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Objective To explore the effects of ApoC3 gene on the severity of hypertriglyceridemiainduced acute pancreatitis (AP).Methods ApoC3 transgenetic mice and C57BL/6J mice AP model was induced by cerulein intraperitoneal injection,and ApoC3 transgenetic mice and C57BL/6J mice injected by normal saline solution in equal volume served as control group.Serum triglyceride and cholesterol were detected,and the pathological changes of the pancreas were observed.RT PCR method was used to examine the changes of the inflammatory factor including IL-1β,IL-6,α-SMA and TNF-α mRNA levels,which reflected the severity of the inflammation.Results Serum triglyceride and cholesterol were higher in ApoC3 transgenetic mice than in C57BL/6J mice [(3.434 ± 0.931) mmol/L vs (0.766 ± 0.120) mmol/L,(2.553 ±0.178) mmol/L vs (1.996 ± 0.080) mmol/L],and the differences were statistically different (P ＜ 0.05).The pathological changes of the pancreas were more severe in ApoC3 transgenetic AP mice than in C57BL/6J AP mice,and the IL-1β,IL-6 and α-SMA mRNA levels in the pancreatic tissue were obviously higher in ApoC3 transgenetic AP mice than in C57BL/6J mice (1.72 ± 0.07vs 0.78 ± 0.09,1.58 ± 0.09vs 0.87 ±0.04,0.83 ± 0.05vs 0.44 ± 0.04),and the differences were statistically significant (P ＜ 0.05),while there was no statistical difference on TNF-αmRNA level (0.70 ± 0.09vs 0.65 ± 0.08,P ＞ 0.05).Conclusions ApoC3 gene could aggravate the severity of the inflammation in hypertriglyceridemia-induced AP.
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Objective To investigate functional role and clinical significance of the methylation in the promoter of TPAP gene in the development and progression of pancreatic cancer.Methods Surgically resected specimen from 68 patients who were pathologically diagnosed as pancreatic cancer in Changhai Hospital from July 2006 to August 2009 were collected.The methylation in the promoter of TPAP gene in tumor and nontumor adjacent tissue was detected by methylation specific PCR.Results The methylation rate of tumor and non-tumor adjacent tissue was (0.214 ± 0.057) ％ and (0.084 ± 0.096) ％,respectively,and pancreatic cancer tissue had significantly higher methylation rate than the adjacent tissue.Hypermethylation of TPAP gene was not correlated with age,gender,tumor differentiation,lymphatic metastasis,serum CEA and CA19-9,but was positively correlated with distant metastasis.Conclusions Hypermethylation in the promoter of TPAP gene may participate in the invasion and metastasis of pancreatic cancer and the hypermethylation of the promoter is closely associated with the tumorigenesis and development of pancreatic cancer.
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Objective To investigate the effects of microRNA-196a(miR-196a) inhibitory sequences transfection on HOXB8 expression in PANC1 cells.Methods PANC1 cells were divided into control group,miR-196a inhibitory sequences group and siRNA control group.Liposomal transfection method was applied to transfect miR 196a inhibitory sequences and siRNA control into PANC1 cells.RT-PCR and Western blot were used to detect the expressions of miR-196a and HOXB8 mRNA and protein.Results After miR-196a inhibitory sequences transfection,when compared with that of siRNA control group,the expression of miR-196a was significantly decreased (0.05 ± 0.054 vs.0.839 ± 0.025,t =3.12,P ＜0.05) ; and the expression of HOXB8 mRNA was significantly increased by 1.57 folds (2.20 ± 0.07 vs.1.29 ± 0.10,t =3.86,P ＜ 0.05),the expression of HOXB8 protein was also obviously increased (0.90 ± 0.03 vs.0.40 ± 0.10,t =3.11,P ＜ 0.05).Conclusions MicroRNA-196a down-regulates the expression of HOXB8.
