Percutaneous kyphoplasty combined with zoledronic acid infusion in the treatment of osteoporotic thoracolumbar fractures in the elderly.

OBJECTIVE: We studied the efficacy of zoledronic acid (ZOL) infusion on radiographic and clinical outcomes after percutaneous kyphoplasty (PKP) for elderly patients with osteoporotic thoracolumbar fractures (osteoporotic vertebral compression fractures [OVCFs]). MATERIALS AND METHODS: We retrospectively analyzed 95 elderly patients (age >65 years) with OVCF. All patients were followed up for 2 years. Thirty-two patients were treated with only once-yearly 5 mg ZOL infusion (ZOL group), 34 patients with only PKP (PKP group) and 29 patients received ZOL infusion 3 days after PKP (PKP+ZOL group). RESULTS: There were no significant differences in the patients' age, gender, body mass index, lumbar spine bone mineral density T-scores, baseline of Visual Analog Scale scores and Oswestry Disability Index scores (P>0.05). The postoperative vertebral heights of patients with OVCF after PKP and PKP+ZOL were 23.70±3.03 and 24.30±3.13 mm, respectively, which were significantly higher than that of patients in ZOL group (P<0.05). The reduction in degrees of kyphotic deformity in the PKP and PKP+ZOL groups were corrected to 8.4° and 8.7°. The bone mineral density T-scores of patients with OVCF in the ZOL group and PKP+ZOL group were significantly higher than that in the PKP group (P<0.05). The Visual Analog Scale and the Oswestry Disability Index scores of the PKP+ZOL and PKP groups were significantly lower than those of the ZOL group (P<0.05). The incidence of recompression vertebral fracture (RVF) in the PKP group was 14.7%, but there was no patient with RVF in the PKP+ZOL group (P<0.05). CONCLUSION: Once-yearly 5 mg ZOL infusion combined with PKP could provide beneficial effects in elderly osteoporotic patients with OVCF.

Cinobufagin Induces Apoptosis in Osteosarcoma Cells Via the Mitochondria-Mediated Apoptotic Pathway.

BACKGROUND/AIMS: Osteosarcoma is a common primary malignant bone tumor that mainly occurs in childhood and adolescence. Despite developments in the diagnosis and treatment of osteosarcoma, the prognosis is still very poor. Cinobufagin is an active component in the anti-tumor Chinese medicine called "Chan Su", and we previously revealed that cinobufagin induced apoptosis and reduced the viability of osteosarcoma cells; however, the underlying mechanism remains to be elucidated. Herein, the present study was undertaken to illuminate the molecular mechanism of cinobufagin-induced apoptosis of osteosarcoma cell. METHODS: U2OS and 143B cells were treated with different concentrations of cinobufagin. Cell viability, colony formation ability and morphological changes were assessed by a CCK-8 assay, a clonogenic assay and light microscopy, respectively. Cell apoptosis was detected by Hoechst 33258 and Annexin V-FITC/PI staining. Reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) were determined by flow cytometry. Glutathione (GSH) levels were detected by a GSH and GSSG assay kit. The levels of apoptosis-related proteins were determined by western blotting, and 143B cells were introduced to establish a xenograft tumor model. The effect of cinobufagin on osteosarcoma was further investigated in vivo. RESULTS: Our results showed that cinobufagin significantly reduced the viability of U2OS and 143B cells in vitro in a dose-and time-dependent manner. In addition, cinobufagin-induced apoptosis in U2OS and 143B cells was concentration-dependent. Moreover, we found that cinobufagin treatment increased the level of intracellular ROS, decreased ΔΨm, reduced GSH and inhibited GSH reductase (GR). The effects of cinobufagin on cell proliferation, apoptosis, ROS generation and ΔΨm loss were dramatically reversed when the cells were pretreated with the thiol-antioxidants NAC or GSH. Moreover, cinobufagin treatment increased the expression of the pro-apoptotic protein Bax and decreased the expression of the anti-apoptitic protein Bcl-2, thus altering the ratio of Bax to Bcl-2. Furthermore, Cinobufagin treatment caused cytochrome c release from the mitochondria to cytoplasm, thus increasing the protein levels of cleaved-caspase family members to induce apoptosis. Ac-DEVD-CHO or Z-LEHD-FMK significantly reduced cinobufagin-induced apoptosis. Finally, a subcutaneous xenograft animal study verified that cinobufagin also significantly suppressed osteosarcoma growth in vivo. CONCLUSIONS: Our present data demonstrated that cinobufagin triggered cell apoptosis in osteosarcoma cells via the intrinsic mitochondria-dependent apoptosis pathway by the accumulation of ROS and the loss of ΔΨm. In an in vivo subcutaneous xenograft model, cinobufagin exhibited excellent tumor inhibitory effects. These results suggest that cinobufagin might potentially be further developed as an anti-tumor candidate for treating osteosarcoma patients in the clinic.

