Objective To observe the effect of simvastatin on bone metastasis of breast cancer in nude mice model.Methods Sixty mice were divided into three groups randomly with 20 in each group.Mice were inoculated with MDA-MB-231 cells into the left cardiac ventricle.After 7 days,mice were treated with either simvastatin,saline,or nothing twice per week for 19 days.The area of osteolytic metastases was subsequently measured in long bones of all mice using an image analysis system.After sacrifice,parathyroid hormone-related protein (PTHrP) concentrations in bone marrow from all mice were determined using a two-site immunoradiometric assay.Osteoclast number expressed per millimeter of tumor/bone interface was assessed using an OsteoMeasure System.Measured data were compared with analysis of variance,and P ＜ 0.05 for the difference was statistically significant.Results The area of osteolytic lesions was significantly lower in mice treated with simvastatin compared with mice receiving saline and no treatment (0.51 ±0.18 mm2 vs 2.13 ± 1.24 mm2 vs 2.29 ± 1.22 mm2 ; F =15.600,P =0.002 ; F =15.673,P =0.001).In addition,treatment with simvastatin decreased the concentrations of PTHrP in bone marrow plasma (0.98 ±0.20 pmol/L vs 2.11 ±0.31 pmoL/L vs 1.99 ± 0.29 pmol/L; F =61.469,P ＜ 0.001 ; F =58.274,P ＜ 0.001) and the osteoclast numbers per millimeter of tumor/bone interface (4.00 ± 1.73 vs 11.40 ±4.93 vs 10.91 ± 3.87 ; F =17.820,P =0.001 ; F =17.184,P =0.002) compared with controls.Conclusion Simvastatin may reduce the production of PTHrP of breast cancer cells,which suppresses the development of destructive bone lesions as well as the growth of breast cancer cells in bone.

Objective To explore and solve the problem of insufficiency and malformation due to cripple hand and defect of digital bone resulted of innate and injured. Evaluation to the methods of bone autografting combinated with flaps to reconstruct the deficient hand and digits. Methods Vascularized iliac bone or metatarsophalangeal joints (MPJ) or toes transplantation incorporated with flaps were used to reconstruct the defected bone in palm or fingers. Combined with tendon absence in such cases should be repaired or restituted by tendon autografting in one stage or by stages. Results Total 16 cases were treated by the methods metioned above. Except 1 of 16 was failure resulted in severe infection, the others were reconstructed successfully. 13 of 16 were followed up 8 months to 2 years, the evalution of the group from motion,sensation and appearance shew that the fine rate of motion was 53.8 percent, the rate of appearance and sensation were both 69 percent. Conclusion Vascularized iliac bone or metatarsophalangeal joints (MPJ) or toes transplantation incorporated with flaps are available to reconstruct the defected bone in palm or fingers, and can reduce disability effectively while obtain a satisfactory outlook and proper functions by those methods.

In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni(+) -nitrilotriacetic acid (Ni(+) -NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni(+) -NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.

ObjectiveTo compare the diagnostic value of X-ray, CT, MRI, and ultrasound in primary synovial osteochondromatosis ( PSO ).Methods The imaging data of X-ray, CT, MRI, and ultrasound of 42 patients with 44 knees with PSO proved by surgery and pathology were retrospectively collected and analyzed. ResultsThe Plain X-ray demonstrated 197 calcific nodules in 28 joints, 96 ossific nodules in 24 joints, and 5 mixed type nodules in 3 joints. Compared with the data of surgery and pathology,36 joints (81.8 ％, 36/44) were diagnosed correctly by X-ray. The CT showed 8 big cartilaginous nodules in 5 joints, 255 calcific nodules in 30 joints, 146 ossific nodules in 28 joints, and 16 mixed type nodules in 7 joints.Twenty-four knees underwent volume rendering technique reconstruction which displayed the quantity, size, shape, and position of non-cartilaginous nodules clearly. Compared with the data of surgery and pathology, 40 joints (90. 9％ , 40/44)were diagnosed correctly by CT. The MRI demonstrated 8 big cartilaginous nodules in 5 joints, 70 small cartilaginous nodules in 4 joints, 248 calcific nodules in 29 joints,146 ossific nodules in 28 joints, and 16 mixed type nodules in 7 joints. All nodules displayed low signal in DWI and there was no enhancement. Compared with the data of surgery and pathology, 43 joints (97.7％,43/44) were diagnosed correctly by MRI. The ultrasound showed 8 big cartilaginous nodules in 5 joints,70 small cartilaginous nodules in 4 joints, 232 calcific nodules in 30 joints, 142 ossific nodules in 28 joints,and 16 mixed type nodules in 7 joints. Compared with the data of surgery and pathology, 43 joints (97. 7％,43/44) were diagnosed correctly by ultrasound. ConclusionsThe less common manifestations of the PSO require multimodality imaging to make the diagnosis. Multimodalities (X-ray,CT, MRI and ultrasound) are particularly useful in fully characterising PSO and to allow for appropriate clinical planning.

