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 research the value of CT perfusion imaging in diagnosing breast tumor .Methods 21 focuses in 17 cases withbreast tumorlike pathological changes diagnosed by molybdenum target examination were scanned by CT perfusion imaging . Cine model,0.5 s/circle ,5 mm in thickness?4,120 kV,60 mA,delayed time 10 s,scan time in all 50 s ,GE 4.2 workstation and perfusion 3 software were used ,the data of creating dynamic picture and the relative informations that image intention changes with time change were analyzed and every parameters related with perfusion were studied and analyzed statistically .Results Blood flow(BF),blood volume(BV),mean transit time(MTT) and permeability surface(PS) were (36.46?17.62)ml?min-1?100 g-1,( 13.76?8.59) ml/100 g,( 28.23?15.75) s and (16.45?12.36) ml?min-1?100 g-1.In cancer group,( 17.35?10.67)ml?min-1?100 g-1,( 4.63?3.47) ml/100 g,( 25.52?12.91) s and ( 3.57?3.36) ml?min-1?100 g-1 in benign lesion group,respectively,there was significant difference between two groups in BF,BV and PS(P

AIM: To investigate the effects of angiotensin II receptor antagonist on remodeling of renal arterioles in hypertension. METHODS: Eighteen 4 weeks old male rats were divided into three groups: Wistar-Kyoto rats (WKY) for normotensive group, and spontaneously hypertensive rats (SHR) for hypertensive group, and SHR treated with losartan orally (15 mg?kg -1 ?d -1 ). The rats were raised to 16 weeks old. The morphometric parameters of the renal arterioles, and the widths of vascular smooth muscle cells (VSMC) and intercellular space were studied on kidney slices by light microscope and electromicroscope respectively, combined with computer-assistant image analysis system. The minimal renal vascular resistance (RVR min ) was studied by isolated kidney perfusion system. RESULTS: The systolic blood pressure of the tail artery, wall thickness, wall area, ratio of wall thickness to inner diameter, width of VSMC of renal arterioles and RVR min were all smaller or lower in losartan group than those of SHR. CONCLUSION: Ang II receptor antagonist losartan can prevent the remodeling of renal arterioles in SHR.

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 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

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 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 .

Site surveys were conducted with a self-designed questionnaire for 4102 urban residents regarding their awareness and utilization rate of service projects and satisfaction (total satisfaction,medical environment,service attitude,content of communication and diagnostic & therapeutic modalities).The influencing factors of satisfaction were analyzed by multiple linear regression analysis.The factors of improving total satisfaction were as follows:①When content of communication,diagnostic & therapeutic modalities,service attitude and medical environment improved 10％,total satisfaction rose by 2.44％,2.50％,2.47％ and 0.98％ respectively; ② Utilization rate rose by 2.33％,1.60％ and 0.85％ respectively if they accepted home care services,rehabilitation services and chronic disease management services ; ③Total satisfaction rose by 0.94％ if they became aware of family planning and reproductive health services.And the factors of reducing total satisfaction were as follows:①total satisfaction declined by 1.68％ if their children accepted preventive vaccination services.②total satisfaction declined by 1.13％ if the respondents maintained their health records.

To explore the current status of job burnout among 1210 staffs at local community health stations and analyze its influencing factors with multiple logistic regression analysis.The regression model of high emotional exhaustion had statistical significance (SE =0.070,P =0.000).The possibility of high emotional exhaustion was negatively correlated with the satisfaction of organizational commitment,rules and regulations,cooperation between colleagues,satisfaction of job nature and family supports.And it had a positive correlation with supports from family and colleagues and satisfaction of welfare.The regression model of high depersonlization had statistical significance (SE =0.072,P =0.000).The possibility of high depersonlization had a negative correlation with organizational commitment,cooperation between colleagues,satisfaction of job nature and smooth communications with leaders and family.And it was positively correlated with compensation,promotion,satisfaction of welfare and smooth inter-colleague communications.The regression model of highly reduced personal accomplishment had statistical significance (SE =0.061,P =0.000).The possibility of highly reduced personal accomplishment had a negative correlation with satisfaction of superiors,job category and communication,age,smooth communications with leaders and family.And it was positively correlated with satisfaction of compensation.

Objective To investigate the killing effect of hematoporphyrin derivative photedynamic therapy (PDT) on cultured human pancreatic cancer cell,and to explore the mechanism of this effect.Methods Biolitec PDT 630 semi-conductor laser therapeutic apparatus was used as the light source.After pancreatic cancer cell PANC1 was incubated 8 h with different concentrations of Photosan(hematoporphyrin derivative) as photosensitizer (0.5mg/L,1 mg/L,2 mg/L,4 mg/L),the cells were given different doses of 630nm laser irradiation(1 J/cm2' 5 J/cm~2,10 J/cm~2 ).The A492 value was determined in each group with MTT method.Cell apoptosis rate was detected by flow cytometry after PDT.Results There was no killing effect when no Photosan was administrated;10 J/cm~2 irradiation had killing effect on PANC1 when Photosan was administrated as 1 mg/L(0.140±0.013 vs 0.213±0.008,P＜0.05);5 and 10 J/cm~2 irradiation all had killing effect on PANC1 when Photosan was administrated as 2 mg/L (0.081±0.024 and 0.049±0.013vs 0.211±0.031,P＜0.05 and P＜0.01 );all doses of irradiation had killing effect when Photosan was administrated as 4 mg/L.There was no significant difference between 5 and 10 J/cm~2 irradiation in term of killing effect.Cell apoptosis rates with 0 or 2 or 4 mg/L Photosan and 10 J/cm~2 irradiation were(13.8±1.8) %,(40.9±1.6)%,(62.5±2.0)%,the difference was statistically significant(P＜0.05).Conclusions Photosensitizer or irradiation alone did not produce PDT effect.With certain dose of photosensitizer and irradiation,the PDT effect increased accordingly.