Objective To analyze the ultra microstructure and histological properties of implanted composites of decalcified bone matrix (DBM) impregnated calcium phosphate cement (CPC) with recombinant human bone morphogenetic protein-2 (rhBMP-2) in bone defect, and evaluate the osteoplastic efficiency of this composites. Methods The rabbit DBM was prepared beforehand. The composites of DBM impregnated CPC with rhBMP-2 was made at 0.2 proportion of DBM. The rabbit's bone defect of femur condyle was filled with implantation of the composites (group A, n=12) or CPC (group B, n=12) or bone cement (group C, n=12). Animals were sacrificed at 6, 12, or 24 weeks after operation, and the implants were examined by histological technique and scanning electron microscopy (SEM). Results In group A, at the 6th week after operation, the interface region between implants and bone tissue became fuzziness, and was crossed by the generated fibers of bone tissue, and filled with woven bone growing inside into the composites; at the 12th week after operation, blood vessels, osteoblasts and new bone were generated inside of composites; at the 24th week after operation, the bone defects were recovered and became solid union, most of the implants were replaced by new bones. In group B, at the 6th week after operation, the interface of CPC and bone tissue was sharp, and was not crossed by new bones; at the 24th week after operation, the bone defects were still filled with materials of CPC and no new bone was found inside CPC. In group C, at the 24th week after operation, the interface of bone cement and bone remained non-union. Higher osteoblastic activity,more neogenetic blood vessels and higher growth rate of woven bone were observed in group A compared with those in group B. Conclusions For bone defect, the implantation of composites with DBM proportion of 0.2 can stimulate the growth of osteoblast, blood vessel and woven bone. It is biodegradable and can be replaced by autogenous bone.

[Objective]To investigate whether fibroblasts from malignant bone tumor could support osteoclast differentiation and bone resorption.[Method]Fibroblasts isolated from 8 fresh tumor samples were cultured in vitro.At passage two,part of these fibroblasts were co-cultured with peripheral blood monocytes.M-CSF,OPG and neutralizing anti-TNF-? antibody was added to the culture respectively.A Transwells system was also applied to observe the secretion of solvable factors in fibroblasts.At day 1,day 14 and day 21,the cultures on coverslips and bone slices were stopped and examined for the formation of tartrate resistant acid phosphatase(TRAP)positive multmucleated cells(MNCs)and lacuna resorption pits respectively.The number of lacuna resorption pits on bone slices,which was a marker of OC activity,was compared between the groups.Another part of the fibroblasts were cultured alone,and RT-PCR was performed to investigate the expression of RANKL and TNF-? mRNA in the fibroblasts.[Result]TRAP positive MNCs and lacuna pits were observed in the co-culture group in the presence of M-CSF,but not in the absence of M-CSE.It was also the case in the Transwell group,even though the number of lacuna pits was resuced.OPG completely inhibited the formation of TRAP positive MNCs and lacuna pits,while anti-TNF-? had not inhibiting effect on this process.RT-PCR results showed that both RANKL and TNF-? mRNA were positive in all the fibroblast involved.[Conclusion]In the presence of M-CSF,fibroblasts from malignant bone tumor are capable of supporting osteoclast formation and activity through expression of RANKL.

[Objective]To investigate the effect of transforming growth factor ?1(TGF-?1) genes transfection on the proliferation and differentiation of chondrocyte by adenovirus vector,and to observe the quality in repair cartilage defect through constructing tissue-engineered cartilage.[Method]Using replicat ion defective adenovirus Adeno-X~(TM) as a carrier,Articular chondrocytes were transferred by high titers level recombinant adenovirus taking TGF-?1 genes.Ultromicrostructure,prolife ration anddifferentiation were observed by light and electron microscope,flow cytometry and so on.Induce expression of exogenous genes coding protein and cartilage cell phenotype were detected by Immunocytochemistry and Northern blot.Gross observation,histology and effect classification were used to evaluate results in repair cartilage defect by combi nation of infected chondrocytes and bone matrix gelatin(BMG).[Result]Immunocytochemistric staining showedthe expression of exogenous gene coding proteins after infection,and procollagen Ⅱ mRNA in cells was detected increasingly by Northern blot.TGF-?1 stimulated proliferation of chondrocytes in primary cultured,and resulted in recruitment of articular chondrocytes into S-G2/M phase.Chondrocytes seeded onto BMG showed high level of proliferation.Transplantation in vivo showed: cartilage defect was repaired satisfactorily by Combination of BMG and infected chondrocytes,and the structure of repair tissue got close to normal articular cartilage.[Conclusion] Articular chondrocytes prolifera tion could be stimulated by adenovirus vector taking TGF-?1 genes,which could be used to constructed tissue-engineered cartilage,and applied in repairing joint cartilage defect.

