Objective To study the effects of concentrated H.pylori culture supernatant(CHCS) on the expressions of COXⅠ mRNA and protein in gastric carcinoma cells,and explore the potential role of H.pylori in the gastric carcinogenesis.Methods After SGC7901 cells were treated by 6 ?l/ml CHCS for 4 h,8 h,12 h respectively,the expression of COXⅠmRNA and protein was detected by RT-PCR and Western blotting.Results The expression of COXⅠmRNA decreased gradually after exposure to CHCS for 4 h.Especially after 8 h,the expression decreased significantly.When treated with CHCS for 12 h,the expression continued to decrease(P

Objective To evaluate the role of mitochondrial pathway in the apoptosis of gastric epithelial cells induced by H. pylori in Mongolian gerbils. Methods A total of 48 Mongolian gerbils were randomly divided into H. pylori infection and without H. pylori infection groups. Eight animals from each group were killed at 1, 3 and 6 months, and histopathological changes in their stomachs were examined. A flow cytometry was used to measure apoptosis, mitochondrial membrane potential and intracellular free Ca2+ . Results The incidences of intestinal metaplasia and dysplasia in the gastric mucosa with H. pylori infection were significantly higher than those without H. pylori infection (P

Objective Using a model of H.pylori infected Mongolian gerbil , we observed the effect of H.pylori and N methyl N’ nitro N nitrosoguanidine (MNNG) on gastric mucosa, in an attempt to clarify the potential role of vitamin C in the prevention of gastric carcinoma. Methods A total of 160 Mongolian gerbils , eight week old, were randomly divided into five groups(each 32 animals): Group A, infected with H.pylori ; Group B, infected with H.pylori followed by MNNG administration; Group C, received MNNG without H.pylori infection; Group D, infected with H pylori followed by administration of MNNG and vitamin C; Group E as control. Eight animals from each group were killed at 12, 24, 36, 48 weeks, and histopathological changes in their stomachs were examined for chronic gastritis, intestinal metaplasia, atypical hyperplasia and adenoma. Results The incidences of intestinal metaplasia and dysplasia in groups A and B were significantly higher than those in the other groups( P

Objective To explore the effects of celecoxib, a selective COX-2 inhibitor, on inducing the apoptosis of gastric cancer cells, and to elucidate the concerning mechanisms. Methods Cell viability was measured by MTT assay. Flow cytometry (FCM) was employed to assay apoptosis, mitochondrial membrane potential and intracellular free Ca 2+. Results After being exposed to celecoxib (25, 50, 100 and 200?mol/L) for 4, 8, 12 and 24h, the proliferation of SGC-7901 cells was strongly inhibited in a dose-time dependent manner. The apoptosis induced by celecoxib was accompanied with the attenuation of mitochondrial membrane potential and the elevation of intracellular free Ca 2+ concentration, suggesting the importance of mitochondria in the apoptotic pathway. Conclusion The mitochondrial pathway may be involved in the apoptosis of gastric cancer SGC-7901 cells induced by celecoxib.

Objective To evaluate the role of mitochondrial pathway in apoptosis of gastric cancer cell line SGC-7901 induced by Helicobacter pylori(H.pylori).MethodsApoptosis was evaluated in SGC-7901 cells by flow cytometry.RT-PCR and Western blotting were used to measure the expressions of genes and proteins associated with mitochondrial pathway.Effect of caspase inhibitors on apoptosis induced by H.pylori strain NCTC11637 was investigated.ResultsNCTC11637 directly induced apoptosis in SGC-7901 cells.Apoptotic rate was 6.30%,11.57%,8.63% and 7.22% respectively at 6,12,24 and 48 h after coculture with H.pylori.H.pylori upregulated the expression of Bax,and induced a time-dependent activation of caspase-9 and-3.Apoptosis was inhibited significantly by pre-incubation with the inhibitors of caspase-9 and-3.Caspase-8 inhibitor reduced H.pylori-induced apoptosis by 20%.ConclusionH.pylori infection upregulates the expressions of Bax mRNA and protein in gastric cancer cell line SGC-7901,and induces activation of caspase-9 and-3.Mitochondrial pathway may be the major one in H.pylori-induced apoptosis in SGC-7901.

