Chlorogenic Acid Inhibited Epithelial-Mesenchymal Transition to Treat Pulmonary Fibrosis through Modulating Autophagy.

Chlorogenic acid (CGA), derived from dicotyledons and ferns, has been demonstrated with anti-inflammatory, anti-bacterial, and free radical-scavenging effects and can be used to treat pulmonary fibrosis (PF). However, the specific mechanism by which CGA treats PF needs to be further investigated. In this study, in vivo experiment was firstly performed to evaluate the effects of CGA on epithelial-mesenchymal transition (EMT) and autophagy in bleomycin (BLM)-induced PF mice. Then, the effects of CGA on EMT and autophagy was assessed using transforming growth factor beta (TGF-ß) 1-induced EMT model in vitro. Furthermore, autophagy inhibitor (3-methyladenine) was used to verify that the inhibitory mechanism of CGA on EMT was associated with activating autophagy. Our results found that 60 mg/kg of CGA treatment significantly ameliorated lung inflammation and fibrosis in mice with BLM-induced PF. Besides, CGA inhibited EMT and promoted autophagy in mice with PF. In vitro studies also demonstrated that 50 µM of CGA treatment inhibited EMT and induced autophagy related factors in TGF-ß1-induced EMT cell model. Moreover, the inhibitory effect of CGA on autophagy and EMT in vitro was abolished after using autophagy inhibitor. In conclusion, CGA could inhibit EMT to treat BLM-induced PF in mice through, activating autophagy.

Particle Deposition in Large-Scale Human Tracheobronchial Airways Predicted by Single-Path Modelling.

The health effects of particles are directly related to their deposition patterns (deposition site and amount) in human airways. However, estimating the particle trajectory in a large-scale human lung airway model is still a challenge. In this work, a truncated single-path, large-scale human airway model (G3-G10) with a stochastically coupled boundary method were employed to investigate the particle trajectory and the roles of their deposition mechanisms. The deposition patterns of particles with diameters (dp) of 1-10 µm are investigated under various inlet Reynolds numbers (Re = 100-2000). Inertial impaction, gravitational sedimentation, and combined mechanism were considered. With the increasing airway generations, the deposition of smaller particles (dp < 4 µm) increased due to gravitational sedimentation, while that of larger particles decreased due to inertial impaction. The obtained formulas of Stokes number and Re can predict the deposition efficiency due to the combined mechanism in the present model, and the prediction can be used to assess the dose-effect of atmospheric aerosols on the human body. Diseases in deeper generations are mainly attributed to the deposition of smaller particles under lower inhalation rates, while diseases at the proximal generations mainly result from the deposition of larger particles under higher inhalation rates.

Characterization and genomic analysis of the novel Aeromonas veronii phage pAEv1812.

Bacteria of the species Aeromonas veronii are generally ubiquitous in different aquatic environments and are capable of causing a variety of diseases in aquatic animals. In this work, the phage isolate pAEv1812, which is lytic to A. veronii strain 1810, was characterized and sequenced. Morphological analysis revealed that pAEv1812 has a symmetrical head and a long non-contractile tail. A one-step growth curve analysis showed that the latent period and burst period of pAEv1812 were approximately 40 min and 80 min, respectively, with a burst size of 190 PFU/infected cell. The genome of phage pAEv1812 is 61,461 bp in length with a G+C content of 61.4%. Eighteen of the 75 putative proteins encoded by this phage have known functions, and there were no rRNA and tRNA genes in the genome. Phylogenetic analysis based on amino acid sequences of the major capsid protein and terminase large subunit suggested that phage pAEv1812 should be included as a member of the genus Chivirus.

Genomic analysis of a novel Aeromonas veronii phage pAEv1810, belonging to the genus Petsuvirus.

