1.Research on the gene expression profile of inducing pancreatic duct stem cells in rats to differentiate into insulin-secreting cells
Kai REN ; Yuerong HUAN ; Jiang WU ; Mengyao HAN ; Guangxian ZHOU ; Pingping SUN ; Mei XIAO
Chinese Journal of Diabetes 2025;33(6):449-461
Objective To investigate the gene expression profile in rat pancreatic ductal stem cells(PDSCs)when induced to differentiate into insulin-secreting cells(IPCs),with the goal of identifying key genes involved in this differentiation process.Methods The expanded PDSCs were categorized into a normal control(NC)group and an induced(Tre)group.PDSCs continued expansion culture in NC group,and cultured in induction medium for 28 days to facilitate the differentiation of PDSCs into IPCs in Tre group.Dithizone staining was employed to morphologically assess whether the cells exhibited a reddish-brown coloration,indicating a positive result.The immunofluorescence staining method was used to detect the expression of insulin(Ins)and PDX1 in the cells following induction.Additionally,ELISA was conducted to measure the Ins release from IPCs,thereby verifying the responsiveness of the induced cells to glucose-stimulated Ins secretion.Concurrently,cells were collected on induction days 0 and 28 for RNA sequencing(RNA-seq),and differentially expressed genes(DEGs)were analyzed and functionally annotated.The analysis revealed that regulatory factor X3(RFX3)was overexpressed in PDSCs,and the impact of RFX3 upregulation on differentiation induction was subsequently verified.Results Compared with NC group,DTZ staining was positive,PDX1 and Ins proteins were expressed,and an increased release of Ins in response to sugar stimulation was demonstrated in the Tre group.RNA-seq analysis identified 4270 DEGs,and functional enrichment analysis utilizing the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases revealed associations with Ins response,positive regulation of Ins secretion,pancreatic endocrine cell development,and overall pancreatic development.Additionally,functionally related genes such as ALDHA2,CREB5,EIF6,FOXO1,RFX3,WNT5a,OGT,GPR39,SMAD6,and TRPM2 were identified,indicating involvement in the cell cycle,TGF-β1 signaling pathway,FOXO signaling pathway,and Wnt signaling pathway in the regulation of the differentiation of pancreatic ductal stem cells(PDSCs)into insulin-producing cells(IPCs).Furthermore,the upregulation of RFX3 can inhibit the expression of TGF-β1 within 72 hours,thereby promoted the formation and release of Ins from insulin-positive cells.Conclusions Multiple genes and signaling pathways associated with pancreatic β-cell function collectively regulate the differentiation of rat PDSCs into IPCs.Notably,the upregulation of RFX3 enhances this differentiation process.
2.Research on the gene expression profile of inducing pancreatic duct stem cells in rats to differentiate into insulin-secreting cells
Kai REN ; Yuerong HUAN ; Jiang WU ; Mengyao HAN ; Guangxian ZHOU ; Pingping SUN ; Mei XIAO
Chinese Journal of Diabetes 2025;33(6):449-461
Objective To investigate the gene expression profile in rat pancreatic ductal stem cells(PDSCs)when induced to differentiate into insulin-secreting cells(IPCs),with the goal of identifying key genes involved in this differentiation process.Methods The expanded PDSCs were categorized into a normal control(NC)group and an induced(Tre)group.PDSCs continued expansion culture in NC group,and cultured in induction medium for 28 days to facilitate the differentiation of PDSCs into IPCs in Tre group.Dithizone staining was employed to morphologically assess whether the cells exhibited a reddish-brown coloration,indicating a positive result.The immunofluorescence staining method was used to detect the expression of insulin(Ins)and PDX1 in the cells following induction.Additionally,ELISA was conducted to measure the Ins release from IPCs,thereby verifying the responsiveness of the induced cells to glucose-stimulated Ins secretion.Concurrently,cells were collected on induction days 0 and 28 for RNA sequencing(RNA-seq),and differentially expressed genes(DEGs)were analyzed and functionally annotated.The analysis revealed that regulatory factor X3(RFX3)was overexpressed in PDSCs,and the impact of RFX3 upregulation on differentiation induction was subsequently verified.Results Compared with NC group,DTZ staining was positive,PDX1 and Ins proteins were expressed,and an increased release of Ins in response to sugar stimulation was demonstrated in the Tre group.RNA-seq analysis identified 4270 DEGs,and functional enrichment analysis utilizing the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases revealed associations with Ins response,positive regulation of Ins secretion,pancreatic endocrine cell development,and overall pancreatic development.Additionally,functionally related genes such as ALDHA2,CREB5,EIF6,FOXO1,RFX3,WNT5a,OGT,GPR39,SMAD6,and TRPM2 were identified,indicating involvement in the cell cycle,TGF-β1 signaling pathway,FOXO signaling pathway,and Wnt signaling pathway in the regulation of the differentiation of pancreatic ductal stem cells(PDSCs)into insulin-producing cells(IPCs).Furthermore,the upregulation of RFX3 can inhibit the expression of TGF-β1 within 72 hours,thereby promoted the formation and release of Ins from insulin-positive cells.Conclusions Multiple genes and signaling pathways associated with pancreatic β-cell function collectively regulate the differentiation of rat PDSCs into IPCs.Notably,the upregulation of RFX3 enhances this differentiation process.
