1.Effect and mechanism of microRNA-146a-5p on high glucose-induced injury of renal tubular epithelial cells
Chenge WANG ; Haimei DU ; Dongsheng YIN
Journal of Clinical Medicine in Practice 2025;29(11):14-19,25
Objective To investigate the effect of microRNA-146a-5p(miR-146a-5p)on high glucose-induced injury of renal tubular epithelial cells by regulating high mobility group A2(HMGA2).Methods Human renal tubular epithelial cells(HK-2)were cultured in vitro and di-vided into different groups according to various treatment methods:normal control group(Con group),high glucose group(HG group),HG+miR-146a-5p group,HG+si-HMGA2 group,HG+miR-146a-5p+pcDNA-HMGA2 group,miR-NC group,miR-146a-5p group,anti-miR-NC group,and anti-miR-146a-5p group.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression levels of miR-146a-5p and HMGA2 mRNA in each group.A dual-luciferase reporter assay was employed to verify the targeting relationship between miR-146a-5p and HMGA2.Western blot was utilized to detect the expression levels of cleaved-caspase3,cleaved-caspase9,and HMGA2 protein in each group.Flow cytometry was applied to determine the apoptosis rate of cells in each group.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflam-matory factors[interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interleukin-8(IL-8)]in each group.The levels of oxidative stress indicators[catalase(CAT),superoxide dismutase(SOD),malondialdehyde(MDA)]in each group were also measured.Results A nucleotide se-quence complementary to miR-146a-5p was found in the 3'-untranslated region(3'-UTR)of HMGA2.The expression level of HMGA2 protein in the miR-146a-5p group was lower than that in the miR-NC group,while was higher in the anti-miR-146a-5p group than that in the anti-miR-NC group(P<0.05).The total apoptosis rate,early apoptosis rate,late apoptosis rate,and the levels of cleaved-caspase3 protein,cleaved-caspase9 protein,HMGA2 mRNA,HMGA2 protein,TNF-α,IL-6,IL-8,and MDA in the HG group were higher than those in the Con group,whereas the levels of miR-146a-5p,CAT,and SOD were lower(P<0.05).In the HG+miR-146a-5p group,these indicators showed opposite trends compared to the HG group(P<0.05).Similarly,in the HG+si-HMGA2 group,the total apoptosis rate,early apoptosis rate,late apoptosis rate,and the levels of cleaved-caspase3 protein,cleaved-caspase9 protein,HMGA2 mRNA,HMGA2 protein,TNF-α,IL-6,IL-8,and MDA were lower than those in the HG group,while the levels of CAT and SOD were higher(P<0.05).In the HG+miR-146a-5p+pcDNA-HMGA2 group,the trends of these indicators were consistent with those between the HG group and the HG+miR-146a-5p group,showing statistically significant differences when compared to the HG+miR-146a-5p group(P<0.05).Conclusion MiR-146a-5p may play a regulatory role in high glucose-induced injury of re-nal tubular epithelial cells by targeting HMGA2.
2.Application of case-based learning guided by ICF in rehabilitation practice teaching
Yuhan KONG ; Guihua TIAN ; Li DU ; Hongwei LI ; Haimei ZHOU ; Dingqun BAI
Chinese Journal of Medical Education Research 2021;20(3):304-307
Objective:To explore the effect of case-based learning (CBL) guided by ICF (International Classification of Functioning, Disability and Health) in rehabilitation practice teaching.Methods:Fifty-eight rehabilitation undergraduates who had practiced in the Department of Rehabilitation Medicine in The First Affiliated Hospital of Chongqing Medical University from February 2019 to December 2019 were randomized into observation group and control group. The observation group adopted ICF-guided CBL method for practice teaching, while the control group adopted traditional teaching methods. At the end of the internship, all the students were assessed in terms of their theoretical knowledge, case analysis and clinical practice ability. At the same time, questionnaires were used to conduct a satisfaction survey on the teaching method, and the results of the two groups were compared and analyzed by ttest and rank-sum test through SPSS 25.0. Results:The scores of the observation group were higher than those of the control group, with statistical significance ( P<0.05) . In the satisfaction survey, the observation group was superior to the traditional teaching method in terms of self-thinking ability and learning initiative, with statistical significance ( P<0.05). Conclusion:ICF-guided CBL method can strengthen students' independent thinking ability and learning initiative, and improve the students' clinical comprehensive ability of rehabilitation treatment.
3.THE EFFECTS OF AECs CONDITIONED MEDIUM ON INDUCING NEURONAL DIFFERENTIATION OF BMSCs
Danni WANG ; Haimei SUN ; Rongping LI ; Juan DU ; Chunli ZHAO ; Deyu ZHENG ; Fengqing JI ; Hui YANG
Acta Anatomica Sinica 1955;0(03):-
Objective To study the effects of amniotic epithelial cells conditioned medium on the differentiation of bone marrow stromal cells into neural cells. Methods Bone marrow stromal cells and amniotic epithelial cells were isolated and cultured in vitro,then the cell surface antigen was detected by flow cytometry and the expressions of nestin and ki67 were detected by immunofluorescence staining method.When the cells were co-cultured with amniotic epithelial cells conditioned medium,the morphological character of cells was observed by inverse phase-contrast microscope,and the expressions of NSE(neurone specific enolase),TH(tyrosine hydroxylase) and DAT(dopamine transporter) were detected by immunofluorescence staining method. Results Amniotic epithelial cells conditioned medium had obvious inductive effect on bone marrow stromal cell's neural differentiation.Conclusion The amniotic epithelial cells conditioned medium may have inductive effect neuron-like cell's differentiation and dopaminergic neuron-like cell's differentiation of bone marrow stromal cells in vitro.

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