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Objective To investigate spliceosome of suppresser of fused(SUFU),a major member of hedgehog signaling pathway in human pancreatic cancer.Methods SUFU fragment was amplified by using reverse transcription and 3' RACE.After sequencing,a new exon was discovered,and then nSUFU was amplified by RT-PCR.nSUFU and SUFU were transfected into SW1990 by liposomes,and then the expressions of SUFU protein encoded by new spliceosome in SW1990 cells and pancreatic cancer tissues were detected by Western blot.Results PCR products by 3'RACE were of 600 bp,after sequencing and comparison with Blast data of NCBI,it was detected that a new exon was inserted between SUFU mRNA isoforml (NM_016169.3) exon 10 and exon 11.After verification with SW1990,it was noted the entire new spliceosome containing new exon was of 1400 bp.SW1990 with nSUFU transfection strongly expressed nSUFU protein,and pancreatic cancer tissues expressed both SUFU and nSUFU protein.Conclusions A new spliceosome of SUFU,which can encode SUFU protein,is present in pancreatic cancer tissue and cell.
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Objective To compare the efficacy and safety of E1B gene-deleted adenovirus (H101)and ethanol in treating advanced pancreatic carcinomas by intratumoral injection combined with intravenous gemcitabine.Methods We constructed an orthotopic nude mouse model of pancreatic carcinoma through cancer cell injection into pancreas.A total of 54 nude mice were randomly allocated to 6 groups to accept H101,ethanol or saline (control) intratumoral injection,combined with or without intravenous gemcitabiein.The animals were sacrificed 4 weeks after the treatment and the pancreatic tumors were collected to determine the size,existence of metastasis,distribution of virus by indirect immunofluorescence and apoptosis in tumor by TUNEL and electron microscope.Results All mice completed the scheduled treatment,while 3 died in 48 hours after ethanol injection resulting in a mortality of 16.7％ (3/18).On the contrary,no mice died in the adenovirus injcction group.The average tumor size in group of H101 intratumoral injection combined with intravenous gemcitabie was significant smaller than that in group of saline injection with or without systemic gemcitabie (P =0.008,0.040,respectively).Similar differences were observed between ethanol intratumoral injection and control groups (P =0.012,0.041).Meanwhile,the H101 was absent in all the other organs except the pancreas,which meant that the selectivity of the H101 was tremcndous.The virus combine gemcitabie group had higher apoptosis rate in tumor (83.2 ± 35.7) ％,determined by TUNEL.Conclusion E1B gene-deleted adenovirus intratumral injection in combination with intravenous gemcitabine treating pancreatic carcinomas is efficient and safe,in spite of its lower effectiveness than ethanol.
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Objective To construct RNAi eukaryotic expressing vectors of human transcription factor glioma-associated oncogene homolog 1 (GLI1) with pGCsi-U6-GFP plasmid and to identify its activity in interfering GLI1.Methods Three GLI1siRNA targeting GLI1 were designed and synthesized according to the GLI1cDNA sequence in GeneBank,and then were cloned into pGCsi-U6-GFP to construct the recombinant plasmids,and transformed into E.coli DH5a,then it was amplified and plasmids were extracted,which were further confirmed by PCR reaction and DNA sequencing,pGCsi-U6-siRNA-C was negative as control wector.Then recombinant plasmids pGCs-U6-GLI1siRNA-1,pGCs-U6-GLI1 siRNA-2,pGCs-U6-GLI1siRNA-3 pGCsi-U6-siRNA-C and a eukaryotic over-expression vector pEGFP-N1-GLI1 were co-transfected into HEK293 cells by Lipofectamine 2000 respectively.The ceils were collected at 48 h after transfection.Semi-quantitative RTPCR and Western Blot were performed to detect the expression of GLI1 mRNA and protein to screen the optimal vector which had the best interfering effect.Results A 369 bp fragment was amplified from all three recombinant plasmids,(pGCs-U6-GLI1 siRNA-1,pGCs-U6-GLI1 siRNA-2,pGCs-U6-GLIlsiRNA-3),showing that synthesized shRNA oligonucleotide fragments were correctly inserted into three recombinant plasmids,which were further confirmed by sequencing.Expression levels of GLIlmRNA and protein in cells in pGCs-U6-GLI1 siRNA-1,pGCs-U6-GLI1 siRNA-2,pGCs-U6-GLI1 siRNA-3 were 0.290 ± 0.011,0.421 ± 0.018,0.373 ±0.018,and 0.318 ± 0.026,0.443 ± 0.021,0.381 ± 0.018,which were significantly lower than those in negative control group (0.834 ± 0.022,0.818 ± 0.024,P =0.000),the inhibitory rates were 65.8 ％,50.7％,55.7％,and 63.9％,48.3％,53.9％.The interfering efficacy of pGCs-U6-GLIlsiRNA-1 was the strongest among the three recombinant plasmids.Conclusions RNAi eukaryotic vectors pGCs-U6-GLIlsiRNA-1,pGCs-U6-GLI1 siRNA-2,pGCs-U6-GLI1 siRNA-3 are successfully constructed and the optimal vector is identified,and this can provide a solid experimental foundation for further functional study of GLI1 gene.
