Objective To study the effect of Bifidobacterium spent culture supernatant (SCS) on adhesion of Pseudomonas aeruginosa to intestinal epithelial cells. Methods The intestinal epithelial cells were cultured,then divided into three groups: control,Pseudomonas aeruginosa adhesion group,SCS and Pseudomonas aeruginosa adhesion group. The effects of SCS on cell viability,the number of adhering and invasive bacteria were evaluated with MTT assay and lysis-counting assay in 1,3,6 h. Results At 3 h after SCS treatment,the amount of Pseudomonas aeruginosa decreased significantly,and the number of adhering and invasive bacteria decreased by 71% and 87% respectively. Conclusion SCS could protect the intestinal epithelial cells by inhibiting the adhesion and invasion of Pseudomonas aeruginosa.

Objective: To observe the clinical effect of midnight-noon ebb-flow day-prescription of acupoints on biliary colic. Methods: 60 cases with biliary colic were randomized into two groups, 30 cases in each group. Day-prescription of five transport points was employed in the treatment group, while point selection based on pattern identification was employed in the control group. Results: The pain-killing rates between the two groups had no statistical differences (P>0.05). Conclusion: Day-prescription of acupoints can obtain similar pain-killing effect with point selection based on pattern identification, but with less points and treatments.

Objective To explore the mechanism of intestinal epithelial cell membrane injury due to Pseudomonas aeruginosa adhesion. Methods The intestinal epithelial cells were cultured and adhered with Pseudomonas aeruginosa . The changes in the viability of the cells and the activity of membranous PLA 2, the calcium content of cell, contents of phospholipid and membrane fluidity were observed. Results At 3rd h after the adhesion of Pseudomonas aeruginosa , the viability of the intestinal epithelial cells decreased significantly, but the activity of membranous PLA 2, the calcium content of cell increased significantly; the contents of phospholipid(PL), phosphatidylinositol(PI) and phosphatidylcholine(PC) in cell membrane decreased gradually, but the membrane fluorescence polarization and microviscosity of intestinal epithelial cell membrane increased significantly. Conclusion The activation of PLA 2 and the degradation of phospholipid due to the overloading of calcium in intestinal epithelial cells after the adhesion of Pseudomonas aeruginosa to intestinal epithelial cells might be the fundamental factors to result in the reduction of membrane fluidity.

Background Posterior capsular opacification (PCO) is a primary cause of blurred vision after extra-capsular cataract extraction (ECCE),and there is a higher incidence of PCO in the patients with diabetes mellitus.Echistatin (Ecs) can suppress the proliferation of lens epithelial cells (LECs) and thereby inhibit the formation of PCO.However,its mechanism and safe dose deserve to study.Objective The aim of this study was to observe the inhibitory effect of different concentrations of Ecs on LECs proliferation in the early stage of PCO in diabetic rabbits and explore the safe dose of Ecs.Methods Diabetic mellitus was induced by injection of 90 mg/kg alloxan via ear vein in 15 New Zealand white rabbis.ECCE was performed in the right eyes of the rabbits.The rabbits were randomized to the control group and 5.0,7.5,10.0 and 15.0 μg/ml Ecs group according to randomized number table method.Ecs of 0.2 ml in above doses was injected into the anterior chamber after ECCE in different concentrations of Ecs groups,and 0.2 ml distilled water was used in the same way in the only diabetic rabbits as the control group.Postopeartive response of ocular anterior segment was observed and PCO was graded under the slit lamp microscope.The corneal and retinal specimens were prepared 10 days after operation for the assay of preliferative cell nuclear antigen (PCNA) expression in LECs by immunochemistry to evaluate the effective dose of Ecs.Regular histopathological examination was performed,and apoptosis of corneal endothelial cells and retinal cells was detected by TUNEL method to assess the safe concentration of Ecs.Results Different degrees of corneal edema and exudation in anterior chamber were seen in the eyes of different groups.The inflammatory response disappeared 3-5 days in the control group and 5.0 μg/ml Ecs group and 7 days in the ≥7.5 μg/ml Ecs groups.PCO was 1-2 grade in the control group and 5.0 μg/ml Ecs group and 0 grade in the ≥ 7.5 μg/ml Ecs groups.The difference in the positive expression level (absorbance,A) for PCNA in LECs was significantly different among the control group and various Ecs groups (F=18.006,P=0.001),and the positive expression level of PCNA in the ≥ 7.5 μg/ml Ecs groups was markedly reduced in comparison with that in the control group (P =0.010,0.001).Hematoxylin and eosin staining showed an normal morphology and order arrangement in corneal endothelial cells and intact structure in retinal internal limiting membrane in the groups.TUNEL assay revealed that the apoptosis values (mean A value) of corneal endothelial ceils and retinal cells in the ≤ 10.0 μg/ml Ecs groups were not significantly changed in comparison with the control group (all at P＞0.05),but the apoptosis values in the 15.0 μg/ml Ecs group were markedly higher than those in the control group (P=0.004,0.018).Conclusions Ecs can inhibit the early PCO in diabetic rabbits and show the optimal effect at the concentrations of 7.5 and 10.0 μg/ml without visible eytotoxicity to eye other tissues.Therefore,these two doses of Ees might be used for the study of long-term therapeutic effectiveness.

