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 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 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 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 .

Objective To investigate the influence of PPARγ excitomotor RSG and ATRA on gastric cancer SGC7901 cell line proliferation in vitro and its potential mechanism study.Methods Human gastric cancer SGC7901 cell line was cultured in vitro.Experiment samples were divided to blank group,10μmol/L ATRA group, 12.5μmol/L RSG group, 25μmol/L RSG group, 10μmol/L ATRA + 25μmol/L RSG group.Proliferation inhibitory effect was determined by MTI assay.Flow cytometry was used to detect cell cycle, H.E stain was used to observed micrography alteration.Expression of PPARγ protein in gastric cancer cells were measured by immunohistochemistry.PPARγ mRNA in gastric cancer cells were measured by RT-PCR.Results ATAR at concentration 10μmol/L, RSG at 12.5 μmol/L and RSG at 25 μmol/L could inhibit the proliferation of SGC7901 cells in a dose-and time-dependent, and when both agents were combined for 72h, growth inhibition ratio was (29.73 ± 0.69) %.Flow cytometry analysis revealed a cell cycle arrest at G1 and S phase, and when both agents combined, S% was (12.87 ± 0.35 )%, cell micrography tended to be normal when both agents combined.Up-regulation of PPARγ protein and PPARγ mRNA expressions were also observed, those effects were enhanced when both agents combined, and grey scale ratio was 0.646.Conclusion The ATRA and RSG could significantly induced growth inhibition of human gastric cancer SGC7901 cell, which may be associated with cell cycle arrest and inducing differentiation, activation of PPARγ protein and PPARγ mRNA expression.Synergistic effect could be caused by the combined use of the two agents.

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 .

Objective To develop user-friendly information education-communication (IEC)materials and guidelines of malaria control for various target groups. Methods The participatory methods were used to make sure participation of target groups at each stages of IEC material development. Results A package consisted of interactive video, 2 posters for primary groups, sto ry booklet for students, manual for activities of "buddy system" for teachers, flipcharts for health staff at grass-root levels, guidelines on communication skill and how to use the first 5kinds of materials, was developed. Conclusion A set of user-friendly IEC materials and guidelines of malaria control has successfully been developed with sufficient participation of target groups.

Objective To investigate the incidence of pancreatic cancer-related depression in Guangzhou,China.Methods A multicenter,prospective survey was conducted,50 patients with pancreatic cancer,60 with liver cancer,50 with esophageal cancer,50 with gastric cancer,52 with colorectal cancer were enrolled from 4 hospitals in Guangzhou between June 2007 and June 2009.Hamilton Rating Scale for Depression-24 (HAMD-24) questionnaire was used to assess the degree of depression.Results The incidence of depression in pancreatic cancer patients was 78% (39/50),which was significantly higher than that among liver cancer patients (60% ,36/60),gastric cancer patients (36%,18/50),esophageal cancer patients(24%,12/50),and colorectal cancer patients(19.2%,10/52,P＜0.05 ).Twelve of 50 patients in pancreatic cancer were reported to have severe depression (24%),which was significantly more than that in liver cancer (10%,6/60),gastric cancer (4%,2/50),esophageal and colorectal cancer (0,P ＜0.05).In pancreatic cancer patients,the incidence of depression was significantly higher in patients with advanced stage (94.3%) than that in early stage (46.7%,P＜0.05).Patients who underwent chemotherapy had high incidence of depression(92.3%)than that of patients who underwent operation (62.5%,P＜0.05 ).Conclusions Compared with other cancers of digestive tract,the incidence of pancreatic cancer-related depression was higher,and its degree was more severe than that of other cancers.

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 .