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Objective To study the efficacy of intra-tumoral injection of different concentrations of ethanol for nude mice with implanted pancreatic cancer and provide evidence for choosing appropriate concentration of ethanol for clinical treatment of pancreatic cancer.Methods A subcutaneous xenograft mouse model of human pancreatic cancer SW1990 was established.Forty-eight nude mice with similar tumor size were randomly divided into 20％,40％,60％,80％,95％ ethanol injection groups and saline injection group.The longest (a) and the shortest diameters (b) of tumor of nude mice were measured.Tumor volume (TV),relative tumor volume (RTV) and the relative rate of tumor proliferation (T/C％) were calculated.Eight days later the nude mice were sacrificed.The tumor tissue was harvested for pathologic examinations.Results RTV in 20％ ethanol injection group was similar that of saline injection group (P =0.212).RTV in 40％,60％,80％ and 95％ ethanol injection groups were significantly lower than that in saline injection group (P ＜ 0.01).RTV was less than 1 and T/C％ was less than 30％ in 60％,80％ and 95％ ethanol injection groups.The values of RTV and T/C％ decreased with the increase of ethanol concentration.RTV in 80％ and 95％ ethanol injection groups were significantly lower than that of 60％ ethanol injection group (P =0.003 and P =0.009).RTV was similar in 80％ and 95％ ethanol injection groups (P =0.819).The pathologic examinations showed no tumor necrosis in saline injection group,while small amounts of necrosis in implanted pancreatic cancer was observed in 20％ and 40％ ethanol injection groups,while a large area of coagulation necrosis could be found in 60％,80％ and 95％ ethanol injection groups.Conclusions Intra-tumoral injection of 80％ ethanol is feasible therapy method for nude mice with human pancreatic cancer xenografts.
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ObjectiveTo assess the effects of DNA methyhransferase 1 ( DNMT1 ) gene silencing on DNMTs activity and methylated CpG sites of hMLH-1 in pancreatic cancer cell line PaTu8988.Methods DNMT1 siRNA and negative control siRNA was constructed by Ambion Company of United States.Then they were transfected into pancreatic cancer cell line PaTu8988 at the concentrations of 15,30 nmol/L,and the cells without transfection was used as the control group.Real-time PCR and Western blotting were applied to detect the DNMT1 mRNA and protein expression,and DNMTs activity was detected by using DNMTs activity assay kit.Change of methylation of CpG island of hMLH-1 was detected by bisulfite sequencing PCR (BSP).The expression of hMLH-1 mRNA was detected by Real-time PCR.ResultsAt 48 h after transfection,Realtime RT-PCR analysis showed that the levels of DNMT1 mRNA in DNMT1 siRNA group ( 15 nmol/L) and DNMT 1 siRNA group (30 nmol/L) were 0.573 ± 0.026 and 0.143 ± 0.044,which were significantly lower than those in control group 1.020 ±0.217 and negative siRNA 15 nmol/L group 0.900 ±0.475,and negative siRNA 30 nmol/L group 0.938 ± 0.327 (P ＜0.05 ).Western blotting analysis showed that the level of DNMT1 protein of DNMT1 siRNA group was also lower than those of negative siRNA and control groups.DNMT activity in DNMT1 siRNA15,30 nmol/L groups was 0.364 ± 0.124and 0.250 ± 0.072,which were significantly lower than those in control group 0.931 ± 0.065and negative siRNA group 0.665 ± 0.055 and 0.472 ± 0.040.DNMT activity was positively correlated with DNMT1 mRNA expression ( r =0.69,P ＜ 0.01 ).DNMT1 RNA interference decreased 8 methylated CpG sites of hMLH-1 to 1 site.Concluslons DNMT1siRNA can specifically inhibit the expression of DNMT1 gene of PaTu8988 and DNMT activity,and can decrease methylated CpG sites of hMLH-1 gene.
