Objective:To study feasibility of preparing artificial antigen by membrane coated with hapten-carrier.To compare the Emodin-BSA membrane antigen immunogenicity and specificity against the liquid antigen.Methods:Emodin-BSA-PVDF membrane was prepared by the method that BSA was coated on PVDF membrane and the BSA was coupled with Emodin-couplint agent derivative.Rats were immunized by subcutaneous implantation.The immunogenicity and antibody specificity were characterized using Emodin-CA or Chrysophanol-CA or Physcion-CA membrane immunoassay. Results: The immunogenicity of Emodin-BSA coated membrane antigen was higher than Emodin-BSA liquid antigen;the specificity for three anthraquinones was almost the same(P>0.05). Conclusion:Emodin antiserum generated using Emodin-BSA coated membrane antigen has a high immunogenicity and specificity to Emodin.The results show it is feasible that membrane coated with hapten-carrier is used as artificial antigen.

Objective:The recombinant shuttle peptide-single chain antibody with neutralizing activity is expected to be obtained by combining the shuttle peptide sequence with rabies single chain antibody and optimizing the expression conditions in E. coli.Methods:In this study, the SO57 single-stranded antibody sequence was recoded and the shuttle peptide of Arg 12 was fused at the N-terminal to construct pET22 (b)-rSO57 expression vector, which was then expressed in E. coli. The growth density OD 600, induction time, induction temperature and inducer concentration were optimized to obtain a higher expression effect. The recombinant protein was purified by immobilized metal chelating chromatography, and the relative affinity of the recombinant single-chain variable fragment (scFv) was determined. The neutralization effect of the recombinant scFv was verified by rSO57/CVS-11 virus strain mixed with infected cells and rapid fluorescent focus inhibition test (RFFIT). Results:The recombinant rSO57 was successfully constructed and induced in BL21 (de3). The optimal expression condition was that when the cell density of OD 600 was 0.8, IPTG of 0.4 mmol/ml was used for induction. After induction at 16 ℃ for 24 hours, rSO57 was expressed in soluble form, accounting for 19.8% of the soluble protein in the cell. After chromatography purification, the recombinant rSO57 with a purity of 84% was obtained. ELISA showed that the relative affinity coefficient Ka of rSO57 was 4.3×10 5. When rSO57 was mixed with CVS-11 strain, the infection rate of cells increased with the increase of dilution, indicating the neutralization activity of recombinant antibodies. Using the RFFIT, 50 μg rSO57 was equivalent to 2.17 IU of rabies neutralizing serum. Conclusions:In this study, the recombinant scFv fusion with shuttle peptide was expressed, which could neutralize rabies virus, in order to prepare specific and targeted antiviral drug carriers for rabies treatment.

Objective To explore the application effect of 3D printing technology applied in nursing teaching of neurosurgery.Methods A total of 76 nursing students who had internship in the neurosurgery of our hospital from July 2015 to March 2016 were randomly divided into the control group and the intervention group by computer random sequence,with 38 cases in each group. Traditional PPT instruction method was applied in the control group,while 3D printing technology was applied in the teaching of intervention group,and it was used to print the 3D models of patients′ skull,intracranial tumor,encephalic blood vessel,etc. Teaching effect of two groups was observed and compared by theoretical examination,self-evaluation and mutual evaluation between nursing students and teachers,and teaching effectiveness.Results The score of theoretical knowledge in the intervention group(90.05±0.74),was higher than that in the control group(82.08±1.04)(t=6.22,P<0.01). The total score of nursing examination in the intervention group was(92.95±0.41),which was higher than that in the control group(80.05±0.87)(t=13.29,P<0.01). Mutual evaluation satisfaction of nursing students and teachers in the intervention group were 92.1% and 97.4%,and they were higher than those in the control group which were 84.2% and 81.6%(P>0.05).Conclusions The application of 3D printing technology in nursing teaching of neurosurgery can effectively improve interns′ comprehensive ability in neurosurgery theoretical knowledge,enhance students′ learning enthusiasm and initiative,and raise the general level of interns.