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INTRODUCTION: Developing effective methods to enhance tumor radiosensitivity is crucial for improving the therapeutic efficacy of radiotherapy (RT). Due to its deep tissue penetration, excellent safety profile, and precise controllability, sonosensitizer- based sonodynamic therapy (SDT) has recently garnered significant attention as a promising combined approach with RT. METHOD: However, the limited reactive oxygen species (ROS) generation ability in the aggregated state and the absence of specific organelle targeting in sonosensitizers hinder their potential to augment RT. This study introduces a fundamental principle guiding the design of high-performance sonosensitizers employed in the aggregated state. Building upon these principles, we develop a mitochondria-targeted sonosensitizer molecule (TCSVP) with aggregation-induced emission (AIE) characteristics by organic synthesis. Then, we demonstrate the abilities of TCSVP to target mitochondria and produce ROS under ultrasound in H460 cancer cells using confocal laser scanning microscopy (CLSM) and fluorescence microscopy. Subsequently, we examine the effectiveness of enhancing tumor radiosensitivity by utilizing TCSVP and ultrasound in both H460 cells and H460 and 4T1 tumor-bearing mice. RESULTS: The results indicate that evoking non-lethal mitochondrial oxidative stress in tumors by TCSVP under ultrasound stimulation can significantly improve tumor radiosensitivity (p <0.05). Additionally, the in vivo safety profile of TCSVP is thoroughly confirmed by histopathological analysis. CONCLUSION: This work proposes strategies for designing efficient sonosensitizers and underscores that evoking non-lethal mitochondrial oxidative stress is an effective method to enhance tumor radiosensitivity.
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Human space activities have been continuously increasing. Astronauts experiencing spaceflight are faced with health problems caused by special space environments such as microgravity, and the investigation of cell injury is fundamental. The development of a platform capable of cell culture and injury detection is the prerequisite for the investigation. Constructing a platform suitable for special conditions in space life science research is the key issue. The ground-based investigation is an indispensable part of the research. Accordingly, a simulated microgravity (SMG)-oriented integrated chip platform capable of 3D cell culture and in situ visual detection of superoxide anion radical (O2â¢-) is developed. SMG can cause oxidative stress in human cells, and O2â¢- is one of the signaling molecules. Thus, a O2â¢--responsive aggregation-induced emission (AIE) probe is designed, which shows high selectivity and sensitivity to O2â¢-. Moreover, the probe exhibits abilities of long-term and wash-free staining to cells due to the AIE behavior, which is precious for space cell imaging. Meanwhile, a chip with a high-aspect-ratio chamber for adequate medium storage for the lack of the perfusion system during the SMG experiment and a cell culture chamber which can integrate the extracellular matrix (ECM) hydrogel for the bioinspired 3D cell culture is fabricated. In addition, a porous membrane is introduced between the chambers to prevent the hydrogel from separating during the SMG experiment. The afforded AIE probe-ECM hydrogel-integrated chip can achieve 3D culturing of U87-MG cells and in situ fluorescent detection of endogenous O2â¢- in the cells after long-term staining under SMG. The chip provides a powerful and potential platform for ground-based investigation in space life science and biomedical research.
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Salt-tolerant proteases with remarkable stability are highly desirable biocatalysts in the salt-fermented food industry. In this study, the undigested autocleavage product of HlyA (halolysin A), a low-salt adapted halolysin from halophilic archaeon Halococcus salifodinae, was investigated. HlyA underwent autocleavage of its C-terminal extension (CTE) at temperatures over 40 °C or NaCl concentrations below 2 M to yield HlyAΔCTE. HlyAΔCTE demonstrated robust stability over a wide range of -20-60 °C, 0.5-4 M NaCl, and pH 6.0-10.0 for at least 72 h. Notably, HlyAΔCTE is the first reported halolysin with such exceptional stability. Compared with HlyA, HlyAΔCTE preferred high temperatures (50-75 °C), low salinities (0.5-2.5 M NaCl), and near-neutral (pH 6.5-8.0) conditions to achieve high activity, consistently with its production conditions. HlyAΔCTE displayed a higher Vmax value against azocasein than HlyA. During fish sauce fermentation, HlyAΔCTE significantly enhanced fish protein hydrolysis, indicating its potential as a robust biocatalyst in the salt-fermented food industry.
