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The nanoelectrosprayer is a key device in the hyphenation of nanoLC-ESI-MS, and its development plays a crucial role in pushing forward the mining depth of biological discovery and industrialization of omics science. In this work, a new type of nanoelectrospray emitter, a rod sprayer, was developed based on microfluidic manufacture. Due to its porous silica structure, the rod sprayer in effect worked as a multinozzle sprayer, which is composed of a bunch of micrometer sized spray channels. Without the need for sophisticated microfabrication equipment, a superclean environment, or a complicated assembling process, such sprayer rods can be facilely fabricated in a mass production style: 3,600 rods with excellent monodispersity have been fabricated in 1 h, and rod sprayers thus made have demonstrated excellent intraday, interday, and interbatch reproducibilities: RSD = 1.9, 4.9, and 6.1%, respectively. The rod sprayer can generate stable electrospray in a wide voltage range from 2.6 to 3.2 kV and flow rates from 50 to 1000 nL/min, covering typical flow rates of subnanoLC, nanoLC, to microLC, and work steadily even under complex matrix environments (e.g., Hank's balanced salt solution containing sodium, magnesium, and calcium ions) without clogging. Meanwhile, the rod sprayers exhibited 200-1800% ionization efficiency enhancement in comparison with commonly used tapered tip emitters, for small molecule drugs, peptides, and proteins, respectively, and provided a broadened linear dynamic range of 4 orders of magnitude. The excellent characteristics of the rod sprayer, together with its small size and mass production capacity, should provide a high quality, high durability, high consistency, and disposable use-supported nanoelectrospray solution for MS-based bioanalyses.
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The aim of this study is to explore the potential targets and molecular mechanism of matrine (MAT) against aging. Bioinformatic-based network pharmacology was used to investigate the aging-related targets and MAT-treated targets. A total of 193 potential genes of MAT against aging were obtained and then the top 10 key genes (cyclin D1, cyclin-dependent kinase 1, Cyclin A2, androgen receptor, Poly [ADP-ribose] polymerase-1 (PARP1), histone-lysine N-methyltransferase, albumin, mammalian target of rapamycin, histone deacetylase 2, and matrix metalloproteinase 9) were filtered by the molecular complex detection, maximal clique centrality (MMC) algorithm, and degree. The Metascape tool was used for analyzing biological processes and pathways of the top 10 key genes. The main biological processes were response to an inorganic substance and cellular response to chemical stress (including cellular response to oxidative stress). The major pathways were involved in cellular senescence and the cell cycle. After an analysis of major biological processes and pathways, it appears that PARP1/nicotinamide adenine dinucleotide (NAD+)-mediated cellular senescence may play an important role in MAT against aging. Molecular docking, molecular dynamics simulation, and in vivo study were used for further investigation. MAT could interact with the cavity of the PARP1 protein with the binding energy at -8.5 kcal/mol. Results from molecular dynamics simulations showed that the PARP1-MAT complex was more stable than PARP1 alone and that the binding-free energy of the PARP1-MAT complex was -15.962 kcal/mol. The in vivo study showed that MAT could significantly increase the NAD+ level of the liver of d-gal-induced aging mice. Therefore, MAT could interfere with aging through the PARP1/NAD+-mediated cellular senescence signaling pathway.
Assuntos
Matrinas , NAD , Camundongos , Animais , NAD/metabolismo , Simulação de Acoplamento Molecular , Poli(ADP-Ribose) Polimerase-1/metabolismo , Envelhecimento , Estresse Oxidativo , Mamíferos/metabolismoRESUMO
INTRODUCTION: The study of peripheral circular RNA (circRNA) expression profile in patients with allergic rhinitis (AR) was absent to date, and we aimed to obtain the circRNA expression profile and identify the candidate biomarker from AR. METHODS: circRNA chip was performed to screen differentially expressed circRNAs in the peripheral blood sample from AR patients and healthy controls. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways were analyzed to further search the function of the differential expressed circRNAs. The real-time quantitative reverse transcription-polymerase chain reaction was further used to verify the candidate circRNA and also analyze its potential correlation with clinical parameters. RESULTS: Significantly up-regulated expression level of hsa_circRNA_404013 in AR was obtained by circRNA chip and further verified in 79 AR patients and 48 healthy controls. hsa_circRNA_404013 was significantly positively correlated with nasal discharge, nasal itching, the total nasal symptoms of AR, and brain-derived neurotrophic factor (BDNF) expression level in peripheral blood. Receiver operating characteristic curve analysis results showed that hsa_circRNA_404013 may be used as peripheral blood circulating marker for the diagnosis of AR with the area under curve of 0.8499 (95% CI: 0.783-0.916). In further bioinformatics analysis, hsa_circRNA_404013 may regulate the expression of BDNF through hsa-mir-182-5p contributing to the pathogenesis of AR. CONCLUSION: The expression profile of circRNAs from the peripheral blood sample of AR patients was obtained. The expression of hsa_circRNA_404013 was significantly up-regulated in the peripheral blood of AR patients, which may be used as a circulating marker for AR patients. Furthermore, hsa_circRNA_404013 may regulate the expression level of BDNF through hsa-mir-182-5p in AR pathogenesis.
