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

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.

AIM: To study the relationship between respiratory syncytial virus (RSV) infection and apoptosis, between RSV infection and expressions of FasL, Fas, Bcl-2 and Bax. METHODS: Apoptotic cells were examined by flow cytometry and transmission electron microscope. Immunohistochemical analysis was used to detect the expressions of apoptosis-associated gene FasL, Fas, Bcl-2 and Bax in A549 cells during RSV infection. RESULTS: Apoptotic index increased at 72 h and 120 h postinfection. Apoptotic cells were detected by transmission electron microscope. High-expressions of FasL, Fas and Bax genes and low-expression of Bcl-2 gene were detected by immunohistochemical staining. CONCLUSION: Apoptosis in A549 cells was induced by RSV infection. This apoptosis may be induced by up-regulating the expression of FasL, Fas, Bax genes and down-regulating the expression of Bcl-2 gene.

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 determine the role of Fas antigen and caspase-8 in modulating apoptosis of osteosarcoma cells induced by bacterial redox protein azurin. METHODS: The human osteosarcoma cell line U2OS was treated with bacterial redox protein azurin (150 mg/L) for 6, 12, 24 and 48 h, respectively. Cell immunohistochemistry and quantitative image pattern analysis were applied for detecting the expression of Fas antigen. Caspase-8 activity was detected using caspase-8 fluorescent assay kit. The apoptotic rate was measured by FCM. RESULTS: Compared with the control group, the expression of Fas antigen and activity of caspase-8 significantly increased in U2OS cells treated with 150 mg/L azurin (P

Objective To investigate feasible immunotherapy strategy using tumor specific cell against osteosarcoma,and to analyze the therapeutic effect of adoptive cellular infusion therapy on osteosarcoma.Methods Decitabine (DAC) was employed as a hypomethylating agent for the treatment in osteosarcoma cell lines HOS and U2OS.After treatment,the expression of cancer-testis antigen (CTA) was evaluated by PCR and Western Blot.In animal studies,human osteosarcoma cell line HOS,which was transfected by luciferase and HLA-A0201 in previous,was inoculated into immune deficient NOD-SCID mice to establish osteosarcoma xenografts.Ex-vivo expanded CTA specific homo CD8+T-ells were labeled with DiR and injected into the mice via the tail vein.In vivo imaging system was utilized to detect the distribution of administrated CD8+ T-cells.In addition,the progression of tumor xenografts was monitored.Moreover,mouse K7M2 osteosarcoma cell line was used to establish animal models in immune competent BALB/c mice.Immune competent models were utilized to evaluate the effectiveness of hypomethylating treatment in regarding to spontaneous immune attack against tumors.Flow cytometry was used to analyze the proportion of intratumoral lymphocytes and the status of these effector antitumor immune cells,and to reveal the effect of hypomethylating treatment in facilitating lymphocyte infiltration and activation.Results The expression of all the evaluated cancer/testis antigens were elevated in HOS and U2OS osteosarcoma cell lines after hypomethylating treatment with DAC.The proliferation of in vitro cultured osteosarcoma cells can be significantly suppressed after at least 5 d treatment with DAC.Besides,DAC alone controlled osteosarcoma cell proliferation.In immune deficient mouse models,hypomethylating pre-treatment resulted in successful T-cell homing to tumor sites.Moreover,the combination treatment with DAC and CTA specific T-cell adoptive transfer significantly suppressed tumor proliferation.In immune competent mouse models,hypomethylating treatment with DAC improved autologous T-cell infiltration into the tumor,and strengthened the activity of intratumoral CD8+ T-cells,elevated the secretion of IFN-gamma,granzyme B and perforin by CD8+ T-cells.Conclusion Hypomethylating treatment is able to suppress osteosarcoma cell proliferation,improve the expression of CTA in osteosarcoma cells,and consequently provide optimal environment for CTA specific T-cell adoptive therapy.

