【Objective】To investigate the expression of nitric oxide synthase 3 (NOS3) and vascular endothelial growth factor (VEGF) in hepatocellular carcinoma (HCC).【Methods】The expression of NOS3 and VEGF were detected by immunohistochemistry in 51 cases of HCC.【Results】The positive of NOS3 and VEGF in HCC were 53% and 63%.The expression rate of NOS3 in the recurrent group was significantly higher than that in the non-recurrent group (P<0.01).The expression rates of VEGF in the group with carcinoma embolus of portal vein and the recurrent group were significantly higher than those in the group without carcinoma embolus and non-recurrent group (P<0.05).The expression of NOS3 was closely related with the expression of VEGF positively in HCC.【Conclusions】NOS3 and VEGF are related with the biological behavior of HCC closely.

Objective To study the expression and significance of survivin and cyclin D 1 in hepatic cellular carcinoma (HCC).Methods The expression of survivin and cyclin D 1 were detected in 61 cases of HCC by immunohistochemistry.Results The expressive rates of survivin and cyclin D 1 in HCC were 44.3% and 39.3%.Both survivin and cyclin D 1 rates in the lower-differentiated group were significantly higher than those in the well-differentiated (P

Objective To construct high-capacity ribosome display single-chain Fv library for selection of high affinity ScFv antibody.Methods We isolate human lymphocyte from peripheralblood(2 normal,3 gastric cancer,3 colonic cancer,1 pancreatic cancer,each 5 mL and 2 newborn,each 2 mL)and extract RNA for cloning whole human heavy chain and light chain gene by RT-PCR.VH and VL were rearranged randomly by SOEing(splicing by overlap extension,SOEing).Finally,the elements for in vitro screening such as T7 promoter and ribosome binding site were introduced while the SOEing products were amplified.Moreover,ribosome display template were verified by blue/white screening and further sequencing.Results We successfully constructed ribosome display ScFv library with a volume of 1.1?1013.Conclusion The construction of high-capacity ScFv library shed light on multiple therapeutic ScFv screening.

Objective To investigate the prevalence, clinical features and complications of gastrointestinal diverticula in Chinese people. Methods 551 patients with gastrointestinal diverticula were analyzed retrospectively. Results Among the 551 patients, 58.6% were over 60 years, and 11.6% were below 40 years. The incidence of esophageal diverticula, gastric diverticula, duodenal diverticula, jejunoileal diverticula and diverticula of the large intestine was 2.5%, 1.8%, 71.7% , 12.7% and 11.3% respectively. There were 6 and 11 patients, 9 and 6 patients, 69 and 271 patients, 1 and 24 patients, and 21 and 33 patients who were diagnosed by endoscopy and double contrast barium examination respectively. Among the 14 esophageal diverticula patients, the incidence of midesophageal diverticula (78.6%) was higher than that in pharyngoesophageal diverticula (11.2%). Among the 62 diverticula of the large intestine patients, the incidence of right side diverticula (56.5%) was higher than that of left side diverticula (38.7%). Almost esophageal diverticula and gastric diverticula were asymptomatic, while about half of the diverticula of the small and large intestine was symptomatic. The incidence of cholelithiasis and pancreatitis in patients with duodenal diverticula was 34.2% and 10.1% respectively. And the incidence of cholelithiasis and pancreatitis in descending segment was significantly higher than that in non descending segment ( P

Objective To study the spiral CT features of gastrointestinal invasion by carcinoma of gallbladder. Methods Eight patients with surgical-pathologically documented gastrointestinal invasion by carcinoma of gallbladder were analyzed retrospectively. All patients underwent plain and contrast-enhanced dual-phase scanning of the abdomen. Oral contrast medium (1.2% Angiografin) was used to fill the gastrointestinal tract before CT scanning. Results There were 2 cases of gastric antrum invasion, 6 duodenal invasion and 3 colonic invasion according to the surgical and pathological findings. Spiral CT correctly diagnosed 2 gastric invasion and 4 duodenal invasion based on several imaging features, like blurring of fat plane, focal wall thickening and luminal narrowing of involved gastrointestinal segments, and mass formation. However CT was unable to diagnose the 3 cases of hepatic flexure of colon invasion. Conclusion CT is valuable for diagnosing upper gastrointestinal tract invasion by carcinoma of gallbladder, yet the diagnosis of hepatic flexure of colon invasion is still difficult.