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MicroRNAs , Rinite Alérgica , Biomarcadores , Fator Neurotrófico Derivado do Encéfalo/genética , Estudos de Casos e Controles , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , RNA/genética , RNA/metabolismo , RNA Circular , Rinite Alérgica/diagnóstico , Rinite Alérgica/genéticaRESUMO
We aimed to study the effects of an ethyl acetate fraction of Physalis alkekengi (PAE) on d-galactose (d-gal)-induced senescence and the underlying mechanism. Firstly, analysis of the phytochemical composition revealed total flavonoids, total phenolics, total saponins, rutin, and luteolin contents of 71.72 ± 2.99 mg rutin equivalents/g, 40.19 ± 0.47 mg gallic acid equivalents/g, 128.13 ± 1.04 mg oleanolic acid equivalents/g, 1.67 ± 0.07 mg/g and 1.61 ± 0.01 mg/g, respectively. The mice were treated with d-gal for six weeks, and from the fifth week, the mice were administered with PAE by gavage once a day for five weeks. We found significant d-gal-induced ageing-related changes, such as learning and memory impairment in novel object recognition and Y-maze, fatigue in weight-loaded forced swimming, reduced thymus coefficient, and histopathological injury of the liver, spleen, and hippocampus. The PAE effectively protected from such changes. Further evaluation showed that PAE decreased the senescence-associated ß-galactosidase of the liver, spleen, and hippocampus, as well as the oxidative stress of the liver, plasma, and brain. The abundance of flavonoids, phenols, and saponins in PAE may have contributed to the above results. Overall, this study showed the potential application of PAE for the prevention or treatment of ageing-associated disorders.
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Comportamento Animal/efeitos dos fármacos , Senescência Celular , Galactose/farmacologia , Transtornos da Memória/tratamento farmacológico , Physalis/química , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/farmacologia , Acetatos/química , Animais , Modelos Animais de Doenças , Masculino , Camundongos , Estresse OxidativoRESUMO
Cucurbitacin B (CuB) isolated from Cucumis melo by using a PC12 cell bioassay system exhibited significant nerve growth factor (NGF)-mimic or NGF-enhancing activity in PC12 and primary neuron cells. It was also demonstrated pro-neurogenesis effects in ICR and APP/PS1 mice and improved memory deficit of APP/PS1 mice. Its possible mechanism includes significant induction of the phosphorylation of glucocorticoid receptor (GR), protein kinase C (PKC), phospholipase C (PLC) and inhibition of cofilin. ChemProteoBase profiling, binding assay and cellular thermal shift assay (CETSA) were used to determine the target protein. Results revealed that CuB could affect actin dynamics as an actin inhibitor but did not bind with GR. The protein level of cofilin in PC12 cells after treating 0.3 µM and different temperatures was significantly higher than that of control group. Other neurotrophic signalling pathways, such as TrkA/TrkB, were analysed with specific inhibitors and Western blot. The inhibitors of TrkA, PLC, PKC, Ras, Raf and ERK1/2 significantly decreased the percentage of PC12 cells with neurite outgrowth and shortened the length of neurite outgrowth induced by CuB. CuB significantly induced the phosphorylation of TrkA, ERK and CREB. The phosphorylation of these proteins was obviously decreased by their specific inhibitors. These results suggest that cofilin is a candidate target protein of CuB in PC12 cells and that the GR/PLC/PKC and TrkA/Ras/Raf/ERK signalling pathways play important roles in the neuroprotective effect of CuB.
