AIM:To detect the expression of runt-related transcription factor gene 2(RUNX2) in human gastric cancer cell line SGC7901 and to investigate the influence of small interfering RNA(siRNA)-mediated silencing of the RUNX2 gene on the proliferation and apoptosis of SGC7901 cells.METHODS:SGC7901 cells were divided into three groups:blank control group,negative control group(transfected with an empty vector),and experiment group(transfected with RUNX2 siRNA).After SGC7901 cells were transfected with RUNX2 siRNA,the mRNA and protein expression of RUNX2 was examined by RT-PCR and Western blot,respectively;cell proliferation was evaluated by MTT assay;and cell apoptosis was detected by ? ow cytometry(FCM).RESULTS:Compared with cells of the blank control group,the expression of RUNX2 mRNA(0.27 ± 0.068 vs 0.45 ± 0.058,F = 75.6,P < 0.01) and protein(F = 123.8,P < 0.001) was down-regulated in cells transfected with RUNX2 siRNA.At 24,48,and 72 h after transfection,the proliferation rates of SGC7901 cells transfected with RUNX2 siRNA were significantly lower than those of non-transfected cells(0.23 ± 0.039 vs 0.32 ± 0.012;0.31 ± 0.037 vs 0.45 ± 0.074;0.52 ± 0.021 vs 0.72 ± 0.006;F = 173.744,14.012,253.145;all P < 0.001).The apoptosis rate of SGC7901 cells transfected with RUNX2 siRNA was signif icantly higher than those of cells of the blank control group and negative control(45.65 ± 0.64 vs 4.46 ± 0.27,4.23 ± 0.33,both P < 0.01).CONCLUSION:RUNX2 expression was detected in SGC7901 cells.SiRNA-mediated silencing of the RUNX2 gene can inhibit proliferation and induce apoptosis in SGC7901 cells.RUNX2 may be a new gene therapy target for gastric cancer.

AIM: To investigate the impact of small hairpin RNA (shRNA)-mediated silencing of the tumor necrosis factor receptor-associated factor 6 (TRAF6) gene on the lipopolysaccharide (LPS)/ toll-like receptor (TLR) 4 signaling pathway in vitro. METHODS: ShRNA sequences targeting the TRAF6 gene were designed, synthesized and used to construct eukaryotic expression plasmids. After transfection of the recombinant plasmids into RAW264.7 cells, cell proliferation was measured by methyl thiazolyl tetrazolium (MTT) assay. Inflammatory cellular models were established by LPS stimulation. Levels of tumor necrosis factor-α (TNF-α), interleukin- 1β (IL-1β) and transforming growth factor-β1 (TGF-β1) in the supernatants, mRNA expression of TRAF6, interleukin-6 (IL-6) and cyclooxygenase- 2 (COX-2), protein expression of TRAF6, and translocation of NF-κB were assayed by ELISA, real-time quantitative PCR and Western blot, respectively. RESULTS: The mRNA and protein expression of TRAF6 was lower in cells transfected with TRAF6-shRNA1 or TRAF6-shRNA2 (79.17% and 68.74%, respectively) compared to other groups. Therefore, cells transfected with pGCsi-TRAF6- shRNA1 or 2 were used for subsequent experiments. TRAF6 knockdown significantly inhibited the proliferation of RAW264.7 cells within 72 h after transfection, reduced the production of pro-inflammatory cytokines and mediators including TNF-α, IL-1β, IL-6 and COX-2, and inhibited NF-κB nuclear translocation. Moreover, TRAF6 knockdown could suppress the release of TGF-β1 at the protein level. CONCLUSION: TRAF6 knockdown can, to some extent, inhibit early inflammatory response stimulated by LPS. TRAF6 may become a potential therapeutic target for many inflammationrelated diseases.