[Objective]To explore the effective treatment method of the articular fracture in distal radius.[Method]Severely comminuted AO type-C3 intra-articular fractures of the distal end of the radius were treated with combined internal and external fixation.The Gartland and Werley system and the Green and O'Brien system were used for comparison of the pre-operative and after operative evaluation.Total articular congruity had been assessed with both clinical rating systems.[Result]At a mean of ninteen months following-up,the mean arc of flexion-extension was 78% of that on the uninjured side and the mean grip strength was 82% of that on the uninjured side.The mean total articular incongruity(the gap plus the step-off)averaged 2 mm,and the radial length was restored to a mean of 11 mm.According to the Gartland and Werley demerit-point system,seventeen of the patients had a good or excellent result.According to the modified Green and O'Brien clinical rating system,eleven had a good or excellent result.Postoperative total articular incongruity had a moderately strong correlation with the outcome as assessed with both clinical rating systems.[Conclusion]Open reduction and combined internal and external fixation of AO type-C3 fractures can restore radiographic parameters to nearly normal values,maintain reduction throughout the period of fracture-healing,and provide satisfactory functional results.

[Objective]To analyze the clinical effect of microwave hyperthermia in limb salvage surgery for malignant bone tumor of extremities. [Methods]From July 1999 to July of 2005, 309 patients with malignant or bone tumors of extremities were treated by heat necrosis of tumor-bearing bone in situ for limb salvage.The most common diagnosis was osteosarcoma. The first step of the traditional limb salvage was en bloc resection of the tumor-bearing bone. For the novel method, the tumor bearing bone was just separated from surrounding normal tissues, and was devitalized by hyperthermia in situ.After re-strengthening the dead bone, its mechanical property became strong enough to support the body weight.[Results]The beyond 3 years survival rate was 60.2% for high-grade malignancy. In great majority of the patients, cosmetic and useful limbs were preserved. The complication rate was lower than that in the literature reports.[Conclusion]The long term experience has proved that the new method has made its way in the field of orthopedic oncology. The applying of hyperthermia for treatment of bone tumors is an effective, simple, and inexpensive method. The oncological and functional results are encouraging. Hyperthermia should deserve more attention than it has in the clinical practice.

[Objective] To analyze the treatment of malignant of highly aggressive bone tumors of pelvis by microwave-induced hyperthermia.[Methods]A novel surgical model was devised:after careful dissection of the tumor-bearing bone from surrounding normal tissues,the microwave antennae array was inserted into the tumor mass for emitting electromagnetic microwave which produced tumor cellular death via thermo-coagulation.No special reconstruction procedure was necessary,excepting the strengthening measures.[Results]From May 1994 to December 2005,152 patients with pelvic malignant or highly aggressive tumors received radical thermotherapy.Among 67 patients with stage IB tumors,48 patients achieved local and systematic control.Among 61 patients with stage IIB tumors,19 patients died from lesion,and the remaining 42 patents did not developed either metastasis or local recurrence after 3 to 11 years.Of 24 patients with pelvic metastatic lesions,11 patients collapsed during six months to three years,and 13 patients still lived without evidence of disease within one to seven years.In the majority of the patients,functional and cosmetic acceptable limbs were reserved.[Conclusion]The results revealed that the novel and greatly simplified method is justified from both oncological and functional standpoints.Hyperthermia should deserve more attention than it has in this field.