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Precursor de Proteína beta-Amiloide/metabolismo , Memória/efeitos dos fármacos , Neurogênese/efeitos dos fármacos , Crescimento Neuronal/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Oligopeptídeos/metabolismo , Triterpenos/farmacologia , Animais , Linhagem Celular Tumoral , Camundongos , Camundongos Endogâmicos ICR , Neuritos/efeitos dos fármacos , Neuritos/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Células PC12 , Ratos , Transdução de Sinais/efeitos dos fármacosRESUMO
The present study was designed to evaluate the effects of matrine (MAT) on scopolamine (SCOP)-induced learning and memory impairment. After successive oral administration of MAT to mice for three days at doses of 0.4, 2, and 10 mg/kg, we assessed improvements in learning and memory and investigated the mechanism of action of SCOP-induced amnesia. Donepezil at a dose of 3 mg/kg was used as a standard memory enhancer. MAT significantly improved SCOP-induced learning and memory impairment in novel object recognition and Y-maze tests at doses of 0.4, 2, and 10 mg/kg. Furthermore, MAT inhibited acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities and decreased oxidative stress in the brain, as evidenced by increased total antioxidant capacity, total superoxide dismutase levels, and catalase activities as well as decreased malondialdehyde levels. Additionally, there was a significant negative correlation between the percentage of spontaneous alternation in the Y maze and AChE activity in the cortex and hippocampus. MAT ameliorated SCOP-induced amnesia by the inhibition of both AChE/BuChE activities and oxidative stress. This study provides further evidence to encourage the development of MAT as a drug for the prevention or treatment of Alzheimer's disease.
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Alcaloides/uso terapêutico , Amnésia/tratamento farmacológico , Antioxidantes/uso terapêutico , Inibidores da Colinesterase/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Quinolizinas/uso terapêutico , Alcaloides/farmacologia , Amnésia/induzido quimicamente , Amnésia/metabolismo , Animais , Antioxidantes/farmacologia , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Catalase/metabolismo , Inibidores da Colinesterase/farmacologia , Masculino , Malondialdeído/metabolismo , Memória/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos ICR , Fármacos Neuroprotetores/farmacologia , Quinolizinas/farmacologia , Escopolamina , Superóxido Dismutase/metabolismo , MatrinasRESUMO
The SAM-pointed domain-containing ETS transcription factor (SPDEF) is an epithelial-specific transcription factor of the E26 transformation-specific (ETS) family, which binds the target gene through the high-affinity sequence of GGAT. It is suggested that SPDEF targets the promoter activity of Forkhead Box M1 (FoxM1), which has been proven to be highly expressed in gastric cancer. We found that SPDEF was overexpressed both at the messenger RNA (mRNA) and at the protein level in human gastric cancer species. The gastric cancer cells transfected with the SPDEF expression plasmid or SPDEF small interfering RNA (siRNA) led to observations on the clone genetics assay that indicated the promotion or the inhibition of gastric cancer cell proliferation, respectively. Both mRNA and protein levels of FoxM1 were regulated by SPDEF in gastric cancer cells and FoxM1 was also overexpressed in the corresponding human gastric cancer species. The overexpression and inhibition of FoxM1 could upregulate and downregulate the mRNA and protein levels of SPDEF expression, respectively. The recovery experiments verified that the overexpression of FoxM1 could at least partially revert both the expression of SPDEF and the proliferation of the cell lines even with the siRNA inhibition of SPDEF. The result of the dual luciferase activity assay showed that SPDEF bound to the promoter of FoxM1 and activated it. FoxM1 might also bind to the promoter of SPDEF to affect its expression. The results were checked in vivo. In conclusion, SPDEF is overexpressed in gastric cancer, which can form a positive regulation loop with FoxM1 to promote gastric carcinogenesis.
