1.Differentiation to renal tubular epithelial-like cells from bone marrow mesenchymal stem cells in vitro
Jianxin WAN ; Zhenhuan ZOU ; Yanfang XU ; Danyu YOU ; Jiong CU ; Yangbin PAN ; Mingbu XIE
Chinese Journal of Nephrology 2009;25(6):476-480
Objective To investigate the differentiation of rat bone marrow mesenchymal stem cells (MSCs) to renal tubular epithelial-like cells under different conditions. Methods MSCs were obtained from rat marrow. MSCs were isolated by gradient density centrifugation and plastic adherence and then purified. Surface markers were identified with flow cytometry after amplification in vitro. The purified MSCs of the third passage were cultured respectively as follows: (1) control group: DMEM medium with fetal bovine serum(FBS). (2) all-trans retinoic acid (ATRA) group: DMEM medium with FBS, ATRA and ischemic reperfusion-injured kidney tissue homogenate. (3)combination group: DMEM medium with FBS, ATRA, ischemic reperfusion-injured kidney tissue homogenate, epidermal growth factor (EGF) and bone morphogenetic protein 7 (BMP-7). After 7 days, the MSCs were collected for alkaline phosphatase (AKP) staining, cytokeratin-18 and E-cadherin immunocytochemical analysis. Results The positive rates of the third passage MSCs in CD44, CD90 and CD29 were 97.8%±0.9%, 96.8%±1.4% and 97.6%±2.4%,respectively, but in CD11b/c and CD34 were only 13.2%±0.6% and 1.2%±0.5%. The MSCs in control group were spindle. The MSCs in ATRA group were round and elliptic. The MSCs in combination group became cobblestone-like cells after 7 days. AKP staining showed that tubular epithelial-like cells from MSCs in control group were negative, some above cells in ATRA group were positive and number of above cells increased in combination group. Compared with negative control group, the ratios of cytokeratin-18 positive cells in ATRA group and combination group were respectively increassed by 29.47%±1.08% and 47.52%±2.13% (all P<0.05), the ratios of E-cadherin positive cells in ATRA group and combination group were respectively increased by 14.88%±2.46% and 36.15%±1.13% (all P<0.05). Conclusion MSCs may differentiate by renal tubular epithelial-like cells under the induction of ischemic reperfusion-injured kidney tissue homogenate and ATRA in vitro, which are further differentiated under the combined induction of EGF and BMP-7.
2.Probing into Possible Factors and Gender Differences of Anemia in Patients with Diabetes Nephropathy based on Bayesian Network Model
Chinese Journal of Health Statistics 2023;40(6):827-831
Objective Probing into possible factors and gender differences of anemia in patients with diabetes nephropathy based on Bayesian network model.Methods 224 patients with diabetes nephropathy who were treated in Ningde City hospital from June 2020 to June 2023 were taken as the research objects,patients were divided into an anemia group(98 cases)and a non anemia group(126 cases)based on whether they were anemic.Comparison of anemia related indicators in diabetes nephropathy patients of different sexes.Comparison of clinical data between two groups of patients with diabetes nephropathy.Multivariate logistic regression was used to analyze the influencing factors of anemia in patients with diabetes nephropathy.Build a Bayesian network model using R software and perform inference and prediction of the model.The effectiveness of the model was verified by receiver operating characteristic and calibration curve.Results Male patients showed significantly higher levels of Hb,RBC,and HCT compared to female patients;MCV,MCH,and MCHC were significantly lower than those in female patients,with statistically significant differences(P<0.05).Sex,course of diabetes,stage of kidney disease,ALB,CHE,eGFR and HbA1c were independent risk factors for anemia in patients with diabetes nephropathy(P<0.05).The results of receiver operating characteristic and calibration curve of training set and verification set show that the Bayesian network prediction model has good discrimination and accuracy.Conclusion Sex,course of diabetes,stage of kidney disease,ALB,CHE,eGFR and HbA1c are independent risk factors for anemia in patients with diabetes nephropathy.There are certain gender differences in anemia related indicators among patients.