RESUMO
Surface enhanced Raman spectroscopy (SERS) is a unique analytical technique with excellent performance in terms of sensitivity, non-destructive detection and resolution. However, due to the randomness and poor repeatability of hot spot distribution, SERS quantitative analysis is still challenging. Meanwhile, snus is a type of tobacco product that can release nicotine and other components in the mouth without burning, and the rapid detection technique based on SERS can reliably evaluate the amount of nicotine released from snus, which is of great significance for understanding its characteristics and regulating its components. Herein, the strategy was proposed to solve the feasibility of SERS quantitative detection based on self-assembled core-shell nanoparticles with embedded internal standards (EIS) due to EIS signal can effectively correct SERS signal fluctuations caused by different aggregation states and measurement conditions, thus allowing reliable quantitative SERS analysis of targets with different surface affinity. By means of process control, after the Au nanoparticles (Au NPs) were modified with 4-Mercaptobenzonitrile (4-MBN) as internal standard molecules, Ag shell with a certain thickness was grown on the surface of the AuNP@4-MBN, and then the Au@4-MBN@Ag NPs were used to regulate and control the assembly of liquid-liquid interface. The high-density nano-arrays assembled at the liquid-liquid interface ensure high reproducibility as SERS substrates, and which could be used for SERS detection of nicotine released from snus products. In addition, time-mapping research shows that this method can also be used to dynamically monitor the release of nicotine. Moreover, such destruction-free evaluation of the release of nicotine from snus products opens up new perspectives for further research about the impact of nicotinoids-related health programs.
RESUMO
BACKGROUND: Long noncoding RNAs (lncRNAs) have been identified as important regulatory factors implicated in a wide array of diseases, including various forms of cancer. However, the roles of most lncRNAs in the progression of gastric cancer (GC) remain largely unexplored. This study investigates the biological function and underlying mechanism of a novel lncRNA, XLOC_004787 in GC. METHODS: The location of XLOC_004787 in GES-1 cells and HGC-27 cells were detected by fluorescence in situ hybridization (FISH) assay. The expression levels of XLOC_004787 were assessed using quantitative real-time fluorescence PCR (qRT-PCR) in various cell lines, including GES-1, MGC-803, MKN-45, BGC-823, SGC-7901, and HGC-27 cells. Functional assays such as Transwell migration, cell counting kit-8 (CCK-8), and colony formation experiments were employed to analyze the effects of XLOC_004787 and miR-203a-3p on cell migration and proliferation. Protein levels associated with GC in these cell lines were examined by Western blotting. The intracellular localization of ß-catenin and P-Smad2/3 was assessed using immunofluorescence (IF) assay. Additionally, the interaction between XLOC_004787 and miR-203a-3p was investigated using a dual luciferase assay. RESULTS: XLOC_004787 was localized at both the cytoplasm and nucleus of GES-1 cells and HGC-27 cells. Compared to normal tissues and GES-1 cells, XLOC_004787 expression was significantly upregulated in GC tissues and cells, with the highest and lowest expression observed in SGC-7901 and HGC-27 cells, respectively. Furthermore, a reduced expression of XLOC_004787 was seen to inhibit migration and proliferation in SGC-7901 cells. Western blotting analysis revealed that a decrease in XLOC_004787 expression correspondingly decreased the expression of N-cadherin, mmp2, mmp9, Snail, Vimentin, ß-catenin, C-myc, Cyclin D1, and TGF-ß, while concurrently increasing E-cadherin expression. This was also associated with diminished expression of P-Smad2/3 in relation to Smad2/3, and reduced P-Gsk3ß expression in comparison to Gsk3ß. Additionally, the nuclear entry of P-Smad2/3 and ß-catenin was reduced by lower XLOC_004787 expression. Amplifying XLOC_004787 expression via pcDNA_XLOC_004787 suggested a potential for cancer promotion. Notably, XLOC_004787 was found to negatively regulate mir-203a-3p expression, with potential binding sites identified between the two. Higher mir-203a-3p expression was observed to decrease migration and proliferation, and enhance E-cadherin expression. Conversely, suppression of mir-203a-3p expression suggested a potential promotion of proliferation and migration in GC cells. CONCLUSIONS: These results suggest that XLOC_004787, found to be upregulated in GC tissues, potentially promotes proliferation and migration in GC cells. This occurs through the activation of TGF-ß and Wnt/ß-catenin signaling pathways and the expression of EMT-related proteins. Additionally, XLOC_004787 may influence cell migration and proliferation by modulating the signaling pathway via the adsorption and inhibition of mir-203a-3p.