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Proteína Forkhead Box M1/metabolismo , Proteínas Proto-Oncogênicas c-ets/metabolismo , Neoplasias Gástricas/metabolismo , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/genética , Proliferação de Células/fisiologia , Proteína Forkhead Box M1/genética , Humanos , Técnicas In Vitro , Regiões Promotoras Genéticas/genética , Proteínas Proto-Oncogênicas c-ets/genética , Neoplasias Gástricas/genética , Análise Serial de TecidosRESUMO
Tetradecyl 2,3-dihydroxybenzoate (ABG-001) has been designed and synthesised as a lead compound to treat Alzheimer's disease, based on structure-activity relationships of gentisides. In this paper, the alkyl chain and ester linkage group of ABG-001 were modified. Consequently, several series of novel gentiside derivatives were designed and synthesised, and their neuritogenic activity was evaluated in PC12 cells. Among all the tested compounds, S-dodecyl 2,3-dihydroxybenzothioate (15d, named as ABG-199) was the most potent; the compound induced significant neurite outgrowth at 0.1 µM, which was comparable to that of nerve growth factor at the optimal concentration of 40 ng/mL and ABG-001 at 1 µM. A brief study on the mechanism of action of ABG-199 revealed that extracellular signal-regulated kinase phosphorylation was involved in ABG-199-induced neurite outgrowth in PC12 cells.
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Hidroxibenzoatos/síntese química , Hidroxibenzoatos/farmacologia , Neuritos/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Hidroxibenzoatos/química , Estrutura Molecular , Neuritos/patologia , Células PC12 , Ratos , Relação Estrutura-AtividadeRESUMO
BACKGROUND: Several biological processes are regulated by miR-200a-3p, including cell proliferation, migration, and epithelial-mesenchymal transition (EMT). In this study we aimed to uncover the diagnostic value and molecular mechanisms of miR-200a-3p in chronic rhinosinusitis with nasal polyps (CRSwNP). METHODS: The expressions of miR-200a-3p were detected by quantitative real-time polymerase chain reaction (qRT-PCR), Zinc finger E-box binding homeobox 1 (ZEB1) levels were examined by qRT-PCR and immunofluorescence staining. The interaction between miR-200a-3p and ZEB1 was predicted by TargetScan Human 8.0 and confirmed by dual-luciferase reporter assays. In addition, the effect of miR-200a-3p and ZEB1 on EMT-related makers and inflammation cytokines was assessed by qRT-PCR and Western blotting in human nasal epithelial cells (hNEpCs) and primary human nasal mucosal epithelial cells (hNECs). RESULTS: We found that miR-200a-3p was downregulated in non-eosinophilic and eosinophilic CRSwNP patients when compared with controls. The diagnostic value of miR-200a-3p in serum is reflected by the receiver operating characteristic curve and the 22-item Sino-Nasal Outcome Test. Bioinformatic analysis and luciferase reporter assay identified ZEB1 as a target of miR-200a-3p. ZEB1 was more highly expressed in CRSwNP than in controls. Furthermore, miR-200a-3p inhibitor or ZEB1 overexpression significantly suppressed the epithelial marker E-cadherin; promoted the activation of vimentin, α-spinal muscle atrophy, and N-cadherin; and aggravated inflammation in hNEpCs. Knockdown of ZEB1 significantly alleviated the cellular remodeling caused by miR-200a-3p inhibitor via the extracellular signal-regulated kinase (ERK)/p38 pathway in hNECs. CONCLUSIONS: miR-200a-3p suppresses EMT and inflammation by regulating the expression of ZEB1 via the ERK/p38 pathway. Our study presents new ideas for protecting nasal epithelial cells from tissue remodeling and finding a possible target for disease.
