[Objective] To study the bone metabolic biochemical index of the aged patients with hip fracture,for better predicting the future risk of the old people' s hip fracture.[Method]50 cases of sufferers(over 60 years old) with hip fracture(28 males,and 22 females) and 30 cases of healthy aged people(15 males,and 15 females) were selected to analyze Ⅰ Collagen crosslinked c-telopeptide(ICTP),deoxypyridino line(Dpd) in urine,and serum bone glaprotein(BGP).[Result](1)The mean level of ICTP and Dpd in urine in aged hip fracture group was higherthan that of the control group(P0.05).[Conclusion]Bone absorbability in the aged hip fracture patients is higher than in the aged healthy people.The analysis of ICTP and Dpd in urine may /might give some reference value in preventing and treatlng aged hip fracture patients.

Background and purpose:The screening of the genes being related closely with the mechanism of osteosarcoma metastasis was a difficult point in the realm of orthopaedics.We screened differential expression gene of human osteosarcoma MG-63 cell sublines with different metastatic capabilities with cDNA microarray,and studied the molecular mechanism of osteosarcoma metastasis.Methods:Total RNA of human osteosarcoma MG-63 cell sublines A1 and A2 was extracted,purified to mRNA and then reversely transcripted to cDNA probe respectively.The cDNA probe of A1 was labelled with Cy3 and the cDNA probe of A2 was labelled with Cy5.The two samples were hybridized with the cDNA microarray.The hybridization signals were scanned by Agilent Scanner and obtained data were analyzed using Ima Gene 3.0 software and Genespring software.Results:222 differential expression genes were found between cell sublines A1 and A2 by analyzing gene expression profile.There were 119 upregulated genes and 103 downregulated genes in cell sublines A1.All differential expression genes belonged to six main function groups and 49 genes of these had very obvious differentce in expression.Conclusions:There were many differently expressed genes between A1 and A2 cell sublines and only part of them were closely associated with mechanism of osteosarcoma metastasis.The technology of cDNA microarray could analyze effectively gene expression profile of human osteosarcoma MG-63 cell sublines, and supply a new approach to study the mechanism of osteosarcoma metastasis

[Objective]To investigate the relationship between degeneration of cartilage and the expressions of Matrix Metalloproteinase 7(MMP-7),Matrix Metalloproteinase 9(MMP-9),Matrix Metalloproteinase 13(MMP-13)and Tissue Inhibitor of Matrix Metalloproteinase 1(TIMP-1)in osteoarthritis.[Method]The histological changes of cartilages by hematoxyllin-eosin staining and immunohistochemical expression of Matrix Metalloproteinase 7(MMP-7),Matrix Metalloproteinase 9(MMP-9),Matrix Metalloproteinase 13(MMP-13)and Tissue Inhibitor of Matrix Metalloproteinase 1(TIMP-1)in osteoarthritis.were studied in 20 osteoarthritis cases and 2 normal controls.All data were statistically analyzed by Mann-Whitney U test and correlation analysis.[Result]The osteoarthritis cartilage underwent fibroplasias and tearing.The quantity of chondrocyte increased and the clustered and hypertrophic cells came into being.Little immunostaining of MMP-7 and MMP-13 was observed in normal cartilage,while their expressions increased in degenerated cartilage(P0.05),superficial layer of the moderate and end-stage osteoarthritis.However,it significantly increased in deep layer(P

[Objective]To prove the feasibility of using the distance between sagittal plane of the spinal process and the enter point to individualize the placing of pedicle screw.[Method]Thirty spine specimen were collected and divided into two groups,data were measured,such as the width of the pedicle,distance between the enter point and anterior border of the vertebra,distance between sagittal plane of the spinal process and the enter point,angle from the longitudinal axis of the pedicle to sagittal axis of the vertebra,angle from the longitudinal axis of the pedicle to vertical line of the operating table.In group one the pedicle screws were placed with the help of the distance between sagittal plane of the spinal process and the enter point,the other by the method advised by Ebraheim.CT scan was applied to evaluate the place of the screws,according to the perforation extent,they were classified into 4 grades:A=totally in the pedicle;B=perforation extent4mm.[Result]The individualized group showed much lower perforation rate than the traditional method group in T3~10,and similar in T1,T2,T11,12.[Conclusion]It can obviously improve the accuracy of the pedicle screw placement to use the distance between sagittal plane of the spinal process and the enter point to localize the enter point,especially when anatomic landmark such as articulationes zygapophysiales and transverse process change.

