ObjectiveTo investigate the expression of ErbB3 and Flotillin-2 in osteosarcoma biopsies and possible clinical pathology significance. MethodsThe tissue biopsies were harvested from 38 osteosarcoma patients and 13 osteochondroma patients between September 2009 and March 2014 for immunohistochemical staining. The ErbB3 and Flotillin-2 expressions were observed in osteosarcoma and osteochondroma biopsies, and the correlation and the relationship to the clinical and pathological features were analyzed. ResultsThe expression levels of ErbB3 and Flotillin-2 in osteosarcoma were significantly higher than those in osteochondroma (P<0.05). The high expressions of ErbB3 and Flotillin-2 had good consistency in osteosarcoma (Kappa=0.434, P=0.000). The high expression of ErbB3 was positively correlated to the clinical stages and lung metastasis (P<0.05), but it was not associated with gender, age, tumor location, and size (P>0.05). The high expression of Flotillin-2 had no correlation with clinical and pathological features (P>0.05). The influence factors of patients' overall survival included clinical stages, lung metastasis, high expression of ErbB3, and tumor size (P<0.05). Only lung metastasis and high expression of ErbB3 were independent factor affecting overall survival (P<0.05). ConclusionThe ErbB3 and Flotillin-2 express highly in osteosarcoma and the high expression has good consisitency. Besides, the high expression of ErbB3 is associated with the clinical stages and lung metastasis, indicating a poor prognosis for osteosarcoma.
ObjectiveTo investigate the effectiveness and technical key points of limb salvage surgery by allotransplantation of cryopreservated vascularized bone in children and adolescents with osteosarcoma. MethodsA retrospective analysis was made on the clinical data of 21 children and adolescents with osteosarcoma receiving limb salvage surgery by allotransplantation of cryopreservated vascularized bone from their relatives between February 2004 and April 2012. There were 13 males and 8 females, aged from 7 to 16 years (mean, 12.6 years). According to Enneking stage system, 15 cases were rated as stage ⅡA and 6 cases as stage ⅡB. The tumors located at the distal femur in 10 cases, at the proximal femur in 1 case, at the proximal tibia in 8 cases, at the proximal humerus in 1 case, and at the distal radius in 1 case. Imaging examination showed that epiphyseal extension of malignant bone tumors in 7 cases. The iliac bone allograft with deep iliac vessels was obtained from their lineal consanguinity. After preservation by a twostep freezing schedule, the iliac bone allograft with deep iliac vessels was implanted into the bone defect area after tumor resection. The size of iliac bone flap was 8.0 cm×3.0 cm×2.0 cm-14.0 cm×5.0 cm×2.5 cm. Reserved joint surgery was performed on 16 cases and joint fusion surgery on 5 cases, and external fixation was used in all cases. The chemotherapy was given according to sequential high-dose methotraxate, adriamycin, and cisplatine before and after operation. ResultsAll 21 cases were followed up from 5 months to 11 years (mean, 6.4 years). At 2 weeks after operation, the erythrocyte rosette forming cells accounted for 56.7%±3.9%, showing no significant difference when compared with that of normal control (58.3%±4.3%) (t=1.56, P=0.13), which suggested no acute rejection. At 4 weeks after operation, single photon emission computerized tomography bone scan indicated that the blood supply of bone graft was rich, and the metabolism was active. At 12 weeks after operation, the digital subtraction angiography showed the artery of iliac bone flap kept patency. X-ray films showed that malunion and non-union occurred at 5 and 6 months after operation in 1 case, respectively. The bone graft healed in the other patients, and the healing time was 3.2-6.0 months (mean, 4.4 months). At last follow-up, American Musculoskeletal Tumor Society (MSTS) score was significantly improved to 26.80±2.14 from preoperative value (17.15±1.86) (t=-4.15, P=0.00). The survival rate was 85.7% (18/21) and the recurrence rate was 9.5% (2/21). ConclusionAllotransplantation of cryopreservated vascularized bone from the relatives provides a new method for the treatment of osteosarcoma in children and adolescents. A combination of allotransplantation and chemotherapy can achieve the ideal treatment effect. The correct cutting, preservation, and transplantation of the donor bone, and indication are the key to improve the effectiveness.
