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【Objective】 To detect the expressions of microRNA（miR）-362 and histone deacetylase 6 （HDAC6） in bone cancer pain （BCP） rats and investigate the analgesic effect of miR-362 and its potential analgesic mechanism. 【Methods】 The BCP model was developed by injecting Walker 256 mammary gland carcinoma cells into bone marrow cavity. Plasmid transfection was used to regulate the expressions of miR-362 and HDAC6. The Van Frey filaments and radiant heat instrument were used to detect the paw withdrawal threshold （PWT） and paw withdrawal latency （PWL）. qRT-PCR was used to detect mRNA expression levels of miR-362 and HDAC6， and Western blotting was used to detect protein expression of HDAC6 and nuclear factor kappa-B p65 （NF-κB p65）. ELISA assay was used to detect the protein levels of interleukin （IL）-1β， IL-6 and tumor necrosis factor α （TNF-α）. Luciferase activity assay was used to determine the relationship between miR-362 and HDAC6. 【Results】 Compared to sham group， the significant decrease of PWT and PWL， decrease of miR-362 and the increase of HDAC6 mRNA and protein in the spinal were detected in BCP group （P<0.05）. Compared to BCP group， the significantly increase of PWT and PWL and decrease of HDAC6 mRNA and protein in the spinal were detected in BCP+LV-miR-362 group （P<0.05）. Compared to BCP+LV-miR-362 group， PWT and PWL significantly decreased in BCP+LV-miR-362+LV-HDAC6 group （P<0.05）. In addition， compared to BCP group， significant decreases of NF-κB p65， IL-1β， IL-6 and TNF-α in spinal were detected in BCP+LV-HDAC6 siRNA group （P<0.05）. Moreover， compared to mimic miR-362+HDAC6 3’UTRMUT group， the luciferase activity significantly decreased in mimic miR-362+HDAC6 3’UTRWT group （P<0.05）. 【Conclusion】 As a key factor regulating the mechanism of BCP through “HDAC6-NF-κB p65” signal pathway in rats， targeting miR-362 may be a novel therapeutic method for BCP.
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Objective To produce the biological calcium citrate cement with fresh oyster shells, and investigate its compressive strength and biocompatibility so as to provide the experimental basis for clinical application of the material.Methods The compressive strength of biological calcium citrate cement was measured and its surface morphology was observed by SEM.The calcium release curve and pH value were measured in the simulated body fluid.Last, its biocompatibility was detected by cytotoxicity test.Results Biological calcium citrate cement produced by 0.33mL/g liquid solid ratio had the maximum compressive strength, and the crystal structure of the material was uniform and orderly.The determination of pH value showed that the degradation and absorption of biological calcium citrate cement did not significantly change the pH value of the body fluid.With gradual degradation of the material, the concentration of Ca2+ in the solution increased gradually.Cytotoxicity test showed that this material had good biocompatibility and no cytotoxicity.Conclusion Biological calcium citrate cement possesses strong compressive strength and good biocompatibility, and it can form a microenvironment with low in alkaline and high in calcium.
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Objective To investigate the effects of inositol hexaphosphate (IP6) on proliferation of human osteosarcoma cell line MG-63 and primary cultured osteoblasts so as to explore the optimal concentration for achieving anti-cancer effects.Methods We primary cultured and identified human osteoblasts.Then we made recovery and normal culture of human osteosarcoma cell line MG-63.We tested the proliferation of two kinds of cell lines under different concentrations of IP6 by MTT to determine the optimal concentration and then detected MG-63 cell cycle and apoptosis by flow cytometry.Results When IP6 concentration was more than 1 mmol/L,IP6 began to inhibit the proliferation of MG-63 cell line in the time-dose dependent manner.When the concentration reached 4 mmol/L,this inhibitory effect was the maximum.When IP6 concentration was 0.5 mmol/L or 1 mmol/L,the proliferation of osteoblasts was not obviously inhibited.When it was 2 mmol/L,the proliferation was slightly inhibited.A concentration of 4 mmol/L caused the apoptosis of osteoblasts.Conclusion IP6 can inhibit the proliferation of osteosarcoma cell line MG-63 and lead to its apoptosis.The optimal concentration is 2 mmol/L for achieving anti-cancer effects.
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ABSTRACT:Objective To study the expressions of adrenomedullin (ADM)andβ-catenin in human osteosarcoma of different differentiation grading and their clinical significance.Methods We chose 33 cases of human osteosarcoma with different pathological grading (14 cases of high differentiation grading and 1 9 cases of low grading)and 10 cases of human osteochondroma to detect the expressions of ADM protein,total (T)β-catenin and phosphorylated (P)β-catenin using immunohistochemical method. Results The expression of ADM was detected in the cytoplasm.In osteosarcomas of higher and lower differentiation grading,the positive rate of ADM was 57.1% (8/14)and 100% (1 9/1 9),respectively,with significant differences (P =0.0030.05).The expression of P-β-catenin was detected in the cytoplasm.In osteosarcomas of higher and lower differentiation grading,the positive rate of P-β-catenin was 35.7% (5/14)and 78.9% (1 5/1 9),with a significant difference (P =0.029 0.05 )or between T-β-catenin and P-β-catenin (P = 0.052 > 0.05 ).Conclusion The expressions of ADM and P-β-catenin proteins and the different distributions of expression of T-β-catenin protein in cytoplasm and/or nucleus are important indicators for judging the pathological grading of osteo-sarcoma.
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BACKGROUND:The age, complication, injury to operation time, and pain level are important factors that affect patients with postoperative mortality, but many scholars disagree. Factors affecting death in elderly patients after femoral head replacement stil need further study. OBJECTIVE:To observe factors related to the survival status of elderly patients within 1 year after receiving femoral head replacement. METHODS:A total of 96 elderly patients receiving femoral head replacement at Department of Orthopedics, the First Affiliated Hospital of Xi’an Jiaotong University from January 2011 to January 2014 were selected. On admission, patients’ age, sex, time from injury to operation, injury to admission time, pain classification, admission hemoglobin, serum albumin content of admission, admission lymphocyte count, and the amount of blood transfusion and preoperative complications were recorded. Fisher test was used to analyze the factors affecting survival status within 1 year after replacement in patients with femoral head replacement using one-way analysis of variance. Multivariate Cox test was utilized to perform multi-factor analysis. RESULTS AND CONCLUSION: The injury to hospital time was > 5 days. Pain classification was grades III and IV. The hemoglobin content was 1 000 mL. Serum albumin content was 7 days was significantly higher than patients with the time from injury to operation < 7 days (P < 0.05). The serum albumin content < 35 g/L and grades III and IV of ASA classification were factors related to patient’s death (P < 0.05). Results confirm that the risk factors for survival status within 1 year of femoral head replacement contain intraoperative volume of blood transfusion, admission to operation time, hemoglobin content, and grading of pain.
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Adrenomedullin (ADM) has wide distribution in the body and has many biological effects,which participates in many physiological and pathological processes.What's more,it is closely related to the degree of malignancy,prognosis and metastasis of tumors.ADM has different effects on different types of cells via one or more signal pathways,so that can product various effects.It also provides a new treatment strategy for preventing or slowing the progress of the cancer.