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Fermentación , Alimentos Fermentados , Cloruro de Sodio , Alimentos Fermentados/microbiología , Cloruro de Sodio/química , Estabilidad de Enzimas , Productos Pesqueros/análisis , Concentración de Iones de Hidrógeno , Halococcus/metabolismo , Proteínas Arqueales/metabolismo , Proteínas Arqueales/química , Péptido Hidrolasas/metabolismo , TemperaturaRESUMEN
Multicomponent macromolecular mixtures often form higher-order structures, which may display non-ideal mixing and aging behaviors. In this work, we first propose a minimal model of a quaternary system that takes into account the formation of a complex via a chemical reaction involving two macromolecular species; the complex may then phase separate from the buffer and undergo a further transition into a gel-like state. We subsequently investigate how physical parameters such as molecular size, stoichiometric coefficients, equilibrium constants, and interaction parameters affect the phase behavior of the mixture and its propensity to undergo aging via gelation. In addition, we analyze the thermodynamic stability of the system and identify the spinodal regions and their overlap with gelation boundaries. The approach developed in this work can be readily generalized to study systems with an arbitrary number of components. More broadly, it provides a physically based starting point for the investigation of the kinetics of the coupled complex formation, phase separation, and gelation processes in spatially extended systems.
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Amyloid fibrils have long been associated with many neurodegenerative diseases. The conventional picture of the formation and proliferation of fibrils from unfolded proteins comprises primary and secondary nucleation of oligomers followed by elongation and fragmentation thereof. In this work, we first employ extensive all-atom molecular dynamics (MD) simulations of short peptides to investigate the governing processes of fibril growth at the molecular scale. We observe that the peptides in the bulk solution can bind onto and subsequently diffuse along the fibril surface, which leads to fibril elongation via either bulk- or surface-mediated docking mechanisms. Then, to guide the quantitative interpretation of these observations and to provide a more comprehensive picture of the growth kinetics of single fibrils, a continuum model which incorporates the key processes observed in the MD simulations is formulated. The model is employed to investigate how relevant physical parameters affect the kinetics of fibril growth and identify distinct growth regimes. In particular, it is shown that fibrils which strongly bind peptides may undergo a transient exponential growth phase in which the entire fibril surface effectively acts as a sink for peptides. We also demonstrate how the relevant model parameters can be estimated from the MD trajectories. Our results provide compelling evidence that the overall fibril growth rates are determined by both bulk and surface peptide fluxes, thereby contributing to a more fundamental understanding of the growth kinetics of amyloid-like fibrils.
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The diminishing effectiveness of existing aminoglycoside antibiotics (AGs) compels scientists to seek new approaches to enhance the sensitivity of current AGs. Despite ongoing efforts, currently available approaches remain restricted. Herein, a novel strategy involving the rational construction of an aggregation-induced-emission luminogen (AIEgen) is introduced to significantly enhance Gram-positive bacteria's susceptibility to AGs. The application of this approach involves the simple addition of AIEgens to bacteria followed by a 5 min light irradiation. Under light exposure, AIEgens efficiently generate reactive oxygen species (ROS), elevating intrabacterial ROS levels to a nonlethal threshold. Post treatment, the bacteria swiftly enter a hypersensitive state, resulting in a 21.9-fold, 15.5-fold, or 7.2-fold increase in susceptibility to three AGs: kanamycin, gentamycin, and neomycin, respectively. Remarkably, this approach is specific to AGs, and the induced hypersensitivity displays unparalleled longevity and heritability. Further in vivo studies confirm a 7.0-fold enhanced bactericidal ability of AGs against Gram-positive bacteria through this novel approach. This research not only broadens the potential applications of AIEgens but also introduces a novel avenue to bolster the effectiveness of AGs in combating bacterial infections.