BACKGROUND:Atlantoaxial fusion is currently the main surgical treatment of atlantoaxial dislocation, but the premise is at the expense of atlantoaxial range of motion, especial y the rotation motion. Restricted non-fusion fixation is a method that can maintain the atlantoaxial stability, while retain the atlantoaxial range of motion. Further research should be performed to compare the biomechanical characteristics between the two methods. OBJECTIVE:To develop a three-dimensional finite element model of atlantoaxial instability, compare and determine the biomechanical properties of posterior atlantoaxial restricted non-fusion fixation system and posterior atlantoaxial screw-rod fixation system. METHODS:A verified intact finite element upper cervical (C0-C3) model was established and analyzed by Simpleware 3.0, Geomagic 8.0, Hypermesh 10.0, Abaqus 6.9, and Rhino 4.0 softwares based on the CT data col ected from a 31-year-old healthy male volunteer. The moment couple of 1.5 N?m was loaded, which made the model movement in flexion-extension, lateral bending, and rotating direction, respectively. The range of motion was recorded and compared with the in vitro biomechanical experimental data to verify the effectiveness of the model. The ranges of motion of the posterior atlantoaxial restricted non-fusion fixation system model and the posterior atlantoaxial screw-rod fixation system model were analyzed using the finite element method under flexion, extension, lateral bending, and axial rotation;meanwhile, stress nephograms of the posterior atlantoaxial restricted non-fusion fixation system model were observed. RESULTS AND CONCLUSION:(1) There were 206 747 elements and 72 500 nodes in the intact model of upper cervical spine (C0-C3) in this experiment, and the range of motion of intact model validated with the reported cadaveric experimental data. (2) The range of motion of the posterior atlantoaxial restricted non-fusion fixation system group was similar to which of the posterior atlantoaxial screw-rod fixation system group in flexion-extension direction. (3) In lateral bending direction, the range of motion of the posterior atlantoaxial restricted non-fusion fixation system model was obviously limited, respectively. The range of motion of the posterior atlantoaxial restricted non-fusion fixation system model was larger than that of the atlantoaxial dislocation model and basical y same as that of the normal atlantoaxial model. (4) As to the rotating direction, the range of motion of the posterior atlantoaxial restricted non-fusion fixation system mainly disappeared at the atlantoaxial segment;by contrast, a majority of rotating motion was stil retained in the posterior atlantoaxial restricted non-fusion fixation system group. (5) The stress concentration occurred in the contact part between the screw and the connecting rod in posterior atlantoaxial restricted non-fusion fixation system model. (6) Results suggest that posterior atlantoaxial restricted non-fusion fixation system is effective and useful for atlantoaxial fixation. It not only restricted atlantoaxial flexion-extension, but also preserved axial rotation and lateral bending at the atlantoaxial joint.

BACKGROUND:Current research has shown that many cellgrowth factors can promote the proliferation and differentiation of epiphysis stem cells, and this regulation is closely related to Sox-9.
OBJECTIVE:To investigate the effects of smal interfering RNA-induced specific silence of Sox-9 gene on the proliferation and apoptosis of epiphysis stem cells.
METHODS:Epiphysis stem cells were isolated from the ring of La Croix with enzyme digestion and purified with magnetic activated cellsorting, identified by immunocytochemistry assay. The cells in good conditions were selected to be transfected by Sox-9 silenced by smal interfering RNA with fluorescent marker, and then were observed under the fluorescent microscope to check the transfection efficiency. Next step, epiphysis stem cells were divided into 2 groups:group one was cultured normal y as control group;group two was transfected by Sox-9 silenced by smal interfering RNA through LipofectamineTM 2000 as experimental group.
RESULTS AND CONCLUSION:The primary epiphysis stem cells were separated and purified, which were of stable growth and tightly attachment. The results of immunohistochemistry and immunofluorescence showed the epiphysis stem cells expressed cel-specific markers, fibroblast growth factor receptor 3. After transfection, reverse transcription-PCR results showed that Sox-9 gene expression in epiphysis stem cellwas inhibited specifical y;3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide results showed that cellproliferation in experimental group decreased significantly compared with the control group (P<0.05), and the cellapoptosis detected by flow cytometry showed that the experimental group increased as compared with the control group (P<0.05). Sox-9 gene plays an important role in regulating the proliferation and apoptosis of rat epiphysis stem cells.

BACKGROUND: Precartilaginous stem cells (PSCs) have strong proliferation ability and differentiation potential,but they are instable and prone to differentiate.Importing exogenous gene could immortalize them and leave phenotype character unchanged.OBJECTIVE: To establish immortalized precartilaginous stem cells (PSCs) from neonatal SD rats in vitro for the further related research about the differentiation mechanism and clinical application of precartilaginous stem cells.DESIGN,TIME AND SETTING: Single sample observation.The study was carried out in the Department of Orthopedics.Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from October 2005 to September 2006.MATERIALS: Neonatal SD rats,irrespective of gender,24-hour old,were used for prepare PSCs.METHODS: By using LipofectamineTM 2000,a gene transfection reagent,plasmid pCMVSV40T/PUR containing the simian virus 40 large T antigene gene (SV40Tag) was transfected into the primary cultured PSCs isolated by immuniomagnetic beads coasted with the second antibody.Colonies were isolated by puromycin selection and expanded by many passages.MAIN OUTCOME MEASURES: Biological character of PSCs; plasmid identification; biological character of transfected cells and identification; RT-PCR; growth curve.RESULTS: Immunomagnetic beads separation system obtains PSCs,which was confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive PSCs.Double restriction enzyme was cut,electrophoresis confirmed pCMV was 3 kb,SV40T was 2.3 kb.A particular anti-puromycin cell clone was acquired,which was confirmed as FGFR-3 positive PSCs.The total RNA was isolated from the positive cell clones,and a 588 bp fragment,which was specific for the SV40T antigene gene,was amplified.The transfected cells were expanded to immortalized cell strain,named as immortalized precartilaginous stem cells (IPSCs).Thepopulation doubling time of IPSCs was (22.98±2.77) hours,no significant effect of subculture,freezing and recovering had been found.CONCLUSION: Precartilaginous stem cells could be isolated from neonatal SD rats,cultured in vitro,and immortalized through the transfection of pCMVSV40T/PUR.