Phage-derived therapies are promising treatments in the fight against bacterial diseases as an alternative strategy nowadays. Species of Aeromonas veronii is an important pathogen causing freshwater fish diseases, the findings on genomic sequences of their bacteriophages are limited. In this work, a lytic bacteriophage capable of specifically infecting A. veronii strain AEv1810 was characterized at the gene level and was designated as pAEv1810. Transmission electron microscopic observation revealed that pAEv1810 belongs to the family of Myoviridae. The genome of phage pAEv1810 consists of 235,066 bp with 38.4% G + C content. Twenty-three of 249 putative proteins encoded by this phage have known functions, and four tRNA genes were found in phage genome. Phylogenetic analyses of RNA polymerase and Terminase large subunit revealed that phage pAEv1810 is closing to those phages classified to the genus Petsuvirus.

Genistein alleviates H2O2-induced senescence of human umbilical vein endothelial cells via regulating the TXNIP/NLRP3 axis.

CONTEXT: Genistein (Gen) has shown protective effects against ageing process. OBJECTIVE: To explore the role of Gen on the senescence of H2O2-induced human umbilical vein endothelial cells (HUVECs) and investigate the possible mechanism. MATERIALS AND METHODS: HUVECs were treated with different concentrations of H2O2 (50, 100, 200 and 400 µmol/L) for 1 h or Gen administration (20, 40, 80 and 160 µg/mL) for 24 h. Functional experiments (cell counting kit-8, ß-galactosidase staining and flow cytometry) were used to detect the effect of Gen on H2O2-induced HUVECs. After HUVECs were transfected with TXNIP overexpression plasmids, the expression of p16, p21, thioredoxin-interacting protein (TXNIP), nucleotide-binding and oligomerization domain-like receptor 3 (NLRP3), cleaved caspase-3 and cleaved caspase-1 in HUVECs were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. RESULTS: H2O2 (200 and 400 µmol/L) inhibited the proliferation of HUVECs. At concentrations of >50 µmol/L, H2O2 induced the cell cycle progression arrests in G1 phase and promoted cell senescence of HUVECs. Gen had no obvious cytotoxicity to HUVECs below 160 µg/mL. H2O2-induced HUVEC senescence and the expression of TXNIP and NLRP3 in HUVECs were down-regulated by Gen (40 and 80 µg/mL). Expressions of TXNIP and NLRP3 in HUVECs were up-regulated by H2O2 but down-regulated by Gen. Overexpressed TXNIP partially reversed the suppressive effect of Gen on H2O2-induced senescence and apoptosis of HUVECs. Expressions of p16, p21, TXNIP, NLRP3, cleaved caspase-3 and cleaved caspase-1 in H2O2-treated HUVECs were inhibited by Gen, while the inhibition as such was partially reversed by overexpressed TXNIP. DISCUSSION AND CONCLUSIONS: H2O2-induced HUVEC senescence was alleviated by Gen via suppressing the TXNIP/NLRP3 axis, which may offer a potential therapeutic approach for improving HUVEC senescence and provide a new direction for the treatment of cardiovascular disease.

Effects of interventional therapy on AQP4 gene expression and neuron apoptosis in rabbits with ischemic brain injury caused by carotid artery stenosis.

Objective: To explore the effects of interventional therapy (IT) on Aquaporin-4 (AQP4) gene expression and neuron apoptosis in rabbits with ischemic brain injury caused by carotid artery stenosis, and to further optimize the therapeutic regime of clinical ischemic brain injury (IBI). Methods: 30 New Zealand rabbits were randomly divided into three groups: Sham group (n=10), IBI group (n=10) and IBI+IT group (n=10). A rabbit model of carotid artery stenosis was established to induce IBI. 12 hours later, interventional therapy was achieved through percutaneous transluminal coronary angioplasty (PTCA) by pumping up the balloon to widen the catheter. The expression of AQP4 mRNA and protein were determined by reverse transcription polymerase chain reaction (RT-PCR) and western blot, respectively. The distributions of rabbit hippocampus and cerebral cortex were detected by Nissl staining. The neuronal function of rabbits in the three groups after surgery was assessed with mNSS score. TUNEL (Terminal Transferase-mediated dUTP Nick End-labelling) staining was used to observe neuron apoptosis in rabbit brain tissue. The expressions of Bax and Bcl-2 protein were detected by western blot. Markers of oxidative stress in rabbit brain tissues were detected by Reactive Oxygen Species ELISA Kits. Morphology of the organelles in rabbit brain tissues was observed with transmission electron microscopy (TEM). Results: The protein and mRNA expressions of AQP4 in rabbit brain tissues were significantly decreased in the IBI group. Nissl staining results showed that IT improved IBI in rabbit hippocampus. In addition, IT significantly ameliorated the neuronal function of rabbits, and reduced the apoptosis and oxidative stress level of neurons in brain tissues. Furthermore, we found that after IT, organelle damage was significantly reduced in rabbit neurons. Conclusion: After receiving IT, AQP4 gene level and neuron apoptosis were significantly reduced in rabbit brain tissues with ischemic brain injury from carotid artery stenosis.

