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The underlying mechanisms that determine gene expression and chromatin accessibility in retinogenesis are poorly understood. Herein, single-cell RNA sequencing and single-cell assay for transposase-accessible chromatin sequencing are performed on human embryonic eye samples obtained 9-26 weeks after conception to explore the heterogeneity of retinal progenitor cells (RPCs) and neurogenic RPCs. The differentiation trajectory from RPCs to 7 major types of retinal cells are verified. Subsequently, diverse lineage-determining transcription factors are identified and their gene regulatory networks are refined at the transcriptomic and epigenomic levels. Treatment of retinospheres, with the inhibitor of RE1 silencing transcription factor, X5050, induces more neurogenesis with the regular arrangement, and a decrease in Müller glial cells. The signatures of major retinal cells and their correlation with pathogenic genes associated with multiple ocular diseases, including uveitis and age-related macular degeneration are also described. A framework for the integrated exploration of single-cell developmental dynamics of the human primary retina is provided.
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Epigenômica , Transcriptoma , Humanos , Transcriptoma/genética , Retina/metabolismo , Neurogênese , Cromatina/genéticaRESUMO
Purpose: Retinal microglia promote angiogenesis and vasculopathy in oxygen-induced retinopathy (OIR); however, its specific molecular mechanism in the formation of retinal angiogenesis remains unclear. The lectin galactoside-binding soluble 3 binding protein (LGALS3BP), a member of the scavenger receptor cysteine-rich (SRCR) domain protein family, is involved in tumor neovascularization, and we therefore hypothesized that LGALS3BP plays an active role in microglia-induced angiogenesis. Methods: The expression of LGALS3BP in microglia was detected by immunofluorescence, RT-qPCR, and western blotting. Functional assays of human umbilical vein endothelial cells (HUVECs) such as migration, proliferation, and tube formation were measured by Transwell, EdU, and Matrigel assays. Angiogenesis-related factors and PI3K/AKT levels were detected by western blotting. The relationship between LGALS3BP and PI3K or HIF-1α was investigated by immunoprecipitation. Results: Our results showed that the expression of LGALS3BP was significantly increased in microglia surrounding neovascularization of the OIR mice and was also upregulated in human microglial clone 3 (HMC3) cells after hypoxia. Moreover, HUVECs co-cultured with hypoxic HMC3 cells showed increased migration, proliferation, and tube formation, as well as levels of angiogenesis-related factor. However, the proangiogenic ability and angiogenesis-related factor expression of HMC3 cells was suppressed after silencing LGALS3BP. LGALS3BP induces the upregulation of angiogenesis-related factors through the PI3K/AKT pathway and then promotes angiogenesis in microglia. Conclusions: Collectively, our findings suggest that LGALS3BP in microglia plays an important role in angiogenesis, suggesting a potential therapeutic target of LGALS3BP for angiogenesis.
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Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Animais , Antígenos de Neoplasias/metabolismo , Biomarcadores Tumorais/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Hipóxia/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Lectinas , Camundongos , Microglia/metabolismo , Neovascularização Patológica/metabolismo , Oxigênio/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de SinaisRESUMO
To establish a method for simultaneously determining the content of four glucosinolates and five flavonoids in leaves of Moringa oleifera via quantitative analysis of multi-components by single-marker(QAMS) and verify the feasibility and applicability of the established method. The glucosinolates and flavonoids were analyzed via Waters Acquity UPLC HSS T_3 column(2.1 mm × 100 mm, 1.8 µm). The gradient elution was carried out with the mobile phase composed of 0.1% formic acid-acetonitrile and 0.3% formic acid at the flow rate of 0.4 mL·min~(-1) and the column temperature of 40 â. The wavelengths for the detection of glucosinolates and flavonoids were 225 nm and 260 nm, respectively. With 4-O-(α-L-rhamnoyloxy)-benzyl glucosinolate and vecenin-2 as internal reference substances, the relative correction factors of four glucosinolates and five flavonoids were respectively calculated for determining the content of the 9 ingredients in leaves of M. oleifera. To verify the accuracy and feasibility of QAMS, we used internal standard method(ISM) and external standard method(ESM) to determine the content of glucosinolates and flavonoids, respectively. The t-test results showed that there was no significant difference in the content of glucosinolates obtained by ISM and QAMS methods or in the content of flavonoids obtained by ESM and QAMS methods. The content of glucosinolates and flavonoids varied among M. oleifera of different varieties and from different producing areas. The total glucosinolates and total flavonoids had the highest content in the Indian variety while the lowest content in the variety â Honghe No. 1'. The established QAMS method is rapid, simple and accurate and can be used for simultaneous determination of glucosinolates and flavonoids in the leaves of M. oleifera. This study provides experimental data for the quality control and utilization of M. oleifera leaves.