Objective To analyze the clinical effect of minimally invasive transforaminal lumbar interbody fusion (MI-TLIF) in combination with unilateral pedicle fixation on lumbar degenerative diseases in the elderly.Methods From June 2011 and June 2013,51 elderly patients with lumbar degenerative diseases who underwent MI-TLIF in combination with unilateral pedicle fixation,including 39 males and 12 females,aged from 60 to 72 years (average 65 years old),were included in this study.All the 51 cases had mono segmental lesion,including 11 cases with the lesion in L3.4,22 cases with the lesion in L4.5,and 18 cases with the lesion in L5-S1.The operation time,blood loss,postoperative drainage and complications were recorded.Postoperative outpatient follow-ups were conducted once every 2-3 months for all cases,and JOA scoring criteria (29 points) of lower back pain were adopted to evaluate the clinical effects.Thin-layer helical CT scanning reconstruction was applied to evaluate the fusion rate of bone grafting.X-ray film of lumbar vertebra AP & LAT was employed to evaluate the intervertebral space height,the internal fixation and the position of cage.Results The operation time ranged between 50-90 min with a mean value of 65 min; blood loss ranged between 80-180 ml (average 110 ml); and postoperative drainage volume ranged between 20-70 ml (average 40 ml).Healing phase I was achieved in 49 cases,while post-operative local cutaneous necrosis around the incision site occurred in 2 case and the wound healed well after wound dressing.Dural laceration without nerve root injury occurred in 2 cases.The follow up in 51 patients ranged between 18 24 months (average 20 months).In terms of JOA scoring,the preoperative score was (9.7± 1.6),the post operative score was (21.4 ± 1.4) 7 d after operation and the score was (25.4 ± 1.2) in the final follow up.During the final follow-up,35 cases were evaluated as excellent,11 cases as good and 5 cases as moderate,the excellent and good rate was 90.2％,and the fusion rate of bone grafting was 88.2％.The intervertebral space height ranged from (9.2±1.5) mm before operation to (11.2±1.3) rnm 7 d after operation,to (11.0±1.2) mm in the final follow-up.There were significant difference in intervertebral space height before and after operation (both P＜0.05),and no difference was found between 7d after operation and the final follow-up (P＞0.05).Looseness,fracture or cage displacement were not observed.Conclusions The clinical effect of MI-TLIF in combination with unilateral pedicle fixation on lumbar degenerative diseases in the elderly is satisfied.It has the advantages of high fusion rate,less trauma,quicker recoverer,higher cost effectiveness and convenience.However,the indications for operation should be restricted and the long-term curative effect remains to be studied in further clinical studies.

Objective To explore the short?term functional outcomes of the reconstruction of the proximal humerus by re?verse shoulder arthroplasty after tumor rescetion. Methods 8 patients who underwent reverse shoulder arthroplasty after tumor resection between January 2013 and December 2014. 5 were female and 3 were male, mean aged was 38 years old (25-61). 2 chon?drosarcomas and 6 giant cell tumors. Enneking stageing of 2 cases with chondrosarcoma were stageⅠB and stageⅡB. 6 giant cell tumors were Campanacci stage 3, meanwhile 3 cases had pathological fractures. The deltoid and axillary nerve were intact in all patients by image analysis before the operation. The proximal humerus was resected according to Malawer typeⅠresection. Then reverse shoulder arthroplasty reconstruction and bone graft was performed. The follow?up was scheduled, and the patient received X?ray examination of the shoulder. The range of motion of the shoulder was measured, the Constant?Murley score and musculoskel?etal tumor society(MSTS) score was recorded. Results The mean duration of the operation was 2.7 h (2-3.5 h). The bleeding in the operation was 510 ml (300-850 ml). The mean length of humerus resection was 8 cm (6-10 cm). The allografts were used in 7 cases and reimplantation after tumor bone deactivation was used in one. The latissimus dorsi transfer were performed in 2 cases. The rotator cuff were resected 1-1.5 cm from the great and lesser tubercles. The follow?up was 13 months (3-26 months). No infec?tion, dislocation, or loosening of prosthesis was found by the last follow?up. The X?ray showed the case who received reimplanta?tion after tumor bone deactivation had achieved bone union 1 year postoperation,7 cases received allograft had still nonunion at the host?graft junction. Bone resorption were found in all cases in different extent but the prosthesis were stable. No local recur?rence of the tumor was found. At last follow?up, active abduction was 155° (100°-175° ) and active forward elevation was 150° (115°-170°) and Constant?Murley score was 76%(68%-87%). The MSTS score was 92%(87%-97%). Conclusion The func?tional outcomes of the reconstruction of the proximal humerus by reverse shoulder arthroplasty after tumor rescetion was satisfied in early period. The reverse shoulder arthroplasty can be used in younger patient, but long?term results need further study.