Objective To evaluate the relationship of systemic immune inflammatory index (SII) with the clinical features and prognosis of osteosarcoma patients. Methods The clinical data of patients with osteosarcoma surgically treated in Fuzhou Second Hospital between January 2012 and December 2017 were retrospectively collected. The preoperative SII value was calculated, which was defined as platelet × neutrophil/lymphocyte count. The best critical value of SII was determined by receiver operating characteristic (ROC) curve analysis, and the relationship between SII and clinical features of patients was analyzed by χ2 test. Kaplan-Meier method and Cox proportional hazard model were used to study the effect of SII on overall survival (OS). The nomogram prediction model was established according to the independent risk factors of patients’ prognosis. Results A total of 108 patients with osteosarcoma were included in this study. Preoperative high SII was significantly correlated with tumor diameter, Enneking stage, local recurrence and metastasis (P<0.05). The median follow-up time was 62 months. The 1-, 3-, 5-year survival rates of the low SII group were significantly higher than those of the high SII group (100.0%, 96.4%, 85.1% vs. 95.4%, 73.7%, 30.7%), and the survival of the two groups were statistically different (P<0.05). Univariate Cox regression analyses showed that tumor diameter, Enneking stage, local recurrence, metastasis and SII were associated with OS (P<0.05). Multiple Cox regression analysis showed that Enneking stage (P=0.031), local recurrence (P=0.035) and SII (P=0.001) were independent risk factors of OS. The nomogram constructed according to the independent risk factors screened by the Cox regression model had good discrimination and consistency (C-index=0.774), and the calibration curve showed that the nomogram had a high consistency with the actual results. In addition, the ROC curve indicated that the nomogram had a good prediction efficiency (area under the curve=0.880). Conclusions The preoperative SII level is expected to become an important prognostic parameter for patients with osteosarcoma. The higher the SII level is, the worse the prognosis of patients will be. The nomogram prediction model built on preoperative SII level, Enneking stage and local recurrence has a good prediction efficiency, and can be used to guide the diagnosis and treatment of clinical osteosarcoma.
Objective To design, construct and select the optimal repl ication-defective recombinant adenovirus mediated short hairpin RNA (shRNA) which is transduced into human osteosarcoma cells to silence c-myc gene expression, and to construct the recombinant adenovirus vector expressing c-myc-shRNA and determine its viral titer. Methods Three pairs of complementary single-stranded ol igonucleotides (ss ol igos) were designed and synthesized, and then they were annealed to create a double-stranded ol igonucleotide (ds ol igos).The ds ol igos were cloned into pENTR/U6 vector to produce the shuttle plasmid pENTR/U6-shRNA, which was transduced into osteosarcoma cells by l iposome after sequencing. The plasmid with good silence effect was chosen by RT-PCR to perform the LR recombination reaction to the adenovirus backbone plasmid. The expression clone was transfected into HEK293A cells to produce repl ication-incompetent recombinant adenovirus mediated shRNA against c-myc whose cytopathic effect was observed and viral titer was determined by the viral particle (VP) method and 50% tissue culture infective dose (TCID50). Results Ds ol igos, which was verified by electrophoresis, was cloned into pENTR/U6 vector to produce pENTR/U6-shRNA shuttle plasmid, which was confirmed to be corrected by sequencing. The optimal plasmid with good silence effect was chosen by RT-PCR from the three pairs of double-stranded ol igonucleotide. By Pac I enzyme, the l inearrization repl ication-defective recombinant adenovirus mediated shRNA was constructed to perform the LR recombination reaction to the adenovirus backbone plasmid. The cytopathic effect and vacuole phenomenon of adenovirus mediated shRNA appeared at 3 days and became obvious at 6 days. The adenovirus virus titer in the first generation was 5.23 × 109 VP/mL, and reached 2.26 × 1012 VP/mL via 3-4 generations’ ampl ification. The viral titer was 10-3.8/0.1 mL determined by VP method and TCID50. Conclusion The recombinant adenovirus mediated shRNA c-myc is constructed in vitro through RNA interference technology.
Objective To investigate the effect of limb salvage on treating osteosarcoma with pathological fracture. Methods From October 2002 to January 2003, 2 cases of osteosarcoma with pathological fracture were treated by limb salvage. Intraarterial chemotherapy was given by subcutaneous implantable delivery system with caffeine. Replacement with prosthesis was performed after 5 times of chemotherapy. Results Two patients were followed up for twenty-four months and 21 months respectively. No infection, aseptic loosening, local recurrence or metastasis occurred, and function recovery of joints was satisfactory. Conclusion Limb salvage can be considered in condition that primary osteosarcoma with pathological fracture can be treated by effective and comprehensive chemotherapy.