Background Dimethyl sulfoxide(DMSO) is a commonly used adjuvant to promote testing drug solubility to prepare multi-levels testing drug concentrations.DMSO is cell type-dependent cytotoxic and its toxicity can interfere the testing drug evaluation.Determining its safe concentration on commonly used cell types is important for ocular drug development.Objective This study was to determine the minimal toxic concentration of DMSO for in vitro ocular cell lines in a simulated drug screening setting.Methods Retinal pigment epithelial (RPE) cells were isolated from one pigmented rabbit and primarily cultured.Human RPE cell strain (ARPE19),scleral fibroblasts line (S75-Fron),human Müller cell line (MIO-M1),human lens epithelial cell line (HLEC),human choroidal melanoma cell line (OCM-1),human umbilical endothelial cell (HUVEC) and human HeLa cell line (HELA) were cultured.Different concentrations of DMSO (1.6％,1.0％,0.8％,0.4％,0.2％ and 0.1％) were prepared with 160 μl DMSO solution and 9.84 ml RPMI1640 (or DMEM/F12 or DMEM) containing 2％ fetal bovine serum.Different concentrations of DMSO were added in medium for 96 hours,and the and viability (absorbance) of the cells was detected using MTS to evaluate the cytotoxicity of DMSO.Results Rabbit primary RPE cells showed the yellow-green fluorescence for cytokeratin(CK) and HMB45 red fluorescence for S100.The viability of the cells was gradually declined as the increase of DMSO dose,showing significant differences in ARPE19,S75-Fron,HLEC,OCM-1,HUVEC and primary RPE cells (all at P＜0.05),and when DMSO concentrations were ≥ 0.8％,the cell viabilities were significantly lower.But no significant difference was found in MIO-M1 cells among different doses of DMSO (F=0.830,P=0.547).The minimal toxic concentration of DMSO for ARPE19,HUVEC,HELA,HLEC,MIO-M1,OCM-1,primary RPE cells and S75-Fron was 0.8％,0.1％,0.8％,＞1.6％,＞1.6％,0.2％,0.2％,0.2％,respectively,and HUVEC was more sensitive to the cytotoxicity of DMSO (P=0.02),and MIO-M1 was the least sensitive to DMSO (P =0.39).The viability of HUVEC and primary RPE cells went down with the increase of DMSO dose,and S75-Fron viability started to decline in 0.1％ DMSO and then stabilize with the higher concentrations until 1.6％ DMSO at which the viability showed further decline.Conclusions The tolerability of ocular cells in vitro to DMSO varies depending on the cell types.The minimal toxic concentration ranged from 0.1％ to 1.6％.The result suggests that a concurrent DMSO control should be set up along with the testing compound.

Objective Retrospectively analyzed the data of patients with delayed bleeding after colorectal polypectomy,summarized the risk factors and treatment methods of bleeding patients,and provided the basis for further prevention and treatment of postoperative delayed bleeding.Methods Collected the clinical data of 1 243 patients who were admitted into the department of gastroenterology of third affiliated hospital of the third military medical university and accepted polypectomy with colonoscopy from January 2014 to December 2016.Divided these patients into the bleeding group and the non-bleeding group according to whether there was delayed bleeding after surgery.The age,size of polypus,location of polypus,postoperative pathology of the two groups were compared and the postoperative treatment of bleeding was evaluated.Results Among the 1 243 patients underwent colorectal polypectomy,there were 14 cases of delayed bleeding,and the incidence was 1.13%.In the bleeding group,there was 1 case of secondary delayed bleeding and 2 cases of bloody stool after hemostasis for the delayed bleeding.Delayed bleeding occurred at (4.73±2.49)days after surgery.The predilection site of of polypus was rectum in the bleeding group (7/14,50%), and the diameter of polypus was (16.65±4.91)mm in the bleeding group,which was lager than (8.07±4.23)mm in the non-bleeding group with statistical difference (P<0.05).The proportion of hypertensive and diabetic patients in the bleeding group was significantly higher than that in the non-bleeding group (P<0.05).Juvenile polyps and tubular adenoma with high grade intraepithelial neoplasia were more common in the bleeding group(P<0.05).The bleeding group achieved good hemostatic effect by purse suture,hemostatic clip,electrocautery or injection hemostasis.Conclusion Older age,hypertension and diabetes,lager size of polypus,rectum polypus,juvenile polyps and tubular adenoma with high grade intraepithelial neoplasia were risk factor for delayed bleeding.In the event of delayed bleeding,different choice of purse suture,hemostatic clip,electrocautery or injection hemostasis according to different wounds can achieve the desired effect.