Objective Evaluate data of 1270 cases with free flap transplant,to find the problems in the process of operation,and then to analyse its causes and how to prevent and solve it,as well as providing reference for clinical colleagues. Methods To study 1270 cases of free flaps,musculocutaneous flap and perforator flap who was treated in our hospital from October 2000 to October 2010 retrospectively. A total of 722 cases of the group were followed-up 6 months to 5 years. To detective and search the problems and imperfection from designing,harvesting,tranfer,to the management and function of donor site after free flap transplantion.And also to analysis the couse of problems and operation failure,discuss the conclude of and to provide advisable measure. Results Total 1270 free flaps were transplanted successfully except 64 can-celled or failured for some reason, the success rate was 95.0％, the postopertive necrosis rate was 3.8％.Seventy-six cases were encountered vascular complications venous crisis in 42,arterial crisis in 38.Fifty-five cases were saved successfully without surgery,and 15 cases survived completely by vessel explorative operation. Five cases were partial necrosis and 7 cases necrosis. The rate of postoperative infection of emergency cases and chronic one were 4.7％ (682 cases)and 8.8％(588 cases) which show the infective incidence of latter was higher than former. In addition, there were many other problems were found such as distal flaps necrosis,contracture,deformation,impairment function of doner site,etc. Conclusion Preventing and management timely to vessle crisis is the key to flap suvive. The principle of dissecting flap should be followed strictly,and control the indications of modified processing during flap harvest,keep the proper flap tension were technical requirements in flap transplantion. Right way of donor site closed and management of insufficient timely were equally important to prevent and solve to complications and dysfunction.

Objective To study the method of solving the complications of large allograft including resorption, nonunion and refracture by means of vascularization and the way of improving bone healing. Methods The bone defect longer than 10 cm of limbs were found in 21 cases, plate or external fixator were used to fix allograft bone, and then vascularied autologous bone or periosteum were transplanted or inserted to the massive allograft In order to vascularizate allograft, improve bone healing and prevent complications of bone resorption and osteolysis. 4 cases were implanted by local vascular bondle in one end. 4 cases were transplanted vascular iliac bone into middle part the allograft bone, 2 cases were into one end. 2 cases were transplanted by vascular fibular bone. 6 cases were used vascular periosteum. 3 cases were used combined methods. Results Twenty-one cases with 10 cm or more bone defect in this group were treated by the method above. 14 cases were achieved primary healing, 6 were healing by farther operation, 1 failure. Complications were found in 7 cases, 4 occur infection in all. All were achieved satisfactory function and outlook by follow-up. Conclusion The method of vascularied autologous bone or periosteum combined with massive allograft are effective to improve bone union, reduce the complication of bone resorption and osteolysis,which proved usefull to treat large bone defect.

