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.

BACKGROUND: Heme oxygenase-1 (HO-1) plays an important role on preventing tissues and organs from oxidant stress injury, which remains currently one of the most active areas of investigation. OBJECTIVE: To study HO-1 over-expression on the survival and fiver function of rats after reduced-size liver transplantation by constructing recombinant adenovims Ad5-HO-1.DESIGN, TIME AND SETTING: Randomized controlled animal study was performed in the Institute of Pediatrics, Children Hospital Affiliated to Chongqing Medical University from September 2003 to March 2005. MATERIALS: Seventy-four SD rats were used to establish in situ reduced-sized liver transplantation models. METHODS: Recombinant adenoviral vector encoding Ha-1 gene (AdS-HO-1) was generatred by molecular biology method, wluch was administered to donors via voln at 48 hours before transplantation. All rats were randomly divided into control group(n=12),saline group(n=12), cobalt protoporphyrin (CoPP)group(n=13),Ad5-HO-1 group(n=13),Ad5-green fluorescent protein(GFP)group(n=12),and Ad5-HO-1+zinc protoporphyrin (ZnPP)group (n=12).Livers of donor wefe harvested and stored for 24 hours at 4℃ in HTK solution. Before it is implantedinto recipient. MA IN OUTCOME MEASURES: Survival rate and liver function; portal vein blood flow monitored 2 hours after transplantation by Color Doppler How Imaging; pathological changes of transplanted liver tissue observed by HE staining;HO-1 activity, tumor necrosis factor-alpha(TNF-α),Bcl-2 and Bax expression detected by immunohistoehemical staining; changes of HO-1,TNF-α,bcl-2 and bax mRNA detected by molecular viewpoint. RESULTS: Survival rate in the Ad5-HO-1 group was significantly higher than that in the saline group at al,7,and 21 days after liver transplantation(P＜0.05).Glutamic pyruvic transaminase decreased in the Ad5-HO-1 group as compared to that in the saline,Ad5-GFP,and Ad5-HO-1+ZnPP groups(P＜0.05);portal venous blood flow significantly increased 2 hours after transplantation(P＜0.05);HO-1 activity also significantly increased(P＜0.05).RT-PCR and immunohistochemical staining showed that HO-1 and bcl-2 expressions increased(P＜0.05),but bax and TNF-αexpressions decreased(P＜0.05).CONCLUSION:Ad5-HO-1 significantly induces high expression of HO-1 increases portal venous blood flow within 2 hours alter liver transplantation, and promotes liver functional recovery so as to prolong survival time of rats after liver transplantion.

Objective To compare the therapeutic effect and complication of immediate and selective interlocked intramedullary nailing in treatment of open tibial shaft fractures. Methods A total of 123 open tibial shaft fractures of 102 cases were treated with immediate fixation with interlocked intramedullary nailing after debridement (Group A) and with selective fixation with interlocking intramedullary nailing after debridement and closing the wound (Group B). Results The follow up for more than half a year showed that all fractures were healed, with excellent joint function of knee and ankle. Of all, 12 cases with skin necrosis and defect but without deep infection were healed after pedicle flap reconstruction. The result of Group A was much better than that of Group B in regard of total operation time, hospital stay, cost, amount of antibiotics used and partial weight bearing. No statistical difference in therapeutic effects and complications was found between two group. Conclusion Infection is not the contra-indication of interlocked intramedullary nailing in open tibial fractures. Immediate operation can be done after thorough debridement.

AIM To study the effects of degreasing-frosting on reducing toxicity and enhancing efficacy of Periplaneta americana L..METHODS The 95％ ethanol extracts from P.americana raw powder and processed products were extracted by petroleum ether,ethyl acetate,n-butanol and water.TLC was applied to comparing the component changes in various fractions.GC-MS was used for analyzing the kinds and contents of liposoluble constituents before and after processing.Shaking flask method combined with UV spectrophotometry method was adopted in the determination of Papp values in octanol-water and different pH values (2.23,3.28,4.22,5.23,6.12,7.24,8.23,9.10,10.15 and pure water) of buffer solutions.RESULTS After the petroleum ether fraction was processed,the colors of TLC spots became much shallower,which changed more significantly with the prolonging of processing time and increase of pressure.The TLC spots in the n-butanol and water fractions were clear without obvious changes.And no TLC spots were found in the ethyl acetate fraction.The kinds of liposoluble constituents were the same before and after processing,whose absolute contents were decreased after processing.Compared with raw powder,the Papp values of minor polypeptides in various processed products at different pH values exhibited significant differences (P ＜0.05,P ＜0.01),which also showed certain differences at the same pH values.CONCLUSION Degreasing-frosting can affect the contents of liposoluble constituents in P.americana and Papp values of minor polypeptides,which may be the mechanism of reducing toxicity and enhancing efficacy.

Objective To investigate the effect of Curcumin on rats with severe acute pancreatitis associated renal injury and explore the possible mechanisms.Methods A total of 72 rats were randomLy divided into sham-operated group(S group, n =24), severe acute panceratitis with renal injury group(M group, n =24) ,Curcumin-treated group(Cur group,n=24).The S and M groups were given 1.5 ml saline through intragastric administration 3 hours before operation,while the Cur group was fed with same amount of Curcumin diluent(200 mg/kg).The pancreas and pancreatic tail-segment was dissociated and the head of pancreas was occlused in rats to form the model,blood vessel forceps was loosed after 3 hours.All the rats were sacrificed at 12 h,24 h, 36 h and 48 h after modeling.The level of ascites, serum amylase, creatinine, blood urea nitrogen were detected and the pathological chang of pancreas was observed under light microscope.Take the right kidney for Superoxide Dismutase (SOD) determination and the expression of hypoxia inducible factor-1αmRNA in the right kidney was detected with realtime polymerase chain reaction (RT-PCR).Results Compared with Cur group, the level of ascites, serum amylase, creatinine, blood urea nitrogen in M group were significantly increased, but the activity of SOD, the express of hypoxia inducible factor-1αmRNA had a significant decline (P＜0.05).The tissue damage of pancreas and the kidneys became more serious.But a better performance was to be found in the function of the Cur group's kidney and pancreas, and the the difference was statistical significance (P ＜0.05).Conclusion Curcumin has a good protective effect on severe acute pancreatitis associated renal injury.It may be through up-regulation expression of HIF-1α mRNA and increase the activity of superoxidase dismutase, then reduce the cell apoptosis and necrosis of the kidney, improve the ability of the kidney to tolerate hypoxia.

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.