Objective To develop a diagnostic test based on indirect immunofluorescence assay(IFA) to detect special antibodies in the serum of SARS patients, thus to provide a reference material for confirmation of the clinical diagnosis of SARS. Methods SARS coronavirus GZ01 and BJ01 strains isolated in our laboratory were used to infect Vero E6 cells. When CPE reached 25%, cells were trypsinized and transferred to 10 well slides in a quantity of 40?l with a cell density of 2?10 7 /ml. After 4 hour incubation at 37℃,the slides were fixed with acetone, and IFA was used to detect antibodies in serum samples, which were obtained from 154 SARS patients and 14 non SARS patients with respiratory disease, as well as 100 healthy volunteers. Results IFA method for detecting antibodies of SARS coronavirus was developed. Sera from one hundred and forty two out of 154 clinically diagnosed patients were IFA positive, with a positive rate of 92 3%. Sera from 14 non SARS patients with respiratory disease and 100 healthy persons were all IFA negative. Conclusion The IFA method we developed was sensitive and specific in detecting SARS antibodies in serum, and was a reliable test for laboratory diagnosis of SARS coronavirus.

BACKGROUND: Patients who suffered total hip replacement are mostconcerned about the survivorship of prosthesis. OBJECTIVE: To evaluate the postoperative curative effect following ce mented and cementless THR with a retrospective method, so as to provideexperience for prolonging the survivoship of prosthesis. DESIGN: Randomized and controlled observation. SETTING: General Center of Orthopaedic Department, General Instituteof Bone Oncology, Tangdu Hospital, Fourth Military Medical University ofChinese PLA. PARTICIPANTS: We admitted 356 patients who underwent THR fromDepartment of Orthopaedics, Tangdu Hospital, Fourth Military Medical U niversity of Chinese PLA between March 1993 and March 2004. Amongthem, 298 were contacted and 105 (108 hips) followed up. The patientsparticipated in the review voluntarily. They were of either gender and haddifferent types of prosthesis. Prosthesis made in China was adopted before2000 and prosthesis made in American STRIKER company after 2000: Prosthesis made in China was made of home-made bone cement; Prosthesisbone cement (import) was provided by American STRIKER prosthesiscompany. Home-made bone cement and import have the same components. Barium was added in both bone cement . The whole operation was con ducted by the physicians who worked in the artificial joint department afterexamination. METHODS: According to informal discussion summary about total hipreplacement of Chinese Journal of Surgery in 1982 and Evaluation Scale ofMayo Total Hip Replacement Curative Effect, we designed follow-up tableby ourselves. Totally 105 (108 hips) patients were followed up, amongthem, 62 (63 hips) were in the cemented group, 43 (45 hips) in the ce mentless group. Pain, function and motion range of the patients and X-raywere evaluated and analyzed respectively. MAIN OUTCOME MEASURES: ① Postoperative pain degrees. ② Postoperative function of hips. ③ Postoperative motion range. ④ width oflight around the prothesis , distance of horizontal or vertical shift of theprosthesis. ⑤ range of ectopic ossification of the prosthesis. ⑥Osteolysisdegree of proximal femur. RESULTS: ①There was no significant difference of lateral femoral painduring follow-up period [Cemented group: 24 hips (38.5%) ,cementlessgroup: 18 hips(40.0% )]. ② Limping appeared in the both two groups ③ There was no significant difference of range of motion above 160° betweentwo groups (Cemented group: 62 hips; cementless group: 44 hips). ④Therewas no significant difference in subsidence of femoral prosthesis and hori zontal or vertical shift of acetabular prosthesis between two groups . ⑤ There was no significant difference of re lative value of femoral proximalbone density between cemented group [57.4(9-118)] and cementless group[72.8( 14-130)]. ⑥There was no significant difference of postoperative cu rative effect, possible survival rate of prosthesis and femoral proximal ex tensive osteolysis of the patients between the two groups. CONCLUSION: Postoperative curative effect of the patients between ce mented group and cementless group are similar, both not obtaining an idealfixed effect. The choice of prosthesis type does not affect the survivorship of prosthesis, but it depends on the age of patients to decide whether rebuilding is necessary or not: Osteolysis is not related to age, gender or prosthesis type of the patients.

