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Article in Chinese | WPRIM | ID: wpr-313813


<p><b>OBJECTIVE</b>To construct Epithelia Membrane Protein 1 gene-deficient in human fetal nucleus pulposus model by lentivirus-mediated RNA interference for building a platform for illustrating the biomechanisms role of EMP-1 during human intervertebral disc degeneration.</p><p><b>METHODS</b>The lentivirus vector with shRNA targeting EMP-1 mRNA was transected into 293FT cells by liposome. Then the lentivirus supernatant was obtained and used for infecting human fetal nucleus pulposus. The expression of GFP was observed under fluorescence microscope after 48 h. The viral particles were collected at 72 h after transfection. The efficacy of gene interference was tested by Western blot and Real-time RT-PCR. Analysis the results of the fluorescent microscope scenes and get the average values of EMP-1/GAPDH by detected the interference efficiency of various interference DNA sequences with western blot and semi quantitative RT-PCR methods.</p><p><b>RESULTS</b>The lentivirns with high titer were obtained and the EMP-1 gene deficient cell strains were obtained. Semi quantitative RT-PCR and Western blot proved the average values of EMP-1/GAPDH decreased from 0.46 to 0.32 and 0.5 to 0.25 (P < 0.01).</p><p><b>CONCLUSION</b>Lentivirus packaging technology can be mastered skillfully. EMP-1 gene-deficient cell models are successfully established.</p>

Fetus , HEK293 Cells , Humans , Intervertebral Disc , Metabolism , Lentivirus , Genetics , Neoplasm Proteins , Genetics , RNA Interference , Receptors, Cell Surface , Genetics , Transfection
Article in Chinese | WPRIM | ID: wpr-297801


<p><b>OBJECTIVE</b>To study the curative effect of surgical treatment of drug-resistant spinal tuberculosis.</p><p><b>METHODS</b>From March 2005 and April 2009, the clinical data of 60 patients with drug-resistant spinal tuberculosis were retrospectively analyzed. Including 36 males and 24 females; aged from 5 to 79 years with an average of 47.3 years. Thirty-four patients had neurological deficits, among them, 2 cases were grade A, 5 cases were grade B, 13 cases were grade C, 14 cases were grade D according to ASIA standard. According to the severity and location of the infection, the patients underwent anterior, posterolateral costotransversectomy or posterior debridement and bone grafting and internal fixation. The antituberculous chemotherapy for a total of 12 to 18 months was guided by conventional and genotypic drug susceptibility testing. Tubercular relapse, neurological function, spinal fusion were observed by ASIA grade, X-ray and CT scan.</p><p><b>RESULTS</b>All cases were followed up from 1 to 5 years with an average of 3.1 years. Recurrence was found in 2 cases who were cured after second operation. 34 cases with neurological deficits recovered totally or partially. X-ray or CT films showed spinal fusion in 57 patients.</p><p><b>CONCLUSION</b>The therapeutic effect of individuall operative options is good in treating drug-resistant spinal tuberculosis after antituberculous chemotherapy based on conventional and genotypic drug susceptibility testing.</p>

Adolescent , Adult , Aged , Antitubercular Agents , Therapeutic Uses , Child , Child, Preschool , Drug Resistance, Bacterial , Female , Humans , Male , Middle Aged , Mycobacterium , Genetics , Radiography , Retrospective Studies , Spine , Tuberculosis, Multidrug-Resistant , Diagnostic Imaging , Drug Therapy , Microbiology , General Surgery , Tuberculosis, Spinal , Diagnostic Imaging , Drug Therapy , Microbiology , General Surgery , Young Adult