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Treatment of naphthenic acids (NAs) in wastewater is necessary due to its high toxicity and difficult degradation. In the heterogeneous Fenton-like advanced oxidation of organic pollutant system, the insufficient accessibility of oxidizing agent and NAs greatly hamper the reaction efficiency. CO2-responsive phase transfer materials derived from polyethylene glycol (PEG)-based deep eutectic solvents were specific targeted at the immiscible-binary phase system. The NAs oxidative degradation process was optimized including the kinds of catalyst (Molecular weight of PEG, constitute of DESs, and dosage.), temperature, flow rate of CO2, et al. With the help of fluorescence properties of catalyst, the hydrophilic-hydrophobic interaction was visual-monitored and further studied. The amphipathic property of PEG-200/Sodium persulfate/Polyether amine 230 (PEA230) greatly reduced the aqueous/organic phase transfer barrier between sodium persulfate and NAs (up to 84 %), thus accreting oxidation rate. The surface tension decreased from 35.364 mN/m to 28.595 mN/m. To control the reaction rate, the CO2 respond structure of amido played an important role. In addition, the interfacial transfer intermediates and oxidation pathways were also explored by nuclear magnetic resonance, flourier transform infrared spectroscopy, surface tension, and radical inhibition experiments. The mechanism of advanced oxidation of NAs catalyzed by CO2-responsive phase transfer catalyst was proposed, which would made up for the deficiency of the system theory of heterogeneous chemical oxidation of organic pollutants.
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The interactive flexible device, which monitors the human motion in optical and electrical synergistic modes, has attracted growing attention recently. The incorporation of information attribute within the optical signal is deemed advantageous for improving the interactive efficiency. Therefore, the development of wearable optical information-electronic strain sensors holds substantial promise, but integrating and synergizing various functions and realizing strain-mediated information transformation keep challenging. Herein, an amylopectin (AP) modified nanoclay/polyacrylamide-based nanocomposite (NC) hydrogel and an aggregation-induced-emission-active ink are fabricated. Through the fluorescence-transfer printing of the ink onto the hydrogel film in different strains with nested multiple symbolic information, a wearable interactive fluorescent information-electronic strain sensor is developed. In the sensor, the nanoclay plays a synergistic "one-stone-three-birds" role, contributing to "lightening" fluorescence (≈80 times emission intensity enhancement), ionic conductivity, and excellent stretchability (>1000%). The sensor has high biocompatibility, resilience (elastic recovery ratio: 97.8%), and strain sensitivity (gauge factor (GF): 10.9). Additionally, the AP endows the sensor with skin adhesiveness. The sensor can achieve electrical monitoring of human joint movements while displaying interactive fluorescent information transformation. This research poses an efficient strategy to develop multifunctional materials and provides a general platform for achieving next-generation interactive devices with prospective applications in wearable devices, human-machine interfaces, and artificial intelligence.
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Conductividad Eléctrica , Hidrogeles , Nanocompuestos , Dispositivos Electrónicos Vestibles , Hidrogeles/química , Nanocompuestos/química , Humanos , Fluorescencia , Resinas Acrílicas/química , Adhesivos/química , ImpresiónRESUMEN
Cancer-cell-specific fluorescent photosensitizers (PSs) are highly desired molecular tools for cancer ablation with minimal damage to normal cells. However, such PSs that can achieve cancer specification and ablation and a self-reporting manner concurrently are rarely reported and still an extremely challenging task. Herein, we have proposed a feasible strategy and conceived a series of fluorescent PSs based on simple chemical structures for identifying and killing cancer cells as well as monitoring the photodynamic therapy (PDT) process by visualizing the change of subcellular localization. All of the constructed cationic molecules could stain mitochondria in cancer cells, identify cancer cells specifically, and monitor cancer cell viability. Among these, IVP-Br has the strongest ability to produce ROS, which serves as a potent PS for specific recognition and killing of cancer cells. IVP-Br could translocate from mitochondria to the nucleolus during PDT, self-reporting the entire therapeutic process. Mechanism study confirms that IVP-Br with light irradiation causes cancer cell ablation via inducing cell cycle arrest, cell apoptosis, and autophagy. The efficient ablation of tumor through PDT induced by IVP-Br has been confirmed in the 3D tumor spheroid chip. Particularly, IVP-Br could discriminate cancer cells from white blood cells (WBCs), exhibiting great potential to identify circulating tumor cells (CTCs).
