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Objective To discuss the effect of H2 on osteoclast differentiation of RAW264.7 cells induced by RANKL and RANKL/TNF-α.MethodsRAW264.7 cells were treated with H2 in the presence of RANKL and RANKL/TNF-α.RAW264.7 cells viability was assessed by CCK-8.Test the Oxidative Stress of the induced RAW264.7.The number of TRAP-positive cells were counted under light microscopy.The levels of cathepsin K (CTK) and matrix metalloprotein-9(MMP-9) mRNA were analyzed by real-time PCR.ResultsH2 can not influence the RAW264.7 cell viability but can lower oxidative stress.The significant difference(P<0.05) indicated that H2 could significantly decrease the number of TRAP-positive MNCs.The significant difference among the 4 groups in CTK and MMP-9 genes (P<0.05) indicated that H2 could down-regulate their mRNA expression.ConclusionH2 can reduce the oxidative stress and inhibit differentiation of RAW264.7 cells into osteoclasts.
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Objective To explore the improvement effect of rehabilitation nursing on joint swelling and pain after total knee arthroplasty.Methods According to the digital table,100 patients with total knee arthroplasty were randomly divided into observation group and control group,50 cases in each group.The control group after operation used routine nursing care,the observation group was given targeted rehabilitation nursing.They were compared with visual analogue scale (VAS),American Knee Society Score (AKSS),range of motion (ROM),length of hospital stay,joint swelling degree.Results Mter treatment,the VAS score of the observation group was lower than the control group[(2.43 ±0.51) points vs.(4.58 ± 0.81) points,t =15.882,P ＜ 0.05],AKSS scores and ROM were higher than those in the control group[(69.94 ± 14.23)points vs.(53.23± 12.91)points,(103.47 ± 14.84)° vs.(91.23 ±12.53) °,t =6.149,4.456,all P ＜ 0.05].The hospitalization time of the observation group was shorter than that of the control group[(14.23± 3.81)d vs.(17.82 ± 4.03) d,t =4.577,P ＜0.05].At 3d and 7d after operation,the swelling degree of the observation group was lower than those of the control group[(1.23 ±0.26)mm vs.(1.97 ±0.38)mm,(0.62 ± 0.21) mm vs.(1.38 ± 0.35) mm,t =11.364,13.166,all P ＜ 0.05].Conclusion The application of rehabilitation nursing after total knee joint surgery can effectively relieve joint swelling and pain,and it is worth to be popularized.
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Objective To determine the lumbar anatomical structure parameters of the lumbar spine posterior column and its sample data of three-dimensional conformation,and based on these data to design the lumber laminar screw dynamic fixation system.Methods 20 human L3 ～ S1 bone specimens,the thickness of the lumber laminar region were measured to obtain the normal anatomical parameters of the lumbar laminar.And 20 healthy lumbar vertebrae L3-S1 for three-dimensional reconstruction were detected by CT scan,which could obtain the structure data of the normal lumbar posterior three-dimensional conformation,and to obtain conformational sample data of three-dimensional space of the posterior structure.Results Lumbar lamina medial 1/3 could serve as a spine laminar screw setting area by anatomical observations and measurements.Lamina thickness of L3-4 on average (6.6 ±0.9) mm,L5 ～ S1average (6.0 ± 0.6) mm.Lumber laminar screw length 6.5 ～ 8.0mm could meet the requirements of bilateral cortical fixation.Conclusion Lower lumbar spinal lamina area can serve as the region to fix the bilateral cortical bone screw,and the optimal length of the laminar screw is 6.5 ～ 8.0rmm.
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BACKGROUND:Neurotrophin-3 is found in the repair of spinal cord injury in the role of the strongest neurotrophic factor.It can effectively promote axonal regeneration through the glial scar tissue in repairing spinal cord injury.OBJECTIVE:To construct recombinant lentiviral vectors for gene delivery of homo sapiens neurotrophin-3(hNT3),and to investigate the expression of hNT3 gene in Schwann cells after transfection.DESIGN,TIME AND SETTING:Observational experiment was performed from June 2007 to March 2008 at the Central Laboratory of Changhai Hospital.MATERIALS:Bilateral sciatic nerves were harvested from 3-day-old Sprague Dawley rats for culture and identification of Schwann cells.Three-plasmid lentivirus systems:pGC-E1-EGFP,pHelper 1.0 and pHelper 2.0 were gained from Shanghai Chemical Technology Co.,Ltd.METHODS:pGC-E1-hNT3-EGFP plasmid was constructed by double restriction enzyme digestion and ligation,and then the plasmid was transformed into E.coli DH5?.Purified pGC-E1-hNT3-EGFP plasmids from the positive clones was confirmed by PCR and sequencing.293T cells were cotransfected with lentiviral vector pGC-E1-hNT3-EGFP,pHelper 1.0 and pHelper 2.0 by Lipofectamine 2000 to produce lentivirus.2 mL recombinant virus complete culture solution was added according to multiplicity of infection=1,4,8,10,12.The titer of virus was tested according to the expression level of enhanced green fluorescent protein.The control groups were Schwann cells and Schwann cells transfected by no-loaded lentivirus.MAIN OUTCOME MEASURES:The lentiviruses were transduced to Schwann cells,and the transfection efficiency was examined by flow cytometry,the overexpression of hNT3 was determined by Real-time PCR and Western blotting.RESULTS:The exogenous gene sequence of the recombinant hNT3 was completely in accordance with that of its open reading frame in GeneBank.The titer of concentrated virus was 5?107 TU/L.After recombinant LV-hNT3 infection,Schwann cells gave off strikingly bright green fluorescence,and the transfection efficiency amounted to 85%(multiplicity of infection=10).Real-time RCR test showed that the hNT3 mRNA were highly expressed in hNT3-Schwann cells,while did not express in the control group.Western blotting showed the hNT3 expression in Schwann cells.CONCLUSION:Construction of hNT3 gene lentiviral vector can transfect Schwann cells leading to an efficient overexpression of hNT3.