BACKGROUND: Mesenchymal stem cells (MSCs) are capable of differentiating into a variety of lineages, including bone, cartilage, or fat, depending on the inducing stimuli, specific growth and differentiation factors.Recombinant human bone morphogenetic protein 2 (rhBMP-2) produced by gene engineering has an obvious osteoinductive activity and can induce undifferentiated mesenchymal cells into cartilage and bone irreversibly, resulting in new bone formation. Basic fibroblast growth factor (bFGF) modulates chondrogenic and osteogenic differentiation of MSCs.OBJECTIVE: To investigate the effect of bFGF and rhBMP-2 on the differentiation and proliferation of cultured rabbit mesenchymal stem cell in order to find out an optimal way of osteogenesis instead of conventional osteogenic supplements (OS).DESIGN: A randomized and controlled trial.SETTING: General Institute of Orthopaedic Oncology, Tangdu Hospital,Fourth Military Medical University of Chinese PLA.MATERIALS: The subjects were rabbit mesenchymal stem cells cultured by the author.METHODS: This experiment was conducted at the Center of Orthopaedic Surgery, General Institute of Orthopaedic Oncology, Tangdu Hospital,Fourth Military Medical University of Chinese PLA from June 2004 to December 2004. ①Rabbit MSCs cultured in vitro were treated with different growth factor (100 μg/L rhBMP-2, 100 μg/L bFGF, 10 μg/L rhBMP-2and 100 μg/L bFGF and OS; ②The proliferation and differentiation of MSCs were observed through activity of MTT, expression of alkaline phosphatase(ALP), and von Kossa staining.MAIN OUTCOME MEASURES: the rate of proliferation and the activity of ALP.RESULTS: ①rhBMP-2 could promote the proliferation and differentiation of MSCs, especially the cell differentiation; ②bFGF could stimulate the proliferation , the cellular proliferation rate increased 100% as compared with control group, and has no effect on differentiation of MSCs , but it could enhance effect on the cell proliferation of rhBMP-2.CONCLUSION: bFGF and rhBMP-2 are effective induction factors for MSCs. Both of them can stimulate the proliferation and differentiation of MSCs in vitro. bFGF and rhBMP-2 exerted a synergetic action in speeding up the pace of osteoinduction and osteogenesis and can be used to differentiate seed cells for tissue engineering bone.

Objective To compare and evaluate the defect-repaired capabilities of human bone morphogenetic protein-2(hBMP-2) gene modified tissue engineered bone in the segmental bone defect model of rabbit's radius.Methods Rabbit's bone mesenchymal stem cells (BMSCs)were transferred with hBMP-2 gene through Adeno-XTM adenoviral expression systems,then seeded onto the compound scaffold of calcium phosphate cemept(CPC)and fibrin glue(FG)to construct a new kind of gene modified tissue engineered bone after proliferation in vitro for three weeks(Group A).Meanwhile,the compound scaffold of calcium phosphate cement(CPC)and fibrin glue(FG),which were seeded by rabbit's bone knesenchyrmal stem cells(BMSCs) after proliferation in vitro for three weeks(group B)and the compound scaffold without cells(Group C)acted as control groups.Then,three kinds of reconstructive modalities were implanted into segmental bone defect of donator rabbit's radius.Besides these three groups,bone defect model of rabbit's radius without treatment(Group D)represented blank group.The defect-repaired capabilities were assessed by gross observation,radiograph,Single Photo Emission Computed Topography (SPECT)and histological analysis in the 4th week,8th week and 12th week after operation.The rates of bone healing in the different groups were compared each other.Results All defects that had been treated with implants(Group A,B,C)exhibited new bone formation and could attain osseous tissue healing 12 weeks after operation,but defects in blank group(Group D)were repaired only by fibrous tissue.The defects in the Group A regenerated more new bone,bridged earlier and stronger than those in the Group B and Group C.The quantity and rate of new bone formation in the Group B and Group C had no significant difference and the rates of bone healing in different groups showed the same results.Conclusion hBMP-2 gene modified tissue engineerod bone have better potential to form new bone and the rate of bone healing in repairing bone defects is higher,so this way is an optimal kind of material for artificial bone graft.