Objective To study the application of small interfering RNA silencing S100A4 protein in human gastric cancer cell BGC-823 proliferation,apoptosis and the effect of chemotherapy sensitivity.Methods Human gastric carcinoma cell line BGC-823 transfection siRNA,RT-PCR detected the changes of mRNA after transfection.Groups divided into interference group,negative control group and normal control group.MTT test determined different concentrations of oxaliplatin in gastric cancer cells and calculated IC50,then draw cell growth curve,TUNEL method to detect apoptosis,RT-PCR tested each cell mRNA changed,Western blot detected the change of the S100A4 protein.All data analysis by SPSS17.0,t test applied,RT-PCR and Western blot results analysis by SPSS17.0,comparing multiple samples by using single factor analysis of variance and LSD test.P ＜ 0.05 was statistically significant.Results RT-PCR results showed that BGC-823 cell transfection,S100A4mRNA expression quantity respectively after 48 hours:(0.674+0.011),(0.652+0.021),(0.345 + 0.040),the interference group and normal control group were statistically significant (P =0.012,P ＜ 0.05) and the negative control group with interference group differences were statistically significant (P =0.000,P ＜ 0.05),and normal control group was no statistically significant difference with the negative control group (P =0.380,P ＞ 0.380);Western blot results showed BGC-823 cell transfection S100A4 expression significantly lowered respectively after 48 hours,there were (0.654 + 0.025),(0.642 + 0.014),(0.317 ± 0.061),the interference group and normal control group was statistically significant (P =0.01,P ＜ 0.05),between negative control group and interference group were statistically significant (P =0.000,P ＜ 0.05),normal control group and the negative control group had no significant difference (P =0.341,P ＞ 0.341).After S100A4-siRNA transfection,gastric carcinoma BGC-823 cell proliferation decreased,TUNEL method showed obviously increase apoptosis,MTY showed that IC5o of oxaliplatin was 56.31 μmol/L,after transfection,IC50 was 0.654 μmol/L.Conclusions This study showed that the siRNA silence S100A4 protein inhibit gastric cancer cell proliferation,induced apoptosis and improved chemotherapy sensitivity of oxaliplatin.S100A4 might be prompt targets for the treatment of gastric carcinoma.

Objective To investigate the siRNA interference of S100A4 mRNA of human pancreatic cancer cell radiosensitivity.Method Cultured human pancreatic BxPC-3,AsPC-1 in vitro,logarithmic phase cells as the experimental object,were divided into three groups:normal control group (without any treatment),negative control group (transfected with negative control fragment),interference group (transfected S100A4 protein fragment siRNA),the chemical synthesis siRNAS100A4 fragment interference of S100A4 mRNA 0,1,2,3,5,7,10 Gy given 6MV X-ray irradiation,the use of clone formation assay,Giemsa stained colony formation rate is calculated and SF2,and the fitting cell survival curve.Results The siRNA interference S100A4 after BxPC-3 cells SF2 value are:the control group 0.68±0.02,negative control group 0.65±0.01.interference group,0.38±0.02,P＜0.05.AsPC-1 cells SF2 value:control group 0.48±The0.02,negative control group 0.47±0.02; interference group:0.37±0.04,P＜0.05.Conclusion siRNA interference S100A4 after increased radiosensitivity of human pancreatic cancer cell lines.

AIM: DNA vaccines expressing protective domain of surface protective antigen A（spaA）of Erysipelothrix rhusiopathiae have been relatively ineffective at generating high-titer, long-lasting, neutralizing antibodies in murine models. METHODS: This paper report using a DNA vaccine expressing a fusion of the spaA protein and various elements, such as a secretion leader sequence from the highly expressed human gene encoding α1-antitrypsin (AAT), a highly soluble and stably folded domain from the rat cartilage oligomerization matrix protein (COMP), and three copies of the complement component, C3d3, to enhance the titers of neutralizing spaＡ-specific antibody. RESULTS: Analysis of titers of the antibody raised in vaccinated mice at different time points indicated that immunizations with the DNA expressing pcDNA3-AAT-COMP-spaAN-3C3d((pcD-ACSC)) had higher titers than pcDNA3-spaA_N(pcD-S) at weeks 4. Furthermore, the immune protective efficacy of the spaA-chimeras was demonstrated by lethal challenge with a virulent homologous strain 1249 against immunized mice. CONCLUSION: These results suggest that using a plasmid vector containing a strong heterologous signal sequence that mediate efficient antigen secretion in vivo and a fused piece of sequence improving antigens solubility, as well as C3d3, genetic adjuvant, could enhance the antibody responses level. This approach might be an efficient way to improve the antibody level of spaA_N DNA vaccination.

