ABSTRACT
<p><b>OBJECTIVE</b>The purpose of the present study was to observe the changes in CD4+CD25+Nrp1+Treg cells after irradiation with different doses and explore the possible molecular mechanisms involved.</p><p><b>METHODS</b>ICR mice and mouse lymphoma cell line (EL-4 cells) was used. The expressions of CD4, CD25, Nrp1, calcineurin and PKC-α were detected by flow cytometry. The expressions of TGF-β1, IL-10, PKA and cAMP were estimated with ELISA.</p><p><b>RESULTS</b>At 12 h after irradiation, the expression of Nrp1 increased significantly in 4.0 Gy group, compared with sham-irradiation group (P<0.05) in the spleen and thymus, respectively, when ICR mice received whole-body irradiation (WBI). Meanwhile the synthesis of Interleukin 10 (IL-10) and transforming growth factor-β1 (TGF-β1) increased significantly after high dose irradiation (HDR) (> or = 1.0 Gy). In addition, the expression of cAMP and PKA protein increased, while PKC-α, calcineurin decreased at 12h in thymus cells after 4.0 Gy X-irradiation. While TGF-β1 was clearly inhibited when the PLC-PIP2 signal pathway was stimulated or the cAMP-PKA signal pathway was blocked after 4.0 Gy X-irradiation, this did not limit the up-regulation of CD4+CD25+Nrp1+Treg cells after ionizing radiation.</p><p><b>CONCLUSION</b>These results indicated that HDR might induce CD4+CD25+Nrp1+Treg cells production and stimulate TGF-β1 secretion by regulating signal molecules in mice.</p>
Subject(s)
Animals , Female , Male , Mice , Calcineurin , Genetics , Metabolism , Cyclic AMP , Metabolism , Dose-Response Relationship, Radiation , Gene Expression Regulation , Radiation Effects , Immunosuppression Therapy , Interleukin-10 , Genetics , Metabolism , Lymphocyte Subsets , Physiology , Neuropilin-1 , Genetics , Metabolism , Phosphoinositide Phospholipase C , Genetics , Metabolism , Protein Kinases , Genetics , Metabolism , Signal Transduction , Transforming Growth Factor beta , Genetics , Metabolism , Whole-Body IrradiationABSTRACT
<p><b>OBJECTIVE</b>To label human insulin-like growth factor-I (hIGF-I) eukaryotic expression vector with green fluorescent protein (GFP) for the repair of articular cartilage defects.</p><p><b>METHODS</b>GFP cDNA was inserted into pcDNA(3.1)-hIGF-1 to construct the co-expression vector with two multiple cloning sites mammalian expression vector under two cytomegalovirus promoters/enhancers respectively. Recombinant pcGI was transfected into NIH 3T3 cells with the help of lipofectamine.</p><p><b>RESULTS</b>Enzyme digestion and agarose gel electrophoresis analysis revealed that pcGI vector contained correct GFP and hIGF-I cDNA. Expression of hIGF-1 and GFP was confirmed in transfected NIH 3T3 cells by immunocytochemical analysis and fluorescence microscopy.</p><p><b>CONCLUSIONS</b>hIGF-I eukaryotic expression vector has been successfully labeled with GFP.</p>
Subject(s)
Humans , Cartilage, Articular , Wounds and Injuries , Cells, Cultured , Eukaryotic Cells , Cell Biology , Physiology , Fibroblasts , Cell Biology , Physiology , Gene Expression Regulation , Genetic Therapy , Methods , Genetic Vectors , Green Fluorescent Proteins , Pharmacology , Immunohistochemistry , Insulin-Like Growth Factor I , Genetics , Therapeutic Uses , Luminescent Agents , Pharmacology , Sensitivity and Specificity , Transfection , Methods , Wounds and Injuries , Diagnosis , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the feasibility of the pedicle screw pilot holes placement in thoracic spine using the spiral wires as the guide pin.</p><p><b>METHODS</b>The pedicle screw pilot holes were drilled within the center of the pedicle and the lateral and medial pedicle walls were violated in 9 human dried thoracic vertebrae. Kirschner wires or spiral wires were separately placed in the holes, and then the posteroanterior and lateral radiographs were taken. The radiographs were evaluated by 3 experienced spine surgeons and 3 young orthopedists. After radiographs were shown to these observers, they combined the posteroanterior and lateral radiographs in each place and determined whether the pedicle screw pilot hole violated the pedicle cortex or not. The results were analyzed by a statistical software.</p><p><b>RESULTS</b>Sensitivity, specificity and accuracy of the method using spiral wires to detect pedicle pilot hole placement were significantly higher than those of using Kirschner wires. With a true posteroanterior radiograph, the sensitivity, specificity and accuracy of the method using spiral wires approximated or attained 100%.</p><p><b>CONCLUSIONS</b>The method of intrapedicular pilot hole placement verification using spiral wires is effective for guiding the accurate placement of pedicle screws.</p>