Objective To explore the effect of pyropheophorbide-a methyl ester-mediated photodynamic therapy (MPPa-PDT) on the apoptosis in human osteosarcoma cell line MG63 and the underlying mechanism. Methods Human osteosarcoma MG63 cells in logarithmic growth phase were divided into 4 groups: blank control group (control group), the MPPa treatment group (MPPa group), the light irradiation group (LED group), and MPPa-PDT treatment group (MPPa-PDT group). MPPa-PDT group and MPPa group were incubated with MPPa (0.75?μmol/ L) for 20 hours in dark condition; control group and LED group were incubated with equal volume of fresh medium for 20 hours in the same condition. After washing with PBS and replacement with fresh culture medium, LED group and MPPa-PDT group cells were exposed to light (4.8 J/cm2) for 120 seconds. After light exposure, all groups were cultured in dark condition again. Then cellular morphology changes were observed by an inverted phase contrast microscopy, endoplasmic reticulum morphology changes were observed by transmission electron microscopy, cellular apoptosis was detected by Hoechst33258 nuclear staining, cell apoptotic rate and the levels of Ca in cells were analyzed by flow cytometry, the expression of p-PERK, C/EBP homologous protein (CHOP), cleaved-Caspase-12 were assayed by Western blot. Results In MPPa-PDT group, the retracted and round cells were observed; Hoechst33258 nuclear staining showed nuclear condensation, fragmentation, and other typical apoptotic morphological changes; the cell apoptotic rate (48.76%±3.54%) was significantly higher than that of control group (5.04%±0.41%), MPPa group (5.33%±0.38%), and LED group (6.48%±0.46%) (P < 0.05); the levels of Ca2+ in cells (485.29±58.77) was also significantly higher than that of control group (97.24±4.77), MPPa group (97.95±6.30), and LED group (101.17±5.26) (P < 0.05); swelling endoplasmic reticulum was observed under transmission electron microscope; the expressions of p-PERK, CHOP, and cleaved-Caspase-12 gradually increased at 1, 3, and 6 hours after treatment respectively, which were significantly higher than those of the other groups (P < 0.05). There was no typical apoptotic morphological changes and endoplasmic reticulum morphological changes in control group, MPPa group, and LED group, and there was no significant difference in the above indexes among 3 groups (P > 0.05). Conclusion MPPa-PDT can significantly induce apoptosis in MG63 cells. The endoplasmic reticulum stress pathway is involved in the MPPa-PDT induced apoptosis.
ObjectiveTo investigate the causes of the complications and prevention strategy by analyzing occurrence of prosthesis-related complications after extensible semi-joint prosthesis replacement for lower limbs osteosarcoma in children. MethodsEleven children with lower limbs osteosarcoma underwent resection of tumor and replacement of the extensible semi-joint prosthesis between May 2006 and October 2012. There were 6 boys and 5 girls, with an average age of 9.3 years (range, 7-12 years). The lesions located at the distal femur in 6 cases, at the proximal femur in 2 cases, and at the proximal tibia in 3 cases. The disease duration was 2-8 months (mean, 3.6 months). According to the Enneking stage, 3 cases were rated as stage ⅡA and 8 cases as stage ⅡB. The pulmonary CT and ECT results showed no pulmonary metastasis or multi spots before operation. All patients received preoperative chemotherapy treatment for 4 times. ResultsPrimary healing of incision was obtained in 10 cases. Infection occurred in 1 case at 1 week after operation, and was cured after symptomatic treatment. Nine patients received postoperative chemotherapy for 12 times, 2 patients for 2 times and 4 times respectively. One case died of multiple metastasis; in 3 cases of pulmonary metastasis, 2 cases died and 1 case survived after resection of metastatic lesion. Eight survival cases received a follow-up of 25-89 months (mean, 42.5 months). Loosening and dislocation of the proximal femoral prosthesis occurred in 1 case, loosening and subsidence of the distal femoral prosthesis in 1 case, subluxation in 1 case, and retraction in 1 case. The incidence of prosthesis-related complications was 50%. Lengthening operation was performed on 3 cases for 1 time, and on 1 case for 2 times. And 4 cases did not undergo lengthening operation. According to Enneking function evaluation standard after malignant tumor limb-salvage surgery, the results were excellent in 1 case, good in 3, fair in 3, and poor in 1 at last follow-up with an excellent and good rate of 50%. ConclusionThe prosthesis-related complications include loosening and subsidence, dislocation, knee instability, and retraction after extensible semi-joint prosthesis replacement for lower limbs osteosarcoma. The prosthesis-related complications can be reduced by the improvement of prosthesis design and manufacture, and the use of intraoperative bone cement, artificial mesh, and postoperative restrictive brace.
