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Objective To prepare specific mouse monoclonal antibodies against Homo sapiens dynein,axonemal, heavy chain 2 (DNAH2). Methods Firstly, recombinant plasmid encoding His tagged immunogen, targeting N-terminal sequence of DNAH2 protein (1-300 aa), in E. coli was constructed. IPTG was used to induce the expression of His-immunogen, which was then purified and immunized in BALB/c mice. Hybridoma cells were obtained through the fusion between myeloma cells and splenocytes isolated from BALB/c mice. Finally, ELISA and Western blot assays were performed to screen the positive hybridoma. Results IPTG was used efficiently to induce the expression of DNAH2 immunogen in E. coli. DNAH2 protein bands were detected in screened positive hybridoma. Conclusion Mouse monoclonal anti-DNAH2 antibody is prepared successfully.
RESUMO
[Objective]To assess the accuracy and value of radiographic technique using computed tomography(CT) measurements in cadaveric specimens during anterior cervical corpectomy in order to define the safe lateral limit of dissection.[Method]The gutter of C4～6 corpectomy on 5 cadaveric samples was filled with radioopaque dye.An antero-posterior X-ray was used to measure the narrowest distance from dye column to the line of uncovertebral joints.A CT scan confirmed the distance from the borderline of the gutter to vertebral arteries.[Result]The distance between the X-ray dye column and the line of uncovertebral joints averaged 4.7 mm in the left and 4.2 mm in the right.While the distance from the borderline of the gutter to vertebral arteries with CT scan averaged 6.1 mm in the left and 5.9 mm in the right.The measured distance by radiographic technique underestimated the true distance.[Conclusion]Intraoperative radiographic technique can estimate the lateral distance between the gutter of corpectomy and vertebral arteries by measuring the distance from the dye column to the line of uncovertebral joints.This technique provides surgeons an additional margin of safety during anterior cervical corpectomy.