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BACKGROUND: Osteosarcoma (OS) is the most common bone cancer mostly seen in people aged 10-25 years. This research aims to clarify the function of hsa_circ_0001982 in osteosarcoma (OS) and its effect on drug resistance, preliminarily exploring its mechanism. METHODS: The expression of hsa_circ_0001982 and miR-143 in OS clinical tissues and cells was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR), MTT, colony formation assay, and transwell assay assessed cell proliferation, colony formation, migration, and invasion, respectively. The targeted relationship of hsa_circ_0001982 and miR-143 was verified by a dual-luciferase reporter assay. RESULT: The expression of hsa_circ_0001982 was significantly increased in OS tissues and cells (MG63), as in well as chemoresistant OS tissues and cells (MG63/Dox). Overexpression of hsa_circ_0001982 promoted proliferation, colony formation, migration, invasion, and multidrug resistance in MG63 cells. By contrast, knockdown of hsa_circ_0001982 markedly reduced the resistance of MG63/Dox cells to doxorubicin (IC50 evidently reduced). Bioinformatic prediction showed that miR-143 was a target miRNA of hsa_circ_0001982, and a dual-luciferase reporter assay proved this. Further experiments revealed that miR-143 expression was notably downregulated in OS tissues, chemoresistant OS tissues, and MG63/Dox cells. Moreover, miR-143 was negatively correlated with hsa_circ_0001982 in OS cells and tissues. CONCLUSION: The regulation of malignant behaviors such as proliferation, invasion, migration, and multidrug resistance of OS cells by hsa_circ_0001982 may be achieved by targeting miR-143. Moreover, hsa_circ_0001982 is a potential target for early diagnosis and targeted therapy of OS.

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OBJECTIVE: MicroRNA (miR)-22 plays crucial roles in malignant tumors and is involved in regulation of chemosensitivity. Additionally, altered expression of circulating miR-22 has been reported in various cancers. This study was designed to investigate plasma miR-22 expression in patients with osteosarcoma (OS) and determine its diagnostic, prognostic, and chemosensitivity prediction value. METHODS: Plasma miR-22 levels in 120 patients with OS and 120 healthy controls were detected by real-time quantitative reverse transcription PCR. Associations of plasma miR-22 expression with the patients' clinicopathological features and prognosis were then assessed. RESULTS: Plasma miR-22 levels in patients with OS were significantly lower than those in healthy controls. Low plasma miR-22 levels were correlated with large tumor size, advanced clinical stages, positive distant metastasis, and poor tumor response to preoperative chemotherapy. Plasma miR-22 could discriminate OS patients from controls and distinguish patients with a good response to therapy from those with a poor response to therapy. Multivariate analysis revealed that low plasma miR-22 expression was a significant independent predictor of unfavorable prognosis. CONCLUSIONS: Altered plasma levels of miR-22 might serve as a novel, noninvasive biomarker for OS diagnosis, prognosis, and chemosensitivity prediction.

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Advanced oxidation protein products (AOPPs) are recognized as novel markers of oxidative stress and contribute to various medical conditions, which are associated with secondary osteoporosis. However, little is currently known regarding the role of AOPPs in the development of secondary osteoporosis. As the commander cells of bone remodeling, osteocytes are involved in the pathogenesis of osteoporosis. The present study aimed to determine the cytotoxic mechanisms of AOPPs on osteocytic MLO­Y4 cells. The results demonstrated that treatment with AOPPs significantly triggered apoptosis of MLO­Y4 cells, in a dose­ and time­dependent manner. Furthermore, exposure to AOPPs induced phosphorylation of c­Jun N­terminal kinases (JNK) and p38 mitogen­activated protein kinases (MAPK). Conversely, N­acetylcysteine inhibited the activation of JNK and p38 MAPK, thus suggesting that the AOPPs­induced activation of JNK/p38 MAPK is reactive oxygen species (ROS)­dependent. In addition, SB203580 and SP600125 suppressed apoptosis, but did not affect ROS production, following AOPPs treatment. Notably, AOPPs also induced a significant upregulation in the expression levels of sclerostin and receptor activator of nuclear factor kappa­B ligand (RANKL) in a JNK/p38 MAPK-dependent manner. These findings provide novel insights into the molecular mechanisms underlying AOPPs­mediated cell death, and suggest that modulation of apoptotic pathways via the MAPK signaling cascade may be considered a therapeutic strategy for the prevention and treatment of secondary osteoporosis.