Manufacture and clinical application of the forearm pronation's assistant tableware in the severely burned.

INTRODUCTION: Even after reconstructive surgery, it is still difficult for patients with severe burns to achieve independent eating activity. In this project, we customized the forearm pronation's assistant tableware to assist in improvement with eating activities. METHODS: From January 2017 to December 2018, 28 patients with severe burns including the hands were recruited. For the patient's independent eating activities, we customized forearm pronation's tableware (forks and spoons). We compared modified Barthel index (MBI) and Visual analogue scale (VAS) of satisfaction under three conditions: no auxiliary tableware, ADL universal cuff, or forearm pronation tableware; to compare the duration and the weight of food spilled during lunch when the patients wore the ADL universal cuff or the forearm pronation's tableware. Differences in MBI (rank data) were tested by the Friedman test, differences in VAS (normal distribution) were tested with One-way ANOVA (Bonferroni), differences in the duration and the weight (normal distribution data) were tested by paired sample t test. RESULTS: After wearing the forearm pronation's assistant tableware, MBI VAS both increased more than when the patients did not wear the auxiliary tableware (all p＜0.05). When the subjects wore forearm pronation tableware, the duration of lunch significantly decreased and the quality of eating activity significantly improved compared to the ADL universal cuff in eating activity (all p＜0.05). CONCLUSION: After wearing the forearm pronation's assistant tableware, the patients with severe burns completely or almost completely accomplished independent eating, the duration was decreased, and during eating activity the quality and the satisfaction were improved. CLINICAL TRIAL REGISTRATION: Chinese Clinical trial registry, ChiCTR1800019963.

Functional Genetic Variation in the 3'-UTRNTRK2 is Associated with Risk of Ischemic Stroke.

BACKGROUND: Stroke is a leading cause of death and disability worldwide. It remains difficult to treat brain injury and improve functional rehabilitation after cerebral ischemia. Brain-derived neurotrophic factor (BDNF) is involved in ischemic stroke (IS) through interactions in the CREB1-BDNF-NTRk2 pathway. In this study, we aimed to determine the association of NTRK2 gene polymorphisms and the effects of intergenetic interactions in the Chinese population. MATERIALS AND METHODS: A total of 400 patients diagnosed with IS and 400 healthy controls were enrolled for genotyping. Detailed sequence-based analysis was predicted through bioinformatical investigation. Polymorphisms associated with miRNA were analyzed by a dual-luciferase reporter assay system. RESULTS: Analysis of clinical characteristics revealed that IS was highly associated with exposure to cigarette smoking, alcohol intake, as well as metabolic diseases, such as diabetes, hypertension, and higher serum triglyceride concentration. Three polymorphisms in NTRK2 located in the 3'-untranslated region (3'-UTR) were genotyped. Logistic regression analysis showed that IS patients with rs11140793, rs7047042, and rs1221 polymorphisms had a higher risk of stroke and indicated a worse short-term recovery. The mRNA level of NTRK2 was suppressed in a mutant genotype compared with wild genotype. The suppression of NTRK2 was induced by the gain-of-binding ability of certain miRNAs through the direct binding of 3'-UTR. CONCLUSION: Our research indicated that, by influencing the expression of NTRK2, the SNPs rs11140793, rs7047042, and rs1221 in the 3'UTR of NTRK2 can be used as risk factors for IS patients.

