ABSTRACT
Background and Objectives@#This study was to investigate the role of microRNA-29a-3p (miR-29a-3p) in human bone marrow mesenchymal stem cells (hBMSCs), and its relationship with steroid-associated osteonecrosis. @*Methods@#and Results: The online tool GEO2R was used to screen out the differentially expressed genes (DEGs) in GSE123568 dataset. Quantitative real time-polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-29a-3p, forkhead box O3 (FOXO3), alkaline phosphatase (ALP), bone gamma-carboxyglutamate protein (OCN) and RUNX family transcription factor 2 (Runx2) in the hBMSCs isolated from the patients with steroid-associated osteonecrosis. CCK-8 assay was executed to measure cell viability; western blot assay was utilized to detect FOXO3, ALP, Runx2, OCN and β-catenin expression. Cell apoptosis and cell cycle were detected by flow cytometry. Immunofluorescence assay was used to detect the sub-cellular localization of β-catenin. Bioinformatics analysis and luciferase reporter gene assay were performed to confirm whether miR-29a-3p can combine with FOXO3 3’UTR. MiR-29a-3p was markedly up-regulated in the hBMSCs of patients with steroid-associated osteonecrosis, while FOXO3 mRNA was significantly down-regulated. Transfection of miR-29a-3p mimics significantly inhibited the hBMSCs’ proliferation, osteogenic differentiation markers’ expressions, including ALP, Runx2, OCN, and repressed the ALP activity, as well as promoted cell apoptosis and cell-cycle arrest. FOXO3 was identified as a target gene of miR-29a-3p, and miR-29a-3p can inhibit the expression of FOXO3 and β-catenin, and inhibition of miR-29a-3p promoted translocation of β-catenin to the nucleus. @*Conclusions@#MiR-29a-3p can modulate FOXO3 expression and Wnt/β-catenin signaling to inhibit viability and osteogenic differentiation of hBMSCs, thereby promoting the development of steroid-associated osteonecrosis.
ABSTRACT
Objective To understand the situations of departments of intensive care units (ICUs) of different level hospitals in Guizhou Province, and to provide directions and evidences for improving quality control in critical care medicine.Methods A county-level hospital and a provincial-level hospital's comprehensive ICU in Guizhou Province were selected to record and analyze and compare the structural indicators, patient admission and transfer, disease distribution, ventilator associated pneumonia (VAP), intravascular catheter related blood stream infection (CRBSI) and catheter-associated urinary tract infection (CAUTI) of the two hospitals' comprehensive ICU in 2017.Results The ICU of the People's Hospital of Suiyang County (county hospital) was found in 2012, and the ICU of the Affiliated Hospital of Guizhou Medical University (provincial hospital) was found in 1994. Until 2017, there were 10 and 46 beds, 6 (all of them hold bachelor's degree) and 18 physicians (6 of them hold PhD, 5 of them hold master's degree, 7 of them hold bachelor's degree), 17 (4 of them hold bachelor's degree, 13 of them hold college degree or graduated from secondary school) and 69 nurses (2 of them hold master's degree, 53 of them hold bachelor's degree, 14 of them hold college degree or graduated from secondary school) in the two ICUs respectively, there were significant differences in the education background of the physicians and nurses between the two ICUs (bothP < 0.01). During 2017, 471 cases were admitted to the ICU of the county hospital while 1633 cases were admitted to the ICU of the provincial hospital. Compared with the ICU of the provincial hospital, the ratio of the patients with acute physiology and chronic health evaluationⅡ(APACHE Ⅱ) ≥ 15 at admission was lower (74.8％ vs. 85.1％,P < 0.01), the ratio of direct admission was higher (30.8％ vs. 17.4％,P < 0.01), the ratio of the patients admitted to the ICU more than once was lower (0.8％ vs. 5.0％,P < 0.01), the ratio of the patients whose the length of ICU stay less than 24 hours was higher (51.6％ vs. 13.7％,P < 0.01), the ratio of the patients whose the length of ICU stay more than 28 days was lower (1.1％ vs. 2.9％, P < 0.05), the ratio of the patients discharged against-advice (25.5％ vs. 20.5％,P < 0.05) was higher, the ratio of the patients transferred to other hospitals was higher (5.1％ vs. 0.3％,P < 0.05), and the ICU mortality was lower (4.0％ vs. 13.9％,P < 0.01) in the ICU of the county hospital. The top three kinds of diseases treated in the ICU of the county hospital were brain injury (27.4％), trauma (19.1％) and toxication (6.8％); while in the ICU of the provincial hospital were brain injury (18.6％), sepsis (16.2％) and severe acute pancreatitis (4.8％). In addition, the incidences of VAP, CRBSI and CAUTI in the ICU of the county hospital were 10.0/1000 ventilator days, 1.4/1000 catheter days, 0.5/1000 catheter days; while in the ICU of the provincial hospital were 5.8/1000 ventilator days, 2.0/1000 catheter days, 3.7/1000 catheter days, respectively.Conclusions There are short of physicians and nurses in the ICU of the provincial and county hospitals in Guizhou Province, and the educational level of the medical staff in the ICU of the county hospital is relatively low. Moreover, there were significant differences in the admissions and treatments and the outcomes of the critically ill patients between the two ICUs. The characteristics of the ICUs of county hospitals should be fully considered when the quality control of critical care medicine and continuing medical education are done.
ABSTRACT
We performed this research mainly to explore the effect of bone sialoprotein (BSP) silence by siRNA on the adhesion ability to bone matrix of bone-seeking breast cancer cells (MDA-MB-231BO). Also we aimed to provide experimental data for prevention and treatment of breast cancer bone metastasis by targeting BSP. We explored the effects of BSP gene silence on characteristics of bone-seeking breast cancer cells: proliferation by MTS[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] assay, bone adhesion ability by a mouse bone adhesion model in vitro, morphology of the cells by SEM, and secretion of transforming growth factor-beta1 (TGF-beta1) and receptor activator of nuclear factor-kappa B ligand (RANKL) by ELISA kits. We performed intra-cardiac injection in nude mice to explore bone metastatic ability of different cell lines. The results showed that knockdown of BSP significantly inhibited the proliferation of MDA-MB-231BO cells and their adhesion to bone matrix. We also observed bone destruction caused by bone resorption around some adhering cells. The appearances of the cells changed in BSP gene silenced group, and the secretion of TGF-beta1 and RANKL decreased. The results showed BSP gene silence can partial inhibition bone metastasis of breast cancer cells in nude mice by X-ray assay and hematoxylin-eosin staining. Based on our research, siRNA-mediated BSP silencing can inhibit proliferation and adhesion to bone matrix of bone-seeking breast cancer cells and change their surface structure, thus inhibits their bone metastatic ability.