Objective To investigate the effect of caffeine and cisplatin induced apoptosis of osteosarcoma cell line(OS-732), and to explore the potential mechanism of caffeine enhancing cytotoxic effect of cisplatin in osteosarcoma cell line. Methods The OS-732 cell line was cultured for 72 hours; treated with caffeine, cisplatin and caffeine combined with cisplatin for 72 hours respectively, the apoptosis rates of OS-732 cell line were analysed by flow cytometry. Mitochondrial transmembranous potentials were measured by cellular rhodamine 123 stain on flow cytometry. Apoptosis was assessed by electron microscope at 80 kV. Results The OS-732 cell line was cultured for 72 hours; treated with caffeine (5.0 mmol/L ), cisplatin (10.0 ?g/ml ) and caffeine (5.0 mmol/L ) combined with cisplatin (10.0 ?g/ml) for 72 hours respectively. The apoptosis rates were 2.50%, 10.62%, 31.62% and 57.44% respectively. The percentage of decline of mitochondrial transmembranous potentials were 8.12%, 26.45%, 17.82% and 38.26% respectively. Electron microscope revealed the characteristic apoptosis alterations,such as shrinking cellular chromatin condensation, crescent nucleus, cytoplasmic vacuoles and so on. Conclusion Caffeine and cisplatin can induce apoptosis of osteosarcoma cell line(OS-732), while the cell line treated with caffeine and cisplatin simultaneously, the apoptosis rate was increased obviously. The induction of apoptosis of osteosarcoma cell line by caffeine may be one of potential mechanism enhancing cytotoxic effect of cisplatin in osteosarcoma cell line.

AIM: To investigate influence of biomimetic nanoapatite coatings of titanium surface on behavior of osteoblasts-like and provide evidence for surface modification and biological effects of titanium implant.METHODS: Biomimetic nanoapatite coatings were developed by functionally modified methods with a combination of topographic,chemical and biomimetic treatments on the surface of titanium(Ti) substrate.The biological behavior and bioactivity of functionally modified SLA implants with chemical and biomimetic treatments(SCB-treated Ti) were investigated to compare with untreated Ti and SLA Ti plates as controls.The cell attachment,proliferation,alkaline phosphotase(ALP) activity,cell morphology and differentiation were evaluated by using MTT,RT-PCR,scanning electron microscopy(SEM) and confocal laser-scanning microscope(CLSM) analysis system.RESULTS: The cell adhesion and proliferation were enhanced on functionalized titanium surface with nano-scale apatite compared to the controls.SEM micrographs also revealed that the osteoblast-like cells spreadly grew along the surface.Cell morphology and differentiation were further observed distinctly by CLSM graphs.Moreover,mRNA expression of alkaline phosphotase on the SCBtreated Ti increased obviously on the twelfth day compared with the controls.CONCLUSION: The in vitro results demonstrate the remarkable improvement on cell adhesion and proliferation of the biomimetic nanoapatite on SCB-treated Ti,which could improve early bone-implant interface bonding ability and be used for orthopaedic/dental implants.

Objective To investigate the aesthetic relations and internal regularities in normal craniofacial stereoscopic architecture. Methods By means of computer aided three dimensional measurement and analysis, a study on morphological features and inter relations of the cranio orbitozygomatic maxilla structures of twenty youths with pretty facial appearance was carried out. Results There were remarkable differences between each craniofacial structure in different sexes, but the ratio indices that represented the inter-relations of different craniofacial structures were closely similar in both sexes. In the group of youths the asymmetry ratios between bilateral craniofacial structures were below 5 % , presenting the perfect symmetry feature. The asymmetry ratios and their standard deviations of the zygomas and angulus mandibulae were relatively large . The results suggested that these two characteristic parts which influence the whole appearance were easily changeable with different individuals. There were close co relations between coronal axis width and sagittal axis protrusion in main structures of craniofacial region. These relationships could sum up in linear regression equations. Conclusion There are internal aesthetic regularities of coordinative ratios and harmonious symmetry in normal three dimensional craniofacial architecture, which determines the pretty appearance of youths. These internal regularities and aesthetic principles should be considered in designing and performing craniofacial aesthetic plastic operation so as to get excellent surgical results.

