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Objective:To explore the effects of conditioned medium of human bone marrow mesenchymal stem cells (BMSCs) on the proliferation, adhesion and differentiation of immortalized human Müller cell line (MIO-M1).Methods:The differentiation was induced in the third-passage BMSCs with osteogenic, chondrogenic and adipogenic medium and identified by alizarin red, alcian blue and oil red O staining, respectively.The expression levels of mesenchymal stem cell markers CD73, CD90 and CD105 and hematopoietic cell markers CD34, CD45 and human leukocyte antigen-DR (HLA-DR) were assayed by flow cytometry.The expressions levels of Müller cell markers SOX9, glutamine synthetase (GS), vimentin and cellular retinaldehyde-binding protein (CRALBP), retinal stem cell markers SOX2, nestin and CHX10, and cell proliferation marker cyclin D3 (CCND3) in MIO-M1 cells were detected by immunofluorescence staining.The MIO-M1 cells were divided into standard medium group, 293T conditioned medium group, and BMSC conditioned medium group and were incubated in the medium according to grouping.The cellular area, circularity, elongation factor and perimeter were analyzed quantitatively.The cell cycle was detected by flow cytometry, and the cell proliferation was determined by neurospora experiment and 5-ethynyl-2'-deoxyuridine (EdU) staining.The expression of vascular cell adhesion molecule 1 (VCAM-1) at protein and mRNA levels in the culture supernatant was detected by enzyme linked immunosorbent assay (ELISA) and quantitative real-time PCR (qRT-PCR), respectively.The expression of retinal neuron markers protein kinase C (PKCα), Rhodopsin, microtubule-associated protein 2 (MAP2) and β-tubulin (Tuj1) was detected by immunofluorescence staining and qRT-PCR.Results:CD73, CD90, CD105 showed an enhanced expression, and CD34, CD45 and HLA-DR showed weakened expression in the BMSCs.The BMSCs differentiated into osteoblasts, chondrocytes and adipocytes.Expression of SOX9, GS, vimentin and CRALBP, SOX2, CHX10, nestin and CCND3 was found in the MIO-M1 cells.Compared with standard medium group and 293T conditioned medium group, MIO-M1 cells cultured in BMSC conditioned medium group changed into an elongated spindle-shaped or multipolar morphology with reduced cell area, increased elongation index and decreased circularity, showing statistically significant differences among them ( F=6.973, 12.370, 6.311; all at P<0.01). There were increased neurospheres formed by MIO-M1 cells in BMSC conditioned medium group compared with standard medium group and 293T conditioned medium group at different time points ( Fgroup=134.300, P<0.001; Ftime=82.910, P<0.001). Compared with the standard medium group and 293T conditioned medium group, the EdU-positive rate and proliferation index of MIO-M1 cells in BMSC conditioned medium group were significantly increased, with statistically significant differences ( F=6.973, 74.110; all at P<0.05); the VCAM-1 protein expression in cell supernatant and the relative expression level of VCAM-1 mRNA in BMSC conditioed medium group were significantly increased ( F=13.720, 7.896; all at P<0.05); the mRNA expression levels of PKCα, Rhodopsin, Tuj1 and MAP2 were higher in MIO-M1 cells of BMSC conditioned medium group under the condition of differentiation ( F=14.490, 5.424, 14.330, 7.405; all at P<0.05). Conclusions:BMSCs conditioned medium can change the morphology of MIO-M1 cells and promote their proliferation, adhesion and differentiation into retinal neurons.