Objective To investigate the effect of S100A4 silencing on tumor related gene COX-2,bcl-2,Surviving,MMP-9 mRNA expressions of pancreatic cancer BxPC-3,AsPC-1 cells,and explore their relationship.Methods Small interfering RNA interfering S100A4 gene (siRNA-S100A4) was applied to transfect human pancreatic cancer BxPC-3,AsPC-1 cells,and nonhomologous siRNA-C was used as negative control,and cells without transfection were used as control group.The expressions of S100A4,COX-2,Survivin,MMP-9,bcl-2 mRNA after interference were detected by using RT-PCR.Results S100A mRNA expressions of BxPC-3's control group,siRNA-C group,siRNA-S100A4 group were 0.661 ± 0.023,0.659 ± 0.043,0.379 ± 0.039,and expressions of COX-2 mRNA were 0.760 ± 0.026,0.830 ± 0.017,0.443 ±0.006,and expressions of Survivin mRNA were 0.948 ± 0.049,0.909± 0.081,0.068 ± 0.006,and expressions of bcl-2 mRNA were 0.462 ±0.018,0.421 ±0.049,0.184 ±0.025,and expressions of MMP-9 mRNA were 0.813 ± 0.008,0.908 ± 0.063,0.246 ± 0.027.S100A mRNA expressions of AsPC-I's control group,siRNA-C group,siRNA-S100A4 group were 0.641 ± 0.042,0.626-± 0.053,0.320 ± 0.081,and expressions of COX-2 mRNA were 0.727 ± 0.021,0.743 ± 0.025,0.560 ± 0.035,and expressions of Survivin mRNA were 0.994 ± 0.032,0.984 ± 0.049,0.063 ± 0.005,and expressions of bcl-2 mRNA were 0.458 ±0.004,0.537 ± 0.046,0.181 ± 0.007; and expressions of MMP-9 mRNA were 0.698 ± 0.011,0.718 ± 0.073,0.199± 0.013.The expressions of S100A,COX-2,Survivin,bcl-2,MMP-9 mRNA in groups with siRNA-S100A4 transfection were significantly lower than those of siRNA-C group and control group (P ＜0.01),but the difference between siRNA-C group and control group was not statistically significant.Conclusions S100A4 plays a role in the pathogenesis of pancreatic cancer through up-regulation of COX-2,Survivin,bcl-2,MMP-9 expressions.

Objective To investigate the effect of rhodiola on severe acute pancreatitis associated renal injury and explore the potential mechanisms.Methods A total of 90 rats were randomly divided into sham-operated group (S group,n =18),severe acute pancreatitis with renal injury group (M group,n =18),low rhodiola dose group (3 g/kg,T1 group,n =18),moderate dose rhodiola group (6 g/kg,T2 group,n=18),high rhodiola dose group (9 g/kg,T3 group,n =18).The S and M groups were given 6 g/kg saline through intraperitoneal injection before operation while the T1 group,T2 group and T3 group were given with 3 g/kg,6 g/kg,9 g/kg dose of rhodiola through intraperitoneal injection,respectively.The pancreas was dissected and the head of pancreas was occluded by blood vessel forceps for 3 hours to make rat model.All the rats were sacrificed at 12 h,24 h,36 h after modeling.The level of ascites amylase,serum amylase,creatinine,blood urea nitrogen,interleukin 1β(IL-1β) and interleukin 10 (IL-10) were detected and the pathological change of pancreas and the left kidney was observed under light microscope.The serum levels of IL-1β and IL-10 were detected by enzyme-linked immunosorbert assay (ELISA).Take the right kidney for superoxide dismutase (SOD) determination and the expression of hypoxia inducible factor-1α (HIF-1α)mRNA in the right kidney was detected by real-time polymerase chain reaction (RT-PCR).Results Compared with the S group,the level of serum amylase,creatinine,blood urea nitrogen and IL-1β in M group increased significantly,but the activity of SOD has a significant decline (P ＜ 0.05).Compared with M group,the level of serum amylase,creatinine,blood urea nitrogen and IL-1 β in T2 group has a significant decline,but the activity of SOD,the express of HIF-1α mRNA and IL-10 has a significant increase (P ＜ 0.05).With the dose of rhodiola increased,the renal and pancreatic function in T2 group had a better performance than T1 group,and the difference was statistical significant (P ＜ 0.05).But compared with T2 group,the renal and pancreatic function in T3 group did not increased significantly (P ＞ 0.05).Conclusions Moderate dose of rhodiola (6 g/kg) has a good protective effect on severe acute pancreatitis associated renal injury.It may be associated with the inhibitory expression of IL-1β,up-regulated expression of IL-10,HIF-1α mRNA,and the increased activity of SOD.So it can then reduce cell apoptosis and renal necrosis,and improve the ability of the kidney to tolerate hypoxia.