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MicroRNAs , Pólipos Nasais , Rinossinusite , Humanos , MicroRNAs/genética , MAP Quinases Reguladas por Sinal Extracelular , Pólipos Nasais/genética , Inflamação/genética , Proliferação de Células , Transição Epitelial-Mesenquimal/genética , Luciferases , Linhagem Celular Tumoral , Movimento Celular , Homeobox 1 de Ligação a E-box em Dedo de Zinco/genéticaRESUMO
Bioinsecticides and transgenic crops based on the bacterial pathogen Bacillus thuringiensis (Bt) can effectively control diverse agricultural insect pests, nevertheless, the evolution of resistance without obvious fitness costs has seriously eroded the sustainable use of these Bt products. Recently, it has been discovered that an increased titer of juvenile hormone (JH) favors an insect host (Plutella xylostella) to enhance fitness whilst resisting the Bt pathogen, however, the underlying regulatory mechanisms of the increased JH titer are obscure. Here, the involvement of N6 -methyladenosine (m6 A) RNA modification in modulating the availability of JH in this process is defined. Specifically, it is found that two m6 A methyltransferase subunit genes, PxMettl3 and PxMettl14, repress the expression of a key JH-degrading enzyme JH esterase (JHE) to induce an increased JH titer, mitigating the fitness costs associated with a robust defense against the Bt pathogen. This study identifies an as-yet uncharacterized m6 A-mediated epigenetic regulator of insect hormones for maintaining fitness during pathogen defense and unveils an emerging Bt resistance-related m6 A methylation atlas in insects, which further expands the functional landscape of m6 A modification and showcases the pivotal role of epigenetic regulation in host-pathogen interactions.
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Bacillus thuringiensis , Mariposas , Animais , Mariposas/genética , Mariposas/metabolismo , RNA/metabolismo , Epigênese Genética/genética , Endotoxinas/genética , Endotoxinas/metabolismo , Endotoxinas/farmacologia , Toxinas de Bacillus thuringiensis/metabolismo , Insetos , Bacillus thuringiensis/genética , Bacillus thuringiensis/metabolismo , Hormônios Juvenis/metabolismo , MetilaçãoRESUMO
Nolinospiroside F is a steroidal saponin isolated from Ophiopogon japonicus (O. japonicus). In this study, we found that nolinospiroside F significantly extends the replicative lifespan of K6001 yeast at doses of 1, 3 and 10 µM, indicating that it has an anti-aging effect. This may be attributed to its anti-oxidative effect, as nolinospiroside F could increase yeast survival under oxidative stress conditions and decrease the level of malondialdehyde (MDA), an oxidative stress biomarker. It could also increase anti-oxidative stress genes, SOD1 and SOD2, expression, and the activity of superoxide dismutase (SOD). It increase the activity of SIRT1, an upstream inducer of SOD2 expression. In sod1 and sod2 mutant yeast strains, nolinospiroside F failed to extend their replicative lifespan. These results indicate that SOD participates in the anti-aging effect of nolinospiroside F. Furthermore, nolinospiroside F inhibited the expression of UTH1, a yeast-aging gene that is involved in the oxidative stress of yeast, and failed to extend the replicative lifespan of uth1 or skn7 mutant yeast cells. SKN7 is the transcriptional activator of UTH1. We also demonstrate that SOD and UTH1 regulate each other's expression. Together, these results suggest that SOD and UTH1 genes are required for and play interactive roles in nolinospiroside F-mediated yeast lifespan extension.
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Nano-resveratrol liposome (RES-LIP) was prepared by the thin film rotary-evaporated method combined with ultrasonication and characterized by transmission electron microscopy (TEM), zeta potential, dynamic light scattering (DLS), and Fourier-transform infrared (FT-IR). The physicochemical stability, in vitro release, antioxidant activity, and cytotoxicity of RES-LIP were studied. Data showed that RES-LIP was a spherical vesicle with a diameter of less than 100 nm, the zeta potential was - 60 mV and the encapsulation efficiency was 86.78%. The physicochemical stability of RES-LIP was determined by Ea, ΔG, ΔH, and ΔS, which suggested that the process of RES-LIP degradation was spontaneous and endothermic. The in vitro release of RES-LIP was pH-dependent, belonged to the Weibull model, and was non-Fick diffusion. The antioxidant activity of RES-LIP was stronger than free resveratrol. The MTT assay and flow cytometry results suggested that resveratrol decreased cytotoxicity after being encapsulated by liposome. The prepared RES-LIP had high encapsulation efficiency, was sustained-release, had low cytotoxicity, was pH-targeted, and had potential usage in food and medicine fields.