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ObjectiveTo investigate the protective effects of p38MAPK inhibitor SB203580 on acute necrotizing pancreatitis associated lung injuries in rats.Methods Fifty-four SD male rates were randomly divided into 3 groups,including control group,ANP group,SB203580 group with 18 rats in each group.ANP was induced by intraperitoneal injection of L-arginine solution. Rats in control group were intraperitoneally injected with same amount of saline.Before ANP induction,the rats in SB203580 group received 10 μmol/L SB203580 dissolved by dimethyl sulfoxide at a dose of 5mg/kg weight via intraperitoneal injection.The rats were sacrificed at 3,6,and 12 h after operation,the serum levels of amylase,TNF-α,IL-6 was determined.Pathological changes of pancreas and lung were observed.The wet/dry (W/D) weight ratio of lung and MPO were measured.CINC mRNA of lungs was determined by RT PCR. Expression of phosphated-p38MAPK (p-p38MAPK) protein was evaluated by Western blotting.ResultsThe serum levels of amylase,TNF-α,IL-6and wet/dry (W/D) weight ratio of lung,MPO activity,CINC mRNA and p-p38MAPK protein expression of lungs were (1035 ±73)U/L,(0.94 ±0.16)μg/L,(4.77 ±0.86) μg/L,3.92 ±0.29,(0.39 ±0.02)U/g,0.28 ±0.04,0.09 ±0.04 in control group at 6 h after operation,and the corresponding values were (5848 ±656) U/L,(3.84 ±0.32)μg/L,(103.54 ± 15.32)μg/L,4.97 ±0.47,(1.03 ±0.08) U/g,0.62 ±0.06,0.52 ±0.14 in ANP group,while they were (4259 ±286) U/L,( 1.64 ±0.21 ) μg/L,(76.56 ± 11.46) μg/L,4.32 ±0.34,(0.78 ±0.05)U/g,0.37 ±0.04,0.27 ±0.08 in SB203580 group.The values in ANP group were significantly higher than those in control group,and the values in SB203580 group were significantly lower than those in ANP group,but they were still significantly higher than those in control group ( P ＜ 0.01 ).ConclusionsSB203580 may attenuate injury of lung and pancreas in ANP by blocking p38MAPK signal transduction pathway,and decreasing the production of inflammatory cytokines.
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Objective To investigate Toll-like receptor-4 (TLR4) protein expression in human pancreatic adenocarcinoma,and to evaluate the relationship between TLR4 protein expression and angiogenesis.Methods Sixty-two surgically resected human pancreatic adenocarcinoma specimens and 35 normal para-cancerous tissues were investigated for TLR4 protein expression by immunohistochemical SP methods,and CD31 antibody was used to mark microvascular endothelial cells and determine the microvessel density (MVD).The correlation among TLR4 protein expression and MVD and clinicopathologic features of pancreatic adenocarcinoma were analyzed.Results TLR4 protein positive expression rate and MVD in human pancreatic adenocarcinoma was 74.2％ (46/62) and 47.3 ± 13.5,respectively,which were significantly higher than those in the normal pancreatic tissue [17.1％ (6/35),12.6 ±4.8; P ＜0.01].TLR4 protein positive expression rate in the cases with lymph node metastasis was 83.8％,which was significantly higher than that in the cases without lymph node metastasis (60.0％,P =0.036).TLR4 protein positive expression rate in the patients with stage Ⅲ and Ⅳ of TNM classification was 85.3％,which was significantly higher than that in the patients with stage Ⅰ and Ⅱ (60.7％,P=0.028).MVD was closely related to tumor size,lymph node metastasis and TNM stage of pancreatic adenocarcinoma (P =0.008,0.036,0.010).There was a strong positive correlation between TLR4 protein expression and MVD (r =0.534,P ＜0.01 ).Conclusions TLR4 protein expression is closely related to the development and progression of human pancreatic adenocarcinoma and its potential mechanism is related to the promotion of tumor angiogenesis.