Objective To discuss the diagnosis, differential diagnosis and therapeutics of the juxtaarticular bone cyst in the acetabulum. Methods Between August 1990 to April 2009, nineteen cases of clinical data of the juxta-articular bone cyst in the acetabulum data were collected from hospital records and analyzed retrospectively. There were 11 males and 8 females, with an average age of 42.3 years (range, 18-59). Seventeen patients felt an aching pain in the hip. The duration of symptoms was from 2 months to 20years with the median 15 months. All the radiographs showed a well-demarcated oval or circular radiolucent defect, close to subchondral bone, and outlined by a thin rim of sclerotic bone. Five cases accompanied with developmental dysplasia of the hip. After curettage of the bone cyst, 14 patients received iliac crest autogenous bone grafts, 3 patients received xenoma spongy bone-graft particles, and 2 patients received artificial bone grafts. Results Fourteen patients were precisely diagnosed as bone cyst by radiograph, and 4 patients were diagnosed by CT and MRI. The major axes of the focus were 2-4 cm in 16 patients, and 4-8 cm in 3patients. The sizes of the focus had no relationship with the symptoms and the course of the disease. Seventeen cases of the focus were located at the anterior superior of the bearing surface of the acetabulum. All patients were successfully followed up from 11 months to 13 years, averaged 6.2 years, and no relapses occurred during the follow-up period in every case. Fourteen patients felt asymptomatic, 3 patients felt uncomfortable in hip occasionally, and 2 patients with developmental dysplasia of the hip felt ameliorated in the initial stage of the therapy, followed by aggravation accompanying the hip joint space narrow. Conclusion Bone graft after thorough curettage of the capsular space was an effective therapy for the juxta-articular bone cyst in the acetabulum, and the anatomic distribution of radiological lesions indicated that the stress concentration maybe the etiological factor of the bone cyst.

Objective To analyze the clinical effect of percutaneous sacroiliac screw internal fixation in treatment of sacroiliac joint dislocation. Methods From June 2002 to August 2006,16 patients with sacroiliac joint dislocation were treated with percutaneous sacroiliac screw internal fixation under C-arm X-ray tomography.There were 10 males and 6 females at age range of 10-58 years(mean 34.3 years).Results The operation lasted for 30-90 minutes(average 50.5 minutes).All patients were followed up for 12-36 months(average 18.3 months).The results of postoperative normotopia,lateral,ingate and egress Xray and CT scanning showed that all the screws were located within S1 and S2 of all,14 patients obtained satisfactory result of reduction,with no infection,nerve injuries,loosening or breakage of the screw fixation.The function and the sensation of the sacroiliac ioint and low extremities recovered to normal. Conclusions Percutaneous sacroiliac screw intemal fixation is an ideally safe and effective way to treat sacroiliac joint dislocation,for it has many advantages such as minimal invasion,reliable fixation,less complication and quick recovery.It is also very necessary to take caudad and cephalad view under an image intensifier during the operation to assure the accuracy of implantation.