ABSTRACT
Objective:To study the effects of different calcium ion concentrations on epithelial mesenchymal transformation (EMT) of human peritoneal mesothelial cell (HPMC) via endoplasmic reticulum stress (ERS).Methods:HPMC cell line HMrSV5 was cultured in vitro and treated in groups. The cells in the control group, high calcium group 1, and high calcium group 2 were treated with medium containing calcium ion concentrations of 1.25, 1.75, and 2.25 mmol/L, respectively. The solvent control group was treated with medium containing 1.25 mmol/L physiological calcium ion concentration and 0.1% dimethyl sulfoxide (DMSO), the high calcium+solvent group was treated with medium containing 2.25 mmol/L calcium ion concentration and 0.1% DMSO, the high calcium+4-phenylbutyric acid (4-PBA) group was treated with medium containing 2.25 mmol/L calcium ion concentration and 1 mmol/L ERS inhibitor 4-PBA, and each group was treated for 48 hours. Morphological changes of cells in each group were observed under light microscope. The expressions of epithelial cell phenotype marker zonula occluden-1 (ZO-1) and mesenchymal cell phenotype marker α-smooth muscle actin (α-SMA) in the cells were observed by immunofluorescence staining. The expressions of EMT marker genes E-cadherin, ZO-1, α-SMA and Vimentin were detected by fluorescence quantitative polymerase chain reaction (PCR). The expressions of ERS marker proteins phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK), phosphorylated eukaryotic initiation factor 2α (p-eIF2α), transcription activating factor 4 (ATF4) and C/EBP homologous protein (CHOP) were detected by Western blotting. Results:Compared with the control group, the morphology of HMrSV5 cells became slender and fibrotic, the fluorescence intensity of ZO-1 increased, and the fluorescence intensity of α-SMA decreased in high calcium 1 and high calcium 2 groups, indicating that the cells transformed from epithelial cells to mesenchyme cells. The mRNA expressions of E-cadherin and ZO-1 were significantly decreased, while the mRNA expressions of α-SMA and Vimentin and the protein expressions of p-PERK, p-eIF2α, ATF4 and CHOP were significantly increased, moreover, the expressions of the above marker genes or proteins in the high calcium 2 group was more obvious than those in the high calcium 1 group [E-cadherin mRNA (2 -ΔΔCt): 0.53±0.05 vs. 0.75±0.09, ZO-1 mRNA (2 -ΔΔCt): 0.42±0.06 vs. 0.69±0.06, α-SMA mRNA (2 -ΔΔCt): 1.81±0.16 vs. 1.32±0.14, Vimentin mRNA (2 -ΔΔCt): 2.05±0.22 vs. 1.48±0.16, p-PERK protein (p-PERK/β-actin): 0.81±0.09 vs. 0.59±0.06, p-eIF2α protein (p-eIF2α/β-actin): 0.87±0.10 vs. 0.50±0.06, ATF4 protein (ATF4/β-actin): 0.93±0.10 vs. 0.72±0.06, CHOP protein (CHOP/β-actin): 0.79±0.09 vs. 0.46±0.04, all P < 0.05]. Compared with the solvent control group, the morphological changes of cells, the expressions of EMT marker genes and ERS marker proteins after high calcium ion concentration of 2.25 mmol/L were consistent with those in the high calcium 2 group than control group. Compared with the high calcium+solvent group, the cell morphology recovered the characteristics of polygonal and pebble-like epithelial cells in the high calcium+4-PBA group, the fluorescence intensity of ZO-1 increased, the fluorescence intensity of α-SMA decreased, and the mRNA expressions of E-cadherin and ZO-1 in the cells were significantly increased [E-cadherin mRNA (2 -ΔΔCt): 0.86±0.09 vs. 0.57±0.04, ZO-1 mRNA (2 -ΔΔCt): 0.81±0.06 vs. 0.48±0.05, both P < 0.05], the mRNA expressions of α-SMA and Vimentin and the protein expressions of p-PERK, p-eIF2α, ATF4 and CHOP were significantly decreased [α-SMA mRNA (2 -ΔΔCt): 1.21±0.13 vs. 1.77±0.15, Vimentin mRNA (2 -ΔΔCt): 1.30±0.14 vs. 1.94±0.20, p-PERK protein (p-PERK/β-actin): 0.38±0.04 vs. 0.92±0.11, p-eIF2α protein (p-eIF2α/β-actin): 0.34±0.05 vs. 1.05±0.13, ATF4 protein (ATF4/β-actin): 0.57±0.06 vs. 0.97±0.11, CHOP protein (CHOP/β-actin): 0.51±0.04 vs. 0.90±0.12, all P < 0.05]. Conclusion:High calcium ion concentrations of 1.75 mmol/L and 2.25 mmol/L promote EMT of HPMC via activating ERS.
