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Objective:To observe the protective effects of Clostridium butyricum and butyrate on pancreas, liver and intestinal mucosa in rats with acute necrotizing pancreatitis (ANP), and to explore the possible mechanism.Methods:Forty Sprage-dawley rats were randomly divided into control group, ANP group，Clostridium butyricum treated group(CB group) and butyrate treated group(SB group), with 10 rats in each group by random number method. The ANP rat models were prepared by retrograde injection of sodium taurocholate into the biliopancreatic duct. Rats in CB and SB group were given intragastic administration of Clostridium butyricum 1×10 9 CFU or sodium butyrate (100 mg/kg) in 10 days once a day before modeling. Serum amylase (SAMY), lypase, ALT, AST, TBil, tumor necrosis factor alpha(TNF-α), IL-6 and HMGB1were measured after 24 h. Protein from intestinal mucosa was extracted and Western Blotting was used to measure expression of tight-junction proteins ZO-1, claudin-1 and occludin. Pancreas and liver tissues were stained with hematoxylin-eosin and scored by pathology. Results:The levels of amylase [(9365.1±716.5), (5947.3±512.0), (6517.7±269.6)U/L], lipase[(8343.7±1041.4), (6600.4±899.7), (6754.4±1046.4)U/L], AST[(560.5±72.7), (432.0±76.2), (429.8±40.5)U/L], ALT[(499.9±65.2), (385.7±46.0), (395.8±45.8)U/L], TBil[(134.2±56.2), (74.3±65.2), (81.3±35.3)U/L], TNF-α[(162.0±14.4), (100.4±6.3), (119.2±12.5)ng/L], IL-6[(161.4±26.0), (104.8±15.2), (105.5±12.7)ng/L], HMGB1[(100.1±6.7), (58.0±7.7), (63.4±7.2)ng/L] in ANP group, CB group and SB group were detected; and the pathological scores of pancreas[(11.2±1.08)，(9.45±1.06), (9.04±0.89)] and liver[(2.89±0.73), (2.09±0.49), (2.12±0.52)] in ANP group，CB group and SB group were higher than those in control group[(100.6±5.20)U/L, (966.5±301.9)U/L，(30.2±6.3)U/L, (27.6±5.9)U/L, (2.4±0.6)U/L, (29.5±4.8)ng/L，(36.9±7.6)ng/L，(35.5±5.7)ng/L，(1.18±0.05)，(0.56±0.09)]. However, those indexes in CB group and SB group were lower than those in ANP group, and the difference was statistically significant (all P<0.05). The expression of ZO-1 in control group, ANP group, CB group and SB group was 1.83, 0.79, 1.25 and 1.16. The expression of claudin-1 in control group, ANP group, CB group and SB group was 0.58, 0.13, 0.43 and 0.37. The expression of occludin in control group, ANP group, CB group and SB group was 1.06, 0.38, 0.82 and 0.79. The expression of TJ proteins in ANP group was significantly lower than that in other groups and the difference was statistically significant (all P<0.05). Conclusions:Clostridium butyricum and metabolites butyrate can alleviate the inflammatory response in ANP rats with liver injury, maintain the function of intestinal mucosal barrier and prevent the liver injury.
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Objective To explore the effect of Clostridium butyricum ( C. butyricum ) and its metabolite butyrate on the function of intestinal mucosal barrier and intestinal flora in acute necrotizing pancreatitis ( ANP) rats with intra-abdominal hypertension ( IAH) . Methods Eighty SD rats were randomly divided into normal control group (A group, n=20), ANP with IAH group(B group, n=20), ANP with IAH and C. butyricum treated group ( C group, n=20 ) , ANP with IAH and sodium butyrate treated group ( D group, n=20). Rats of C and D group were given intragastric administration of C. butyricum 1 × 109 CFU once a day or 100 mg/kg sodium butyrate once a day from 10 days before modeling. Sodium taurocholate injection method via pancreatobiliary ducts was used to establish ANP with IAH rat model, and the intra-abdominal pressure was measured by direct puncture of left lower belly 24 h after modeling. Blood samples were collected for detecting serum amylase(AMY), tumor necrosis factor alpha (TNF-α), diamine oxidase( DAO ) , lipopolysaccharide ( LPS ) and D-Lactate, and the pathological changes of terminal ileum was observed. Real-time quantitative PCR was used to detect the populations of 6 bacteria in ileum mucosa. Results The levels of AMY, TNF-α, LPS,DAO, D-Lactate and ileum mucosa score were obviously higher in B, C and D group than those in A group, but the number of piobiotic flora in ileum mucosa was lower than that in A group, while the number of pathogenic bacteria was higher than that in A group. The levels of LPS, DAO, D-Lactate and ileum mucosa pathological score were lower in C group and D group than those in B group, but the number of piobiotic flora in ileum mucosa was lower than that in B group, while the number of pathogenic bacteria was higher than that in B group. All the differences above were statistically different (P<0.05). Conclusions C. butyricum and butyrate can maintain the function of intestinal mucosal barrier in ANP rats with IAH, and also readjust the imbalance of intestinal flora.
