly lower in UC patients than in normal controls(P＜0.05),however,there was no relation between Smad3 expression and lesion extent(P＞0.05).There was a negative correlation between the expression of Smad3 and histological grade(r=-0.283,P＜0.05).The expression of Smad7 was significantly higher in UC patients than in normal controls,and its expression in active disease was higher than that in clinical remission(Z=2.097,P=0.036).There was a positive correlation between the expression of Smad7 and histological grade(r_s=0.453,P=0.000),and no relation between Smad7 expression and lesion extent(r_s=0.066,P=0.614).The statistical analysis showed a negative correlation between Smad3 expression and Smad7 expression(r=-0.420,P＜0.05).Conclusion The abnormal expressions of Smad3 and Smad7 are correlated with pathogenesis of UC.Furthermore.Smad7 may serve as marker for disease activity of UC.

Objective To prospectively compare the efficacy of omeprazole and esomeprazole on H. pylori eradication related to cytochrome P450 (CYP) 2C19 polymorphism in Chinese Han poputafion. Methods A total of 240 patients with peptic ulcer were randomly assigned to receive either EAC (amoxicillin, clarithromyein and esomeprazole) treatment or OAC (amoxicillin, clarithromycin and orneprazole) treatment for one week with 120 each. Then, the patients were sustained treated with esomeprazol or omeprazole for 3 weeks. The endoscopic evaluation was performed 2 weeks after treatment. At the end of the treatment, a carbon-13 (13C) urea breath test (13C-UBT) was performed to determine H. pylori status. Polymerase chain reaction and restriction fragment length polymorphism were used to detect CYP2C19 genotypes including extensive metabolizer (EM), poor metabolizer (PM), homozygous (homEM) and hetEM. Results Two hundred and twenty five of 240 patients completed the study. H. pylori eradication was 79.2% with OAC regimen and 88.3% with EAC regimen by intention to-treat (ITT) analyses without difference (P>0.05). Whereas H. pylori eradication was 87.20/00 with OAC regimen and 91.4% with EAC regimen by per-protocol (PP) analyses without difference (P>0. 05). However, both ITT and PP analysis showed that there was significant difference in H. pylori eradication between EAC regimen (91.9% and 97.1%) and OAC regimen (71.8% and 77.8%) in patients with homEM genotype (P= 0.037 and P=0.028). Two weeks after treatment, the percentage of ulcer healing was 79.2% or 81.9% in EAC group and 69.2% or 76.1% in OAC group by ITT or PP analysis, respectively (P> 0.05). Low side effects were noted in EAC or OAC groups (3.3% vs. 7.5%,P>0.05). Conclusion The high eradication will be achieved by esomeprazole-based triple regimen which is superior to omeprazole-based triple regimen in treatment of patients with homEM genotype.

Objective To investigate the relationship between TNF-α, IFN-γand intestinal muco-sal permeability in a mouse colitis model and its inhibiting effect by balsalazide. Methods Forty-five C57BL/6J mice were divided randomly into five groups. Normal group was only fed with distilled water, DSS group and balsalazide groups at doses of 42, 141,423 mg/kg were both fed with 5% DSS. Balsalazide was given by intragastric administration. At the end of the experiment, colon tissue was collected for assessment of histological index(HI) and the MPO activity. Small intestinal mucosa was collected for assessment of the content of TNF-α and IFN-γ,transmission electron microscope(TEM), and detection of permeability by Ev-arts blue method. Results Compared with normal group, DSS group mice all manifested severe weight loss associated with hematocbezia and diarrhea, HI score, and the colon MPO activity and the content of TNF-α and IFN-γ were increased significantly. Intestinal mucosa showed a thinning of microvillous carpet, with de-curtated and broaden junctional complex and enlarged intercollutar space under TEM observations. The amount of Evans blue permeated into intestinal wall was obvious. Compared with DSS group, the HI score, the MPO activity and the content of TNF-α and IFN-γ were decreased by balsalazide. The amount of Evans blue permeated into intestinal wall was less. Ileal microvillous carpet was ameliorated dose dependently by balsalazide. Conclusion In DSS-induced colitis model, the change of the content of the TNF-α and IFN-γ, was accordance with the increase of intestinal mucosal permeability while balsalazide can significantly amelio-rate intestinal mucosal permeability by its anti-colitis effect.

AIM: To investigate the effects of melatonin (MT) on the immunological function in colon of rats immunological colitis. METHODS: The rats colitis was produced by enema with trinitrobenzene sulfonic acid and ethanol. The experiment groups included normal group, model group, 5-aminosalicylic acid (5-ASA) group (100 mg?kg~(-1)), MT groups (2.5,5.0,10.0 mg?kg~(-1)), and treated intracolon with saline, saline, 5-ASA and MT respectively (once per day, from the d7 after the establishment of colitis model to the end of the experiment). The content of interleukin (IL-1, IL-2) and tumour necrosis factor (TNF)-? in rats colon were detected; The inflammatory colon were homogenatized and incubated with lipopolysaccharides (LPS), and designed as control group, model group, 5-ASA group (100?mol/L), MT groups(0.01, 0.1, 1.0 mmol/L). The content of IL-1, IL-2, TNF-?, nitric oxide (NO), lactic dehydrogenase (LDH), myeloperoxidase (MPO) and malondiadehyde (MDA) in the supernant were detected. RESULTS: In model group, the contents of IL-1, IL-2 and TNF-? in colon elevated remarkably and MT depressed this effectively. In the inflammatory colon homogenates of rats colitis, the content of IL-1, IL-2, TNF-?, NO, LDH, MPO and MDA elevated remarkably, Pretreation with MT depressed the content of IL-1, TNF-?, LDH, MPO and MDA effectively. MT (1.0mmol/L) also reduced the production of NO obviously. CONCLUSION: The abnormal immunological function is shown obviously in rats colitis. MT normalizes this change in vivo and in vitro and attenuates the mucosal damage.