Objective:To study influencing factors for in‐stent restenosis (ISR) during one year in patients with coro‐nary heart disease (CHD) after coronary sirolimus‐eluting stent (SES) implantation .Methods :According to results of coronary angiography (CAG) ,a total of 275 patients ,who hospitalized in our department from Jan 1st ,2012 to Dec 30th ,2013 and have received SES implantation and reviewed CAG after one year ,were divided into non‐ ISR group (n=247) and ISR group (n=38) .Clinical characteristics were compared between two groups ,and Logistic regression analysis was used to analyze influencing factors for ISR .Results:Compared with non‐ISR group ,there were significant rise in percentages of occlusion lesions (17. 9% vs .31. 9% ) ,multiple overlapping stents (16. 7% vs . 31.9% ) ,and significant reduction in percentage of stent post‐dilatation (34.9% vs .10.6% ) in ISR group ,P<0.05 or <0. 01 ;Logistic regression analysis indicated that coronary occlusion lesion was a risk factor (OR :2. 855 ,95%CI :1.197~6.808 ,P=0.018) ,and post‐dilatation was a protective factor (OR :0.198 ,95% CI :0.057~0.691 , P=0.011) for ISR occurrence .Conclusion:Multiple overlapping stents and coronary occlusion lesions increase one‐year in‐stent restenosis rate ;stent post‐dilatation can reduce one‐year in‐stent restenosis rate ;coronary occlusion le‐sions is a risk factor , and stent post‐dilatation is a protective factor for restenosis during one‐year after coronary drug‐eluting stent implantation .

AIM: To observe the effects of aspirin and prostaglandin E_2 (PGE_2) on the cell viability and cell cycle in SW1990 human pancreatic carcinoma cell lines, and to investigate the mechanisms of aspirin-induced growth inhibition and cell cycle arrest. METHODS: After incubated with aspirin or PGE_2 and their combination, the viability of SW1990 cells was measured by MTT assay. The levels of intracellular PGE_2 were determined by ELISA. The effects of aspirin or PGE_2 on cell cycle were investigated by flow cytometry (FCM). The expression of p21~ Wafl/cipl and p27~ Kipl/pic2 (the cyclin-dependent kinase inhibitors) were analyzed by Western blotting. RESULTS: Aspirin could inhibit the growth of cells and level of intracellular PGE_2 in a dose-dependent manner. Aspirin enhanced the expression of p21~ Wafl/cipl and p27~ Kipl/pic2 and induced cell cycle arrest at G_0/G_1 phase. PGE_2 increased the cell viability of SW1990 cells. However, it couldn't antagonize the changes of cell viability and cell cycle that induced by aspirin. CONCLUSIONS: The inhibitory effects of aspirin on growth and cell cycle of pancreatic carcinoma cells might not be mediated by a COX-dependent pathway completely. Cell cycle arrest induced by aspirin might be associated with up-regulation of p21~ Wafl/cipl and p27~ Kipl/pic2 .

AIM:To establish an effective and rapid method to develop transplanted subcutaneous pancreatic carcinoma by inducing PANC-1 cells into nude mice, and then use this mouse model to evaluate the tumor-homing and gene-silencing effects of siRNA-loading nanoparticles in vivo.METHODS:Different numbers of PANC-1 cells in 100 μL or 300 μL PBS were inoculated subcutaneously into the right flank of BALB /c (nu/nu) mice.When the tumor volume reached 100 mm 3 , siRNACY 5.5 nanoparticles were injected through the mouse tail vein to perform in vivo imaging assay.Be-sides, the mice were randomly divided into 3 treatment groups treated with PBS, scrambled control RNA nanoparticles and siKras nanoparticles, respectively.The protein expression of Kras was detected by Western blotting and immunohistochemi-cal staining.RESULTS:After inoculated with 1 ×10 7 PANC-1 cells in 300 μL PBS, all mice developed tumors within 2 weeks.The in vivo results showed that siRNA-loading nanoparticles accumulated in the tumor tissues and exerted gene si-lencing effect.CONCLUSION:In the present study, an effective and rapid method was established for PANC-1 cells to induce transplanted subcutaneous pancreatic carcinoma in nude mice within 2 weeks, which is suitable for in vivo imaging and treatment evaluations as a reproducible and reliable way for the further experiments .