Construction of a plasmid for overexpression of human circadian gene period2 and its biological activity in osteosarcoma cells.

Beyond their established role in the mammalian circadian clock, recent studies have confirmed that the circadian genes have been implicated in tumor onset and progression. Currently, the biological effects of circadian genes on osteosarcoma cells' proliferation and migration are not well understood. Period2 (Per2) is one of the core circadian genes that act as master regulators of development and is frequently dysregulated in several cancers. However, the effects of human Per2 (hPer2) on the biological behavior of osteosarcoma cells are rarely reported. In the present study, to address the expression of hPer2 in osteosarcoma cells, the pEGFP-N1-hPer2 eukaryotic expression vector was constructed and transfected into cultured MG63 cells using Lipofectamine™ 2000. The overexpression of hPer2 in MG63 cells was verified by qRT-PCR and Western blotting, respectively. Finally, we investigated the effects of hPer2 protein overexpression on MG63 cells' viability, cycle, apoptosis, and invasive ability. In conclusion, the recombinant pEGFP-N1-hPer2 plasmid had been constructed successfully and expressed effectively in MG63 cells. Furthermore, results also showed that the viability, proliferation, and invasive abilities were suppressed, and the apoptosis was enhanced in MG63 cells. This preliminary study provides ground work for further research on the roles of circadian gene hPer2 in osteosarcoma cells MG63 and would offer promise for the development of novel therapeutic strategies in the treatment of osteosarcoma.

[Characteristics of poloxamer thermosensitive in situ gel of dexamethasone sodium phosphate].

Thermosensitive in situ gel is a novel drug delivery system which can form gel in situ after injection of the polymer solution into the body and releases the drug in a controlled manner, thus provides a promising strategy for localized drug delivery. The aim of the present work is to investigate the characteristics including gelation temperature, sol-gel transition temperature (T(s-g)), gel strength, stable viscosity, erosion and drug release behavior of the thermosensitive in situ gel which are composed of different concentrations of poloxamer Pluronic F127 and F68. The gelation temperature was determined by tube-reverse method. Rheological measurements were carried out to evaluate T(s-g), stable viscosity and gel strength. Erosion of the gels and release of dexamethasone sodium phosphate (DSP) from the gels were investigated by membrane-free method and HPLC. Increased F127 concentration in gel decreased the gelation temperature, T(s-g) as well as erosion of the gel and drug release rate, while viscosity and gel strength rose accordingly. However, increased F68 in gel could lead to the opposite result. The poloxamer solution below T(s-g) is Newtonian fluid with comparatively low viscosity, but shows the characteristics of the pseudoplastic fluid when temperature rises near to T(s-g). Drug release was controlled by the erosion of the gel matrix, and both of them followed the zero-order kinetics. An optimized formation containing 22.5% F127 and 2.5% F68 showed more desirable characteristics which meet the clinical requirements and is of potential in future clinical therapy.

[A new method of oxygen delivery to preventing and relieving the anoxia injury--an experimental study and clinical application of hyperoxygen compound sodium chloride solution in treatment of burn shock].

OBJECTIVE: To evaluate the clinic effects and prognosis of hyperoxygen compound sodium chloride solution (HCSCS) in treatment of burn shock. METHODS: (1) Severe burned patients with 50% - 69% TBSA were enrolled and randomly divided into 2 groups, treated by HCSCS and CSCS respectively. Indices such as PaO(2), Hb, Hct and complication were observed. (2) Severe burned patients with more than 70% TBSA were enrolled and indices above and lactate were investigated as well. RESULTS: (1) The cases treated with HCSCS showed relatively stabilization during shock stage, little complication, less liquor volume and remarkable higher oxygen pressure. (2) Severe burned patients with more than 70% TBSA had high level of PaO(2) and normal lactate and less complication. CONCLUSION: The early application of HCSCS is a relatively effective method in treatment of burn shock and favourable prognosis.