ObjectiveTo explore the effectiveness and safety of high-intensity focused ultrasound (HIFU) for bone tumors, so as to provide a reference for clinical decision. MethodsPubMed, EMbase, The Cochrane Library, CNKI and VIP databases were systematically searched for clinical effectiveness and safety studies of HIFU for bone tumors up to August 2014. Study selection, data extraction and quality assessment were applied independently by two reviewers, and then RevMan 5.1 software was used for conducting meta-analysis. If the data cannot be synthesized, the research outcome was described with a qualitative analysis. ResultsA total of 10 case series including 257 patients (157 males, 100 females) were included. The current evidence indicated that overall survival rates for all primary bone malignancy at 1-, 2-, 3- and 5-year were 89.8%, 72.3%, 60.5% and 50.5%, respectively. For the patients with clinical stage Ⅱb, the rates were 93.3%, 82.4%, 75% and 63.7%, respectively. For those with clinical stage Ⅲ, the rates were 79.2%, 42.2%, 21.1% and 15.8%, respectively. The local recurrence rate of HIFU for bone tumors was 7% to 9%, and recurrences at 1-, 2-, 3- and 5-year were 0%, 6.2%, 11.8% and 11.8%, respectively. The amputation rate was 2% to 7%. The adverse reaction rate was 27.2% (70/257), and among them the main was mild skin burn (21/257, 8.2%), followed by I degree burns (16/257, 6.2%), nerve damage (10/257, 3.9%) and fracture (6/257, 2.3%). ConclusionHIFU provide an alternative choice for patients with bone malignancy, with a certain effectiveness and safety. However, high-quality, large-scale randomized controlled trials or cohort studies which may focus on vary kinds of tumors, clinical stage and site of lesions are urgently needed, so that clinicians can use sufficient evidence for their clinical decision-making.
As the most common primary malignant bone tumor in children and adolescents, osteosarcoma has the characteristics of high malignancy, easy metastasis and poor prognosis. The recurrence, metastasis and multi-drug resistance of osteosarcoma are the main problems that limit the therapeutic effect and survival rate of osteosarcoma. Among them, lung metastasis is often the main target organ for distant metastasis of osteosarcoma. In recent years, people have paid attention to the signaling pathway of the occurrence and development of osteosarcoma and made in-depth studies on its mechanism. A variety of relevant signaling pathways have been constantly clarified. At present, there is still a lack of systematic and multi-directional exploration and summary on the signaling pathway related to the pulmonary metastasis of osteosarcoma. This paper explores the new direction of targeted therapy for osteosarcoma by elucidating the relationship between the signaling pathway associated with osteosarcoma and the pulmonary metastasis of osteosarcoma.
Objective To investigate the clinical application and early results of combined epiphyseal preservation and autograft bone transfer for limb salvage in children with osteosarcoma. Methods Between March 2010 and March 2011, 3 children with osteosarcoma were treated with epiphyseal preservation and autograft bone transfer. There were 1 boy and 2 girls with the age of 10-14 years. The disease duration was 2 weeks to 3 months. The tumors were rated as type II according to San-Julian radiological classification and as type IIB according to Enneking surgical classification. The locations were the distal femur in 1 case and the proximal tibia in 2 cases. The surgical technique included preoperative neoadjuvant chemotherapy, excision of part of the epiphysis, preservation of subarticular epiphysis, external fixation with Ilizarov apparatus and transport osteogenesis treatment. Safe excision border was confirmed by histological examination. Postoperative observation included the wound healing, local recurrence or distant metastasis, length, speed, alignment, and regeneration of transplanted bone, the length and function of the affected limb. Results The postoperative histological examination proved the safe surgical margin in all 3 patients. The incision healing by first intention was obtained at distal femur in 1 case and by second intention at proximal tibia in 2 cases at 4 weeks after changes of dressing. Three patients were followed up 12, 18, and 24 months, respectively. There was no local recurrence or distant metastasis during follow-up. Two cases had pin-tract infection at 10 months after external fixation and were cured after changes of dressing and antibiotics administration. The length of transplanted osteogenisis was 18.0, 9.5, and 16.0 cm, respectively. The speed of lengthening was 2.57, 2.07, and 1.20 cm/month, respectively. One patient had alignment deviation during lengthening, which was adjusted under anaesthesia. Bony healing was achieved at 8 months after lengthening end in 2 cases and external fixation was removed; 1 patient had poor bone growth and was given retraction for promoting bone growth. At last follow-up, the length of the affected limbs was 1.0-1.5 cm shorter than that of normal limbs, but no abnormalities was observed at donor sites. The affected knee flexion reached 90° and 120° in 2 patients and poor knee function was observed in 1 patient for external fixation. Conclusion The combined epiphyseal preservation and transport osteogenesis technique can be used for bone defect repair by lengthening the residual bone. It is a promising limb salvage treatment for children with osteosarcoma.