AIM:To synthesize a safe , efficient and targeted nanoparticulate carrier for siRNA delivery to pan-creatic cancer cells .METHODS: Iron oxide nanocrystal with carboxylic acid group-polyethyleneimine ( IONP-PEI ) was synthesized and investigated as a nonviral carrier of siRNA to the pancreatic cells .The size, surface and charge using zeta potential were characterized .The perfect charge ratio between amino groups of IONP-PEI and phosphate groups of siRNA ( N/P) was determined by the transfection efficiency detection , gel retardation assay and MTS assay .An antibody-directed nonviral vector , scFvCD44v6-IONP-PEI nanoparticle attaching to the cancer-associated CD44v6 single-chain variable frag-ment, was constructed as a cancer-targeting nanocarrier for siRNA delivery .Prussian blue staining and immunofluorescent staining were performed to detect the distribution of scFv CD44v6-IONP-PEI/siRNA complexes in the cells .The transfection efficiency , fluorescence intensity and the expression of KRAS at mRNA and protein levels in the cells transfected by IONP -PEI/siRNA and scFv CD44v6-IONP-PEI/siRNA were detected by flow cytometry , fluorescence microscopy , real-time PCR and Western blotting, respectively.RESULTS:The mass ratio of IONP to PEI was 0.75.The suitable ratio of N/P was 20. The averaged size and surface zeta potential of IONP-PEI/siRNA in deionized water were (51.3 ±2.2)nm (diameter) and (21.73 ±8.07)mV, respectively.Red fluorescence was seen in both targeting and nontargeting groups , which clearly re-vealed the intracellular distribution of siRNA and delivery agents .Transfection efficiencies in targeting and nontargeting groups were (89.75 ±1.81)%and (59.87 ±4.52)%, respectively.Down-regulation of the KRAS mRNA in Panc-1 cells transfected with siKRAS by scFvCD44v6-IONP-PEI and IONP-PEI was up to (34.02 ±6.15)%and (51.09 ±6.70)%, re-spectively .The protein level of KRAS was lower in targeting group than that in nontargeting group .CONCLUSION:scFvCD44v6-IONP-PEI is a safe and efficient nanoparticulate carrier for gene delivery .It is more effective to transfer siRNA into the cells and mediate gene silencing effect in vitro than the nontargeting group .

AIM:To investigate the preparation techniques and anti-tumor effects both in vitro and in vivo of a novel nanoparticles control-releasing preparation of 5-fluorouracil(5-FU)by intravenous injection.METHODS:With polylactic acid(PLA)as marix materials,we adopted ultrasound emulsification method to prepare PLA enveloped 5-FU nanoparticles(5-FU-NPs).Scanning electricity microscopy was used to observe the morphology of 5-FU-NPs and laser optical scattering experiment was conducted to determine its diameter distribution.The drug-carrying capacity(ratio)of the nanoparticles was determined by means of high-power liquid chromatography(HPLC)and MTT test was used to observe cytotoxicity in vitro.The anti-tumor effects were determined at different dosages,frequencies of taking drugs in vivo.RESULTS:Scanning electron microscopy showed that the 5-FU-NPs were globular particles with smooth surface in an average particle diameter of 191.9 nm with a normal distribution,and the drug-carrying capacity of 5-FU-NPs was 15.2%.5-FU-NPs had the same anti-cancer effect as unenveloped drug in vitro and showed typical dose-effect relationship.Compared to naked 5-FU,5-FU-NPs presented significant difference(P