3.Application of an intervention plan based on unplanned readmission risk model in the rehabilitation of patients with acute myocardial infarction complicated with cardiogenic shock after percutaneous coronary intervention
Sujuan WANG ; Yuerong REN ; Xiaohui FU ; Hao CHEN ; Hongwen MA
Chinese Critical Care Medicine 2021;33(4):487-490
Objective:To study and analyze the application effect of intervention plan based on unplanned readmission risk model (LACE) in the rehabilitation of patients with acute myocardial infarction (AMI) complicated with cardiac shock (CS) after percutaneous coronary intervention (PCI).Methods:Ninety-three patients with AMI complicated with CS who received PCI in Tianjin Union Medical Center from January 2019 to December 2020 were enrolled. The patients were divided into LACE intervention group ( n = 46) and routine intervention group ( n = 47) according to the different nursing intervention methods. The patients in the routine intervention group received routine interventions, including drug care, diet care, psychological care, health education and telephone follow-up, while those in the LACE intervention group were assessed for the risk of LACE, and then intervention measures were formulated according to the score of LACE index, including strengthening risk awareness, life behavior, daily life ability, self-care ability, health recovery and health needs. The follow-up period in both groups was 3 months. The changes of cardiac function, incidence of adverse cardiac events, readmission rate, self-management ability after intervention and quality of life level before and after intervention were compared between the two groups. Results:There was no significant difference in cardiac function or quality of life before intervention between the two groups. After intervention for 3 months, the cardiac function and quality of life in the two groups were improved as compared with those before intervention. The left ventricular ejection fraction (LVEF) in the LACE intervention group was significantly higher than that in the routine intervention group (0.533±0.076 vs. 0.492±0.072, P < 0.05), the left ventricular end diastolic diameter (LVEDD) was significantly lower than that in the routine intervention group (mm: 47.09±7.01 vs. 53.23±7.15, P < 0.01), and the World Health Organization Quality of Life-brief (WHOQOL-BREF) score was also significantly higher than that in the routine intervention group (66.32±6.19 vs. 55.79±7.26, P < 0.01). The scores of self-management ability in the coronary heart disease self-management scale (CSMS) after intervention in the LACE intervention group were significantly higher than those in the routine intervention group (adverse hobbies score: 17.37±3.24 vs. 14.21±2.73, symptoms score: 14.82±3.11 vs. 10.56±2.65, emotional cognition score: 16.17±2.83 vs. 12.95±2.41, first aid score: 11.85±1.94 vs. 10.62±1.56, disease knowledge score: 15.58±2.73 vs. 12.68±2.61, daily life score: 17.80±2.61 vs. 14.33±2.36, treatment compliance score: 11.47±1.84 vs. 8.56±1.23, all P < 0.01). The incidence of adverse cardiac events and readmission rate in the LACE intervention group were significantly lower than those in the routine intervention group (10.87% vs. 29.79%, 4.35% vs. 17.02%, both P < 0.05). Conclusion:The intervention plan based on LACE risk model can effectively promote postoperative rehabilitation of patients with AMI complicated with CS after PCI, and also help to improve patients' self-management ability and quality of life, which is worthy of clinical promotion and application.

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