Assuntos
MicroRNAs , RNA Longo não Codificante , Neoplasias Gástricas , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/genética , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , beta Catenina/genética , beta Catenina/metabolismo , Glicogênio Sintase Quinase 3 beta/genética , Glicogênio Sintase Quinase 3 beta/metabolismo , Hibridização in Situ Fluorescente , Linhagem Celular Tumoral , Proliferação de Células/genética , Via de Sinalização Wnt/genética , Caderinas/genética , Movimento Celular/genética , Regulação Neoplásica da Expressão GênicaRESUMO
As a new form of nicotine introduction for novel tobacco products, the interaction of nicotine salt with biological macromolecules may differ from that of free nicotine and thus affect its transport and distribution in vivo. Hence, the mechanism underlying the interaction between 2,6-dihydroxybenzoic acid nicotine salt (DBN) and human serum albumin (HSA) was investigated by multi-spectroscopy, molecular docking, and dynamic simulation. Experiments on steady-state fluorescence and fluorescence lifetime revealed that the quenching mechanism of DBN and HSA was dynamic quenching, and binding constant was in the order of 10^4 L mol-1. Thermodynamic parameters exhibited that the binding was a spontaneous process with hydrophobic forces as the main driving force. Fluorescence competition experiments revealed that DBN bound to site I of HSA IIA subdomain. According to the results of synchronous fluorescence, 3D fluorescence, FT-IR spectroscopy, circular dichroism (CD) spectroscopy, and molecular dynamics (MD) simulation, DBN did not affect the basic skeleton structure of HSA but changed the microenvironment around the amino acid residues. Computer simulations positively corroborated the experimental results. Moreover, DBN decreased the surface hydrophobicity and weakened the esterase-like activity of HSA, leading to the impaired function of the latter. This work provides important information for studying the interaction between DBN as a nicotine substitute and biological macromolecules and contributes to the further development and application of DBN.
Assuntos
Simulação de Dinâmica Molecular , Albumina Sérica Humana , Sítios de Ligação , Dicroísmo Circular , Humanos , Hidroxibenzoatos , Simulação de Acoplamento Molecular , Nicotina , Ligação Proteica , Albumina Sérica Humana/metabolismo , Espectrometria de Fluorescência , Espectroscopia de Infravermelho com Transformada de Fourier , TermodinâmicaRESUMO
Circular RNAs (circRNAs) are implicated in keloidogenesis and development. We aimed to investigate the role of a new identified phosphodiesterase 7B-derived circRNA (hsa_circ_0002198; henceforth named as PDE7B) in human keloid fibroblasts (HKFs) and to further confirm its mechanism via competing endogenous RNA (ceRNA) network. Transcriptional and translational levels of circPDE7B, microRNA (miR)-661, fibroblast growth factor 2 (FGF2), cleaved caspase3, B-cell lymphoma (bcl)-2, and bcl-2-associated X protein (bax) were detected by real-time quantitative PCR and western blotting. Relationship among circPDE7B, miR-661, and FGF2 was confirmed by bioinformatics algorithm, dual-luciferase reporter assay, RNA immunoprecipitation, RNA pull-down assay, and Spearman's rank correlation analysis. Cell progression was measured by cell counting kit-8 assay, 5-ethynyl-2-deoxyuridine assay, transwell assays, and flow cytometry. Expression of circPDE7B was upregulated in human keloid tissues and HKFs, accompanied with miR-661 downregulation and FGF2 upregulation. High circPDE7B accelerated proliferation, migration, and invasion, and inhibited apoptosis. These effects were paralleled with increased bcl-2 and decreased cleaved caspase3 and bax. Moreover, low circPDE7B played opposite effects to high circPDE7B. Restoring miR-661 could suppress HKFs progression, while blocking miR-661 could facilitate that. Notably, miR-661 was directly sponged by circPDE7B and then directly governed FGF2 gene expression. Deleting miR-661 and re-expressing FGF2 both abrogated the suppression of circPDE7B knockdown in HKFs progression. In conclusion, circPDE7B might contribute to HKFs progression via functioning as ceRNA for miR-661, suggesting a novel circPDE7B/miR-661/FGF2 pathway underlying keloid formation and treatment.