Objective To observe clinical curative effect of the FLAG regimen combined donor lymphocyte infusion after granulocyte colony stimulating factor(G‐CSF) mobilization(G‐DLI) ,for the acute myeloid leukemia (AML) of allogeneic Peripheral blood hematopoietic stem cell trans‐plantation (allo‐HSCT) after recurrence of hematology .Methods For the patients with recur‐rence after allo‐HSCT ,giving the FLAG regimen chemotherapy when the WBC dropped to the lowest point ,followed by giving G‐DLI that infusion peripheral blood stem cell from the original donors ,to observe curative effect and survival situation .And searched the literature review through the PubMed etc .Results Through FLAG regimen combined G‐DLI ,3 cases of relapse after transplan‐tation again obtained complete remission (CR) .Case 1 :disease‐free survival (DFS) was 13 month and overall survival(OS) was 23 months after G‐DLI .The patient has been the central recurrence and remission in bone marrow ,he was dead after 23 months due to multipleorgan function failure .He occurred Ⅱ acute GVHD in Skin and Ⅰ acute GVHD in liver after G‐DLI and obtained effective control ,not chronic GVHD .Case 2 :DFS and OS were 12 months and 13 months ,as bone marrow relapse again and giving up treat‐ment ,so died a month later .Respectively ,he has limitations chronic GVHD in skin after G‐DLI .Case 3:DFS was 16 months after G‐DLI since the disease‐free survival ,had limitations GVHD in skin that was control for given small dose of immunosuppressive drugs .Conclusion Joint FLAG scheme and G‐DLI may be one of the effective treatment of postoperative recurrence of allo‐HSCT .

In order to obtain three isoforms of apolipoprotein E (apoE), the cDNA encoding apoE3 was obtained by RT-PCR from normal human liver tissue. Site-directed mutagenesis was used to obtain the cDNAs encoding apoE2 and apoE4 isoforms. The 3 cDNAs were subcloned into vector pGEM-3Z and verified by DNA sequencing. The expression recombinant which can express the target protein as a (His) 6-tagged fusion was constructed by subcloning apoE cDNA into vector pT7-PL. The purified proteins were gained by Ni-NTA column. The SDS-PAGE results revealed the 6 His fusion proteins (apoE2, apoE3 and apoE4) were correctly expressed and purified successfully.

In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni+-nitrilotriacetic acid (Ni+-NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni+-NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.

BACKGROUND: Peak bone mass and standard deviation (SD) in different regions are varied, which have great influence on diagnosis of osteoporosis. To establish a complete database can provide accurate evidence for osteoporosis diagnosis. OBJECTIVE: To investigate the influence of bone mineral densities (BMD) and their SD of young people on the detective rate of osteoporosis in general population. DESIGN, TIME AND SETTING: Investigation analysis was performed at Beijing, Shanghai, Guangzhou, Nanjing, Jiaxing and Chengdu between January 1997 and December 1999. PARTICIPANTS: 11418 subjects from related 6 centers of BMD reference database in China were investigated and analyzed using prospective and retrospective methods, including 3 666 males, and 7 752 females aged 20-90 years. Of them, 2385 were from Beijing, 1178 from Guangzhou, 1404 from Shanghai, 2938 from Nanjing, 1425 from Chengdu, and 2088 from Jiaxing. The subjects were selected from community investigation, physical examination volunteers. METHODS: BMD of the lumbar spine (L2-4) and the hip in 11, 418 subjects from the related 6 centers in China was measured with GE-Lunar dual energy X-ray absorptiometry (DXA) and the BMD reference database was established. The accuracy rate of the inner machine was 0.3%-0.7%, and the accuracy of different machines averaged 1.1%. MAIN OUTCOME MEASURES: Lumbar BMD distribution of different age groups from 6 centers; influence of young people's BMD and its SD on detective rate of osteoporosis. RESULTS: Different BMD and SD were found in the individual subject from 6 centers, and the maximum differences were 0.098 g/cm2 and 0.027 g/cm2. With mean BMD and SD of the individuals from 6 centers as references, different T-scores and the detective rates of osteoporosis derived from the T-scores were found in the same group. The detective rate increased by 1.6% when BMD of the young people increased by 0.01 g/cm2 (positive correlation), but the detective rate decreased by 4% when SD increased by 0.01g/cm2 (negative correlation). CONCLUSION: Changes in BMD and SD of the young people can influence the detective rate of osteoporosis. To achieve comparability for the detective rate of osteoporosis in different centers, the specific reference database should be established for the same race, the same area, and the same bone densitometry machine. The T-score should be determined with the normal BMD and SD of the young, people as the reference database.