MITF protects against oxidative damage-induced retinal degeneration by regulating the NRF2 pathway in the retinal pigment epithelium.

Oxidative damage is one of the major contributors to retinal degenerative diseases such as age-related macular degeneration (AMD), while RPE mediated antioxidant defense plays an important role in preventing retinopathies. However, the regulatory mechanisms of antioxidant signaling in RPE cells are poorly understood. Here we show that transcription factor MITF regulates the antioxidant response in RPE cells, protecting the neural retina from oxidative damage. In the oxidative stress-induced retinal degeneration mouse model, retinal degeneration in Mitf+/- mice is significantly aggravated compared to WT mice. In contrast, overexpression of Mitf in Dct-Mitf transgenic mice and AAV mediated overexpression in RPE cells protect the neural retina against oxidative damage. Mechanistically, MITF both directly regulates the transcription of NRF2, a master regulator of antioxidant signaling, and promotes its nuclear translocation. Furthermore, specific overexpression of NRF2 in Mitf+/- RPE cells activates antioxidant signaling and partially protects the retina from oxidative damage. Taken together, our findings demonstrate the regulation of NRF2 by MITF in RPE cells and provide new insights into potential therapeutic approaches for prevention of oxidative damage diseases.

Very Short-Term Surface Solar Irradiance Forecasting Based On FengYun-4 Geostationary Satellite.

An algorithm to forecast very short-term (30-180 min) surface solar irradiance using visible and near infrared channels (AGRI) onboard the FengYun-4A (FY-4A) geostationary satellite was constructed and evaluated in this study. The forecasting products include global horizontal irradiance (GHI) and direct normal irradiance (DNI). The forecast results were validated using data from Chengde Meteorological Observatory for four typical months (October 2018, and January, April, and July 2019), representing the four seasons. Particle Image Velocimetry (PIV) was employed to calculate the cloud motion vector (CMV) field from the satellite images. The forecast results were compared with the smart persistence (SP) model. A seasonal study showed that July and April forecasting is more difficult than during October and January. For GHI forecasting, the algorithm outperformed the SP model for all forecasting horizons and all seasons, with the best result being produced in October; the skill score was greater than 20%. For DNI, the algorithm outperformed the SP model in July and October, with skill scores of about 12% and 11%, respectively. Annual performances were evaluated; the results show that the normalized root mean square error (nRMSE) value of GHI for 30-180 min horizon ranged from 26.78% to 36.84%, the skill score reached a maximum of 20.44% at the 30-min horizon, and the skill scores were all above 0 for all time horizons. For DNI, the maximum skill score was 6.62% at the 180-min horizon. Overall, compared with the SP model, the proposed algorithm is more accurate and reliable for GHI forecasting and slightly better for DNI forecasting.

Tagging Functional Polymorphism in 3' Untranslated Region of Methylene Tetrahydrofolate Reductase and Risk of Ischemic Stroke.

BACKGROUND/AIMS: The association between genetic polymorphisms in the exon or untranslated region of the methylenetetrahydrofolate reductase gene (MTHFR) and the risk of human ischemic stroke (IS) has been well-documented. In this study, we focused on a polymorphism previously screened by high-throughput analysis and on its potential function in patients with IS Methods: This hospital-based case-control study was conducted in 400 patients and 400 healthy volunteers. Genotyping was conducted using TaqMan probes. Potential interactions were predicted by multiple bioinformatics analysis. Relative expression levels of MTHFR were detected confirmed by dual-luciferase assay. RESULTS: The MTHFR polymorphism rs142884651 was significantly associated with a decreased risk of IS compared with the wild-type GA genotype (P = 0.02) and AA genotype (P = 0.001). According to bioinformatics analysis and luciferase assay, this polymorphism could attenuate the binding of let-7f and miR-196a (P = 0.021 and 0.019, respectively) leading to the accumulation of MTHFR and degradation of serum homocysteine, and resulting in a better IS outcome. CONCLUSION: This study demonstrated that the MTHFR rs142884651 polymorphism is associated with a decreased risk of IS and may act as a biomarker of short-term outcome in patients with IS.