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Objective To investigate the alternation of insulin -like growth factor -Ⅱ( IGF-Ⅱ) gene promoter P4 methylation status in hepatocellular carcinoma ( HCC) and explore its relationship with expression of P4 mRNA levels.Methods Liver specimens of 43 patients with HCC and normal liver specimens of 9 control patients were collected in operation .Tissue DNA and total RNA were extracted from these specimens .IGF-Ⅱ P4 methylation status and P4 mRNA expression levels were detected .Results (1)The incidence of IGF -ⅡP4 methyl-ation in HCC group was significantly lower than that in normal liver specimens (16.28%vs 88.89%,χ2 =19.12,P<0.01).(2)The expression level of IGF -ⅡP4 mRNA in HCC group was significantly higher than that that in normal liver specimens[(0.96 ±0.74) vs (0.25 ±0.19),t=5.48,P<0.01].(3)In HCC group,the IGF-Ⅱ P4 mRNA expression level with hypomethylation gene was significantly higher than that without hypomethylation gene [(1.18 ± 0.76) vs (0.32 ±0.27),t=5.28,P<0.01].Conclusion The hypomethylation alternation of IGF -Ⅱ P4 gene promoter which is accomplished by up -regulate P4 mRNA expression has a close relationship with HCC .
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Objective To observe the traditional Chinese medicine (TCM) and umbilical treatment on diarrhea-predominant irritable bowel syndrome(IBS-D)with mental disorders. Methods Thirty-seven cases with IBS-D were randomized into a treatment group(33 cases) and a control group(31 cases). The treatment group was given Shugan-Jianpi TCM decoction and pepper umbilical dressing. The control group was given Pinaverium Bromide Tablets and Flupentixol and Melitracen tablets. After having been treated for 4 weeks, Hamilton Anxiety Scale(HAMA), Hamilton Depression Scale(HAMD), IBS Symptom Severity Scale(IBS-SSS), IBS quality of life questionnaire (IBS-QOL) were determined. Results After the treatment, HAMD, HAMA, IBS-SSS in the treatment group and the control group all significantly decreased (the treatment group t=14.280, 13.530, 18.847,the control group t=11.514, 9.424, 13.829, P<0.01), and IBS-QOL increased markedly(t=-16.848, -11.712, P<0.01). Compared with HAMA, IBS-SSS and IBS-QOL in treatment group, the control group showed statistical difference(t=-3.183, -2.549, 3.904, P<0.01 or 0.05). The total effective rate of the treatment group was 94%higher than 75%of the control group(χ2=4.727, P<0.05). Conclusion Shugan-Jianpi TCM decoction and pepper umbilical dressing can improve the clinical symptoms and psychological symptoms and improve the quality of life of IBS-D patients.