BACKGROUND: The precartilaginous stern cells are limited regarding in vitro proliferative capacity, but the immortalized cell lines can provide a large number of stable immortalized cells, and simian virus 40 large T antigen gene (SV40Tag) is one of gene fragments which are commonly used and effective in vitro immortalized ceils. OBJECTIVE: To construct human immortalized precartilaginous stem cells (IPSCs) using human precartilaginous stem calls induced by SV40LTAg gane. METHODS: The human immortalized precartilaginous stem calls were isolated from aborted fetus and purified with enzyme digestion and immunomagnetic beads screening method. By using liposome-mediated gene transfection technology, plasmid pCMVSV40T/PUR containing SV40Tag was transfected in primary embryonic precartilaginous stem cells, while non-transfected cells sewed as negative controls. Positive clones were cultured to observe the cell morphology and the passage recovery, to calculate cell survival rata and population doubling time, to drew call growth curve. Immunofluorescence cytochemistry was used to detect the expression of IPSCs fibroblast growth factor receptor 3, the expressions of SV40Tag and fibroblast growth factor receptor 3 in the human precartilaginous stem cells were determined by RT-PCR. RESULTS AND CONCLUSION: Morphology of human IPSCs seemed coincidence with primary human precartilaginous stem cells. The survival rate of human IPSCs was not influenced by subculture, freezing and recovery, but the survival rate was descended in the human precartilaginous stem cells at the 6~(th) and 10~(th) passages (P < 0.01). Compared with cells at the 6~(th) and 10~(th) passages, the proliferation of human IPSCs was greater, with short population doubling time and high growth rate (P < 0.01). The immunofluorescence showed that fibroblast growth factor receptor 3 was positive in human IPSCs at the second passage, and the RT-PCR results of fibroblast growth factor receptor 3 revealed a specific amplification band at 400 bp,.while that of SV40Tag revealed at 560 bp. No band was seen in the primary cells. It is indicated that SV40Tag human IPSCs can be constructed successfully using immunomagnatic bead screening technology and liposome transfection technique.

Objective To evaluate the safety and effectiveness of balloon kyphoplasty combined with physiotherapy in the treatment of osteoporotic vetcrbral compression fractures (OVCFs). Methods A retrospective review was of 12 OVCF cases ( including 16 fracture vertebrae) treated with balloon kyphoplasty was performed. Each patient had also been treated with anti-osteoporotic medication and the Rehabilitation of Osteoporosis Program-Exercise (ROPE) protocol, and each had received a six-month follow-up visit. The following parameters were recorded before the operation and 1 day and 6 months afterward : fracture recurrence, the severity of pain before and after treatment, and the change of vertebral height. The severity of pain was evaluated by using a visual analogue scale (VAS). Results During the treatment, no complication or adverse event was found. The average VAS values preoperation, postoperation and at the 6-month follow-up were 7.6±1.3, 2.1±1.2 and 2.6±1.4, respectively. The average changes in vertebral height preoperation, postoperation and at the 6-month follow-up were 49.2±18% , 74.3 ±14% and 69.8±16% respectively. All of the measures evaluated improved noticeably after the combined treatment. Conclusion Balloon kyphoplasty combined with anti-osteoporotie medication and physiotherapy showed good effects in the treatment of osteoporotic veterbral compression fractures.

Objective To study the impacts of dynamic compressive stress on the mRNA expression of osteopontin ( OPN ),runt related gene 2 ( Runx2 ),osteocalcin ( OC ),osterix,alkaline phosphatase (ALP) and bone morphogenetic protein 2 (BMP-2) in the osteoblasts of Sprague-Dawley (SD) rats. Methods Osteoblasts extracted from skull periosteum tissue of neonatal SD rats were digested using trypsin and collagenase （Ⅰ）,then were subcultured and amplified in vitro.ALP staining and alizarin red staining were performed to identify the purified cells.The cells were treated with compressive stress at 20,50 or 100 mmHg for 24 h.The expression levels of OPN,Runx-2,OC,osterix,ALP and BMP-2 were measured and quantitatively analysed using a real-time quantitative polymerase chain reaction. Results Under 20 mmHg of dynamic compressive stress the expression levels of OPN,Runx2,OC,osterix,ALP and BMP-2 all were elevated compared with the control group.The peak expression oecured under 50 mmHg pressure. The expression levels did not change significantly compared with the control group under 100 mmHg pressure. Conclusions Moderate dynamic compressive stress can promote the expression of OPN,Runx-2,OC,osterix,ALP and BMP-2 mRNA in osteoblasts,which might be an important mechanism for promoting the union of fractures.