Objective: To construct an adenoviral vector containing cDNA of EWS-FLI1 and detect its expression in peripheral blood mononeuclear cell(PBMC).To Investigate the antitumor immunity in vitro of the EWS-FLI1 gene modified-dendritic cells.Methods: The EWS-FLI1 cDNA in plasmid Pec1/ EWS-FLI1 was digested and subcloned into the shuttle plasimid padtrack-cmv.The shuttle plasmid and the bone plasmid pADeasy-1 were cotransformed into BJ5183 cells.The recombinant plasmid was generated by homologous recombination in BJ5183 cells.The positive clone was obtained by digestion and electrophoresis.Transforming the recombinant plasmid into "293 cells" by lipofectamine method.Adenoviruses with high titer and purity were obtained by amplifying in the"293 cells" on a large scale and ultra-centrifugation in CsCL step gradient solutions.The cytotoxic activity of stimulated T cells to Ewing sarcoma cells was detected by ~(51)Cr release assay.Results: PCR showed that the adenovirus contained EWS-FLI1(cDNA.) After the PBMC were transfected by Ad EWS-FLI1,the EWS-FLI1 mRNA was detected by(RT-PCR.) The antigen-specific CTL was induced successfully by the EWS-FLI1 gene modified-DC.The vigorous antigen-specific CTL response against A673 cells was detected by ~(51)Cr release assay.The killing percentage was 35.18%?0.0128% at effector-target ratio 40∶1,which was more efficient than that of the control.Conclusion: The recombinant adenovirus was successfully constructed and could efficient express EWS-FLI1 in PBMC.After T lymphocytes were stimulated by DCs modified with EWS-FLI1 gene,the specific CTL response against Ewing's sarcoma cell line A 673 in vitro was observed successfully.~(51)Cr release assay showed that there was significant difference between the experimental group and the control group.

[Objective]To summary wound related complications and risk factors of sacral tumor surgery,and discuss the prevention and treatment strategy in order to decrease surgical risk and prevent the related complications.[Methods]From August 1997 to June 2008,302 patients with sacral tumors who underwent operations were retrospective studied to review complication types,risk factors and treatment strategies of complications.[Results]Wound nonunion,deep infection,cerebrospinal fluid leakage,viscera injuries and severe bleeding were the most common complications of peri-operation stage.Tumor diameters larger than 10cm,surgery and radiation history were the major risk factors of wound nonunion,deep infection and viscera injuries.Severe bleeding happened in 11 patients,and the major risk factors were tumor larger than 10cm,tumor locating at S2 or higher and en bloc surgery.Surgery and radiation history were the main reasons of cerebrospinal fluid leakage.[Conclusion]Understanding the risk factors of complications of sacral tumor surgery are helpful to judge and prevent complications after surgery.Some measures of prevention and treatment are effective to decrease the surgical risks.

[Objective]To retrospectively study all the low grade central osteosarcoma(LGCO) cases treated in our center in about a past decade.[Method]Thirteen cases with LGCO treated from January 1998 to June 2008 were studied.The mean age at diagnosis was 31-year-old.The anatomical location of the lesions was similar to that of a conventional osteosarcoma.Pain and mass were common complaints with a mean duration of 16 months before diagnosis.Eight cases accepted the first operation in our center,while the other 5 cases had undergone operations in other hospitals with an intralesional resection in four and a marginal resection in one.Eleven patients had an Enneking stage I disease and two had a stage III disease.The first choice for patients with stage I disease was wide resection.[Result]In two patients with local recurrence,the postoperative pathological diagnoses were fibrosarcoma and malignant fibrous histocytoma respectively.The metastatic lesions were found to be a LGCO in both patients with stage III disease.The average follow up duration was 29 months.Lung metastasis occurred in one stage I patient 15 months after operation.Gama-knife was used to get rid of the lung lesions and then systemic chemotherapy was given.Now he was still alive with disease 27 months after the first operation.One of the 2 patients with stage III disease died of the disease 26 months after operation and the other one was failed to follow up 6 months after operation.The other 10 patients were alive without disease at the last follow-up.For those who received limb salvage operations,the mean MSTS 93 score was 79%.[Conclusion]For most patients with LGCO,wide resection of tumor is preferred.Good oncologic and functional results could be expected after a limb salvage operation without chemotherapy.Insufficient resection of tumor would lead to recurrence.Long time follow up is compulsory.High grade sarcomas would be found in some of the local recurrent lesions.Metastasis would occur in few patients and the metastatic lesions may still be a LGCO.