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Objective To investigate the effect of mild hyperthermia on chemoresistance of gemcitabine in the pancreatic cancer cell line PANC1.Methods The PANC1 cell was treated with gemcitabine ( 1 μmol/L) for 1 h,then was heated at 37℃,42℃ and 45℃ for 1 hour,and was cultured for 0 h,24 h,48 h.Cell growth was analyzed by CCK-8 assay.Cell apoptosis was analyzed by Annexin Vfluorescein isothiocyanate ( FITC)/propidium iodide (PI) staining.Results The proliferation of cells at 42℃ and 45℃ for 24 h and 48 h were significantly lower than that at 37℃ (0.96 ± 0.05,0.88 ± 0.03vs 1.05 ±0.02;1.28 ±0.04,0.94 ±0.04vs 1.49 ±0.09;t =4.367,25.120,P ＜0.05).The proliferation of cells at 45℃ was significantly lower than that at 42℃ (t =3.348,11.732,P ＜0.05).The cell apoptosis rates at 37℃,42℃,45℃ after 48 h were (7.125 ±0.064)％,(9.985 ±0.615)％,(14.845 ± 1.987)％,the difference among the 3 groups was statistically significant (t =10.320,9.832,4.575,P ＜0.05).Conclusions Mild hyperthermia can reduce chemoresistance of gencitabine in pancreatic cancer cell PANC1.
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ObjectiveTo detect the expressions of β-catenin protein in different pancreatic tissues ( NP,PanINs,IPMNs and PDAC) and evaluate its significance during the carcinogenesis of PDAC.Methods The expression of β-catenin protein in 12 samples of normal pancreatic tissues,52 samples of IPMN (IPMA 20 foci,IPMB 13 foci,IPMC 19 foci),PanINs 118 foci (PanIN-1 73 foci,PanIN-2 29 foci,PanIN-3 16 foci),50 cases of PDAC was determined by using immunohistochemistry. The correlation between β-catenin expression and clinicopathologic characteristics of PDAC was analyzed. Results β-catenin was mainly expressed in cell membrane of NP,the quantity was 4.38 ± 2.11 ; in PanINs,PDAC and IPMN,β-catenin membrane expression was significantly decreased or absent,the β-catenin membrane expressions of PanIN-1,PanIN-2,PanIN-3,PDAC,IPMA,IPMB,IPMC were 5.22 ±2.21,2.24 ±2.31,1.44 ±1.37,2.71 ±2.08,4.85 ±2.28,4.15 ±2.51,2.68 ±2.75.The cytoplasm expression of PanIN-1 was 12.3％ (9/73),while the nuclear expression was 0 ; and the corresponding values were 34.5％ (10/29) and 3.4％ ( 1/29 ) in PanIN-2; 43.8％ (7/16) and 12.5％ (2/16) in PanIN-3; 44.0％ (22/50) and 10.0％ (5/50) in PDAC.The IHCS of β-catenin membrane expression decreased with the severe tissue atypia along the progressive multistage.The β-catenin membrane expression was significantly associated with tumor size,neural infiltration and lymphatic metastasis ( P ＜ 0.05 ).Ectopic cytoplasm expression was significantly associated with tumor size (P＜0.05 ). Ectopicnuclear expression was significantly associated with tumor differentiation (P＜0.01).The membrane or ectopic cytoplasm expression of β-catenin was significantly associated with postoperative survival.ConclusionsAbnormal Wnt/β-catenin signal activation induces decreased β-catenin membrane expression and increased ectopic expression,which is an important mechanism of pathogenesis and development of PDAC,and promotes the growth and metastasis of PDAC.The expression of β-catenin was associated with postoperative survival.
