Objectives:To study the effects of gene therapy with tissue type plasminogen activator(t PA)cDNA on the formation of thrombo embolism in vascular anastomotic sites. Methods:①The cDNA encoding t PA was amplified by RT PCR using the isolated total RNA as the template from the Bowes melanoma cells.②Recombinant plasmid pAdCMV t PA was cotransfected into 293 cells with pJMa 17 ,and the infectious but replication deficient AdCMV t PA was generated.③The rats were randomly divided into the control and treatment groups.11 0 nylone medical suture was applied to perform rat carotid artery end to end anastomoses.In the treatment group,AdCMV t PA solution was injected into the vascular anastomotic site while AdCMV (no containing t PA DNA) solution was injected into the control group. By means of RT PCR and chromogenic plasmin substrates,the following results were obtained. Results:①The t PA cDNA was successfully cloned and its eukaryotic expressing vector was constructed.②When the isolated RNA was performed with RT PCR,1.69 kb band appeared in the treatment group while the band could not be found in the control group.The t PA activity could be detected postoperatively on the 1st,2 nd,3 rd,4 th,5 th,6 th,7 th,10 th and 13 th day of the treatment,but could not be detected in the control group. Conclusions:The t PA gene can produce t PA having biological activity at anastomotic sites, possibly prevent the formation of thrombus embolism effectively and develop the anastomotic patency.

Objective To investigate clinical results of the treatment of posterior shoulder instability in a combined way of bone block procedure and posterior capsulorrhaphy. Methods The double contrast CT arthrography was performed for 5 patients suffering from posterior shoulder instability so as to detect the pathology. The treatments combining bone block procedure and posterior capsulorrhaphy were carried out according to the pathological conditions of bone and soft tissue. Results The double contrast CT arthrography showed posterior bone defects, posterior glenoid labrum tear and enlargement of posterior capsular cavity. After the bone block procedure and posterior capsulorrhaphy, no patients complained shoulder instability in the follow-up of 5 to 38 months. Conclusions Many pathological factors may induce posterior shoulder instability, which should be corrected by an appropriate operation rather than a “standard”method of operation. For patients involving several pathological factors, a combined operation may be needed in order to correct various pathological changes.

Objective To investigate the radiological characteristics and the relationships between the parameters by X?ray and direct magnetic resonance arthrography (dMRA) in non?completely dislocated dysplasia hips. Methods We retro?spectively reviewed patients (above 13 years old) with dysplasia hips from August 2009 to August 2012. These patients were classi?fied as Hartofilakidis typeⅠand typeⅡ. 188 patients (274 hips) involved 28 males (30 hips) and 160 females (244 hips), average aged 27.3 years (range 13-47 years). Standard pelvic A?P, bilateral 65° oblique X?rays, and dMRA were conducted in each pa?tient. Lateral central?edge angle (LCE), anterior central?edge angle (ACE), femoral neck?shaft angle (FNSA), and tonnis acetabu?lar index (AI) angle were measured based on the X?ray. The dysplasia hips were classified into 4 grades (GradeⅠ:CE 11°-20° , GradeⅡ:CE 1°-10°, GradeⅢ:CE-9°-0°, and GradeⅣ:CE≤-10°). Femoral neck anteversion angle (FNA), labral tear, labral inversion, and labral cyst was observed on dMRA films. Variance analysis and rank correlation were applied to find the relation?ship between CE grades and the relevant parameters by using SPSS 19.0. Results LCE has negative correlation with AI and grade of osteoarthritis, and has positive correlation with age, ACE. However, there was no relationship between FNSA and FNA. There was significantly different in Shenton's line, labral tear, labral inversion, cartilage lesion, among different grade of LCE. Conclusion LCE is an important parameter used in evaluating for dysplasia hips without complete dislocation. LCE can reflect age of onset, ACE, AI angle and degree of joint damages. Evaluation for the severity of non?completely dislocated dysplasia hips can be conducted according to grades of CE.