ABSTRACT
【Objective】 To investigate the correlation of serum exosomal microRNA-301a (miR-301a) with cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy. 【Methods】 A total of 211 patients with diabetic nephropathy treated with peritoneal dialysis from June 2019 to June 2020 in the First Hospital Affiliated of Hebei North University were selected as study subjects. Serum exosomal miR-301a was detected by real-time fluorescence quantitative polymerase chain reaction. The patients were divided into high miR-301a group and low miR-301a group based on the median of miR-301a; the clinical data of the two groups were compared. The correlation of miR-301a with high-sensitivity C-reactive protein (hs-CRP) was analyzed by Spearman. Linear regression was applied to analyze the factors associated with the effect of miR-301a. The patients were followed up for two years. Kaplan-Meier and Log-Rank were conducted to compare the cumulative incidence of cardiovascular and cerebrovascular events between the two groups, and COX regression and restricted cubic spline were used to analyze the level-effect relationship between miR-301a and cardiovascular and cerebrovascular events after peritoneal dialysis. 【Results】 Thirty-seven cases (17.54%) of cardiovascular and cerebrovascular events occurred during follow-up. The hs-CRP level and dialysis duration were lower in low miR-301a group than in high miR-301a group (P<0.05). There was a positive correlation between miR-301a and hs-CRP (rs=0.237, P=0.001). Linear regression analysis showed that hs-CRP was independently associated with miR-301a (P<0.05). The cumulative cardiovascular and cerebrovascular event rate in low miR-301a group was 3.70% (4/108), which was lower than that in high miR-301a group [32.04% (33/103), P<0.001]. COX regression analysis showed that high serum albumin level was an independent protective factor for cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy,while high hs-CRP level and miR-301a >1.46 were independent risk factors for cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy. Restricted cubic spline fitting COX regression analysis showed a non-linear relationship between miR-301a and cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy (P<0.05). 【Conclusion】 hs-CRP is independently associated with miR-301a in diabetic nephropathy peritoneal dialysis patients. High miR-301a level suggests a high risk of cardiovascular and cerebrovascular events after peritoneal dialysis in diabetic nephropathy.
ABSTRACT
Most of the existing near-infrared noninvasive blood glucose detection models focus on the relationship between near-infrared absorbance and blood glucose concentration, but do not consider the impact of human physiological state on blood glucose concentration. In order to improve the performance of prediction model, particle swarm optimization (PSO) algorithm was used to train the structure paramters of back propagation (BP) neural network. Moreover, systolic blood pressure, pulse rate, body temperature and 1 550 nm absorbance were introduced as input variables of blood glucose concentration prediction model, and BP neural network was used as prediction model. In order to solve the problem that traditional BP neural network is easy to fall into local optimization, a hybrid model based on PSO-BP was introduced in this paper. The results showed that the prediction effect of PSO-BP model was better than that of traditional BP neural network. The prediction root mean square error and correlation coefficient of ten-fold cross-validation were 0.95 mmol/L and 0.74, respectively. The Clarke error grid analysis results showed that the proportion of model prediction results falling into region A was 84.39%, and the proportion falling into region B was 15.61%, which met the clinical requirements. The model can quickly measure the blood glucose concentration of the subject, and has relatively high accuracy.