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Objective To investigate the expression of DNMT3b and its correlation with clinicopathological parameters of pancreatic cancer.Methods The expressions of DNMT3b protein in 12 pancreatic cancer tissues and para-cancerous tissues were detected by Western blot.The expressions of DNMT3b protein in 59 pancreatic cancer tissues were detected by immunohistochemistry.The association of DNMT3b expression and clinicopathological parameters of pancreatic carcinoma were analyzed.Results Western blot results showed the expressions of DNMT3b protein in pancreatic cancer tissues and para-cancerous tissues were 0.69 ±0.13and 0.14 ±0.03,and the protein level of DNMT3b in pancreatic cancer tissues were significantly higher than that in para-cancerous tissues (t =4.464,P ＜0.05 ).Immunohistochemistry examination showed that the positive rates of DNMT3b protein were 59％ in pancreatic cancer tissues and negative in para-cancerous tissues.DNMT3b expression was positively correlated with the TNM stage and lymph node metastasis.Conclusions DNMT3b is highly expressed in pancreatic cancer tissues,and it is related with malignant biological behaviors of pancreatic cancer cells.
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ObjectiveTo study the role and mechanism of resisitin in prophylactic and therapeutic treatments of rosiglitazone,a specific peroxisome proliferator-activated receptor-γ(PPARγ) ligand,in rats with severe acute pancreatitis (SAP) and pancreatitis-associated pulmonary injury.MethodsThe levels of amylase ( AMY ),Resistin,TNF-α,IL-1 β and C reactive protein (CRP) in blood plasma,lung myeloperoxidase ( MPO ) activity,pancreas/body weight ratio and lung wet/dry weight ratio were evaluated.Pancreatic and pulmonary pathology were observed.The expression of resistin in pancreas was detected byimmunohistochemistry.The gene expression of resistin mRNA was investigated by real-time PCR.ResultsBoth prophylactic and therapeutic treatments with rosiglitazone could obviously ameliorate the levels of AMY,resistin,TNF-αt,IL-1β and CRP ( all P ＜ 0.01 ).Compared with the control group,both prophylactic and therapeutic treatment groups were higher( all P ＜ 0.01 ).The prophylactic treatment group was not different from the therapeutic treatment group.Both prophylactic and therapeutic treatments with rosiglitazone could significantly reduce pancreas/body weight ratio,pancreatic pathology,MPO,pulmonary pathology ( all P ＜ 0.01 ).Compared with the SAP group,the expression of resistin mRNA in the prophylactic and therapeutic treatment groups were obviously decreased.ConclusionRosiglitazone could obviously ameliorate pancreatitis and pulmonary injury induced by L-arginine.