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Objective To retrospectively investigate the clinical features and incidence trends of hypertriglyceridemic acute pancreatitis (HLAP) in Guangzhou First People's Hospital for 20 years.Methods The medical records of 1 362 patients,who were admitted to Guangzhou First People's Hospital with acute pancreatitis during January 1991 to December 2010,were reviewed and 99 patients met the HLAP criterion.These patients were divided into four groups according to years (1991 ～ 1995,1996 ～2000,2001 ～2005,and 2006 ～2010).The incidence and clinical features were compared among the 4 groups.Results Among the 99 patients,there were 61 males and 38 females,the mean age was (44 ± 12) years old,and young male was predominant in HLAP.HLAP accounted for 7.3％ of all AP patients,and the incidence was increased 1.5 times during the 20 years period (5.4％,5.7％,6.7％,and 8.3％ respectively).The serum amylase level was (513.3 ±462.7) mmol/L,TG level was (12.7 ± 7.0) mmol/L,Ranson score was (1.2 ± 1.1),CTSI score was 2.2 ± 1.1,incidence of SIRS was 36.4％,and the incidence of MODS was 18.2％,and the incidence of pancreatic pseudocyst was 5.1％,but the incidence and mortality of SAP was not changed with time.The incidence and mortality of SAP of HLAP were significantly higher than those of acute biliary pancreatitis (20.2％ vs 8.5％,6.1％ vs 2.8％).The rate of blood purification increased from 0 to 10.7％,and the hospital stay was decreased from 26 d to 14 d,and the difference between the two groups was statistically significant (P ＜ 0.05).Conclusions Incidence of HLAP is increased gradually,with a trend towards more severe disease,but the prognosis is improved.
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Objective To explore the effects of low dose amitriptyline (AMT) on gastrointestinal function and its adverse effects in healthy volunteers.Methods In this randomized,double-blind,placebo-controlled cross-over study,28 healthy volunteers were divided into AMT with placebo group (n=14) and placebo with AMT group (n=14).The former took AMT for seven days at first stage,followed with a 14 days wash out stage and then took placebo for seven days at second stage.Patients of the latter group took medication in reverse order.The dose of medication was 12.5 mg three times per day.The subjects underwent drinking-ultrasonography test and lactulose hydrogen breath test before taking medication and between the seventh and eighth day of first and second stage.The data were analyzed by two stages cross-over analysis,Wilcoxon signed-rank test and chi-square test.Results The results of drinking-ultrasonography test showed that there were no significant differences in proximal gastric cross-sectional area between AMT and placebo after drinking 200,400,600 and 800 mL water (all P＞ 0.05).After drinking 800 mL water,there were no significant differences in gastric liquid emptying rate between AMT and placebo at the fifth and the tenth minutes (both P＞0.05).After drinking 600 mL and 800 mL water,the visual analogue scale (VAS) of AMT was significantly lower than that of placebo (2.98±0.85 vs 3.57±0.94,Z=4.412,P＜0.01; 4.57±0.98 vs 5.57±0.82,Z=4.170,P＜0.01).The results of lactulose hydrogen breath test revealed that orocecal transit time of AMT was obviously longer than that of placebo ((109.29±29.68) min vs (96.61±23.90) min,F=9.918,P＜0.01)).The common adverse effects were mild sleepiness,bitter taste and dry mouth.Conclusions Low dose AMT can prolong orocecal transit time and improve gastric sensitivity,but can not significantly affect proximal gastric accommodation and gastric liquid emptying.The adverse effects are mild and the safety is good.
