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.

Aim To study the effect of ginsenoside Rg1(Rg1)on cardio myocyte hypertrophy induced by prostaglandin F2?(PGF2?),and to probe primarily into its mechanism.Methods The cultured neonatal rat cardiomyocyte hypertrophic response and the antihypertrophic effects of Rg1 were observed by measuring the cell diameter,protein content and the expression of atrial natriuretic factor(ANF) mRNA,which was assayed by real-time PCR.For mechanism studies,the intracellular free calcium concentration(i) in cultured cardiomyocytes was measured by using Fura-2/AM as a fluorescent indicator,nitric oxide(NO) metabolite level in the culture medium was tested by using Nitric Oxide Synthase Detection Kit.Results PGF2? 0.1 ?mol?L-1 caused the increases in the cardiomyocyte cell diameter,protein content and the expression of ANF mRNA.It could increase the i in cultured cardiomyocytes.Rg1 15.6、31.2、62.4 ?mol?L-1 could concentration-dependently inhibit the cardiomyocyte hypertrophy induced by PGF2?,and the cell diameter of cardiomyocyte treated by PGF2? was decreased by 18.4%、32.7%、43.8%(P

ObjectiveTo investigate whether fatty liver disease is a risk factor for lacunar infarction. MethodsA retrospective analysis was performed for the clinical data of 457 patients with lacunar infarction (lacunar infarction group) and 120 control patients, who were hospitalized in our hospital from 2007 to 2017, to analyze whether fatty liver disease is a risk factor for lacunar infarction. The chi-square test was used for comparison of categorical data between groups, and the t-test was used for comparison of continuous data between groups. Univariate and multivariate regression analyses were used to screen out the risk factors for lacunar infarction. ResultsThe lacunar infarction group had a significantly higher incidence rate of fatty liver disease than the control group (60.39% vs 39.17%, χ2=17.379, P＜0.001). The multivariate regression analysis showed that after adjustment for age, sex, smoking, drinking, hypertension, and diabetes, fatty liver disease was an independent risk factor for lacunar infarction (odds ratio ［OR］=1.96, 95% confidence interval ［CI］: 1.21-3.02, P=0.003). There was a significant interaction between obesity and fatty liver disease (P=0.001). Non-obese fatty liver disease was an independent risk factor for lacunar infarction (OR=3.29, 95% CI: 1.55-7.23, P＜0.001); however, in the obese subgroup, obese fatty liver disease was not an independent risk factor for lacunar infarction(P=0532)，Age(OR=6.67,95%CI:1.98~121.56,P＜0001), hypertension (OR=638,95%CI:512~12.06,P＜0001) were independent risk factors for lacunar infarction. ConclusionNon-obese fatty liver disease is an independent risk factor for lacunar infarction.

10.3969/j.issn.1000-3606.2013.06.019

Objective To construct a multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector and identify it preliminarily. Methods According to recombinant pcDNA3-p30-ROP2 restriction sites,HBV HBsAg gene sequences of primers were designed and synthesized to amplify target fragment,and then cloned into pcDNA3-HbsAg-p30-ROP2 expression vector. Af-ter sequencing,it was identified finally by restriction enzyme digestion and other molecular biology techniques. Results HBV HBsAg gene segment was amplified by PCR and the multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector was constructed and identified to be correct as theoretical values. The PCR and restriction enzyme digestion results showed that HBsAg and p30-ROP2 gene in recombinant plasmid were confirmed by DNA sequencing. Conclusion The multi-gene recombinant pcD-NA3-HBsAg-p30-ROP2 expression vector is successfully constructed.

Objective To compare the positioning accuracy between personalized polyurethane foam with wing boards and negative pressure vacuum bag in radiotherapy for lung cancer using kilovoltage cone beam computed tomography(CBCT). Methods Thirty-nine patients with lung cancer who received chest radiotherapy in our hospital from 2015 to 2016 were enrolled as subjects. In those patients, 20 were immobilized by negative pressure vacuum bags(VB group)and the others by personalized polyurethane foam with wing boards(PF group).CBCT images were acquired weekly and registered with planning CT images. Setup errors in the left-right, superior-inferior, and anterior-posterior directions, three-dimensional(3D) displacement vector,and setup time were recorded. The margin of the planning target volume(PTV)was calculated using the Van Herk formula(2.5∑+0.7σ). Between-group comparison was made by paired t test. Results The PF group had a significant smaller setup error in the y-direction than the VB group (2.54±1.79 vs.3.19±2.14 mm,P=0.03),while there were no significant differences in setup errors in the x-or z-direction between the two groups(1.80± 1.48 vs. 1.90± 1.41 mm, P=0.46;2.14± 1.75 vs. 2.25± 1.75 mm,P=0.35). There were no significant differences in rotational setup errors in the Rx-,Ry-,or Rz-direction between the two groups(1.15°±0.76°vs. 1.15°±0.85°, P=0.50;0.71°±0.60°vs. 0.72°±0.43°, P=0.45;0.62°±0.54° vs. 0.46°±0.30°,P=0.06). The PTV margins in the x?,y?,and z?directions were expanded by 5.56, 8.57, and 7.02 mm, respectively, in the PF group, and by 5.62, 9.27, and 7.23 mm,respectively,in the VB group. The proportion of patients with 3D displacement vectors larger than 5 mm was 40% in the PF group and 45% in the VB group.Conclusions For patients undergoing radiotherapy for lung cancer,personalized polyurethane foam with wing boards can,to a certain extent,reduce the setup error in the superior-inferior direction and PTV margin expansion.[Key words] Lung neoplasms/radiotherapy; Polyurethane foam; Vacuum bag; Setup errors;Margin