Objective To investigate the radiosensitivity of silencing N-Ras by RNA interference in hepatoma carcinoma cell MHCC-97.Methods N-Ras RNA interference (RNAi) vector was constructed by using pcDNA 6.2-GW/EmGFP-mir plamid.The RNAi effect was detected by RT-PCR,Western bolt,immunohistochemisty and MTT method.Survival curve for each cell line were obtained by measuring the clone forming abilities of irradiated cell populations.Results After silencing the N-Ras by RNAi,The expression level of N-Ras mRNA,N-Ras protein,immunohistochemisty were decreased 96.9% ±0.159%(t=40.377,P＜0.05),89.8%±0.012% (t=31.595,P＜0.05),90%,respectively,and The survival of hepatoma carcinoma cell MHCC-97 line were inhibited 21.9% (F = 4.63,P < 0.05).Which have significant difference in statistics.The SER of hepatoma carcinoma cell MHCC-97 line after interference was 1.15.Conclusions RNAi targeting silence N-Ras may increase the radiosensitivity of hepatoma carcinoma cell MHCC-97 line.

Objective Nonsyndromic cleft lip with or without cleft palate(NSCL/P)is a common craniofacial birth defect which results in lifelong medical and social consequences.Although Asians have the highest birth prevalence of oral-facial clefts,the majority of gene mapping studies of cleft lip with or without cleft palate(CL/P)have been in European or Ameriean Caucasians.Therefore,the obiective of this study was to evaluate association between transforming growth factor alpha(TGF-α)gene BamH Ⅰ polymorphism and NSCL/P in Chinese.Methods 107 patients with NSCL/P and 136 healthy controls were examined for TGF-α/BamH Ⅰ genotypes.TGF-α/BamH Ⅰ typing was carried out by digesting the locus specific polymerase chain reaction amplified products with alleles specific BamH Ⅰ restriction enzyme(PCR-RELP).Resuits A1 allele frequency was 0.06 and A2 allele frequency was 0.94 in the controls.A1 allele frequency was 0.14 and A2 allele frequency was 0.86 in patients with NSCL/P(x2=8.27,df=1,P＜0.05).A1 allele frequency was 0.17 and A2 allele frequency was 0.83 in the bilateral cleft lip with or without cleft palate.A1 allele frequency was 0.13 and A2 allele frequency was 0.87 in the unilateral cleft lip with or without cleft palate(x2=0.36,df=1,P＞0.05).There was no statistically significant between the case with family history and the case without family history(x2=0.34,df=1,P＞0.05).Conclusions The above data demonstrate that there is evidence for the association of TGF-α polymorphism with development of NSCL/P in Chinese.

Objective To investigate the xenotransplantation model of fetal pig skin precursor tissue and its development after transplantaion. Methods Porcine skin precursor tissue was obtained from the embryo of gestation day 56 (E56), and made into microskin or stamp skin graft. The microskin was transplanted to the dorsal wound in BALB/c nude mice, then covered with human corpse skin. The stamp skin graft was imbedded subcutaneously into the back of nude mice, and microskin was injected subcutaneously into the auricles of nude mice. Their growth and development were observed and they were examined by HE staining at 6th and 12th week after transplantation respectively. Two-sample t test was used to analyze the size of newly grown skin tissue. Results Porcine skin precursor tissue graft in three models above survived and continued growth after transplantation, and growth ability of the dorsal wound transplantation model was significantly stronger than that of the auricle model. Epidermis and hypodermis were detected in newly grown skin tissues. Hair follicles, a few of sebaceous glands, but no sweat glands were observed in auricle model, while many sebaceous glands and sweat glands were observed in the dorsal wound model. Conclusion Transplantation of microskin to dorsal wound is the optimal model of investigating the xenotransplantation of fetal pig skin precursor tissue and its development after transplantion.

AIM:To investigate the mechanism of endothelium-derived microparticles(EMP)-induced endothelial dysfunction and the role of superoxide anion(O-?2) in EMP-induced endothelial dysfunction.METHODS:EMP were isolated from human umbilical vein endothelial cells stimulated with plasminogen activated inhibitor-1.(1) Cultured bovine aortic endothelial cells(BAEC) were divided into 3 groups and pretreated with nothing in group 1,EMP(1?108/L) in group 2,EMP(1?108/L) + L-nitroarginiemethylester(L-NAME,1 mmol/L) in group 3 for 30 min and A23187(5 ?mol/L) stimulated O-?2 generation was determined by superoxide dismutase(SOD)-inhibitable ferricytochrome C reduction.(2) Facialis arteries(60-150 microns) were isolated from C57BL/6 mice and divided into 4 groups.The vessels were pretreated with nothing in group 1,EMP(1?108/L) in group 2,EMP(1?108/L) + SOD(2?105 U/L) in group 3,EMP(1?108/L) + polyethylene glycolated-SOD(PEG-SOD,2?105 U/L) in group 4 for 10 min and acetylcholine(ACH)-induced vasodilation was measured.RESULTS:(1) EMP significantly increased O-?2 generation in BAEC culture,which was prevented about 50% by pretreating the BAEC with L-NAME.(2) EMP significantly impaired ACH-induced vasodilation.SOD could not restore EMP-impaired ACH-induced vasodilation and PEG-SOD showed partial restoration of vasodilation.CONCLUSION:These data indicate that at least some EMP-induced endothelial dysfunction occurs by inducing intracellular O-?2 generation.It may provide a theoretical evidences in finding a multiple treatment including removal of O-?2 in the future.

