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 clarify great benefit to prevention of wound infection as well as reduction of economic expenditures by a new method of connecting latex gloves and cigarettes in peritoneal drainage. Methods We randomly selected 121 cases of patients with peritoneal pyogenic infection, divided them into the latex gloves group (64 eases)and the common dressing group (57 cases). The incision infection rate and eco-nomic cost in two groups were comparately studied. Results The latex gloves group's incision infection rate was 10.94% (7 / 64), the common dressing was 31.58% (18/57). Each patient of the latex gloves group spent less cost about 15.70 yuan,changed dressing 5.3 times less than the common dressing group. Conclusions Connecting latex gloves and cigarettes in peritoneal drainage is a new method of reducing economic expandi-tures and the chances of incision infection. Clinical application of this method should be fully prospected.

Objective To summarize the role of nuclear factor kappa B(NF-?B) in the occurrence and progression of various sorts of liver injury.Methods Literatures on the structures,property of molecular biology and function of NF-?B,as well as its relationships with liver injury were collected and reviewed.Results NF-?B was an important nuclear factor existed in cells widely distributed in most cell types.The activation of NF-?B was induced by various sorts of liver injury.The activated NF-?B could affect the liver injury by regulating cytokines,adhesion molecules,and activating factor involving in immunologic reaction,inflammatory reaction and the apoptosis.Conclusion NF-?B plays an important role during the occurrence and progression of liver injury,and may become a new target in the treatment of liver injury.

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

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 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.