Objective:To approach the function mechanism of Danhuangfang(DHF)promoting the liver cell regenerate on the liver partial excision of liver regeneration model in rats.Methods:To adopt the liver partial excision of liver regeneration model in rats,carry on the intervention with DHF,examine the correlation factor which close liver regeneration expression of PC3 mRNA,c-fos mRNA,LRF-1 mRNA by gel electrophoresis and RT-PCR,observe the effectiveness which DHF influence liver regeneration.Results:DHF could enhance the hepatic tissue expression of PC3 mRNA,c-fos mRNA,LRF-1 mRNA in rats liver regeneration model,the DHF group had the significance difference comparing with the model group;the DHF effected the hepatic tissue expression of LRF-1 mRNA which not to be remarkable.Conclusion:DHF can possibly promote the liver cell multiplication through enhancing the hepatic tissue expression of PC3 mRNA,c-fos mRNA.

Objective:To explore the effects and probable mechanism of Danhuangfang(DHF) on rats with laboratory acute hepatic failure.Methods: The rats were injected with thioacetamide(TAA) 600mg/kg twice at same time for two days to induce the model of acute hepatic failure rat except the normal group injected with the normal saline hypodermically.The rats were treated with the normal saline,DHF low,middle and high dosages,PHGF respectively every day for 3 days before TAA were injected.To observe the mortality of the rats in 48 hours,and 48 hours later,the blood were collected to measure the indexes of ALT、AST、TBil、tumor necrosis factor(TNF-α)、interleukin-6(IL-6),and at the same time,pathological morphologies of hepatic tissues were observed respectively.Results: The DHF treatment group was super to controlled group in the mortality and the indexes reducing ALT、AST、TBil、TNF-α、IL-6,improving liver tissue pathological changes of rats with acute hepatic failure.Comparing with model group and control group,there are significant difference(P<0.05 or P<0.01).Conclusion: The DHF have resistance against the rat with acute hepatic failure by TAA,it related to decrease TNF-α the important medium of acute hepatic failure and IL-6.

Objective:To observe the effect of Rhubarb on expression of signal regulatory protein α1 in hepatic regenera-(ting) rats.Methods:Twenty four rats were randomly divided into normal group,model group,Rhubarb group and PHGF(lyophilized bovine transfer factor) control group.Medication rats were given 1 ml/100g Rhubarb or PHGF injection respectively 3 days before hepatectomy once a day.At the same time,the rats of normal group and model group were hypodermically injected with the 0.85% salt solution.Except the normal group,70% of liver including medial and left lobes were sectioned.Forty eight hours later,all animals were sacrificed,hepatic tissues were fixed and embedded into paraffin and sectioned.The expression of signal regulatory protein α1 in hepatic tissue was determined by immunohistochemistry.Results: Rhubarb can obviously proliferation signal regulatory protein α1 in hepatic regenerating rats comparing with model group and control group,there were significant difference(P<0.01).Conclusion:Rhubarb has the function of regulation the expression of signal regulatory protein α1 in hepatic regenerating rats,improve liver regeneration.

[Objective] To study the effect of Rhubarb on liver damage and liver regeneration in thioacetamide-induced fulminant hepatic failure(FHF) in rats.[Methods] The rats were randomly divided into normal group,model group,Rhubarb group and lyophilized bovine transfer factor control group. Medicines were given 1 ml/100 g respectively 3 days before hypodermically injected with thioacetamide(TAA) once a day.At the same time,the rats were hypodermically injected with TAA 600 mg·kg~(-1) twice for two days to induce the model of FHF except those in the normal group which were injected with the 0.85 % salt solution hypodermically.Forty-eight hours later,the blood was collected to measure ALT,AST,TB and parameters of liver regeneration such as liver mitotic index and proliferation cell nuclear antigen.[Results] Rhubarb ould obviously decrease ALT,AST,TB and improve liver mitotic index and proliferation cell nuclear antigen of rats with FHF,which has significant difference compared with model group and control group(P0.05 or P0.01).[Conclusion]Rhubarb can prevent the development of FHF and improve liver regeneration.