Assuntos
Fator 2 de Crescimento de Fibroblastos/biossíntese , Fibroblastos/metabolismo , Queloide/metabolismo , MicroRNAs/metabolismo , RNA Circular/metabolismo , Transdução de Sinais , Regulação para Cima , Linhagem Celular , Fator 2 de Crescimento de Fibroblastos/genética , Humanos , Queloide/genética , MicroRNAs/genética , RNA Circular/genéticaRESUMO
Surface enhanced Raman spectroscopy (SERS) is a highly sensitive analytical detection technique that provides unique chemical and structural information on target molecules. Here, simultaneous extraction and SERS detection of nicotine for the rapid and reliable identification of nicotine released from snus products were performed based on a nano-Au assembly hierarchy structure in the capillary. Based on this strategy, the time evolution of the concentrations of nicotine released from the snus products was measured. Through comparison of the intensities of the spectral peaks of the symmetrical breathing of the pyridine moiety of nicotine molecules, with the prolongation of time, the concentration of nicotine released decreased significantly, which is helpful for establishing a method for the rapid evaluation of the processing and selection of excipients of snus products, and provides a new idea for further study of the production of snus pouches and related tobacco products. Moreover, based on data fitting, it can be calculated that the concentration of nicotine in the extraction presented an obvious quadratic relationship with time, and the release of most of the nicotine in the snus pouch, which is held through the gums and palate, was basically completed after â¼15 min. Such destruction-free simultaneous measurements of snus products are opening up new perspectives for further research about the impact of nicotinoids on smokers' health and cessation programs.
Assuntos
Produtos do Tabaco , Tabaco sem Fumaça , Humanos , Nicotina , Fumantes , Análise Espectral RamanRESUMO
The extensive relevance of nanoparticles arouses the requirement for manufacturing although the predictable technique are frequently perilous and energy saving. In the current study, zinc oxide nanoparticles manufactured from Allium cepa avert UVB radiation interceded irritation in human epidermal keratinocytes (HaCaT cells). In the current study, the zinc oxide nanoparticles (ZnO-NPs) was synthesized from the extract of A. cepa. The optimized ZnO-NPs hence attained and was enumerated and exemplified by UV visible spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), scanning electron microscope (SEM) and EDAX impending analysis. In addition, amalgamated ZnO-NPs were experienced for cell viability (MTT), formation of reactive oxygen species (ROS), apoptosis, and antioxidant and lipid peroxidation (TBARS) levels. Also, we explored the effect of A. cepa ZnO-NPs in molecular level by evaluating the inflammatory and apoptotic markers, in which ZnO-NPs reinstated the interleukins 6, 10 and related signaling molecules like iNOS, COX-2 levels. Ultimately, ZnO-NPs induce apoptotic markers (Bax, Bcl-2) and also recommended that ZnO-NPs might aggravate cancer cell apoptosis in HaCaT cells.
Assuntos
Epiderme/metabolismo , Queratinócitos/efeitos dos fármacos , Queratinócitos/efeitos da radiação , Cebolas/metabolismo , Raios Ultravioleta/efeitos adversos , Óxido de Zinco/química , Óxido de Zinco/farmacologia , Animais , Linhagem Celular , Humanos , Inflamação/etiologia , Inflamação/metabolismo , Inflamação/patologia , Inflamação/prevenção & controle , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Espaço Intracelular/efeitos da radiação , Queratinócitos/citologia , Queratinócitos/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Protetores contra Radiação/química , Protetores contra Radiação/metabolismo , Protetores contra Radiação/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Óxido de Zinco/metabolismoRESUMO
Five new polyphenolic derivatives, sepiumols A-E (1-5), were isolated from the root barks of Periploca sepium. Their structures were elucidated by interpretation of NMR spectroscopic and mass spectrometric data. Compounds 1, 3 and 5 were found to exhibit significant antifungal activity, particularly for 3 with the remarkable activity against Gibberella saubinetii and Alternaria longipes with MIC values of 1.56 and 3.13⯵g/mL (ketoconazole: 0.78⯵g/mL), respectively. In addition, compounds 1, 3 and 5 also displayed significant antibacterial activity against methicillin-resistant Staphylococcus aureu with MIC values of 12.50-25⯵g/mL (ciprofloxacin: 0.78⯵g/mL).
Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Periploca/química , Polifenóis/farmacologia , Alternaria/efeitos dos fármacos , Antibacterianos/isolamento & purificação , Antifúngicos/isolamento & purificação , Gibberella/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Estrutura Molecular , Compostos Fitoquímicos/isolamento & purificação , Compostos Fitoquímicos/farmacologia , Casca de Planta/química , Raízes de Plantas/química , Polifenóis/isolamento & purificaçãoRESUMO
Nicotine in tobacco is harmful to health and the environment, so there is an environmental requirement to remove nicotine from tobacco and tobacco wastes. In this study, the biotransformation of nicotine by Rhodococcus sp. Y22 was investigated, and three metabolites (NIC1, NIC4 and NIC5) were isolated by column separation, preparative TLC and solid plate's method, respectively. NIC1 was identified as 6-hydoxynicotine based on the results of NMR, MS, HPLC-UV and HRESIMS analysis; NIC4 was a novel compound and identified as 5-(3-methyl-[1,3]oxazinan-2-ylidene)-5H-pyridin-2-one based on the results of NMR, MS and UV analysis; NIC5 was identified as nicotine blue based on the results of NMR and MS analysis. Meanwhile, two metabolites NIC2 and NIC3 were identified as 6-hydroxy-N-methylmyosmine and 6-hydroxypseudooxynicotine by HRESIMS analysis, respectively. According to these metabolites, the possible pathway of nicotine degradation by Rhodococcus sp. Y22 was proposed. The nicotine can be transformed to nicotine blue through two pathways (A and B), and 6-hydroxy-N-methylmyosmine is the key compound, which can be converted to 6-hydroxypseudooxynicotine (pathway A) and 5-(3-methyl-[1,3]oxazinan-2-ylidene)-5H-pyridin-2-one (pathway B), respectively. Moreover, the encoding gene of nicotine dehydrogenase, ndh, was amplified from Rhodococcus sp. Y22, and its transcriptional level could be up-regulated obviously under nicotine induction. Our studies reported the key metabolites and possible biotransformation pathway of nicotine in Rhodococcus sp. Y22, and provided new insights into the microbial metabolism of nicotine.
Assuntos
Proteínas de Bactérias/genética , Nicotina/metabolismo , Rhodococcus/crescimento & desenvolvimento , Proteínas de Bactérias/metabolismo , Biodegradação Ambiental , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Redes e Vias Metabólicas/efeitos dos fármacos , Metabolômica , Nicotina/análogos & derivados , Nicotina/isolamento & purificação , Nicotina/farmacologia , Rhodococcus/genética , Rhodococcus/metabolismo , Nicotiana/químicaRESUMO
Phytochemical investigations on the ethanol extract of the twigs of Garcinia tetralata resulted in the isolation of three new biphenyls, tetralatabiphenyls A-C (1-3), along with three known biphenyl derivatives (4-6). Structural elucidations of 1-3 were performed by spectroscopic methods such as 1D and 2D NMR spectra, in addition to high-resolution mass spectra. Compounds 1-6 were also evaluated for their anti-tobacco mosaic virus (anti-TMV) activity. The results showed that compound 3 showed high anti-TMV activity with inhibition rate of 31.1%. Compounds 1, 2, and 4-6 also showed modest anti-TMV activities with inhibition rates in the range of 18.9-24.5%, respectively.
Assuntos
Antivirais/isolamento & purificação , Antivirais/farmacologia , Compostos de Bifenilo/isolamento & purificação , Compostos de Bifenilo/farmacologia , Garcinia/química , Vírus do Mosaico do Tabaco/efeitos dos fármacos , Antivirais/química , Compostos de Bifenilo/química , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Caules de Planta/químicaRESUMO
Two new xanthones (1-2), together with four known ones (3-6), were isolated from whole herb of Swertia elata. Their structures were elucidated by spectroscopic methods including extensive 1D- and 2D-NMR techniques. Their anti-tobacco mosaic virus (anti-TMV) activity test revealed that 1-6 showed weak anti-TMV activities with inhibition rate in the range of 15.2-28.8% at the concentration of 20 µM.