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Objective To evaluate the feasibility and efficacy of betadine solution irrigation of gastrointestinal tract for infection prevention during the procedure of natural orifice transluminal endoscopic surgery(NOTES).Methods Twelve female porcine were divided into control group(n =4)to receive lavage with 500 ml normal saline and experimental group(n =8)to undergo lavage with 500 ml normal saline followed by 200 ml betadine solution.Fluid from gastrointestinal tract(5 ml)were collected before and after lavage,and after NOTES for culture.Endoscopy was performed 24 hours after NOTES to observe possible existence of inflammation,ulcer or bleeding.The animals were sacrificed 3 weeks after NOTES to explore intra-peritoneal adhesions,abscesses and other infections.Results One swine died of diaphragmatic injury and the other 11 animals successfully survived for 3 weeks.In trans-gastric approach,the average bacterial load of the fluid was 17.5 x 103 CFU/ml before lavage.In control group,the average bacterial load of the fluid was 2.5 × 103 CFU/ml after lavage and 5.5 × 103 CFU/ml after NOTES,while those in experimental group were 0 CFU/ml and 7.5 CFU/ml,respectively.In trans-colonic approach,the average bacterial load of the fluid before lavage was 76.2 × 103 CFU/ml.In control group,the average bacterial load of the fluid was 19.5 × 103 CFU/ml after lavage and 21 × 103 CFU/ml after NOTES,while those in experimental group were 2.25 × 103 CFU/ml and 1 × 103 CFU/ml,respectively.No inflammation,ulcer or bleeding were observed by endoscopy at 24 hours after NOTES.More adhesion and abscess were found in the control group than in the experimental group.In experimental group with trans-colonic approach,only one case of adhesion was observed.Conclusion It is effective and feasible of using betadine solution irrigation of gastrointestinal tract in infection prevention during the procedure of NOTES.However,further clinical studies assessing the effectiveness and safety are still necessary.
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Objective To evaluate the influence of Helicobacter pylori (Hp) eradication on clinical manifestations, endoscopic features and pathological findings of chronic gastritis. Methods This was a multiple-center, prospective and randomized cohort study. Patients with non-atrophy chronic gastritis from January 2009 to December 2010 were randomized into 3 groups as Hp positive group with eradication, Hp positive group without eradication and Hp negative group. Clinical manifestations, endoscopic findings and pathologic changes of inflammation were compared before and after administration of gastric mucosal protective agent for 8 weeks. Results A total of 211 patients were recruited. Changes of symptom score, endoscopic erosion and mucosal inflammation were significantly different before and after treatment in 3 groups. The decrease in symptom scores of eradication group was ( 3.56 ± 1.37 ), which was significantly higher than that of non-eradication group (2. 80 ± 1.30, P ＜0. 01 ). The decrease of mucosal inflammation and inflammatory activity scores in eradicate group was 1.08 ± 1.34 and 1.42 ± 1.09, respectively, which were also significantly higher than those of the eradication group (0. 49 ± 1.47 and 0. 61 ± 1.34, P ＜0. 01 ). But the improvement of endoscopic erosion in 2 groups showed no significant difference. There were no significant differences in these variables between non-eradication group and Hp-negative group ( P ＞ 0. 05 ). Conclusion For chronic non-atrophic gastritis patients with positive Hp infection, combination of mucosal protective agents and Hp eradication can achieve better improvement in symptoms and gastric inflammation repair.
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ObjectiveTo detect the Alu expression in the stool of patients with pancreatic cancer and investigate its value in the diagnosis of pancreatic cancer.MethodsStool samples were obtained from patients with pancreatic cancer (PC) ( n =41 ),chronic pancreatitis (CP) ( n =27 ) and healthy subjects ( n =23 ),the DNA was extracted from the stool and the expression of Alu repetitive sequences was subjected to quantitative analysis by the real-time PCR.ResultsThe expressions of Alu repetitive sequences in PC,CP,and healthy subjects were (5.17 ± 0.99 ),( 3.79 ± 0.94),(0.28 ± 0.35 ) rig/g,and the difference among the three groups was statistically significant (P ＜0.05).The AUC of PC was 74.8％ with the 95％ CI 0.661 ～0.835,and the sensitivity,specificity was 75.6％ and 67.1％,respectively.ConclusionsAlu repetitive sequences are highly expressed in the stool of patients with pancreatic cancer,and it is of value in the diagnosis of pancreatic cancer.