Objective To study PPARγ inhibitor(GW9662) ,on colon cancer SW480 cell proliferation and apoptosis intervened by Nimesulide(N) in vitro ,in order to investigate the role of PPARγpathway in colon cancer cell proliferation inhibition and apop‐tosis promotion induced by Nimesulide .Methods Cells were divided into 4 groups ,namely :the control group ,GW9662 group (GW9662 0 .1 ,0 .5 ,1 .0 ,5 .0μmol/L) ,N group ,GW9662+N group .MTT assay and FCM were used to determine proliferation ,ap‐optosis and cell cycle of SW480 cells .And the expression of PPARγ,p21Waf1 ,p27Kip1 ,Bcl‐2 ,Bax ,VEGF proteins were measured by Western‐blot .Results N inhibited SW480 cells proliferation in a time‐dependent manner (P< 0 .01) .During a special range , GW9662 attenuated effect of nimesulide inhibiting SW480 cells proliferation in a dose‐and time‐dependent manner .The results of FCM showed :the apoptosis rates of SW480 cells had no statistical change between GW9662 group and control group(P>0 .05) . Cell apoptosis rate of group N increased significantly ,compared with control group(P<0 .01) .The apoptosis rates of SW480 cells incubated with Nimesulide and GW9662 dropped significantly compared with Nimesulide alone (P<0 .01) .Above results showed that GW9662 could attenuate the effect of nimesulide on cell apoptosis and cell cycle .The results of Western‐blot :Compared with the control group ,the expression of PPARγ,p21Waf1 ,p27Kip1 ,Bax protein were up‐regulated significantly in nimesulide group(P<0 .05 or P<0 .01) ,but Bcl‐2 and VEGF were down‐regulated significantly(P<0 .01) .Compared with the nimesulide group ,the expres‐sion of PPARγ,p21Waf1 , p27Kip1 and Bax protein were down‐regulated obviously in GW9662+N group(P<0 .05 or P<0 .01) .Corre‐spondingly ,Bcl‐2 and VEGF were up‐regulated obviously(P<0 .05) .Conclusion N could effectively inhibit SW480 cell prolifera‐tion and induce its apoptosis .PPARγpathway may play an important role in proliferation inhibition and apoptosis induced by Nime‐sulide in colon cancer cell .

ObjectiveTo review the clinical characteristics of patients with gastric mucosa-associated lymphoid tissue(MALT) lymphoma and to investigate the effects of Helicobacter pylori (Hp) eradication therapy in these patients.MethodsClinical data of ninety-five patients with gastic MALT lymphoma were reviewed with respect to their clinical manifestations,endoscopic features,histopathological features and Hp infection.The follow-up data of patients treated by Hp eradication therapy were analyzed.The survival curve was calculated with Kaplan-Meier,while the predictive factors for resistance to Hp eradiation were then analyzed by using Cox proportional hazards model.ResultsThe clinical manifestations and endoscopic features of 85 patients with gastric MALT lymphoma were non-specific.Hp positive was found in 97.9％ (93/95) patients at presentation,and 36 patients were treated by Hp eradication with a median follow-up duration of ( 58.1 ± 29.9) months.Total remission was achieved in 94.4％ ( 34/36),including 24 complete remission 10 partial remission.Therapy failure occurred in 2 patients.3-year survival rate of patients treated by Hp eradication were 86.2％ (25/29). Cox multivariate analysis showed that age ( ≥60 years),multiple lesions and non-superficial lesions were independent predictors of resistance to Hp eradication therapy.ConclusionGastric MALT lymphoma is associated with Hp infection.Eradication of Hp can effectively induce remission in these patients.

