Cigarette smoke extract-induced inflammatory response via inhibition of the TFEB-mediated autophagy in NR8383 cells.

Objective: Chronic pulmonary inflammation caused by long-term smoking is the core pathology of COPD. Alveolar macrophages (AMs) are involved in the pulmonary inflammation of COPD. The accumulation of damaged materials caused by impaired autophagy triggers inflammatory response in macrophages. As a key transcription regulator, transcription factor EB (TFEB) activates the transcription of target genes related autophagy and lysosome by binding to promoters, whereas it is unclarified for the relationship between inflammatory response induced by cigarette smoke extract (CSE) and TFEB-mediated autophagy. Thus, we investigated the role of TFEB-mediated autophagy in inflammatory response induced by CSE in NR8383 cells, and to explore its potential mechanism. Methods: Based on cell viability and autophagy, cells treated with 20% concentration of CSE for 24 h were selected for further studies. Cells were divided into control group, chloroquine (CQ, the autophagy inhibitor) group, CSE group, CSE + rapamycin (the autophagy inducer) group and CSE + fisetin (the TFEB inducer) group. The levels of tumor necrosis factor α (TNF-α), interleukin 1ß (IL-1ß), and IL-6 in supernatant were detected by ELISA kits. The protein expressions were tested by western blot. The intensity of fluorescence of Lysosome-associated membrane protein 1 (LAMP1) and TFEB was detected by immunofluorescence. Lyso-Tracker Red staining was applied to detect the lysosome environment. Results: CSE inhibited the cell viability, increased the contents of TNF-α, IL-1ß, IL-6, the ratio of LC3II/I, and the level of P62 protein. Besides, CSE decreased the fluorescence intensity of LAMP1 protein and Lyso-Tracker Red staining, as well as the ratio of nucleus/cytosol of TFEB protein. Activating autophagy with rapamycin alleviated CSE-induced inflammatory response. The activation of TFEB via fisetin alleviated CSE-induced autophagy impairment and lysosomal dysfunction, thus alleviated inflammatory response in NR8383 cells. Conclusion: CSE-induced inflammatory response in NR8383 cells, which may be related to the inhibition of TFEB-mediated autophagy.

Structural characterization and mast cell stabilizing activity of Red-edge tea polysaccharide.

The potential anti-allergic properties of tea have been demonstrated in studies supporting theanine and catechin. However, research on tea polysaccharides' anti-allergic properties has been limited. In this study, we extracted red-edge tea crude polysaccharide (RETPS) and evaluated its anti-allergic activity using the mast cell, passive cutaneous anaphylaxis, and passive systemic anaphylaxis models. We purified RETPS using the DEAE-52 cellulose column, analyzed its composition and structural characteristics, and compared the anti-allergic properties of different polysaccharide fractions. The purified components RETPS-3 and RETPS-4 displayed higher galacturonic acid content and lower molecular weight (106.61 kDa and 53.95 kDa, respectively) compared to RETPS (310.54 kDa). In addition, RETPS-3 and RETPS-4 demonstrated superior anti-allergic activity than RETPS in mice's passive cutaneous and systemic allergic reactions. Our findings provide evidence of the anti-allergic potential of tea polysaccharides and offer a theoretical foundation for developing tea polysaccharides as a functional anti-allergic food product.

Effects of Hydrophobicity Treatment of Gas Diffusion Layers on Ice Crystallization in Polymer Electrolyte Fuel Cells.

The unassisted cold-start capability of polymer electrolyte fuel cells (PEFCs) remains challenging for large-scale automotive applications. Various studies have shown that the freezing of produced water at the cathode catalyst layer (CL) and gas diffusion layer (GDL) interface blocks the oxidant gas and leads to a cold-start failure. However, the impact of GDL properties, including substrate, size, and hydrophobicity, on the freezing behavior of supercooled water is yet to be thoroughly investigated. We use differential scanning calorimetry to perform non-isothermal calorimetric measurements on untreated and waterproofed GDLs (Toray TGP-H-060, Freudenberg H23). By conducting a large number of experiments (>100) for each type of GDL, we obtained the corresponding distribution of onset freezing temperature (Tonset) and found noticeable sample-to-sample variations in both untreated and waterproofed GDLs. Furthermore, ice crystallization is affected by GDL wettability, coating load, coating distribution, and GDL size, whereas the impact of the GDL substrate and saturation level is not apparent. The Tonset distribution allows for predicting the capability of PEFC freeze-start and the freezing probability of residual water at a given subzero temperature. Our work paves the way for GDL modifications toward the improved cold-start capability of PEFC by identifying and avoiding the features that systematically trigger the freezing of supercooled water with high probability.