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Objective:To explore the effect of therapeutic communication on infusion safety, disease uncertainty, coping styles, anxiety and depression in patients with emergency infusion.Methods:A total of 126 patients treated with infusion patients in emergency department in Jiangsu Provincial People's Hospital, the First Affiliated Hospital of Nanjing Medical University from July 2019 to December 2019 were selected and divided into two groups by random digits table method with 63 cases in each group. Patients in the control group received routine nursing, and patients in the intervention group received therapeutic communication nursing intervention on this basis. The incidence of adverse events of infusion during the intervention of the two groups of patients was observed. The disease uncertainty, coping styles, anxiety and depression were evaluated by Mishel's Uncertainty in Illness Scale (MUIS), Medical Coping Mode Questionnaire (MCMQ), Self-rating Anxiety Scale (SAS) and Self-rating Depressive Scale (SDS) before and after intervention.Results:The final collection of 109 patients with complete questionnaire, the recovery rate was 86.51% (109/126), including 55 cases in the intervention group, 54 cases in the control group. The incidence of adverse events of infusion was 16.36%(9/55) in the intervention group and 29.63%(16/54) in the control group, and there was significant difference( χ2 value was 5.057, P<0.05). There was no significant difference in the score of MUIS, MCMQ, SAS, SDS before the intervention between the two groups ( P>0.05). After the intervention, the uncertainty, unpredictability, lack of information and complexity dimension of the intervention group patients' MUIS scores were respectively (17.76 ± 2.49), (12.03 ± 2.51), (11.82 ± 2.12), (11.74 ± 2.24), which were lower than those in the control group (24.72 ± 2.94), (16.31 ± 2.27), (16.13 ± 2.51), (15.39 ± 2.31), the differences were significant( t values were -13.346- -8.375, P<0.05). The confront score of the intervention group patients' MCMQ was (19.13 ± 2.62) higher than that in the control group(13.79 ± 1.96), the avoidance and yield scores were respectively (8.71 ± 1.34), (9.81 ± 1.17), which were lower than those in the control group (14.57 ± 1.93), (15.12 ± 1.86), the differences were significant( t values were 12.031, -18.441, -17.875, P<0.05). The scores of SAS and SDS in the intervention group were (29.43 ± 3.62), (27.67 ± 3.11) respectively, which were lower than those in the control group (37.44 ± 5.31), (40.12 ± 4.92), the differences were significant( t values were -9.216, -15.821, P<0.05). Conclusions:Therapeutic communication can reduce the risk of emergency infusion, reduce the uncertainty of patients to the disease, improve patients' countermeasures to the disease, relieve patients' anxiety and depression, and improve the nursing quality of emergency infusion patients.
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Objective To study the effects of concentration of glucose on the proliferation of bone marrow mesenchymal stem cells (BMMSCs).Methods BMMSCs passaged to the 4th to 6th were inoculated and cultured.The control group was treated with DMEM/F12 (1∶1) standard medium,and the basal glucose concentration was 5.5 mmol/L.The experimental group was treated with DMEM/F12 (1∶1) high-glucose medium with the concentration of 20.5,25.5,30.5 and 35.5 mmol/L respectively,and continuously cultured for 15 d.The absorbance (A) values of each group once per day were measured by thiazolyl blue (MTT) colorimetry.Results Compared with the control group,the absorbance was increased at the 2nd,3rd,and 5th days in the 20.5 and 30.5 mmol/L groups,and the differences were statistically significant (all P＜0.05).For the 25.5 mmol/L group,the absorbance was increased at the 2nd,3rd,5th and 8th days,and the differences were statistically significant (all P＜0.05).For the 35.5 mmol/L group,the absorbance was increased at the 2nd,3rd,5th and 8th days and decreased at the 8th day,and the differences were statistically significant (all P＜0.05).The results of 20.5,25.5,30.5 and 35.5 mmol/L groups were further compared one another.The results showed that the absorbance at the 4th and 12th days in the 35.5 mmol/L group were lower than that of the 20.5 mmol/L group,and the differences were statistically significant (all P＜0.05).The absorbance at the 11th and 14th days in the 30.5 and 35.5 mmol/L groups were lower than that of the 20.5 mmol/L group,and the differences were statistically significant (all P＜0.05).Conclusions High-glucose environment can promote the proliferation of BMMSCs in a short time.However,with the prolongation of culture time,this effect is gradually weakened because of the inhibiting effect of high-glucose environment to the cell proliferation.
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BACKGROUND:Studies have shown that the finite element method could preferably simulate the biomechanical analysis for the object with complicated structures and irregular shapes. The similarities for the finite element model have great influences on the results of the analysis. However, to construct an ideal model is the most time-consuming and complicated portion for the finite element analysis. OBJECTIVE:To construct a finite element model for the maxilary first molar and the periodontal tissue, and to provide a basis of biomechanical researches of the maxilary first molar. METHODS: A volunteer with complete mandibular dentition and healthy periodontal tissue was selected in this study. Cone-beam CT was scanned. The images were saved as DICOM format. These images were imported to the medical modeling software Mimics. The surface model for the maxilary first molar and the alveolar bone was constructed. The model was then imported to GiD for pre-processing. Thus, the complete three-dimensional finite element model for the maxilary first molar and the periodontal tissue was constructed. RESULTS AND CONCLUSION:A finite element model for bilateral maxilary first molar, periodontal ligament and maxilary alveolar bone was constructed, including 896 035 nodes and 4 881 067 elements. This model has restored the geometric shape and the structure of the research object. This study successfuly constructed finite element models of maxilary first molar and the periodontal tissue, which can be a basis of biomechanical researches for the maxilary first molar and the periodontal tissue under the effect of different clinical orthodontic forces.