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Medicamentos de Ervas Chinesas , Moringa oleifera , Cromatografia Líquida de Alta Pressão , Flavonoides/análise , Glucosinolatos/análiseRESUMO
Triethylamine (TEA) is a widely used volatile organic chemical, which is harmful and can cause headache, dizziness, and respiratory discomfort. Developing an efficient sensor to detect trace amounts of TEA is significant for industrial and healthcare monitoring. In this work, SnO2 with a three-dimensional ordered macroporous structure (3DOM) was prepared through a polymethylmethacrylate sphere template route. The TEA sensing performance of the 3DOM SnO2 was enhanced through Pt loading. Aberration-corrected high-angle annular dark-field scanning transmission electron microscopy images and X-ray absorption fine-structure analysis indicate that Pt on the 3DOM 0.20% Pt/SnO2 surface mainly exists in the state of atomic dispersion, which results in more active sites, higher Hall mobility and active oxygen contents, and lower response energy barriers. The 0.20% Pt/SnO2 sensor has a low operating temperature of 80 °C and a low limit of detection (0.32 ppb). Because of the uniform adsorption of TEA on the atomically dispersed Pt, the 3DOM Pt/SnO2 sensor exhibits high selectivity.
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BACKGROUND: Viral resistance to existing inhibitors and the time-dependent effectiveness of neuraminidase inhibitors have limited the number of antivirals that can be used for prophylaxis and therapeutic treatment of severe influenza infection. Thus, there is an urgent need to develop new drugs to prevent and treat influenza infection. OBJECTIVE: The aims of this study was to design and synthesize a novel series of 2-ureidonicotinamide derivatives and evaluate their anti-IAV activities. Furthermore, we predicted the abilities of these compounds to inhibit the PA-PB1 subunit and forecasted the docking poses of these compounds with RNA polymerase protein (PDB ID 3CM8). METHODS: The novel designed compounds were synthesized using classical methods of organic chemistry and tested in vitro for their abilities inhibiting RNP and against influenza A virus. In addition, the 23 synthesized molecules were subjected to the generated pharmacophore Hypo1 to forecast the activity target PA-PB1 subunit of RNA polymerase. The ADMET pharmacokinetic parameters were calculated by the ADMET modules in Discovery Studio 2016. The docking results helped us demonstrate the possible interactions between these compounds with 3CM8. RESULTS: The synthesized 2-ureidonicotinamide derivatives were characterized as potent anti-influenza inhibitors. The target compounds 7b and 7c demonstrated significant antiviral activities and could be considered as novel lead compounds of antiviral inhibitors. In addition, compound 7b revealed suitable ADME properties expressed and might be a significant RNA polymerase inhibitor targeting the PA-PB1 subunit based on the predictable results and the docking results. CONCLUSION: This study revealed a novel series of compounds that might be useful in the search for an effective drug against the influenza virus.
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Vírus da Influenza A , Influenza Humana , Orthomyxoviridae , Antivirais/química , RNA Polimerases Dirigidas por DNA , Inibidores Enzimáticos/farmacologia , Humanos , Influenza Humana/tratamento farmacológicoRESUMO
Only the dried stigma of the saffron, a flower deemed as the most valuable spice globally, is utilized for industrial production. Hence, there exists a growing interest in utilizing saffron floral bio-residues. The anti-hyperuricemic activity of a flavonoid extract from saffron floral bio-residues was assessed in potassium oxonate-induced hyperuricemia mice. In addition, an ultra-high performance liquid chromatography-triple quadrupole mass spectrometry method was established and validated to determine the pharmacokinetics of five main flavonoids and three phase-II metabolites in rat plasma after oral administration of the flavonoid extract for the first time. Compared with pharmacokinetic parameters of kaempferol-3-O-sophoroside, the most abundant flavonoid in the extract, and its aglycone kaempferol, we observed that coexisting compounds significantly reduced the absorption, accelerated the excretion of kaempferol-3-O-sophoroside, while significantly increasing the absorption and prolonging the residence time of kaempferol in the flavonoid extract. These results suggest the promising potential of the flavonoid extract from saffron floral bio-residues as an anti-hyperuricemic agent. Kaempferol was absorbed in plasma at high concentrations owing to the biotransformation of kaempferol glycosides in vivo.