Rapid and sensitive determination of ascorbic acid based on label-free silver triangular nanoplates.

In this study, a new method for the detection of ascorbic acid (AA) was proposed. It was based on the protective effect of AA on silver triangular nanoplates (Ag TNPs) against Cl- induced etching reactions. Cl- can attack the corners of Ag TNPs and etch them, causing a morphological shift from triangular nanoplates to nanodiscs. As a result, the solution changes color from blue to yellow. However, in the presence of AA, the corners of Ag TNPs can be protected from Cl- etching, and the blue color of the solution remains unchanged. Using this effect, a selective sensor was designed to detect AA in the range of 0-40.00 µM with a detection limit of 2.17 µM. As the concentration of AA varies in this range, color changes from yellow to blue can be easily observed, so the designed sensor can be used for colorimetric detection. This method can be used to analyze fruit juice samples.

[Effect of electroacupuncture on pulmonary neuroendocrine cells and secretion of neuroactive substances in lung of COPD rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Zusanli" (ST36) and "Feishu" (BL13) on the activation and secretion of calcitonin gene-related peptide (CGRP) and 5-hydroxytryptamine (5-HT) of pulmonary neuroendocrine cells (PNECs) and inflammatory response in rats with chronic obstructive pulmonary disease (COPD), so as to explore its underlying mechanisms in treating COPD. METHODS: Male SD rats were randomly divided into normal control, COPD model and EA groups, with 7 rats in each group. The COPD model was established by forced inhale of cigarette smoke for 1 h in a self-made box (1 m×1 m×1 m in volume), twice daily for 12 weeks. EA (4 Hz/20 Hz, 1-3 mA) was applied at bilateral ST36 and BL13 acupoints for 30 min, once a day for 14 consecutive days. The pulmonary function including the forced vital capacity (FVC), forced expiratory volume at 0.1 second (FEV0.1), FEV0.3, FEV0.1/FVC and FEV0.3/FVC was detected using a lung function analyzer for small animals. The lung tissue was sampled for observing histopathological changes by using H.E. staining, for observing expression and distribution of PNECs by Grimelius silver staining, and for detecting the immunoactivity (integrated optical density) of CGRP and 5-HT by using immunohistochemistry. The contents of CGRP, 5-HT, tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and transforming growth factor-ß1 (TGF-ß1) in the bronchoalveolar lavage fluid (BALF) and lung tissue were detected by ELISA, and the correlations between TNF-α and CGRP, IL-1ß and CGRP, TNF-α and 5-HT, and IL-1ß and 5-HT levels were analyzed. The mRNA and protein expression levels of nerve fiber markers of CGRP and purinergic receptor P2X ligand gated ion channel 3 (P2X3) which dominate PNECs in the lung tissue were detected by real-time fluorescence quantitative PCR and Western blot, respectively. RESULTS: Compared with the normal control group, the levels of FVC, FEV0.1, FEV0.3, and the ratios of FEV0.1/FVC and FEV0.3/FVC were significantly decreased (P<0.05, P<0.01), while the immunoactivity of PNECs, CGRP and 5-HT, the contents of CGRP, 5-HT, TNF-α, IL-1ß and TGF-ß1 in the BALF and lung tissue, and the expression levels of CGRP and P2X3 mRNAs and proteins in the lung tissue significantly increased in the COPD model group (P<0.01, P<0.05). Following EA intervention, both the increased and decreased levels of all the indexes mentioned above were reversed (P<0.05, P<0.01) except FEV0.3. H.E. staining showed severe deformed bronchial lumen with thickened wall and alveolar septum, and obvious inflammatory cell infiltration and reduced number of alveolar lumen fusion in the COPD model group, which was mild in the EA group. A positive correlation was found between TNF-α and CGRP, IL-1ß and CGRP, TNF-α and 5-HT,IL-1ß and 5-HT levels in both BALF and lung tissues (P<0.01). CONCLUSION: EA at ST36 and BL13 can improve lung function and reduce inflammatory response in COPD rats, which may be related to its function in inhibiting the activation of PNECs and release of neuroactive substances.