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 investigated the biological procedure of allograft decalcified bone matrix(DBM)and bone cement(BC)combined with bovine bone morphogenetic protein (bBMP)used for the repair of femoral defect caused by microwave－ induced hyperthermia in dogsby 99mTc－ MDP bone scintigraphy.Method The canine femoral defect(length 25mm,width 10mm)was caused by microwave－ induced hyperthermia(50℃ ,20minutes)and the composite material was implanted .Then the canine femurs were examined by 99mTc－ MDP bone scintigraphy respectively at different postoperative time and the results were compared with that of X－ ray photography and histological observation.Bone cement was implanted in the other femur as a contrast.Results It could be observed at the first and the second month that the radioisotope was gathered in the place where the composite material was implanted and the amount of radioisotope gathered in was the most abundant at the third month and it was lasted to the fourth month. That of the sixth month was decreased to that of the second month.The radiation count of the first, the second, the third the fourth and the sixth month were 93.9± 12.7, 110.7± 16.4,222.1± 24.0,201.3± 26.9 and 111.6± 20.7 respectively,and the count of the third month and the fourth month were more than that of the first, the second and the sixth month(P<0.01).Conclusion The composite material could be remodeled easily and the new bone could be formed by the induction of bBMP. So it could be merged with the normal bone.While the 99mTc－ MDP bone scintigraphy is the object and reliable index to determine the biological procedure of the composite material in dogs.

Objective:To explore caspase-6's effect on the apoptosis of osteosarcoma cell line SOSP 607. Methods: The expression level of caspase-6 in the osteosarcoma cell line SOSP-9067 was examined by RT-PCR method. The adeno-virus adv5 vector with caspase-6 gene was constructed and transferred into the osteosarcoma cell line SOSP-9607 by lipo-fection. The cell survival rate after transfection was assayed by MTT method. The cell morphological changes were observed by microscope and electron microscope, the apoptosis of transferred cells were examined by gel electrophoresis. Results:No expression of caspase-6 was examined in the osteosarcoma cell line SOSP-9607. A high expression of caspase-6 was identified by RT-PCR after the transfection. The cell growth curve declined after transferring caspase-6. Electrophoresis of DNA displayed the apoptosis ladder. Conclusion: Transferring caspase-6 into the osteosarcoma line SOSP-9607 may inhibit the growth of the osteosarcoma cell line SOSP-9607 and this effect might be achieved by inducing apoptosis.

Objective To investigate the anatomic structure of the Chinese people,develop the procedure of minimally invasive total hip arthroplasty through the anterolateral intermuscular approach and investigate its clinical outcome.Methods Three fresh adult cadavers(6 hips)were used for study of the anatomic construction of the anterolateral intermuscular approach in Chinese people.Sixteen patients were treated with minimally invasive total hip arthroplasty through anterolateral intermuscular approach.The clinical results and operation technique were recorded.Results The anterolateral intermuscular approach was a triangle muscular interval slightly parallel to the femur.The medial-superior angle of the triangle muscular interval consists of the anterior border of gluteus medius and tensor fascia lata muscle with juncture of muscles,where the inferior branch of superior gluteal nerve entered into tensor fascia lata muscle.The average incision length was 8.8 cm(7-10 cm),with mean blood loss of 350 ml(250-550 ml).The patients took out-of-bed activity 3-5 days after operations.During operations,anterior border injury in deep portion of the gluteus medius muscle was observed in seven patients and the injured muscles were trimed or repaired.All patients were followed up for 18-39 months(averaged 27.7 months).Most of the patients had excellent location of the phantoms,except that one acetabulum had a little pitch angle and two acetabulums had a little abduction angle.No complication was observed.The mean Harris scores of hip for all patients was increased from preoperative(39.1±6.7)points to(80.6±11.3)points on six month,(88.7±9.6)points on 12 month and(91.4±13.5)points on 24 months(11 patients).No patient suffered from gluteus medius muscle weakness during the follow-up.ConclusionAnterolateral intermuscular approach has the advantages of simple anatomic construction,small incision,little operative injury,muscle sparing and fast recovery without separate muscle or tendon and is suitable for the Chinese patients.Exact incision and special operative instruments should be emphasized to avoid the increase of acetabular pitch angle and abduction angle.

Objective To discuss the mid?to long?term results and possible prognostic factors of Bernese periacetabu?lar osteotomy for adolescents and young adults with developmental dysplasia of the hip. Methods Bernese periacetabular os?teotomy was performed on 162 patients (171 hips) with hip dysplasia from August 1997 to July 2009. Follow?up data was ac?quirable completely in 123 cases (137 hips), including 30 males (33 hips) and 93 females (104 hips). Age of surgery was 12-48 years (average 27.1 years). The X?ray radiographs and Harris scores were recorded preoperatively and at the last follow?up. The lateral center?edge angle (LCE), acetabular incline angle (AI), continuity of Shenton's line were also recorded. The grades of the hip osteoarthritis were classified by T?nnis classification. Insufficient corrections of osteotomy, excessive corrections of osteotomy, nonunion of the pubis, nerve injuries, and vascular injuries were recorded as complications. Results All the con?tactable cases was followed up again in June and July in 2014. Follow?up duration was 5-17 years (mean, 8.1 years). The LCE angle improved from 7.13° ± 8.86° preoperatively to 30.17° ± 14.98° at the final follow?up. The T?nnis acetabular incline angle decreased from 27.84° ± 12.85° preoperatively to 7.06° ± 10.58° at the final follow?up. The continuity of Shenton's line was in?tact in 106 cases postoperatively, compared with 87 cases preoperatively. Harris score improved from 83.34±9.82 preoperative?ly to 92.79±7.05 at the last follow?up. There was significant correlation between preoperative Harris score and Harris score at the last follow?up. Early complications were observed in 15 cases (15 hips), along with 17 cases (17 hips) of progressions of hip osteoarthritis. Survival rate of the hip at the last follow?up was 97.8%. Conclusion Bernese periacetabular osteotomies can significantly improve bone coverage and joint function of hip dysplasia patients with low rate of short?term complications. Further?more, there was little osteoarthritis progression observed, with good survival rate of the joint in mid?to long?term follow?up. Lower pre?operative Harris scores and higher pre?operative T?nnis classifications are important prognostic factors for post?operative pain.

