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Objective The relationship between calcified nanoparticles (CNPs) and the formation of urinary stones is drawing increasing attention and the specific mechanisms involved. This study aims to investigate the mechanisms of the formation of kidney stone caused by CNPs. Methods A total of 48 rats were randomly and equally divided into a CNPs group (each rat was injected with 2 mL CNPs through the tail vein to establish a rat kidney stone model of CNPs), and a control group (injected the same amount of sterile isotonic saline instead of CNPs). We compared the expression levels of autophagy-related proteins, such as Beclin-1 and LC-3, the formation of autophagosomes and calcium salt crystals in renal tissues at time points of 3h, 6h, 12h, 24h, 1w, 2w, 4w and 8w in two groups. Results The relative expression levels and positive cells of Beclin-1 and LC-3 in CNPs group at 3h,6h,12h,24h, 1w, 2w, 4w, 8w were significantly higher than those in the control group (P< 0.05), and reached the highest value at 24 (P< 0.05). The number of autophagosomes at 24h, 1w, 2w, 4w, and 8w in the CNPs group ((2.83±0.32), (3.00±0.26), (3.70±0.44), (3.90±0.98), (4.70±0.51)/HP, respectively) were significantly higher than those in the control group (0.73±0.15)/HP (P <0.05). The scores of calcium salt crystals in the CNPs group at 2w, 4w, and 8w significantly increased compared to the control group (P <0.05). The calcium salt crystal formation score ((0.92 ± 0.98) points) was positively correlated with the expression intensities of Beclin-1 and LC-3 ((6.78 ± 4.25), (2.61 ± 2.57), respectively) (r = 0.843, 0.628, P <0.05), which was positively correlated with the number of autophagosomes (2.53 ± 1.41) (r = 0.923, P <0.001). Conclusion CNPs may damage renal tubular epithelial cells, and induce immediate autophagic activity, also increase expression of autophagy-related proteins and autophagosome formation, which will promote the formation and aggregation of calcium salt crystals in renal tubules to some extent.
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The literatures were searched and summarized about the autophagic operating mechanism on the TCM benefiting Qi and promoting blood circulation for the diabetes and diabetic complications. The study found that Chinese herbs with the function of benefiting Qi and promoting blood circulation could motivate the tissues's autophagy, which had good the rapeutic effect on diabetes and diabetic complications.
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Objective:To study B cells proliferation and activation induced by tumor derived-autophagosomes ( TDA). Methods:Splenocytes of mice were incubated with 10μg/ml TDAs in vitro ( with the 10μg/ml whole cell lysate as the control).At the 3rd day,the expression of MHCⅠ,Ⅱand co-stimulate molecules including CD86,CD40 were detected by flow cytometry,and B cells proliferation was detected at the 5th day.B cells were selected from spleen of mice using anti-CD43 dynabeads,and incubated with 10μg/ml TDA in vitro ( with the same concentrated whole tumor cell lysate as the control).At the 7th day,IgM in the supernatant were tested by ELISA.Results:When compared with whole cell lysate stimulated B cells,TDA efficently stimulated B cells division in vitro ( TDA group:28.6%, Whole cell lysate group:4.4%) , and significantly up-regulated the expression of MHC class Ⅰ,Ⅱ and co-stimulatory molecules (CD86 and CD40) on B cells,and enhanced the levels of total IgM secretion in vitro.Conclusion:Tumor-derived autophagosomes ( TDA) ,as a vacuole antigen vector, could stimulate B cells proliferation, activation, and increased IgM secretion in vitro.