Objective To investigate the influence of Zhenqing Recipe(ZQR)and Ligustri Lucidi Fructus(LLF)on diabetic rats and its possible mechanism.Methods The model of type 2 diabetic rats was established by feeding a high-sucrose-high-fat diet and injecting a low dose of Streptozotocin in Wistar rats.The model rats were randomly divided into three groups: diabetic model,ZQR-treated,and LLF-treated groups for 8-weeks treatment.The normal Wistar rats were as a normal control group.Results The level of fasting blood glucose in ZQR and LLF groups was decreased compared with model group(P < 0.01,0.05,respectively).Both ZQR and LLF markedly reduced serum triglycerides(P < 0.01,0.05,respectively),and increased the insulin sensitivity index(P < 0.05).Histopathology revealed that ZQR and LLF reduced pancreatic damage.Immunohistochemistry evaluation showed that the percentage of insulin positive cells in pancreatic island was higher than model group(P < 0.01,0.05,respectively).The mRNA and protein expression of SREBP-1c in pancreas were significantly decreased in ZQR and FLL group(P < 0.01).Conclusion ZQR has therapeutic effect on type 2 diabetes,it ameliorates the histopathologlcal changes of pancreas,protects β cells,improves insulin resistance,and attenuates the expression of SREBP-1c.This study also provides the anti-diabetic evidence of FLL even its effects are weaker than ZQR.

Objective Non-invasive detection is the focus of intense research in diagnosis of acute renal allograft rejection currently. Urine protein is considered the cue to reflect the pathological changes in kidney disease. In this study, we explored the urine markers for early acute renal allograft rejection. Methods The urine protein of two patients with acute renal allograft rejection were examined by 2D gel electrophoresis and bioinformatics. We adopted pH 4-7 ready strip IPG and stained the gel with Sypro-Ruby. The digitized 2D maps of urine protein were quantitatively analyzed using 2D-analysis software packages. By analyzing the differential expressions of proteome between different time points (1, 2, 3 days before acute rejection and 7, 14, 21 days after acute rejection), 30 protein spots were selected and analyzed by MALDI-TOF-MS/MS. Results We obtained 2D gel electrophoresis maps of urine protein of the patients with acute renal allograft rejection, which are of good reproducibility and resolution. Sixteen protein spots were identified, resulting in thirteen corresponding proteins. Out of these proteins, we screened three proteins (alpha-1-antichymotrypsin, tumor rejection antigen gp96, Zn-Alpha-2-Glycoprotein) closely related to acute rejection. Conclusion The urine protein spots on 2D gel electrophoresis maps for the patients with acute renal allograft rejection were of obvious difference when detected at different time points of acute rejection. Alpha-1-antichymotrypsin, tumor rejection antigen gp96 and Zn-Alpha-2-Glycoprotein might be the candidate protein markers to diagnose acute renal allograft rejection after renal transplantation.

Objective: To detect the expression of basic fibroblast growth factor(bFGF) in oral mucosa with ulcer in rabbits. Methods: 72 New Zealand rabbits(with the weight of 3 000-3 500 g) were randomly divided into control group,model group,and treatment group(n = 24). 1,3,5 and 7 d after treatment buccal mucous membrane tissues of the rabbits were respectively taken from the 3 groups. The models of oral ulcer were examined by HE staining. The expression of bFGF mRNA was detected by RTPCR. The expression of bFGF protein was detected by immunohistochemistry. Results: The oral ulcer model of the rabbits was successfully established. Both RT-PCR and immunohistochemistry analyses showed that 1-7 d after treatment the expression levels of bFGF mRNA and protein were higher in treatment group than in model group(P ＜ 0. 05) and control group(P ＜ 0. 05),3-7 d after treatment were higher than in model group(P＞ 0. 05). Conclusion: bFGF may be a new therapeutic target for oral ulcer.

Objective To investigate the miRNA expression profiles in rectal cancer tissues and their associations with clinical pathological stage, depth of tumor invasion, and lymph node metastasis, and to evaluate the potential of miRNA as diagnostic and prognostic markers of rectal cancer. Methods Human miRNA microarray was used to profile miRNA expression in rectal cancer tissues and matched adjacent normal tissues (n=71). The up-regulated miR-93-5p and down-regulated miR-27a-3p were screened out, and the top differentially expressed miRNA were validated by quantitative real-time polymerase chain reaction ( qRT-PCR) . The relationship between the expression of miRNA and clinical parameters was analyzed by ANOVA and Spearman correlation. Results The expression of miR-27a-3p was down-regulated in miRNA microarray, but was up-regulated in qRT-PCR analysis;the data were relatively discrete. The expression of miR-93-5p was up-regulated in both miRNA microarray and qRT-PCR analysis;the expression level of miR-93-5p in rectal cancer tissues was 3165 times that in adjacent normal tissues ( P=00058);the expression level was correlated with tumor volume ( P= 0004 ) , and was positively correlated with the level of carcinoembryonic antigen ( CEA) before treatment ( P=0001) and the number of lymph nodes metastases (rs=0534, P=0005). Conclusions There is a differential miRNA expression pattern between rectal cancer tissues and matched adjacent normal tissues. The miR-93-5p is highly up-regulated in rectal cancer tissues and may serve as a diagnostic and prognostic marker of rectal cancer.