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BACKGROUND Early metastasis of osteosarcoma (OS) is highly lethal and responds poorly to drug and radiation therapies. MicroRNAs (miRNAs) are a class of small noncoding RNAs that modulate gene expression at the post-transcriptional level. However, the detailed functions of specific miRNAs are not entirely understood. The aim of the present study was to investigate the role of miR-184 as a mediator of drug resistance in human osteosarcoma. MATERIAL AND METHODS qRT-PCR was used to analyze the expression level of miR-184 in OS cell line U-2 OS and MG-63 treated with doxorubicin. MiR-184 agomir or miR-184 antagomir was transferred into cells to regulated miR-184. The target of miR-184 was predicted by TargetScan and confirmed by luciferase reporter assay. Bcl-2-like protein 1 (BCL2L1) expression was detected by Western blot. Cell apoptosis was determined by Annexin V staining and analysis by flow cytometry. RESULTS Doxorubicin induced time-dependent expression of miR-184 in OS cell line U-2 OS and MG-63. Luciferase reporter assay identified BCL2L1 as the direct target gene of miR-184. Furthermore, doxorubicin reduced BCL2L1 expression, which was reversed by miR-184 overexpression and further decreased by miR-184 inhibition in OS cells. In addition, miR-184 agomir reduced doxorubicin-induced cell apoptosis, whereas miR-184 antagomir enhanced apoptosis in OS cells, suggesting that up-regulation of miR-184 contributes to chemoresistance of the OS cell line. CONCLUSIONS Our data show that miR-184 was up-regulated in OS patients treated with doxorubicin therapy and leads to poor response to drug therapy by targeting BCL2L1.

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BACKGROUND: Large segmental bone defects caused by trauma, infection, or bone tumor resection are difficult to cure and have been a problem in the field of bone repair for decades. The objective of this study was to discuss the efficacy of combined therapy of free periosteum and bone allograft in treating bone defects and to provide a theoretical basis for clinical application of this therapy. MATERIAL/METHODS: A unilateral tibia cortical defect model in New Zealand white rabbits was established according to Girolamo method. Total 48 rabbits were randomized into 3 groups: a simple bone defect group (n=16), an autogenous bone graft group (n=16), and a periosteum and bone allograft combined therapy group (n=16). The efficacy was evaluated by imaging inspections and scoring, HE staining, and RT-PCR in postoperative weeks 2, 4, 8, and 12. RESULTS: The results of imaging and histopathological inspections in the study indicated that in postoperative weeks 4, 8, and 12 the experimental and control groups had statistically significant differences in Lane-Sandhu radiographic scoring and relative bone density when compared with the simple bone defect group (P<0.05). The RT-PCR results suggested that the expression of SPP-1, BMP-2, and VEGF in the experimental group was higher than in the control group (P<0.05) and the expression of Col Ia1 in the control group was higher than in the experimental group (P<0.05). CONCLUSIONS: Efficacies of the combined therapy (periosteum combined with bone allografting) and the criterion standard therapy (autogenous bone grafting) are equivalent in treating bone defects in New Zealand white rabbits.

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The effect of an electrical double layer (EDL) on microchannel flow has been studied widely, and a constant bulk electric conductivity is often used in calculations of flow rate or pressure drop. In our experimental study of pressure-driven micropipette flows, the pipette diameter is on the same order of magnitude as the Debye length. The overlapping EDL resulted in a much higher electric conductivity, lower streaming potential, and lower electroviscous effect. To elucidate the effect of overlapping EDL, this paper developed a simple model for water flow without salts or dissolved gases (such as CO(2)) inside a two-dimensional microchannel. The governing equations for the flow, the Poisson, and Nernst equations for the electric potential and ion concentrations and the charge continuity equation were solved. The effects of overlapping EDL on the electric conductivity, velocity distribution, and overall pressure drop in the microchannel were quantified. The results showed that the average electric conductivity of electrolyte inside the channel increased significantly as the EDL overlaps. With the modified mean electric conductivity, the pressure drop for the pressure-driven flow was smaller than that without the influence of the EDL on conductivity. The results of this study provide a physical explanation for the observed decrease in electroviscous effect for microchannels when the EDL layers from opposing walls overlap.