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Antioxidantes , Lipossomos , Resveratrol , Antioxidantes/farmacologia , Antioxidantes/química , Espectroscopia de Infravermelho com Transformada de Fourier , Portadores de Fármacos/químicaRESUMO
NiTiO3-BiOBr heterostructured photocatalysts were constructed via precipitation, calcination and hydrothermal treatments. Various characterizations demonstrated that BiOBr nanosheets were decorated on NiTiO3 nanoparticals, forming porous rod-like heterojunctions. Compared with independent NiTiO3 and BiOBr, the composites with optimal BiOBr content presented highly improved visible-light photocatalytic efficiency. The degradation rates of Rhodamine B (RhB) and tetracycline (TC) reached 96.6% in 1.5 h (100% in 2 h) and 73.5% in 3 h, which are 6.61 and 1.53 times those of NiTiO3, respectively. The result is an improved photocatalytic behavior from the formation of heterojunctions with a large interface area, which significantly promoted the separation of photogenerated carriers and strengthened the visible-light absorption. Based on the free radical capture experiments and band position analysis, the photodegradation mechanism of type-II heterojunction was deduced. This study provides a new way to fabricate highly efficient NiTiO3-based photocatalysts for degrading certain organics.
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Excessive presynaptic glutamate release after cerebral ischaemia leads to neuronal death mainly through excessive calcium entry of N-methyl-D-aspartate receptors (NMDARs). Our recent study reported that cerebroside can open large-conductance Ca²âº-activated K⺠(BKCa) channels. The present study evaluated the effects of cerebroside-A (CS-A), a single molecule isolated from an edible mushroom, on brain injury after focal or global ischaemia in adult male mice and rats. We herein report that treatment with CS-A after 60-min middle cerebral artery occlusion dose-dependently reduced the cerebral infarction with at least a 6-h efficacious time-window, which was partially blocked by the BKCa channel blocker charybdotoxin (CTX). Treatment with CS-A after 20 min global cerebral ischaemia (four-vessel occlusion) significantly attenuated the death of pyramidal cells in hippocampal CA1 area, which was also sensitive to CTX. CS-A, by opening the BKCa channel, could prevent excessive glutamate release after oxygen-glucose deprivation (OGD). In addition, CS-A could inhibit NMDAR Ca²âº influx, which did not require the activation of the BKCa channel. Furthermore, CS-A blocked the OGD-induced NMDAR-dependent long-term potentiation in hippocampal CA1 region. These findings indicate that treatment with CS-A after stroke exerts potent neuroprotection through prevention of excessive glutamate release and reduction of Ca²âº influx through NMDARs.
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Cálcio/metabolismo , Infarto Cerebral/prevenção & controle , Cerebrosídeos/uso terapêutico , Ácido Glutâmico/metabolismo , Fármacos Neuroprotetores/uso terapêutico , Receptores de N-Metil-D-Aspartato/fisiologia , 6-Ciano-7-nitroquinoxalina-2,3-diona/farmacologia , Análise de Variância , Animais , Isquemia Encefálica/prevenção & controle , Infarto Cerebral/etiologia , Cerebrosídeos/química , Charibdotoxina/farmacologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Interações Medicamentosas , Estimulação Elétrica , Antagonistas de Aminoácidos Excitatórios/farmacologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Glucose/deficiência , Hipocampo/efeitos dos fármacos , Hipocampo/fisiologia , Hipóxia/prevenção & controle , Técnicas In Vitro , Infarto da Artéria Cerebral Média/complicações , Infarto da Artéria Cerebral Média/tratamento farmacológico , Potenciação de Longa Duração/efeitos dos fármacos , Potenciação de Longa Duração/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , N-Metilaspartato/farmacologia , Fármacos Neuroprotetores/química , Neurotoxinas/farmacologia , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Sais de Tetrazólio , Valina/análogos & derivados , Valina/farmacologiaRESUMO
This study used a replicative lifespan assay of K6001 yeast to screen anti-aging food factors in commercial flavonoids. Hesperidin derived from the Citrus genus extended the lifespan of yeast at doses of 5 and 10 µM as compared with the control group (p<0.01, p<0.01). Reactive oxygen species (ROS), real-time PCR (RT-PCR), and lifespan assays of uth1 and skn7 mutants with the K6001 background were used to study the anti-aging mechanisms in yeast. The results indicate that hesperidin significantly inhibits the ROS of yeast, and UTH1 gene expression, and that SKN7 gene are involved in hesperidin-mediated lifespan extension. Further, increases in the Sir2 homolog, SIRT1 activity, and SOD gene expression were confirmed at doses of 5 (p<0.01) and 10 µM (p<0.05). This suggests that Sir2, UTH1 genes, and ROS inhibition after administration of hesperidin have important roles in the anti-aging effects of yeast. However, the aglycon hesperetin did not exhibit anti-aging effects in yeast.