Objective To assess the relationship between serum vitamin D and autoimmune thyroid disease.Methods Subjects included total 520 persons receiving regular health examination,and serum calcium,phosphorus,parathyroid hormone (PTH),thyroid peroxidase autoantibody (TPOAb) and 25-dihydroxy vitamin Ds was measured.The incidence of 25-dihydroxy vitamin D3 deficiency (≤30 μ g/L)was observed.The relationship between 25-dihydroxy vitamin D3 deficiency and autoimmune thyroid disease was analyzed.Results The serum 25-dihydroxy vitamin D3 of all the subjects was (24.47 ± 7.21) μ g/L,and the incidence of 25-dihydroxy vitamin D3 deficiency (≤30 μg/L) was 61.15％ (318/520),and the positive rate of TPOAb was 21.54％ (112/520).The proportion of TPOAb ＞ 50 kU/L or ＞ 100 kU/L in subjects with 25-dihydroxy vitamin D3≤30 μ g/L was higher than that in subjects with 25-dihydroxy vitamin D3 ＞ 30 μg/L [25.79％(82/318) vs.19.80％(40/202) and 9.43％(30/318) vs.4.46％(9/202)],and there was significant difference (P ＜ 0.05).The relationship between 25-dihydroxy vitamin D3 and TPOAb was assessed and showed significant inverse correlation (r =-0.13,P ＜0.05).Conclusions Vitamin D deficiency is very common in the population,and autoimmune thyroid disease is related with vitamin D deficiency,which may has impact on the body's immune regulation.Specific mechanism and whether vitamin D supplementation can intervene and treat autoimmune thyroid disease need further study.

Objective To assess the role of the different pancreatic islet β cell function index in the evaluation of glucose metabolism in different duration of type 2 diabetes mellitus (T2DM).Methods Normal glucose tolerance subjects without diabetes family history (NC group,48 cases) and T2DM patients (182 cases) were enrolled.The T2DM patients were divided into three groups:less than 5 years group (DM ＜5 group,74 cases),5-10 years group (DM5-10 group,51 cases) and more than 10 years group ( DM ＞10 group,57 cases).Oral glucose tolerance test (OGTT) and insulin release test were taken in all groups.Insulin resistance index (HOMA-IR) and whole body insulin sensitivity index [ISI(Matsuda)] were used to estimate insulin sensitivity,and early insulin secretion index ( △ I30/ △ G30) and glucose disposition index (DI) were used to evaluate the function of pancreatic islet β cell.Results HOMA-IR was increased and ISI (Matsuda) was decreased in DM ＜5 group,DM5-10 group and DM ＞10 group compared with those in NC group [HOMA-IR:8.78 ± 7.12,8.08 ± 3.67,7.84 ± 5.08 vs.4.76 ± 3.43;ISI(Matsuda):46.78 ± 29.00,36.71 ± 16.67,38.86 ±21.72 vs.61.13 ± 32.08,P ＜ 0.05],however,there was no significant difference among DM ＜5 group,DM5-10 group and DM ＞10 group.△ I30/ △ G30 and DI were decreased in DM ＜5 group,DM5-10 group and DM ＞10 group compared with those in NC group [ △ I30 △ G30:( 68.41 ± 361.52 ),(4.31 ± 3.42 ),(7.70 ± 5.78 ) mU/mmol vs.(92.65 ± 309.29) mU/mmol;DI:0.0421 ± 0.0123,0.0412 ± 0.0123,0.0363 ± 0.0116 vs.0.1151 ± 0.0236,P ＜ 0.05 ],and there was no significant difference in △ I30 / △ G30 among DM ＜5 group,DM5-10 group and DM ＞10 group,however,DI was decreased in DM＞10 group compared with that in DM＜5 group and DM5-10 group (P＜0.05).ConclusionsHOMA-IR,ISI (Matsuda),△I30/△G30 are not sensitive to evaluate the insulin resistance of different duration.DI can reflect the glucose utilization of pancreatic islet β cell earlier and the ability to regulate blood sugar steady state changes.