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Objective To explore the depression and anxiety status of patients with refractory irritable bowel syndrome (IBS)and its subtypes.Methods In this large sample,multi-center,prospective survey(Registration number in Chinese Clinical Trial Registry:ChiCTR-TRC-12001969),the patients who conformed to the Rome Ⅲ criteria for IBS were recruited from the department of gastroenterology of three hospitals in Guangdong province from June 2012 to September 2012.And 100 healthy subjects from the examination center of hospitals were recruited at the same time.The 17-item Hamilton Depression Rating Scale(HAMD) and the 14-item Hamilton Anxiety Rating Scale(HAMA) were used to analyze the depression and anxiety,respectively.Results 9802 patients were collected from department of gastroenterology,and a total of 1128 (11.5 ％) patients were diagnosed as IBS.601 of questionnaires were collected,including 142 (23.6％) of refractory IBS and 459 (76.4％) of non-refractory IBS.The incidences of depression,anxiety,depression and anxiety and the HAMD,HAMA scores were higher in refractory IBS group than those in the non-refractory IBS and control group,((66.2％ vs 23.3％ vs 9.0％),P＜0.05 ;(65.5％ vs 28.5％ vs 10.0％),P＜0.05;(58.5％ vs 17.9％ vs 5.0％),P＜0.05;(9.77 ±5.16)vs(5.48 ±4.03)vs(1.66±2.50),P＜0.05;(9.75 ±5.08)vs(5.91 ±3.80)vs(2.26 ±2.68),P＜0.05).And the incidences of moderate-severe depression and moderate-severe anxiety were higher in refractory IBS group than those in the non-refractory IBS and control group ((8.5％ vs 1.3％ vs 0),P＜0.05; (15.5％ vs 3.7％ vs 0),P＜0.05).The incidences of depression,anxiety,depression and anxiety and the HAMD scores were not obviously different among IBS-D,IBS-C,IBS-M and IBS-U subtypes of refractory IBS (P ＞ 0.05).However,the HAMA scores of IBS-D and IBS-M were higher than that of IBS-U((10.10 ± 5.03) vs(7.55 ± 3.22),P ＜ 0.05 ; (12.08 ±6.89) vs (7.55 ± 3.22),P ＜ 0.05).Conclusion The high incidences of depression,anxiety and depression and anxiety,and the serious degrees of depression and anxiety existed in the patients with refractory IBS.And these are the important causes for refractoriness in these patients.
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Objective To evaluate the effects of infliximab (TNF-α monoclonal antibody ) on intestinal barrier injury in ANP complicated with MODS in a rat model.MethodsThirty SD rats were randomly divided into sham operation group (SO),ANP group and infliximab treatment group.Sodium taurocholate (4.5％) was injected into the pancreatic duct to induce ANP complicated with MODS model.Infliximab (8 mg/kg) was injected via tail vein in 6h after modeling in infliximab group.Same amount of 0.9％ NS was injected into the pancreatic duct in SO group.After 24 h of modeling,all rats were sacrificed,intestine and pancreas samples were collected for pathologic examination.The blood samples were harvested.The serum levels of amylase,TNF-α,diamine oxidase( DAO),D-lactate,and the rate of carbon propelling in ileum were measured.ResultsThe serum levels of amylase were ( 1125 ± 331 ),( 11024 ± 2203 ),( 545 ±30) U/L in SO group,ANP group and infliximab group; the serum levels of TNF-α were (12.1 ± 4.0),(107.6 ± 18.5),(75.8 ±5.9) U/L; the pathological scores of pancreas were 2.25 ±0.38,14.10 ±0.22,3.93 ± 0.67,the difference among the 3 groups was statistically significant ( P ＜ 0.05 ).The pathological scores of intestine were 2.29 ± 0.32,6.61 ± 0.58,3.91 ± 0.41 ; the DAO levels were ( 87.88 ± 34.51 ),(146.30 ±12.99),(115.00 ± 18.58) ng/ml; the D-lactate levels were (1.50 ±0.49),(2.32 ± 0.35),(2.02 ± 0.25 )mmol/L; and the rates of carbon propelling in ileum were (0.64 ± 0.04 )％,(0.28 ±0.08)％,(0.52 ±0.09)％,the difference among the 3 groups was statistically significant (P ＜0.05).ConclusionsInfliximab can effectively prevent dysfunction of intestinal barrier and improve motility in ANP rats.