Objective To determine the pathway of macrophage metalloelastase (MME)generate active angiostatin by decomposing plasminogen and its effect on inhibiting growth of tumor and microvessel density (MVD) in vivo in mouse models. Methods The recombined plasmid pEGFPC1-MME was constructed. Thirty mice were subcutaneously inoculated with CT-26 cells that were stably transfected with pEGFP-C1-MME (MME-transfected group), 30 with CT-26 cells transfected with empty vector pEGFP-C1 (vector-transfected group) and 30 with CT-26 cells (non-transfected group). Radioiodination and radioisotope tracer were used to explore the pathway of angiostatin generation in vivo. Results SDS-PAGE electrophoresis analysis revealed that, in the PAGE gel contained the protein with molecular weights of 35 000 and 38 000, radioactivity in MME-transfected group was significantly higher than vector-transfected and non-transfected groups (P = 0. 00).Western blotting analysis demonstrated two bands containing 35 000 and 38 000 fragments in three groups. Quantification of the protein signals by image analysis revealed that the levels of 35 000 and 38 000 fragments were obviously increased in MME-transfected group (9.32±1.52 and 5.61±2.24,respectively) than those in vector-transfected (2.47 ± 0.23 and 0. 67 ± 0. 12, respectively) and nontransfected (1.21±0. 69 and 0. 86 ± 0.44, respectively) groups (P= 0.00). The average value of MVD and fluorescent express of vascular endothelial growth factor (VEGF) were lower in MMEtransfected group when compared with those in vector-transfected and non-transfected groups (P =0.00). The average tumor size in MME-transfected group was small in comparison with vectortransfected and non-transfected groups (P= 0.00). Conclusions MME is demonstrated to be one of matrix metalloproteinase that closely related with angiostatin production and has inhibitory effect on tumor growth in tumor-bearing mice.

Objective To investigate the change of serum leptin and ghrelin levels in patients with active ulcerative colitis (UC) and their clinical significance.Methods Fifty-four patients with active UC and 25 normal controls were enrolled in this study,and serum levels of leptin and ghrelin were determined by enzyme-linked immunosorbent absorption.The correlations between serum leptin/ghrelin and the main clinicopathological features of patients with active UC were analyzed.The nutritional status of these patients were assessed by using the Nutritional Risk Screening 2002 ( NRS 2002).Results The serum levels of leptin and ghrelin in patients with active UC were significantly higher than those in normal controls [leptin:(245.51 ± 129.72) vs.( 183.80 ±59.67) pg/ml,P =0.029; ghrelin:(1477.96 ±501.86) vs.(1188.00±474.51) pg/ml,P=0.041]; and both of them were correlated with the disease severity ( leptin:r =0.715,P =0.019 ; ghrelin:r =0.667,P =0.027),but were not correlated with the clinical type,the extent of lesion,and the endoscopic grading (all P ＞ 0.05).In the UC patients with nutritional risk,the serum levels of leptin and ghrelin were also significantly higher than those in subjects without nutritional risk [ leptin:(332.82 ± 172.92) vs.( 194.15 ± 49.71 ) pg/ml,P =0.026 ; ghrelin: ( 1848.60 ±326.48) vs.(1259.93±459.24) pg/ml,P=0.021].Conclusion Patients with active UC have remarkably higher serum leptin and ghrelin levels,which is associated with the disease severity and the nutritional risk.

Objective:To analyze the expression of TNF-related apoptosis-inducing factor TRAIL,c-FLIP and caspase-8 in peripheral blood mononuclear cells of patients with rheumatoid arthritis at different period,in order to provide a experimental basis that treating and looking for a more effective way and method.Methods:mRNA expression of TRAIL,c-FLIP and caspase-8 were detected by Real-time PCR from peripheral blood mononuclear cells;TRAIL,c-FLIP and caspase-8 were checked by Western blot from peripheral blood mononuclear cells.Results:The mRNA expression level of TRAIL,c-FLIP and caspase-8 from PBMC at different groups of RA:TRAIL mRNA in group M was 3.26±0.78 which was much higher than healthy controls(1.30±0.20) (P=0.028),and those of group L and group H (1.56±0.37 and 1.83±0.26 respectively) was slightly higher than controls(P<0.05).C-FLIP mRNA in low activity group (L),middle activity group (M) and high activity group (H) were 1.27±0.28,1.32±0.34 and 1.93±0.40 re-spectively,which was higher than that of healthy controls (1.08 ± 0.12) (P=0.035,0.034,0.030).The relative level caspase-8 mRNA in group L,group M,group H were(2.77 ±0.97),(4.52 ± 0.85),and(2.13 ± 0.44)which was higher than healthy controls (1.04±0.13) (P=0.023,0.012,0.032).The protein level of TRAIL,c-FLIP and caspase-8 in PBMC from different groups of RA patients:The expression of TRAIL in group M was significantly increased than group L,H and control(P<0.05) with no difference in group L.c-FLIP protein in all protein expressed in group M quantity highest,significantly higher than other groups.Caspase-8 expression in the group M and H was significantly enhanced than control (P=0.003,0.001 ) with no difference in group L (P> 0.05). Conclusion:During the different active stages of RA,in peripheral blood mononuclear cells,the expression of TRAIL,c-FLIP and caspase-8 are increased overall trend,possible can provide certain experimental reference for clinical therapy.