MITF acts as an anti-oxidant transcription factor to regulate mitochondrial biogenesis and redox signaling in retinal pigment epithelial cells.

There is increasing evidence that the mechanisms protecting the retinal pigment epithelium (RPE) against oxidative stress are important for preventing retinal degenerative diseases. Little, however, is known about these mechanisms. Here we show that MITF, a transcription factor responsible for RPE development and function, regulates redox signaling by acting through PGC1α, a master regulator of mitochondrial biogenesis. Mitf deficiency in mice leads to significantly higher levels of reactive oxygen species (ROS) in both RPE and retina, suggesting that Mitf dysfunction might lead to oxidative damage in the RPE and, by extension, in the retina. Furthermore, overexpression of MITF in the human RPE cell line ARPE-19 indicates that MITF up-regulates antioxidant gene expression and mitochondrial biogenesis by regulating PGC1α and protects cells against oxidative stress. Our findings provide new insights into understanding the redox function of MITF in RPE cells and its potential contribution to prevention of RPE-associated retinal degenerations.

Regulation of cell proliferation in the retinal pigment epithelium: Differential regulation of the death-associated protein like-1 DAPL1 by alternative MITF splice forms.

Vertebrate eye development and homoeostasis critically depend on the regulation of proliferation of cells forming the retinal pigment epithelium (RPE). Previous results indicated that the death-associated protein like-1 DAPL1 cell autonomously suppresses RPE proliferation in vivo and in vitro. Here, we show in human RPE cell lines that the pigment cell transcription factor MITF regulates RPE cell proliferation by upregulating DAPL1 expression. DAPL1 regulation by MITF is, however, mediated predominantly by (-) MITF, one of two alternative splice isoforms of MITF that lacks six residues located upstream of the DNA-binding basic domain. Furthermore, we find that the regulation of DAPL1 by MITF is indirect in that (-) MITF stimulates the transcription of Musashi homolog-2 (MSI2), which negatively regulates the processing of the anti-DAPL1 microRNA miR-7. Our results provide molecular insights into the regulation of RPE cell proliferation and quiescence and may help us understand the mechanisms of normal RPE maintenance and of eye diseases associated with either RPE hyperproliferation or the lack of regenerative proliferation.

DAPL1, a susceptibility locus for age-related macular degeneration, acts as a novel suppressor of cell proliferation in the retinal pigment epithelium.

The retinal pigment epithelium (RPE) forms a monolayer at the back of the vertebrate eye and is fundamental to retinal function and homoeostasis. During early development, RPE cells undergo rapid proliferation, but in the adult, they remain normally nonproliferative throughout life. Nevertheless, under pathological conditions such as in proliferative vitreoretinopathy or after retinal ablation, mature RPE cells can re-enter the cell cycle and form nodules or multiple cell layers. Here we show that Dapl1, whose human homolog represents a susceptibility locus for age-related macular degeneration (AMD), is highly up-regulated in quiescent but not proliferating RPE cells and that experimental overexpression of DAPL1 in proliferating RPE cells inhibits their proliferation. Consistent with this observation, the percent of Ki67-positive cells is significantly higher in E11.5 Dapl1 knockout mouse embryos compared to age-matched controls. In adult Dapl1-/- mice, which survive without showing any overt pathology, RPE overgrowth leads to multiple cell layers and/or cellular nodules. The antiproliferative effect of DAPL1 is associated with an increase in CDKN1A protein levels. Reduction of CDKN1A by siRNA in DAPL1-overexpressing RPE cells in vitro partially restores cell proliferation. Hence, we show that DAPL1 is a novel regulator of RPE cell proliferation that is important for the maintenance of the RPE as a monolayer. The findings suggest that DAPL1 dysregulation may be involved in abnormal RPE-related proliferative diseases and corresponding retinal dysfunctions in humans.