Objective To evaluate the guidance value of endoscopic ultrasonography (EUS) and CT scan in preoperative clinical staging for diagnosis and treatment of esophageal cancer .Methods 68 patients with esophageal cancer were randomly divided into EUS group and CT group using a random numbers table(34 cases in each group) .Patients in EUS group were examined by EUS , patients in CT group were examined by CT scan ,and staged according to the TNM (2003) staging system ,and were compared with surgical pathologic findings .Results The accuracy rates of T staging by EUS were 0(0/2) for Tis ,75 .0% (3/4) for T1 ,75 .0% (6/8) for T2 ,86 .7% (13/15) for T3 ,80 .0% (4/5) for T4 ,and the totle accuracy rate was 76 .5% (26/34) for T ;those of N staging were 71 .4% (5/7) for N0 ,75 .0% (9/12) for N1 ,0(0/11) for N2 ,0(0/4) for N3 ,and the totle accuracy rate was 41 .2% (14/34) for N .The accuracy rate of T staging by CT scan were 0(0/1) for Tis ,33 .3% (2/6) for T1 ,28 .6% (2/7) for T2 ,78 .6% (11/14) for T3 ,83 .3% (5/6) for T4 and the totle accuracy rate was 58 .8% (20/34) for T ,the difference was statistically significant com-pared with the EUS group(P<0 .05);those of N staging were 77 .8% (7/9) for N0 ,76 .9% (10/13) for N1 ,66 .7% (4/6) for N2 , 50 .0% (3/6) for N3 and the totle accuracy rate was 70 .6% (24/34) for N ,the difference was statistically significant compared with the EUS group (P<0 .05) .Conclusion The accuracy rate of EUS are higher for diagnosis in esophageal cancer and preoperative T staging .The accuracy rate of CT scan are higher for the preoperative N staging .EUS combined with CT scan has great significance for choosing ideal therapy plan for esophageal cancer ,and for estimating prognosis of esophageal cancer .

Objective To study the relationship between copeptin,ischemia modified albumin (IMA) and the degree of myocardial injury in patients with acute carbon monoxide poisoning (ACOP).Methods A total of 110 ACOP patients with different degree of poisoning were selected as poisoning group and included mild poisoning 22 cases (mild group),moderate poisoning 50 cases (moderate group),severe poisoning 38 cases (severe group),and 30 healthy subjects were selected as control group.The cardiac troponin Ⅰ (cTnⅠ),IMA,copeptin level was detected at 2 h,7 d after admission in poisoning group and at admission in control group.According to with or without complications,poisoning group was divided into complications group (26 cases) and non-complications group (84 cases).cTnⅠ,IMA,copeptin level was compared among groups.Results At 2 h after admission,IMA was decreased and copeptin was increased in mild,moderate,severe group[(62.50 ± 2.17),(59.04 ± 3.10),(56.01 ± 8.85) kU/L and (2.82 ± 0.73),(7.31 ±0.95),(13.08 ± 1.96) μg/L],there was statistical difference compared with control group [(67.23 ± 1.40) kU/L and (0.87 ±0.19) μg/L](P＜0.05),and there was significant difference between moderate group and mild group,severe group and mild group,moderate group (P ＜ 0.05);there was no statistical difference in cTnⅠ among groups.At 7 d after admission,there was no significant difference in IMA,copeptin among groups (P ＞ 0.05) ; cTnⅠ was increased in mild,moderate,severe group [(1.80 ± 0.17),(2.34 ±0.46),(2.60 ±0.54) μg/L],and there was statistical difference compared with control group [(1.27 ±0.28) μg/L] (P ＜0.05),and there was significant difference between moderate group and mild group,severe group and mild group,moderate group (P＜ 0.05).IMA,copeptin at 2 h,7 d after admission in complications group was higher than that in non-complications group [(54.62 ± 1.53) kU/L vs.(57.89 ± 4.02) kU/L,(60.65 ± 3.61) kU/L vs.(66.84 ± 1.78) kU/L and (13.88 ± 1.45) μ g/L vs.(6.99 ± 3.39) μ g/L,(6.65 ± 1.82) μ g/L vs.(2.47 ± 0.61) μ g/L](P＜ 0.05).cTnⅠ at 7 d after admission in complications group was higher than that in non-complications group [(3.10 ± 0.22) μ g/L vs.(1.87 ± 0.27) μ g/L] (P ＜ 0.05).Correlation analysis showed that copeptin was negatively correlated with IMA in patients with different degree of poisoning (r =-0.560,P ＜ 0.01).Conclusions The combined detection of IMA and copeptin has important clinical value to the early diagnosis and prognosis in evaluating the prognosis of ACOP myocardial injury.There is important guidance for early clinical drug application.

Objective To isolate and purify VacA protein secreted by Helicobacter pylori or recombinant VacA , and to investigate the effect of VacA-induced cell vacuolar change and apoptosis .Methods VacA proteins were separated and pu-rified from the culture supernatant of H.pylori ( ATCC26695 ) or from the split products of genetically engineered bacteria (pQE30-VacA-E.coli M15) expressing recombinant VacA.The VacA protein obtained was acidified and then incubated with AGS cells for 24 h at different final concentrations of 5 and 10 ng/ml before the vacuolar change and apoptosis of AGS cells were detected via microscopy and flow cytometry assay , respectively .Results H.pylori-secreted VacA and recombi-nant VacA were successfully separated and purified .The H.pylori-secreted VacA significantly induced the vacuolar change and apoptosis of AGS cells (P<0.01) while the recombinant VacA did not.Conclusion H.pylori-secreted VacA protein can effectively induce cell vacuolar change and apoptosis, but recombinant VacA can not, suggesting that the purified VacA protein secreted by H.pylori can be used to explore VacA-induced pathogenesis.