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Objective To observe the changes of inflammation cytokines in pancreas tissues of experimental acute necrotizing pancreatitis (ANP) rats treated with rhubarb.MethodsThirty-three rats were randomly divided into sham operation group,ANP group and rhubarb treatment group with 11 rats in each group.ANP model was induced by retrograde injection of 3％ sodium taurocholate into the biliopancreatic duct,and jejunostomy was performed.Rats in rhubarb treatment group were fed with 1 g/ml rhubarb at the dose of 1 ml/kg body weight via jejunum route.Thirty-six hours later the rats were sacrificed.The serum level of amylase was measured; part of the pancreatic tissue was harvested for routine pathologic examination; total RNA was extracted from pancreatic tissue.The expression of IL-6,IL-8,and TNF-α mRNA was measured by real-time PCR.ResultsAfter ANP induction,the serum level of amylase was significantly increased,and pancreatic tissue necrosis,bleeding and inflammatory cell infiltration were observed,which was consistent with changes of ANP.The expressions of IL-6,IL-8,and TNF-α mRNA were 0.29 ±0.13,0.35 ±0.15,1.09 ±0.32 in sham operation group,while they were 2.23 ±0.49,2.26 ±0.51,5.24 ±0.59 in ANP group,which were significantly higher than those in sham operation group ( P ＜ 0.05 ).The corresponding values were 0.97 ±0.30,1.02 ±0.34,2.59 ±0.36 in rhubarb treatment group,which were significantly lower than those in ANP group,but they were still significantly higher than those in sham operation group ( P ＜ 0.05 ).ConclusionsRhubarb lavage via jejunum route can decrease the expressions of IL-6,IL-8,and TNF-α,therefore attenuate the pancreatic pathologic injuries.
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Objective To investigate the effect of α-tocopherol on fibrosis of chronic pancreatitis (CP) rat and explore its mechanism.MethodsMale Wistar rats were randomly divided into control group,acute necrotizing pancreatitis (ANP) group,α-tocopherol group.CP was induced by dibutyltindich loride ( 8 mg/kg) infusion into the tail vein.Gastric lavage of α-tocopherol (800 mg/kg body weight,daily) was started 24 hours after dibutyhindich loride infusion for 4 weeks.The rats in ANP and control group received 0.6 ml salad oil gastric lavage.The rats were sacrificed 4 weeks later.Pancreatic tissue was harvested for histological examination and collagen staining,and measurement of the levels of hydroxyproline and malondialdehyde (MDA) of the pancreas were performed.The mRNA expression of transforming growth factor (TGF)-β１ was measured by real time PCR.ResultsAfter gastric lavage for 4 weeks,the pancreatic tissue inflammation,fiber deposition and abnormal structure in rats of α-tocopherol group were greatly reduced.The levels of MDA and hydroxyproline in rats of α-tocopherol group were significantly lower than those in ANP group [ (0.40 ±0.20) vs (1.07 ±0.41) nmol/100mg,(402.49 ±27.62) vs (664.92 ±29.04) μg/g,P＜0.05].The expressions of TGF-β1 mRNA in rats of o-tocopherol group were significantly lower than those in ANP group (2.24 ± 0.89 vs 3.35 ± 0.66,P ＜ 0.05 ).Conclusions Tocopherol gamma can improve pancreatic inflammation and fibrosis by reducing the oxidative stress level and down-regulating the expression of TGFβ1mRNA in rats with CP.
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Objective To quantitatively analyze the K-ras gene mutation at codon 12 in pancreatic cancer tissues and the relationship between K-ras gene mutation and clinicopathological parameters. Methods Quantitative detection of K-ras gene at codon 12 in 93 pairs of pancreatic cancer and adjacent tissues were performed by using PNA-mediated PCR clamping with two different fluorescence labeled probes. The quantity of mutation was expressed by percentage of mutation. The percentage of K-ras gene mutation = the copy of K-ras mutation/(copy of wild type K-ras + copy of K-ras mutation) × 100%. Results The percentage of mutation of K-ras gene at codon 12 in pancreatic cancer and adjacent tissues were 83.9% and 65.6%, and the difference was statistically significant(P ＜ 0. 05); and the quantity of mutation were (13.385 ± 1. 745) % and (2. 246 ±0. 728) %, and the difference was also statistically significant(P ＜ 0. 05). The quantity of mutation of K-ras gene at codon 12 was not associated with clinicopathological parameters. Conclusions The percentage of K-ras gene mutation, as well as the quantity of K-ras gene mutation was different in pancreatic carcinoma and adjacent tissues.