Objective:To establish the acute rejection model of orthotopic liver transplantation in rat(ROLT) and observe the basic pathophysiologic changes of acute rejection.Methods:The rats were randomly divided into three groups:control group,isotransplantation group(LEW-LEW),and allotransplantation group(LEW-BN).Recipients were sacrificed on 3,5,7,and 10 days postoperatively and liver tissues and blood samples were collected.Recipient survival rate,histopathological and ultrastructural characteristics were observed.ALT,TBIL,and Alb were measured with automatic biochemical analyser.IL-2 content in serum was assayed by ELISA.Results:There was no rejection in isotransplantation from Lewis to Lewis rat resulted and 14-day survival rate reached 100%.The IL-2 concentration in serum was at normal level.On the contrary,all recipients in allotransplantation group from LEW to BN rat died among 14 days postoperatively,and hepatic histological examination showed typical acute rejection on 7 days.Liver function was severely impaired,which was indicated by significant increase of ALT and TBIL levels and apparent decrease of Alb level.The IL-2 concentration in serum was continuously increased and reached its peak value on 7 days postoperatively.Conclusion:An acute rejection experimental model of liver transplantation in rat could be stably established using Lewis rat as donor and BN rat as recipient.

Objective:To study the role of NF- ?B in the mechanism of liver injury following intestinal perforations due to abdominal firearm wound. Methods:A total of 42 Chang-Bai piglets were assigned randomly into 7 groups: control group and wounded 1, 2, 4, 8, 12, 24 hours group.The model of intestinal perforations due to abdominal firearm wound was established in wounded groups. Hepatic NF-?B and TNF-? content was measured with immunohistochemical staining and image analysis in all groups. Hepatocyte apoptosis indexes and serum ALT levels were also determined at the same time. The alterations of hepatic tissue were observed under light microscope. Results: Levels of hepatic NF-?B activity in wounded groups were significantly elevated compared with control group, and two peaks appeared in 1 h group and 8 h group, respectively (P

Objective:To investigate the changes of gastric tissue COX2 after intestinal perforations due to abdominal firearm wound.And to study its relationship with plasma endotoxin levels and pathological change of gastric tissue.Methods:A total of 42 Chang-Bai piglets were assigned randomly into 7 groups:control group and wounded 1,2,4,8,12,24 hours group.The model of intestinal perforations due to abdominal firearm wound was established in wounded groups.Gastric tissue COX2 activity was measured with immunohisto-chemical staining and image analysis in all groups.The plasma endotoxin levels were measured by chromogenic limulus amebocyte lysate test.The alterations of gastric tissue were observed under light microscope in all groups.Results:The expressions of COX2 of gastric tissue in wounded groups were significantly increased compared with control group (P

Objective:To study the role of TNF-?in the mechanism of hepatocyte apoptosis induced by intestinal perforations due to abdominal firearm wound. Methods:A total of 42 Chang-Bai piglets were divided randomly into 7 groups: control group and wounded 1, 2 , 4, 8, 12, 24 hour groups.The model of intestinal perforations due to abdominal firearm wound were established in wounded groups. Levels of plasma endotoxin were measured using ehromogenic limulus amehoeyte lysate test.Hepatic TNF-? content was measured with immunohistochemical staining and image analysis in all groups. Hepatocyte apoptosis indexes and serum TNF-? levels were determined at the same time. Results: Levels of plasma endotoxin, serum TNF-?, hepatic TNF-? content and hepatocyte apoptosis indexes in wounded groups were all significantly elevated compared with control group(P