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Proteínas de Choque Térmico/genética , Hesperidina/farmacologia , Proteínas de Membrana/genética , Proteínas Mitocondriais/genética , Espécies Reativas de Oxigênio/antagonistas & inibidores , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/efeitos dos fármacos , Sirtuína 2/genética , Superóxido Dismutase/genética , Flavonoides/farmacologia , Expressão Gênica , Proteínas de Choque Térmico/metabolismo , Proteínas de Membrana/metabolismo , Proteínas Mitocondriais/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Sirtuína 2/metabolismo , Superóxido Dismutase/metabolismo , Fatores de TempoRESUMO
Two new cerebrosides, termitomycesphins G and H, were isolated from the edible Chinese mushroom, Termitomyces albuminosus (Berk.) Herm., and exhibited neuritogenic activity against PC12 cells. Their structures and absolute stereochemistry were elucidated by spectroscopic methods and by a comparison of the specific rotation of the hydrogenated products from termitomycesphins H and C. These cerebrosides possessed a unique modification by a hydroxyl group at the middle of the long-chain base, like earlier congeners termitomycesphins A-F. Termitomycesphin G with a 16-carbon-chain fatty acid showed higher neuritogenic activity than that of termitomycesphin H with an 18-carbon-chain fatty acid. This effect was observed within the termitomycesphins, suggesting that the chain length of the fatty acyl moiety played a key role in the neuritogenic activity.
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Processos de Crescimento Celular/efeitos dos fármacos , Cerebrosídeos/isolamento & purificação , Glucosilceramidas/isolamento & purificação , Neuritos/efeitos dos fármacos , Termitomyces/química , Animais , Processos de Crescimento Celular/fisiologia , Cerebrosídeos/química , Cerebrosídeos/farmacologia , China , Ácidos Graxos/farmacologia , Glucosilceramidas/química , Glucosilceramidas/farmacologia , Espectroscopia de Ressonância Magnética , Microscopia de Contraste de Fase , Neuritos/fisiologia , Neuritos/ultraestrutura , Células PC12 , RatosRESUMO
OBJECTIVE: We aimed to discover potential circulating genes and non-coding molecules (micro RNA [miRNA] and circular RNA [circRNA]) in CD4+ T cells in relation to seasonal allergic rhinitis (SAR). METHODS: Microarray data of GSE50223 were obtained from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) during and outside the pollen season were analyzed using R software and by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway analyses. The protein-protein interactions, modules, miRNAs targeting DEGs, merged miRNA-DEG networks, and circRNAs targeted with miRNAs were further analyzed. RESULTS: We identified 211 DEGs during the pollen season and eight DEGs outside the season, of which only MMP12, NR4A2, and CD69 were differentially expressed both during and outside the pollen season. DEGs during the pollen season were enriched in the GO categories 'neutrophil degranulation', 'neutrophil activation involved in immune response', 'neutrophil mediated immunity', and 'neutrophil activation'. A significant module was identified with key nodes of CDK6 and hsa-miR-29b-3p. Six significant circRNAs were also identified. CONCLUSIONS: Some genes, miRNAs, and circRNAs in CD4+ T may play vital roles in SAR and may thus be potential targets for the prevention and treatment of SAR.
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MicroRNAs , Rinite Alérgica Sazonal , Linfócitos T CD4-Positivos/metabolismo , Biologia Computacional , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Circular , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Rinite Alérgica Sazonal/genética , Linfócitos T/metabolismoRESUMO
The anti-aging effects of phloridzin on the yeast Saccharomyces cerevisiae were investigated by employing a replicative lifespan assay of the K6001 yeast strain. After administrating phloridzin at doses of 3, 10, and 30 µM, the lifespan of the yeast was significantly prolonged in comparison with the untreated group (p<0.01, p<0.001). To determine the mechanism of action, anti-oxidative experiments and ROS assay were performed. Phloridzin improved the viability of the yeast dose-dependently under oxidative stress by 7.5 mM H(2)O(2), and a low dose of phloridzin inhibited ROS of the yeast. Further, SOD1, SOD2, and Sir2 gene expression was examined by reverse transcription-polymerase chain reaction (RT-PCR), and was found to be significantly increased. Finally, superoxide dismutase (SOD) and SIRT1 activity assays showed that phloridzin notably increased the activity of SOD and SIRT1. These results suggest that SOD and Sir2 have important roles in phloridzin-regulated lifespan extension of yeast, and potentially anti-aging effects for mammalian cells via SIRT1.