Objective To clarify the temperature curve of the irradiation target area,its adjacent tissue and the whole body during extracorpereal microwave irradiation, then to compare and optimize different irradiation models. Methods Different parts of the chest of adult New Zealand white rabbit were irradiated using different extracorporeal microwave irradiation models. The temperature of the irradiated skin, the subcutaneous and deep parts, the adjacent tissues and the anus was measured. The experiment was bi-factor and multi-level designed according to the repeatedly measured data and the rabbits was divided into group a,b,c and d. Results The increase rate of the surface temperature in the dorsal lung was similar between group d and group b1(F=10.04,P<0.01). However,the increase rate of the surface temperature in the ventral lung of group d was lower, and the mean temperature of this site measured 10 minutes later was also lower than group b1(F=10.04,P<0.01). The increase rate of the rectal temperature of group d was higher,and the mean rectal temperature tested 10 minutes later was also higher than group b1(F=7.04,P<0.01). Conclusions Multi-array irradiation could achieve satisfactory irradiation depth and appropriate therapeutic temperature. Well controlled extracorporeal microwave irradiation under is an ideal thermotherapy method.

BACKGROUND: Composite with different ratios has various elastic noduli, and this can cause stress protection in implanted body so as to influence on biological integration between composite and host bone and plerosis of bone defection.OBJECTIVE: To measure elastic modulus of tissue engineering bone made of composite between true bone ceramic (TBC) granule and bone cement (BC) and provide experimental data for repairing bone defection at various sites.DESIGN: Controlled study.SETTING: Military Institute of Bone Tumor, Tangdu Hospital of the Fourth Military Medical University of Chinese PLA.MATERIALS: The experiment was carried out in the Military Institute of Bone Tumor, Tangdu Hospital of the Fourth Military Medical niversity of Chinese PLA from August 2002 to March 2003. Ten Kunming mice,weighing 10-15 g, of both genders, were selected in this study. Fresh longtubular cortical bone of 1-year calf, 6 samples of spongy bone at inferior extremity and 6 samples at iddle femur of fresh corpse were also selected in this study.METHODS: Bovine bone morphogenetic protein (bBMP) was xtracted to measure osteogenic-induced activity and make TBC granule. bBMP was mixed with TBC as the ratio of 1:25, and then with BC as the ratios of 0:10, 4:6, 5:5,6:4 and 7.5:2.5. In addition, elastic modulus of tissue engineering bone made of TBC-BC composite was measured and compared with elastic modulus of femur and spongy bone of normal adult males.MAIN OUTCOME MEASURES: Measurement and comparison of elastic modulus.RESULTS: Elastic modulus of composite containing 60% TBC (Ratio between TBC and BC was 6:4) was no significantly different from that of spongy bone at inferior extremity of femur of normal adult males (P ＞ 0.05),but other comparisons of every two subjects were significant difference [femur of the adults: (6.216 7±0.222 9) Mpa; spongy bone: (1.351 7±0.306 9) Mpa;TBC/BC (0:10): (5.710 0±0.166 3) Mpa; TBC/BC (4:6): (3.510 1±0.205 0) Mpa;TBC/BC (5:5): (2.004 1±0.150 0) Mpa; TBC/BC (6:4): (1.501 8±0.005 7) Mpa;TBC/BC (7.5:2.5): (0.900 4±0.025 1) Mpa, P ＜ 0.01].CONCLUSION: Elastic modulus of composite containing 60% tissue engineering bone made of TBC-BC composite is similar to that of spongy bone at inferior extremity of femur of adult males. The composite can be used to repair bone defection near by articular facet so as to prevent from articular degeneration.