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Objective To compare the treatment effects of Qingyi Huoxue decoction and infliximab on acute necrosis pancreatitis ( ANP) complicated with MODS in a rat model. Methods 4.5% sodium taurocholate was injected into the pancreatic duct to induce the ANP complicated with MODS model. The ANP rats were randomly divided into 3 groups, ANP group (ANP), Qingyi Huoxue decoction treatment group ( QG) , infliximab treatment group (IG). Rats in infliximab group received infliximab injection at a dose of 8 mg/kg body weight via tail vein 6 h after the ANP induction. The ANP and QG received normal saline and Qingyi Huoxue decoction (20 ml/kg) via gastric lavage 4 h before and 3 h, 9 h after ANP induction. After 24 h, all rats were sacrificed, the serum levels of amylase, total bilirubin, Cr, TNF-α, diamine oxidase ( DAO) , intra-abdominal pressure (IAP) and the rate of carbon propelling rate in ileum were measured. The pancreas samples were collected for pathological examination. The pathological score of pancreas was calculated. Results The pathological scores in ANP, QG, IG were 13.8 ±0.8, 6.1 ±0.4, 3.9 ±0.6, and the difference was statistically significant (P <0.05). The serum levels of amylase, total bilirubin, Cr, TNF-α were significantly decreased. In ANP, QG, IG the serum levels of DAO were (186.3 ± 10.2 ) , ( 134.6 ± 14.3 ) , ( 149.1 ± 16.3) U/L; the carbon propelling rates in ileum were (53 ±0.1)% , (89 ±0.1)% , (61 ±0.1)% ; the IAPs were (11.8 ±1.5), (4.1±0.8), (5.8 ±1.2) mmHg(1 mmHg=0.133 kPa). The DAO and IAP in AG, IG were significantly decreased when compared with that in ANP group, but the carbon propelling rates in ileum was significantly higher than that in ANP group, and the difference was statistically significant ( P < 0.05). In addition, the carbon propelling rates in ileum in QG were higher than that in IG, and IAP and DAO levels were lower than that in IG, and the difference was statistically significant (P <0.05). Conclusions The Qingyi Huoxue decoction and infliximab were significantly effective in the treatment of ANP rats complicated with MODS. But the effects of Qinyi Huoxue decoction on promoting gastrointestinal motility, reducing the IAP and improving the intestinal barrier function were better than those of the infliximab.
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Objective To investigate the time-course change of intra-abdominal pressure (IAP), TNF-α in rat with acute necrotizing pancreatitis (ANP) and its mechanism. Methods 80 SD rats were randomly divided into ANP group and control group. ANP group was induced by retrograde injection of 4.5% sodium taurocholate into the pancreatic duct in SD rats, while control group was induced by injection of saline at the same dosage. Eight rats in each group were killed at 1 h, 3 h, 6 h, 12 h and 24 h, respectively. The serum levels of amylase, TNF-α were determined;IAP, amount of abdominal ascites, pancreatic pathologic changes were evaluated. Results The levels of amylase in ANP group increased progressively with time, which were 32-folds higher than those in control group at 24 h. The serum levels of TNF-α in ANP group reached the peak at 6 h, and were significantly higher than those in the control group. The amount of abdominal ascites increased post-ANP induction and reached the peak at 24 h, which was 4.7-folds higher than that in the control group. The IAP significantly increased 1 h after induction, which was 3 times as high as the control group in 3 h group and 9 times in 12 h group. Pathologic injuries deteriorated progressively and reached the peak at 24 h. Conclusions Significant IAP occurred in ANP rats and reached the peak at 12 h, TNF-α reached the peak at 6 h, therefore IAP may be the secondary response to inflammatory reaction.
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Objective To explore the dose-effect relationship and optimal concentration of sodium taurocholate (NaTc) to establish acute necrosis pancreatitis (ANP) complicating MODS in a rat. Methods 280 SD rats were randomly divided into control group (n =40) and 3.5%, 4%, 4.5%, 5% ANP group (n = 60, respectively). ANP groups were induced by retrograde injection of NaTc solution into the pancreatic duct of rats. Rats in each model group were sacrificed at 1, 3, 6, 12 and 24 h after the model induction. Serum amylase, serum total bilirubin, creatinine, blood gas analysis, and pathological changes of pancreas were determined. Survival rate at 24 h after the model induction was observed in other 20 ANP rats in each group. Results In each groups, the serum amylase level, creatinine, blood gas analysis and pathological changes all increased corresponding to the increase of NaTc, and there was significant dose-effect relationship and time-effect. In 4.5% group, the dysfunction of liver and kidney reached the peak at 6 h,while the respiratory dysfunction reached its peak at 12 h. Its incidence of MODS was 52.5%, which was significantly higher than those in 3.5%, 4.0% group (15.0% and 37.5%) ;and 24 h survival rate of 4.5% group was 65%, which was significantly higher than that in 5% group (30%). Conclusions 4.5% NaTc solution may be the optimal concentration to establish the ANP model, which had higher incidence of MODS and 24 h survival rate.