Objective To investigate the mechanism of mastoparan-1 (MP-1) antagonizing lipopolysaecharide (LPS) in vitro. Methods The affinity of MP-1 for lipid A was assayed by biosensor, and the neutralization of MP-1 on LPS (2 μg/L) was detected by kinetic turbidimetric limulus test. After exposing fluorescin isothiecyanate (FITC) labeled LPS (FITC-LPS) to MP-1 at different concentrations (5, 10, 20, 40 μmol/L), the binding of FITC-LPS to murine RAW264.7 cells was analyzed by laser scanning confocal microscopy. The influence of MP-1 on TLR4 expression in RAW264.7 cells stimulated by LPS (100 μg/L) was detected by immunoeytochemieal staining. The expressions of TLR4, TNF-α and IL-6 at the gene and protein level were detected by RT-PCR and ELISA after exposing LPS (100 μg/ L) stimulated RAW264.7 cells to MP-1 at different concentrations. The effect of MP-1 on the viability of RAW264.7 cells was detected by MTT assay. Results MP-1 had high affinity to lipid A and could neutralize LPS. MP-1 at 10 μmol/L significantly inhibited not only binding of FITC-LPS to RAW264.7 (P < 0.05), but also protein and gene expressions of TLR4, TNF-α and IL-6 in LPS stimulated RAW264.7 cells in a dose-dependent manner (P < 0.05). No toxic effect of MP-1 on the viability of RAW264.7 cells was found (P > 0.05). Conclusions MP-1 inhibits cell viability mediated by LPS, which may be related to its neutralization of LPS and inhibition of binding of LPS to RAW264.7 cell membrane receptors.

Objective To study the changes and the role of MCP -1,IL -8,VEGF,NO,NOS in the T2DM patients with different types of CHD.Methods According to the result of coronary arteriongraphy and clinical symp-toms,and the diagnostic code of T2DM established by Chinese Medical Association diabetology branch in 2007, 126 patients of T2DMwith CHD were chosen and divided into two groups:ACS +T2DM group (A group,74 cases) and SAP +T2DMgroup (B group,52 cases),in addition,50 healthy people were chosen as control group.The levels of MCP -1,IL -8,VEGF were measured by the method of ELISA.The level of NO was measured by the method of nitrate reductase and NOS activity was measured by the method of spectrophotometer.Then,the results were analyzed. Results The levels of MCP -1 and IL -8 in A group and B group were[(115.98 ±39.57)pg/mL,(98.76 ± 31.55)pg/mL],[(131.22 ±42.83)pg/mL,(115.75 ±40.37)pg/mL],which were all higher than those in group C [(75.63 ±23.69)pg/mL,(68.53 ±37.85)pg/mL,t =4.12,2.26,3.78,2.21,all P <0.05)],but the VEGF [(167.87 ±54.98)pg/mL,(128.38 ±36.99)pg/mL)],NO[(46.89 ±12.92)μmol/L,(51.66 ±12.49)μmol/L)]and NOS [(39.04 ±5.19)U /mL,(40.56 ±7.03)U /mL)]were lower than those in C group [(90.21 ± 32.06)pg/mL,(64.05 ±13.58)μmol/L,(47.82 ±5.93)U /mL;t =3.05,3.17,2.43,2.79,2.49,2.25,all P <0.05].The MCP -1,IL -8 levels in A group were higher than those in B group(t =3.13,2.89,all P <0.05),but the level of VEGF and NO were lower than that in B group(t =3.04,2.95,all P <0.05),NOS in A group was lower than that in B group,but there was no statistically significant difference between the two groups(t =1.06,P >0.05). MCP -1 was positively correlated with Il -8,VEGF (r =0.35,0.33,all P <0.01),and it had negative correlation with NO (r =-0.24,P <0.05).Conclusion Inflammatory factor and oxidative stress both participate in the T2DM with different types of CHD,it relates with the degree of CHD.