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Crocus , Hiperuricemia , Administração Oral , Animais , Cromatografia Líquida de Alta Pressão/métodos , Crocus/química , Flavonoides/análise , Camundongos , Extratos Vegetais/química , RatosRESUMO
Structural and biochemical studies elucidate that PAN may contribute to the host protein shutdown observed during influenza A infection. Thus, inhibition of the endonuclease activity of viral RdRP is an attractive approach for novel antiviral therapy. In order to envisage structurally diverse novel compounds with better efficacy as PAN endonuclease inhibitors, a ligand-based-pharmacophore model was developed using 3D-QSAR pharmacophore generation (HypoGen algorithm) methodology in Discovery Studio. As the training set, 25 compounds were taken to generate a significant pharmacophore model. The selected pharmacophore Hypo1 was further validated by 12 compounds in the test set and was used as a query model for further screening of 1916 compounds containing 71 HIV-1 integrase inhibitors, 37 antibacterial inhibitors, 131 antiviral inhibitors and other 1677 approved drugs by the FDA. Then, six compounds (Hit01-Hit06) with estimated activity values less than 10 µM were subjected to ADMET study and toxicity assessment. Only one potential inhibitory 'hit' molecule (Hit01, raltegravir's derivative) was further scrutinized by molecular docking analysis on the active site of PAN endonuclease (PDB ID: 6E6W). Hit01 was utilized for designing novel potential PAN endonuclease inhibitors through lead optimization, and then compounds were screened by pharmacophore Hypo1 and docking studies. Six raltegravir's derivatives with significant estimated activity values and docking scores were obtained. Further, these results certainly do not confirm or indicate the seven compounds (Hit01, Hit07, Hit08, Hit09, Hit10, Hit11 and Hit12) have antiviral activity, and extensive wet-laboratory experimentation is needed to transmute these compounds into clinical drugs.
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Adenosina Trifosfatases/química , Endonucleases/química , Inibidores Enzimáticos/química , Influenza Humana/enzimologia , Adenosina Trifosfatases/antagonistas & inibidores , Adenosina Trifosfatases/ultraestrutura , Domínio Catalítico/efeitos dos fármacos , Desenho de Fármacos/tendências , Endonucleases/antagonistas & inibidores , Endonucleases/ultraestrutura , Humanos , Influenza Humana/tratamento farmacológico , Influenza Humana/virologia , Ligantes , Modelos Moleculares , Simulação de Acoplamento Molecular , Relação Quantitativa Estrutura-AtividadeRESUMO
In order to better utilize saffron floral bio-residues(SFB), a qualitative and quantitative analysis of flavonoids in SFB was conducted using UPLC-MS and UPLC, respectively. On the one hand, 50 flavonols and 5 anthocyanins were putatively characte-rized by using UPLC-Q-TOF-MS. On the other hand, an UPLC method was established for determining the fingerprint of SFB as well as testing the main flavonoids kaempferol-3-O-sophoroside and delphinidin-3,5-di-O-glucoside. Contents of kaempferol-3-O-sophoroside and delphinidin-3,5-di-O-glucoside of 10 batches of samples were 44.21-58.73 mg·g~(-1) and 2.11-6.37 mg·g~(-1), respectively, and the similarities of 10 batches were more than 0.99. In addition, the color of the samples was digitized by using electronic eye technology, and it was found that the color of the samples was significantly correlated with the content of delphinidin-3,5-di-O-glucoside. The richness of flavonoids in SFB indicated its potential for development and utilization, and the large variation in anthocyanin content among samples from different regions suggested that more attention should be paid to the methods of sample pretreatment and storage.
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Crocus , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Flavonoides , Flores , Espectrometria de Massas em TandemRESUMO
We developed a simple and controlled method to synthesize FeCo2O4@MnO2 core-sheath nanoarchitecture (CSN) grown on Ni foam. Ultrathin FeCo2O4 nanoflakes with an average thickness of 10 nm served as the scaffold to deposit the MnO2 nanosheets. The MnO2 nanosheets were able to vertically grow on FeCo2O4 nanoflakes to form a sheath via a hydrothermal reaction. The nanocomposites' thickness could be tailored from 80 nm-550 nm by changing the reaction times. Electrochemical measurements demonstrated that FeCo2O4@MnO2 CSN with an optimal thickness of about 400 nm achieved an areal capacitance of 3.077 F cm-2 at 2 mA cm-2, which is much higher than individual FeCo2O4 nanoflakes (0.295 F cm-2) and MnO2 nanosheets (1.065 F cm-2). An aqueous asymmetric supercapacitor (ASC) was assembled using FeCo2O4@MnO2 CSN as its positive electrode and activated carbon (AC) as its negative electrode. The FeCo2O4@MnO2⫽AC ASC exhibited a capacitance of 0.538 F cm-2 at 5 mA cm-2 with a potential window of 1.65 V, and an excellent cycling stability (99.1% retention even after 5000 cycles). Furthermore, the maximum energy density and power density of FeCo2O4@MnO2⫽AC ASC was 0.203 mWh cm-2 at 3.44 mW cm-2 and 28.6 mW cm-2 at 0.061 mWh cm-2, respectively.