AIM:Many studies have documented an anabolic effect of hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, statins, on undisturbed bone. Reports of their effects on fractured skeleton were limited. A study was therefore conducted to check the effects of statins on fracture healing. METHODS: Simvastatin (10 mg?kg-1?d-1) was injected subcutaneously to tissue overlying the site of fractured tibiae of ovariectomized rats for a treatment period of 5 d. Vehicle reagent was used as control. Healing quality was evaluated at 1, 2 and 4 weeks after fracture. RESULTS: Compared with vehicle group, callus cross section area in simvastatin treated rats were significantly enlarged by 21.3% (P

Objective To discuss the diagnosis, treatment and prognosis of stage Ⅲ osteosarcoma. Methods A retrospective analysis of 23 patients with stage Ⅲ osteosarcoma of extremities from December 1989 to December 2003 was studied. There were 9 females and 14 males, aging from 16 to 31 years with a mean of 22.4 years. 15 patients presented with lung metastases, 7 with bone metastases (including 5 of jumping metastases and 2 of osteosarcomas) and 1 with lung and bone metastases simultaneously. Patients received chemotherapy followed by resection of primary and metastatic lesions and additional chemotherapy. Results After preoperative chemotherapy, lung metastases disappeared in 1 patient, whereas in 1 with lung and bone metastases simultaneously, the lesion remained surgically unresectable because of new metastases after removal of the primary lesion. In 2 patients with osteosarcoma, primary lesion could only be removed, lung metastasis appeared in 2 of 5 patients with jumping metastases respectively after the removal of primary lesion and jumping metastases. 16 patients with lung metastases received thoracotomy and resection of the lung metastatic lesions, and 6 of them received a second thoracotomy because of a second lung metastasis inclunding 3 cases with extra-pulmonary metastases without any additional treatment. The tumor necrosis rate was not found obviously different between primary lesions and metastatic lesions. Of the 23 patients who achieved a mean 74.6 months follow-up (range, 5-168 months), 9 remained continuously free of disease, 4 relapsed with new metastases, and 10 died of tumors. The outcome of the Cox model proportional hazard regression showed the relation of the number of the metastases and the prognosis were significant(P

Objective To discuss morphology and biomechanics of the sacrum fractures so as to provide scientific basis for corresponding clinical treatment. Methods A total of 10 fresh pelvis specimens were collected for dynamic impact test and static destruction test, in the former one of which, the dynamic parameters were measured to make sure the dynamic characters of the fractures. Meanwhile, the fractures of the sacrum wing, the sacral foramina and the sacrum edge were made decalcification, slice of paraffin wax and staining (Masson, Mallory, HE) in order to make a cytological observation of the tissue. Results (1) The form of sacrum fracture or acetabulum fracture, crista iliaceis fracture was relevant to the impact energy. Low impact energy usually caused fractures of the ilium, acetabulum or sacroiliac crest. High impact energy resulted in following three kinds of fractures, just as the classification of Denis: sacral ala fractures belonged to typeⅠfractures, sacral hiatus fractures to type Ⅱ fractures and central vertebral canal fractures to type Ⅲ fractures. All three types of fractures might involve lateral or bilateral nerve roots. (2) There was a significant mechanic difference in regard of the mechanism of both dynamic destruction and static destruction of pelvis, ie, not only the limit pressure differed but also the former increased rapidly with the higher rate of the strain. The clash energy beyond 25 J would beget the cleft fractures of the sacrum via sacrum hiatus and even involve the nerve roots. The clash energy under 20 J usually resulted in fractures of the ilium and the sacrum. The clash energy between 20 J and 25 J more easily caused type Ⅰ fractures. While fracture of the ilium and the acetabulum would happen most in static destruction. (3) The cross section of the sacrum was cracked and the bone board of Haversian system is brittle, as led to separation of bone board and malposition of a few cross bone boards. Conclusions Under dynamic state, the sacrum fractures mostly belong to type Ⅰ and type Ⅱ (Denis classification of sacral fractures), usually involving the nerve roots. The sacrum fracture is relevant to the microstructure, the distribution of the bone trabecula, the osseous lacuna and the Haversian system of the sacrum. The fractures of the ilium and the acetabulum more frquently appear in static state, with slight wound of peripheral tissues.