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Flavonoides/farmacologia , Malus/química , Fenóis/farmacologia , Florizina/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/genética , Proteínas Reguladoras de Informação Silenciosa de Saccharomyces cerevisiae/genética , Sirtuína 2/genética , Superóxido Dismutase/genética , Animais , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Humanos , Estresse Oxidativo/efeitos dos fármacos , Polifenóis , Espécies Reativas de Oxigênio/metabolismo , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/metabolismo , Superóxido Dismutase/metabolismo , Fatores de TempoRESUMO
Due to its unique structure, core-shell material has presented significantly improved chromatographic performance in comparison with conventional totally porous material. This has been well demonstrated in the analytical column format, e.g. 4.6 mm i.d. columns. In the proteomics field, there is always a demand for high resolution microseparation tools. In order to explore core-shell material's potential in proteomics-oriented microseparations, we investigated chromatographic performance of core-shell material in a nanoLC format, as well as its resolving power for protein digests. The results show core-shell nanoLC columns have similar van Deemter curves to the totally porous particle-packed nanoLC columns. For 100 µm i.d. capillary columns, the core-shell material does not have significantly better dynamics. However, both core-shell and totally porous particle-packed nanoLC columns have shown high efficiencies: plate heights of ~11 µm, equivalent to 90000 plates per meter, have been achieved with 5 µm particles. Using a 60 cm long core-shell nanoLC column, 72000 plates were realized in an isocratic separation of neutral compounds. For a 15 cm long nanoLC column, a maximum peak capacity of 220 has been achieved in a 5 hour gradient separation of protein digests, indicating the high resolving power of core-shell nanoLC columns. With a standard HeLa cell lysate as the sample, 2546 proteins were identified by using the core-shell nanoLC column, while 2916 proteins were identified by using the totally porous particle-packed nanoLC column. Comparing the two sets of proteomics data, it was found that 1830 proteins were identified by both columns, while 1086 and 716 proteins were uniquely identified by using totally porous and core-shell particle-packed nanoLC columns, respectively, suggesting their complementarity in nanoLC-MS based proteomics.
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Cromatografia Líquida/métodos , Proteômica/métodos , Cromatografia Líquida de Alta Pressão/métodos , Células HeLa , Humanos , Tamanho da Partícula , PorosidadeRESUMO
OBJECTIVE: To investigate the inflammatory pattern in terms of inflammatory cells and cytokines expression in children with adenoid hypertrophy (AH) and coexistent allergic rhinitis (AR). STUDY DESIGN: A cross-sectional analytical study. PLACE AND DURATION OF STUDY: Department of Otolaryngology-Head and Neck Surgery, Shandong Provincial ENT Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong P. R. China, from October 2018 to August 2020. METHODOLOGY: A sample of 102 children with AH, who underwent adenoidectomy were enrolled. They were divided into two groups of AH, alone and AH with AR (AH+AR). A routine complete blood count, and the number of eosinophils in adenoid tissue was measured using hematoxylin-eosin staining. The tissue expression of cytokines was carried out using real-time quantitative PCR. RESULTS: Forty-eight children (47%) were diagnosed with AR. The number and percentage of eosinophils in peripheral blood and adenoid tissue were statistically (p <0.05) higher in the group of AH+AR than AH alone. Furthermore, in patients with AH+AR, the mRNA expression levels of IL-12 and IFN-γ decreased, while IL-4, IL-8, IL-18, IL-33, H2R, LTR1, LTR2 and GCR all increased in adenoid tissue. CONCLUSION: The pathological mechanism underlying adenoid hypertrophy in children with comorbid allergic rhinitis can be involved with predominant tissue eosinophilia and type 2 (or Th 2) inflammation. Key Words: Adenoid hypertrophy, Allergic rhinitis, Inflammatory features, Cytokines, Eosinophils.