Expression and purification of CDNF and preparation of its polyclonal antibodies / 中国免疫学杂志

Objective: To obtain purified and functional CDNF-his recombinant protein and prepare its polyclonal antibodies.Methods:Preparation of recombinant CDNF-his was carried out in HEK 293 T cells with pVR1012-CDNF-his successfully constructed transfected into them.Then,the recombinant protein was purified by Ni-NTA immunoaffinity chromatography.The purity was analyzed by SDS-PAGE and the protein′s identity was tested by Western blot.MTT was used to verify the biological function of the protein purified.New Zealand white rabbits were immunized with purified CDNF-his protein for preparation of polyclonal antibodies.Results:pVR1012-CDNF-his expressed successfully in HEK 293 T cells.The purity of protein was up to more than 90%after purification.MTT showed that CDNF-his was able to protect PC 12 cells from damage by 6-OHDA.The polyclonal antibody was detected at the end of animal immunizing process.Conclusion: A method to express and purify protein using HEK 293T cell and following Ni-NTA immunoaffinity chromatography has been built.CDNF-his with biological activity is obtained based that.Finally, polyclonal antibodies of CDNF were generated successfully.

Study on MRI sequences in the diagnosis of articular cartilage injury of the knee joint / 中国基层医药

ObjectiveTo compare the accuracy of the four sequences of MRI in the diagnosis of articular cartilage injury of the knee and to investigate the advantage and shortage of the four sequences. In order to evaluate the most adequate sequence in detecting articular cartilage injury of the knee. Methods36 knee joints of 31 patients,underwent MRI scanning prior to arthroscopy included T1WI-TSE,T2 WI-TSE-SPIR, PDW-TSE-SPIR,3D-WATSC. Divided each knee side of every cartilage into five parts. Investigated and diagnosed the 180 articular cartilage surface according to Recha criteria by detecting and compared with arthroscopic examination respectively. The accurate criteria of the four sequences of MRI was sensitivity, specificity, accuracy. The criteria of correlation was the value of Kappa.ResultsCompared with the arthroscopic result which was the gold standard, the sensitivity, specificity, accuracy and Kappa value of the T1 W-TSE sequence was 63.8％ ,95.3％ ,78.8％ and 0. 403 respectively;The sensitivity,specificity ,accuracy and Kappa value of the T2WI-TSE-SPIR sequence was 56.3％ ,97.6％ ,76.1％ and 0. 353 respectively.The sensitivity,specificity,accuracy and Kappa value of the PDW-TSE-SPIR sequence was 79. 8％, 93.0％ ,86. 1％and 0. 642 respectively; The sensitivity, specificity, accuracy and Kappa value of the 3D-WATSC was 96. 8％,88. 3％ ,92.8％ and 0. 812 respectively. ConclusionThe accuracy and Kappa values of Fat-suppressed three-dimensiunal fast spoiled gradient-echo sequence (3D-WATSC)was superior to other sequences in the diagnosis of articular cartilage injury of the knee. The correlation between 3D-WATSC and arthroscopy in detecting cartilage injury was remarkable. 3D-WATSC had been deemed the most adequate sequence in detecting articular cartilage injury of the knee.