Objective To explore the recurrence rate, the complications and functional status of 25 patients with grade I chondrosarcoma of long bones treated by intralesional curettage and electrocauterization, and to determine the feasibility and effi?cacy of this method for grade IA chondrosarcoma of long bones. Methods Twenty?five eligible patients treated in our hospital from May 2003 to December 2011, were collected in this study with a mean age of 49 years (range, 28-72 years). According to En?neking staging system, all the lesions were staged as IA. Patients received surgery of the lesion before were excluded. The involved bones were femur (13 patients), tibia (4 patients), and humerus (8 patients). During the operation, a large elliptical cortical window about the size of the longest dimension of the lesion was made to ensure the thorough exposure of the lesion and avoid inadequate curettage. The lesion was curettaged thoroughly after the cortical window was made, then the high speed bur drill was applied to clear away a thin layer of the reactive bone shell. After a thorough lavage of the cavity, electrocauterization was done alongside the cavity wall slowly twice. Allograft or artificial bone was used to fill in the cavity to enhance bone healing. If mechanical property of the long bone was endangered by the cortical window and the surgical procedure, plate and screws were applied to strengthen the bone to avoid post?operative fracture. Results All the 25 patients were followed up regularly. The period of follow?up was from 38 months to 142 months, with a mean time of 80 months. Calcification was observed clearly in all the lesions. The length of the le?sions varied from 4 cm to 11 cm, with an average length of 7 cm. Nineteen patients received a plate and screws fixation because the mechanical property of the affected bone, while the rest received no internal fixation. Deep infection occurred in 1 patient (4%, 1/25) two months after operation. Thorough debridement and packing of bone cement with antibiotics was done to control the infec?tion. The infection has been controlled till the latest follow?up at 62 months after operation. Local recurrence occurred in one pa?tient thirteen months after the primary operation. Tumor resection, mega?prosthesis replacement was applied to the patient. The pa?tient has been disease free till now for 67 months. Mean Musculoskeletal Tumor Society (MSTS) 93 functional score of all patients was 93%(range, 83%-100%). The five?year survival rate of these twenty?five patients was 100%. Conclusion Intralesional cu? rettage together with the application of high?speed bur drill and electrocauterization is an applicable method for grade I chondrosar?coma of long bones. The excellent functional result and low recurrence rate make it a favorable option for the selected cohort of pa?tients.

Surgical treatment for bone and soft tumors of pelvis and sacrum presents a big challenge, because of the complex anatomy of sacropelvic region, large tumor volume at presentation, rich blood supply to the tumor and visceral involvemen, et al. Therefore, surgical excision and reconstruction are technically difficult for sacropelvic tumors. Extensive intraoperative haemorrhage could be life-threatening, and this issue remains a major concern. How to effectively control bleeding during surgery is critical for successful operation and patient's favorable prognosis. Some previous attempts, such as interventional selective internal iliac artery embolization or manual ligation through an additional anterior approach, were tested to be ineffective. Inspired by the success of resuscitative endovascular balloon occlusion of the aorta (REBOA) which resemble an endovascular tourniquet for traumatic hemorrhagic shock, some researchers have applied this techinique to control surgical bleeding during pelvic or sacral tumor resection.The authors have performed REBOA for more than 1 500 sacropelvic tumr surgeries since 2003 in Peking University People's Hospital. The patient age, the diameter of femoral artery and aorta, atherosclerosis, as well as tumor location, volume and expansion and blood suppy, have to be thoroughly evaluated prior to REBOA administration. Admittedly, the application of REBOA do reduce intraoperative bleeding, shorten the operation duration, improve the safety of surgery, yet some complications were observed including local hematoma at the puncture site, acute arterial thrombosis, femoral artery pseudoaneurysm or occlusio, et al. The purpose of this study is to review the literature on REBOA administration in pelvic and sacral tumors excision, with the focus on its indications, performing procedure, the safety and efficacy, and complications. Moreover, in order to popularize the clinical application of aortic balloon occlusion in the future, we summarize our experience of abdominal aortic balloon occlusion over 10 years.

Objective:To explore the role and value of indocyanine green (ICG) fluorescence imaging in the surgery of bone and soft tissue tumors.Methods:A total of 38 patients with bone and soft tissue tumors, including 17 males and 21 females, aged 36±21 years (range 5-75 years), who underwent resection with application of ICG-based NIR imaging were retrospectively analyzed. ICG was intravenously administrated at a dose of 2 mg/kg on the day before surgery. Intraoperative NIR imaging was performed to confirm the tumor fluorescence and to detect residual tumor after tumor resection. The NIR imaging was post hoc analyzed to explore the influence factor of tumor stain rate and SBR value.Results:Tumor staining with ICG was successful in 34 of the 38 patients, with an overall staining rate of 90%. The ICG tumor stain rate was not influenced by different pathology types, malignant or benign pathology, the reception of neoadjuvant chemotherapies or not, and the length of time between drug administration and surgery ( P>0.05). The median SBR of 34 patients with successful tumor staining was 2.9 (2.3, 5.7). Different pathological types, malignant or benign pathology, whether to receive neoadjuvant chemotherapy, preoperative ICG administration time, preoperative dose of denosumab administration in giant cell tumor of bone patients and tumor response to neoadjuvant chemotherapy had no significant effects on SBR ( P>0.05). After tumor resection, a total of 57 pieces of tissue with residual fluorescence signals were detected and resected under the fluorescence guidance, 30 of which were pathologically confirmed to contain residual tumor lesions, with an overall accuracy of 53%. The accuracy of intralesional resection was significantly higher than that of en bloc resection (71% vs. 16%, χ 2=15.51, P=0.000). Conclusion:A high percentage of bone and soft tissue tumors can be stained with ICG. The tumor stain of ICG was stable and not easily influenced by external factors. This technique was useful to detect residual tumors, especially after piecemeal resection.