ABSTRACT
To investigate the microRNA (miR)-150 expression level in human osteosarcoma cell lines (Saos-2, MG-63) and its function in cell proliferation, and to explore the potential molecular mechanisms. Methods: MiR-150 expression levels in human osteosarcoma cell lines (Saos-2, MG-63) and normal osteoblast cell line (NHOst) were detected by relative quantitative real-time PCR (qRT-PCR). MiR-150 was overexpressed in Saos-2 and MG-63 cells by lentivirus infection. Cell proliferation rates were monitored by MTS assay. RUNX2 and β-actin protein levels were examined by Western blot. Inhibitory effect of miR-150 on binding RUNX2 3'-UTR was detected by Dual-Luciferase assay. Results: MiR-150 expression level is lower in human osteosarcoma cell lines (Saos-2, MG-63) compared to the normal osteoblast cell line (NHOst) (0.23±0.02 and 0.32±0.03 vs 1.00±0.02), which showed statistical significance (P<0.01). After lentivirus infection, miR-150 level increased in Saos-2 (P<0.01) and MG-63 cells (P<0.01). Overexpression of miR-150 decreased cell proliferation and RUNX2 protein level in Saos-2 and MG-63 cells. The binding of miR-150 to RUNX2 3'-UTR decreased luciferase activity by 69% in Saos-2 cells (P<0.05) and 59% in MG-63 cells (P<0.05). Administration of exogenous RUNX2 recovered the cell proliferation in miR-150 overexpressed Saos-2 and MG-63 cell lines (P<0.01). Conclusion: MiR-150 inhibites proliferation in human osteosarcoma cell lines through binding to RUNX2 3'-UTR, resulting in the reducion of RUNX2 protein level.
Subject(s)
Humans , 3' Untranslated Regions , Actins , Metabolism , Bone Neoplasms , Cell Line, Tumor , Cell Proliferation , Genetics , Core Binding Factor Alpha 1 Subunit , Genetics , Pharmacology , Physiology , Gene Expression Regulation, Neoplastic , Genetics , MicroRNAs , Pharmacology , Osteoblasts , Physiology , Osteosarcoma , GeneticsABSTRACT
Objective To study the change of expression of c-fos and c-jun when using IN-1 alone or combination with NT-3 after spinal cord injury. Methods 120 adult health Sprague-Dawley (SD) rats were random divided into three groups, including control group, IN-1 group, and IN-1 combination with NT-3 group. All rats were killed at the scheduled time and its myeloid tissues were taken out. In each group, the expression of c-fos and c-jun gene was detected by using reverse transcription- polymerase chain reaction technique ( RT-PCR ). Result The transcriptional levels of c-fos decreased and c-jun increased when using IN-1 alone, and the levels changed more when using IN-1 combination with NT-3. The peak of c-fos reached to 0. 974 ±0. 126 in control group, 0. 834 ±0. 047 in IN-1 group, and 0. 698 ±0. 052 in IN-1 combination with NT-3 group, and the peak of c-jun reached 0. 642 ±0. 048, 0. 712 ±0. 050, and 0. 814 ±0. 041, respectively. Conclusion One of the mechanisms of IN-1 and NT-3 to protect the spinal cord might be through inhibiting the expression of c-fos and enhancing the expression of c-jun.
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Objective To observe the changes of calcitonin gene-related peptide(CGRP) -immunoreactive positive nerve fibers innervation in bone tissue of femoral heads during the pathological process of early steroid-induced avascular necrosis of the femoral head (SANFH), and explore its significance.Methpathological group of SANFH was induced.Immunohistochemical technique was used and the changes of CGRP-immunoreactive positive nerve fibers innervation in weight bearing area of the femoral heads during the pathological process of early SANFH were observed.Result The number of CGRP-immunoreactive nerves increased first and then decreased ( Peaked at 6 weeks, 10.28 ± 0.66 ), but it was more than that ofnormal control group.There was significant difference between two groups ( P ＜ 0.01 ).Conclusion There were changes in the distribution of CGRP-immunoreactive positive nerves fiber during the process of bone repair after SANFH.CGRP-immunoreactive positive nerves fiber might take part in the process of bone repair in SANFH.
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@#ObjectiveTo investigate the early diagnostic value of serum level of myelin basic protein (MBP) on spinal cord injury (SCI).Methods54 rabbits were divided randomly into slight, moderate and severe SCI groups, and SCI degrees of each group were judged by cortical somatosensory evoked potential, histopathologic examination and Tarlov′s score. ELISA was used to determine the serum levels of MBP before and after SCI.ResultsThe serum levels of MBP of rabbits in three groups were significantly different at 24th, 48th and 72nd hour after injured (P<0.05), but there were not differences at 7th day (P>0.05).ConclusionThe serum level of MBP can be used as a early diagnostic marker for SCI.
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Objective To analyse the application effect of intramedullary interlocking nail in the treatment of femoral and tibial fracture,to discuss the key of the mini-invasive operation.Methods 110 cases of femoral and tibial fracture treated by intramedullary nail from May,1997 to Oct,2001 were reviewed,78 cases were treated by mini-invasive open-reduction and internal fixation,32 cases were treated by intermal fixation without open-reduction.Results All cases were followed-up ,15 cases were delayed at the healing time,but all cases were healed in six months.Conclusion The application of intramedullary nail in the treatment of femoral and tibial frature by mini-invasion has advantages of less trauma,strong fixation,high healing rate,early motion can be obtanied ,but the key of the operation must be grasped.
ABSTRACT
Objective: To report a newly designed method for treating supracondylar fracture of the humerus,and to inquire into the problems of cubitus varus deformity and joint stiff.Methods:Based on a review of anatomy,a new method which called tension band with sphenoid-shaped osteotomy through posterior approach was designed and applied for 46 cases of patients in clinic.The clinical effect was compared with effect of crossing Kirchner's wire.Results:The tension band with sphenoid-shaped osteotomy was a better method and can abide by anatomical and biomechanical principles.After follow-up for 12-36 months,curative effect was estimated as follow:30 cases were excellent,14 cases good and 2 cases poor according to Home's evaluation.Conclusions:It is a better methods in treating supracondylar fracture of the humerus following advantages including simple,reliable fixation and early mobilization etc.