The Pretreatment of Xiaoqinglong Decoction Alleviates Inflammation and Oxidative Damage and Up-Regulates Angiotensin-Converting Enzyme 2 in Lipopolysaccharide-Induced Septic Acute Lung Injury Rats.

Xiaoqinglong decoction (XQLD), a classic prescription of Traditional Chinese Medicine, has already been used clinically to cure acute lung injury (ALI), but its mechanism remains unclear. This subject aimed to explore the preventive role of XQLD in septic ALI rats besides its effects on angiotensin-converting enzyme (ACE)2 and its downstream factors. After, respectively, administrated with different concentrations of XQLD (6.25 g/kg/d, 12.5 g/kg/d, 25 g/kg/d) for 5 days and dexamethasone (DEX, 1 mg/kg) for 0.5 h, the rat models of ALI were established by intraperitoneal injection of lipopolysaccharide (LPS, 5 mg/kg) for 24 h. All rats were evaluated by lung function test, arterial blood gas analysis, morphological observation, lung wet/dry (W/D) ratio, and the lung injury score. The levels of malonaldehyde (MDA), superoxide dismutase (SOD), interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, and angiotensin (Ang) (1-7) in the lung were measured through biochemical and ELISA kits. The expressions of angiotensin-converting enzyme (ACE)2, mitochondrial assembly receptor (MasR), and nuclear factor (NF)-κB in lung tissue were detected by qRT-PCR and western blotting. Positive reaction cells of MasR were observed by immunohistochemistry. The results show that XQLD significantly ameliorated septic lung injury including edema and hemorrhage, as well as improved pulmonary function and arterial blood gas. Furthermore, XQLD markedly decreased the levels of IL-1ß, TNF-α, MDA, and NF-κB while increased the levels of SOD, Ang (1-7), ACE2, and MasR in septic ALI rats. Pearson correlation showed that the expressions of ACE2 were inversely related to IL-1ß, TNF-α, MDA, and NF-κB and positively correlated with SOD contents. Our data indicated that XQLD pretreatment alleviated inflammation and oxidative damage in septic ALI rats, which might be related to the up-regulation of ACE2-Ang (1-7)-MasR axis and inhibition of the NF-κB pathway.

A Flower-Shaped Thermal Energy Harvester Made by Metamaterials.

Harvesting thermal energy from arbitrary directions has become an exciting theoretical possibility. However, an exact 3D thermal energy harvester is still challenging to achieve for the stringent requirement of highly anisotropic and symmetrical structures with homogenous materials, as well as absence of effective characterization. In this Communication, a flower-shaped thermal harvesting metamaterial is originally promoted. Numerical simulations imply that heat flux can be concentrated into the target core and a temperature gradient turns out to be more than two times larger than the applied one without obvious distortion or perturbation to the temperature profile outside the concentrator. Temperature transitions of the actual device are experimentally measured to validate the novel structure with consistency of the simulated results with original methods. With ultraefficiency independent of geometrical size, the flower-shaped thermal harvester facilitates multiple scale energy harvesting with splendid efficient and might help to improve thermoelectric devices efficiency in a totally new perspective.

Nanostructured high-performance dielectric block copolymers.

A new type of insulating-conductive block copolymer was synthesized by metathesis polymerization. The copolymer can self-assemble into unique nanostructures of micelles or hollow spheres. It exhibits a high dielectric constant, low dielectric loss, and high stored/released energy density due to the strong dipolar and nano-interfacial polarization contributions.