AIM: To study whether caspase-3,8 is activated during azurin-induced apoptosis in U2OS cells. METHODS: AnnexinV /PI method was used to detect apoptosis. The changes of procaspase-3 were analyzed by Western blot, the changes of caspase-3 mRNA were detected by semi-quantitative RT-PCR, and caspase-3 relative activity was determined by colorimetric assay. RESULTS: After U2OS cells were treated with 0, 25, 50, 100, 200, 500 mg/L azurin for 24 h, respectively, the level of procaspase-3 protein decreased and the level of caspase-3 mRNA increased as azurin concentration increased. When the cells were treated with 100 mg/L azurin for 6, 12, 24, 48 h , respectively, the caspase-3 activity began to rise from 6 h,reached the peak at 24 h,and was still higher than the control group at 48 h ( P

AIM: To investigate the synergistic induction of apoptosis in rhabdomyosarcoma cells by the combination of TRAIL or TRAIL gene with cisplatin. METHODS: Rhabdomyosarcoma cells were treated with TRAIL, (Ad/GT)-TRAIL, cisplatin, respectively or the combination for 3 days. The cytotoxicity was observed by MTT assay. The apoptotic rates and the expression rates of Fas protein were measured by flow cytometry (FCM). The expression of cFLIP mRNA was determined by RT-PCR. RESULTS: Rhabdomyosarcoma cells were treated with Ad/ GT-TRAIL and TRAIL (100.0 ?g/L), the cytotoxicity index were 52.5% and 43.5%, the percentage of apoptotic cells were 12.95% and 10.26%, respectively. Combined with cisplatin, the cytotoxicity index and the percentage of apoptotic cells were increased significantly (P

AIM: To down-regulate expression of c-myc through antisense therapy and to investigate its effect on the sensitivity of osteosarcoma MG-63 cells to cisplatin-induced apoptosis. METHODS: The recombinant adenovirus (Ad-Asc-myc) encoding antisense c-myc fragment was constructed and transfected into osteosarcoma MG-63 cells in vitro in order to down-regulate the expression of c-myc, and the change in the sensitivity to cisplatin-induced apoptosis was observed. MTT, Western blot, RT-PCR, flow cytometry (FCM) and electron microscope were used to evaluate tumor cell proliferation in vitro, genes expression related to apoptosis regulation and effects on the sensitivity of osteosarcoma MG-63 cells to cisplatin-induced apoptosis. RESULTS: Ad-Asc-myc down-regulated the expression of c-myc protein after transfected MG-63 cells for 48 h, combined with the treatment of 2.0 mg/L cisplatin for 2 h inhibited tumor cell proliferation in vitro by 38.0%. RT-PCR revealed that Ad-Asc-myc down-regulated the expression of Bcl-2 and up-regulated the expression of Bax. No appreciable change was observed in the expression of E_2F-1. FCM showed that Ad-Asc-myc induced apoptosis in intransfected cells, and rendered it more sensitive to cisplatin. CONCLUSION: Antisense c-myc is able per se to induce apoptosis and sensitize osteosarcoma cells to cisplatin-induced apoptosis.

Objective To explore clinical effect of reconstruction for chronic Achilles tendon rupture of Kuwada IV type with flexor hallucis longus (FHL) harvested using a minimally invasive technique.Methods The data of 35 patients with chronic Achilles tendon rupture of Kuwada IV type was retrospectively analyzed who were treated by FHL which was harvested using a minimally invasive technique from July 2006 to June 2011.There were 21 males and 14 females,with the age from 23 to 71 years (average,42.1 years).All patients were unilateral injury.MRI showed Achilles tendon rupture fissures 6.0-9.2 cm.The local appearance and function recovery on postoperation was observed,and all patients were assessed with the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and Leppilahti Achilles tendon repair score.Results Thirty-two patients were followed up for 18 to 72 months,with an average of 33.2 months.Except for 1 patient whose wound healed after six weeks through resuture immediately for the wound dehiscence occurred in the ten days,other patients' wound healed smoothly.The average of AOFAS ankle-hindfoot score had increased from 51.92±7.08 preoperatively to 92.56±6.71 postoperatively.Leppilahti Achilles tendon repair score had increased from 72.56±7.43 preoperatively to 92.58±5.1 postoperatively.Twenty-seven cases were excellent,good in 3,and fair in 2,with the total excellent and good rate 93.8％ (30/32).No case of the sural nerve and tibial nerve injury,plantar painful scar,plantar outside nerve injury.MRI of Achilles tendon showed even signal without signal of tear and cystic degeneration.Conclusion Reconstruction for chronic Achilles tendon rupture of Kuwada IV type with FHL harvested using a minimally invasive technique offers a desirable outcome in rapid postoperative recovery,high strength in tenodesis,fewer complications.