RESUMO
The present study was aimed to establish a modified method for culturing mouse renal proximal tubular epithelial cells (TECs). Renal cortex was isolated from mouse kidney and scissored into pieces. TECs were separated by digesting scissored renal cortex in type II collagenase combined with strainer filtration, and then cultured in DMEM. The morphology of TECs was observed under inverted microscopy. The cell proliferative ability was assessed by flow cytometry, and cell viability was analyzed by CCK-8 assay. The purity of TECs was identified by immunofluorescence. Immunofluorescence observation showed that more than 95% cells were epithelial marker CK18 positive and more than 90% cells expressed renal proximal TECs marker proteins, Villin, AQP1, and SGLT2. The cells could be subcultured for about 5 times. The cell proliferative ability declined following the repeated passage. This study introduced a modified efficient method for culturing highly purified mouse renal proximal TECs.
Assuntos
Células Epiteliais/citologia , Túbulos Renais Proximais/citologia , Cultura Primária de Células , Animais , Células Cultivadas , Citometria de Fluxo , CamundongosRESUMO
This paper proposes an advanced Reinforcement Learning (RL) method, incorporating reward-shaping, safety value functions, and a quantum action selection algorithm. The method is model-free and can synthesize a finite policy that maximizes the probability of satisfying a complex task. Although RL is a promising approach, it suffers from unsafe traps and sparse rewards and becomes impractical when applied to real-world problems. To improve safety during training, we introduce a concept of safety values, which results in a model-based adaptive scenario due to online updates of transition probabilities. On the other hand, a high-level complex task is usually formulated via formal languages, including Linear Temporal Logic (LTL). Another novelty of this work is using an Embedded Limit-Deterministic Generalized Büchi Automaton (E-LDGBA) to represent an LTL formula. The obtained deterministic policy can generalize the tasks over infinite and finite horizons. We design an automaton-based reward, and the theoretical analysis shows that an agent can accomplish task specifications with the maximum probability by following the optimal policy. Furthermore, a reward shaping process is developed to avoid sparse rewards and enforce the RL convergence while keeping the optimal policies invariant. In addition, inspired by quantum computing, we propose a quantum action selection algorithm to replace the existing [Formula: see text]-greedy algorithm for the balance of exploration and exploitation during learning. Simulations demonstrate how the proposed framework can achieve good performance by dramatically reducing the times to visit unsafe states while converging optimal policies.
RESUMO
BACKGROUND: Peritoneal fibrosis (PF) is a common complication of peritoneal dialysis (PD), but a specific and sensitive biomarker for PF is lacking. The present study aimed to determine the use of effluent decoy receptor 2 (eDcR2) as a biomarker for PF in PD patients. METHODS: PD patients (n = 248) were recruited, and peritoneal specimens were collected at PD initiation (n = 30) and cessation (n = 33). Enzyme-linked immunoassay was used to measure eDcR2 and the eDcR2 appearance rate (eDcR2-AR) was calculated. The levels of DcR2 mRNA and protein were determined. The correlation of eDcR2 level with peritoneal function, histological parameters and DcR2 expression were analysed. Receiver operating characteristic (ROC) analysis was used to assess the diagnostic performance of eDcR2 for PF, which was defined as a submesothelial thickness 150 µm or more. Co-localisation of DcR2 with a mesothelial marker, fibroblast markers and fibrotic markers were determined. RESULTS: The eDcR2-AR level correlated with PD duration, D/P Cr values, peritoneal Kt/V and peritoneal injury scores, especially submesothelial thickness (r = 0.638, p < 0.001). DcR2 was primarily expressed in peritoneal fibroblasts, and co-localised with α-SMA, vimentin, collagen I and fibronectin, but not with E-cadherin. Peritoneal DcR2 expression had a positive correlation with eDcR2-AR. ROC analysis indicated eDcR2 had an area under the curve of 0.907 for detection of PF (sensitivity: 78.6%, specificity: 100%) and the best cut-off value was 392.5 pg/min. CONCLUSION: The eDcR2-AR level is a potential biomarker for assessing PF in PD patients. Effluent DcR2 was mainly derived from peritoneal fibroblasts and DcR2-positive cells may accelerate PF, suggesting that it may be a potential therapeutic target.
Assuntos
Diálise Peritoneal , Fibrose Peritoneal , Humanos , Fibrose Peritoneal/diagnóstico , Fibrose Peritoneal/etiologia , Fibrose Peritoneal/metabolismo , Diálise Peritoneal/efeitos adversos , Peritônio/metabolismo , Biomarcadores/metabolismo , Curva ROCRESUMO
A practical and efficient base mediated synthesis of free 3-aminoindazoles has been developed from the reaction of nitriles with hydrazines, which successfully overcomes the difficulty of using aromatic hydrazines as substrates and allows for the synthesis of a wide range of N-aryl substituted free 3-aminoindazoles in moderate to excellent yields under mild conditions in one-pot. This finding provides a rapid and useful strategy for the synthesis of various functionalized 3-aminoindazole derivatives.
RESUMO
Based on the four TM images of Naiman Banner in Inner Mongolia in 1975, 1985, 1995 and 2005, the extent and relative rate of land use change were used as the indices to analyze the regional difference of land use/cover change in the farming-pasturing zone of Naiman Banner, and the indices abundance and importance value were adopted to analyze the spatial distribution features of land use/cover change in the study area. The results showed that from 1975 to 2005, the types of land use/cover became diversified. The annual change rate was high, and the regional difference was significant. In the northern alluvial plain sub-area, woodland area increased rapidly and largely, while sandy land area decreased obviously. The changing speed of the areas of meadow and sandy land was the fastest, while that of residential area was the slowest. The main forms of land conversion were the conversion from sandy land to cropland and woodland. In the middle sandy land sub-area, sandy land had a wide distribution. The changes of other land use types were comparatively small, and the main form of land conversion was the inter-conversion between cropland and sandy land. In southern loess sub-area, cropland was the dominant land use type and had the smallest change, meadow and sandy land changed most quickly, and the conversion from meadow to cropland and woodland was the most important land conversion form. Natural factors determined the principal characteristics of land use structure in each sub-area of the study area, and artificial factors determined the changing trends of each land use type in each sub-area.