Subject(s)
Humans , Algorithms , Blood Glucose , Neural Networks, ComputerABSTRACT
The use of non-invasive blood glucose detection techniques can help diabetic patients to alleviate the pain of intrusive detection, reduce the cost of detection, and achieve real-time monitoring and effective control of blood glucose. Given the existing limitations of the minimally invasive or invasive blood glucose detection methods, such as low detection accuracy, high cost and complex operation, and the laser source's wavelength and cost, this paper, based on the non-invasive blood glucose detector developed by the research group, designs a non-invasive blood glucose detection method. It is founded on dual-wavelength near-infrared light diffuse reflection by using the 1 550 nm near-infrared light as measuring light to collect blood glucose information and the 1 310 nm near-infrared light as reference light to remove the effects of water molecules in the blood. Fourteen volunteers were recruited for
Subject(s)
Humans , Blood Glucose , Diabetes Mellitus , Nonlinear DynamicsABSTRACT
Objective To investigate the effect of Niaoduqing Granule on the quality of life and the activa-tion of TGF-β1/Smad signalling in chronic renal failure rats received peritoneal dialysis. Methods Thirty SD rats were used to establish peritoneal dialysis models and were randomly divided into the model group(group M),the low dose group(group L),and the high dose group(group H).The dosage of group L and group H was 2 and 4 g/(kg·d),respectively. The body mass,the number of independent activities,the index of renal hypertrophy, Scr,BUN,experssion of TGF-β1 and p-Samd2/3 were determined. Results The body weight growth rate was shown decreased as follows:group N>group H>group L>group M.The number of spontaneous activity of group M decreased significantly than those in other groups. Scr,BUN and kidney hypertrophy index were significantly different among the four groups.Compared with group M,BUN,Scr,kidney hypertrophy index,levels of TGF-β 1 and p-Samd2/3 were significantly decreased in group L and H. Levels of TGF-β1 and p-Samd2/3 were signifi-cant different between group L and H. Conclusions Niaoduqing Granule can improve the quality of life and re-duce the level of serum creatinine and urea nitrogen in chronic renal failure rats received peritoneal dialysis,and the mechanism may be associated with the regulation of TGF-β1/Smad signaling pathway.
ABSTRACT
Objective To explore the effects of losartan potassium and simvastatin combination therapy on oxidative stress indicators in diabetic patients on peritoneal dialysis through 12 weeks observation. Methods Diabetic patients with end-stage nephropathy (n=80) who were treated with continuous ambulatory peritoneal dialysis were randomly divided into two groups:control group who received routine treatment (n=40), treatment group who were given losartan potassium 50 mg, once per day and simvastatin 20 mg, once every night (n=40). HbA1C, Insulin dosage, Oxidative stress indicators(SOD, GSH-PX, MDA and Hcy)were compared between two groups before and after peritoneal dialysis. Results There was no significant difference of HbA1C between the 2 groups before and after treatment(P>0.05). The insulin doses increased be?fore dialysis in both groups after CAPD treatment. It is lower in the treatment group than that in the control group ( P<0.05). The expression levels of GSH-PX and SOD in treatment group were higher while the expressions of Hcy and MDA were lower after treatment. The expressions of GSH-PX and SOD were higher while the expressions of Hcy and MDA were lower in treat?ment group than those in control group when comparing the same time point(P<0.05). GSH-PX expression level was lower while the expressions of MDA and Hcy were higher after dialysis than those before dialysis in control group ( P<0.05). Con?clusion Losartan potassium combined with simvastatin treatment can improve curative effect and oxidative stress indicators in diabetic patients on peritoneal dialysis.