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Neoplasias , Fotoquimioterapia , Humanos , Fármacos Fotosensibilizantes/farmacología , Fármacos Fotosensibilizantes/química , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismo , Apoptosis , Mitocondrias/metabolismo , Colorantes/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The formation of amyloid fibrils is a complex phenomenon that remains poorly understood at the atomic scale. Herein, we perform extended unbiased all-atom simulations in explicit solvent of a short amphipathic peptide to shed light on the three mechanisms accounting for fibril formation, namely, nucleation via primary and secondary mechanisms, and fibril growth. We find that primary nucleation takes place via the formation of an intermediate state made of two laminated ß-sheets oriented perpendicular to each other. The amyloid fibril spine subsequently emerges from the rotation of these ß-sheets to account for peptides that are parallel to each other and perpendicular to the main axis of the fibril. Growth of this spine, in turn, takes place via a dock-and-lock mechanism. We find that peptides dock onto the fibril tip either from bulk solution or after diffusing on the fibril surface. The latter docking pathway contributes significantly to populate the fibril tip with peptides. We also find that side chain interactions drive the motion of peptides in the lock phase during growth, enabling them to adopt the structure imposed by the fibril tip with atomic fidelity. Conversely, the docked peptide becomes trapped in a local free energy minimum when docked-conformations are sampled randomly. Our simulations also highlight the role played by nonpolar fibril surface patches in catalyzing and orienting the formation of small cross-ß structures. More broadly, our simulations provide important new insights into the pathways and interactions accounting for primary and secondary nucleation as well as the growth of amyloid fibrils.
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Amiloide , Péptidos , Amiloide/química , Péptidos/química , Solventes , Conformación Proteica en Lámina beta , Movimiento (Física) , Péptidos beta-Amiloides/química , Fragmentos de Péptidos/químicaRESUMEN
Discriminatively visualizing mitochondrial and lysosomal dysfunction is crucial for an in-depth understanding of cell apoptosis regulation and relative biology. However, fluorescent probes for the separate visualization of lysosomal and mitochondria damages have not been reported yet. Herein, we have constructed a fluorescent probe [2-(4-hydroxystyryl)-1,3,3-trimethyl-3H-indol-1-ium iodide (HBSI)] for labeling mitochondria and lysosomes in dual emission colors and discriminatively imaging mitochondrial and lysosomal damage in two different sets of fluorescent signals. In living cells, HBSI targeted both lysosomes and mitochondria to give green and red emission, respectively. During mitochondrial damages, HBSI immigrated into lysosomes, and the red emission decreased. During lysosomal damage, HBSI immigrated into mitochondria, and the green emission decreased. With the robust probe, the different damaging sequences of mitochondria and lysosomes under different amounts of H2O2 and chloral hydrate have been revealed. The sequential damage of lysosomes and mitochondria during cell apoptosis induced by rotenone, paclitaxel, and colchicine has been discovered. Furthermore, the regulation of mitochondria, lysosome, and their interplay during autophagy was also observed with the probe.
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Apoptosis , Peróxido de Hidrógeno , Peróxido de Hidrógeno/metabolismo , Autofagia , Lisosomas/metabolismo , Mitocondrias , Colorantes Fluorescentes/toxicidad , Colorantes Fluorescentes/metabolismoRESUMEN
With the development of the petrochemical industry, a large amount of naphthenic acids in petrochemical wastewater was accumulated in the environment, causing serious environmental pollution. Most of the commonly used methods for the determination of naphthenic acids have the characteristics of high energy consumption, complicated pretreatment, long detection cycle, and the need to send samples to analytical laboratories. Therefore, it is essential to develop an efficient and low-cost field analytical method for rapidly naphthenic acids quantify. In this study, nitrogen-rich carbon quantum dots (N-CQDs) based on natural deep eutectic solvents (NADESs) was successfully synthesized by a one-step solvothermal method. The fluorescence property of carbon quantum dots was used to achieve the quantitative detection of naphthenic acids in wastewater. The prepared N-CQDs showed excellent fluorescence and stability, showed a good response to naphthenic acids and a linear relationship in the concentration range of naphthenic acids from 0.03 to 0.09 molâ§L-1. The effect of common interferents in petrochemical wastewater on the detection of naphthenic acids by N-CQDs was investigated. The results showed that N-CQDs had good specificity for the detection of naphthenic acids. N-CQDs was applied to the naphthenic acids wastewater, and the concentration of naphthenic acids in the wastewater was successfully calculated according to the fitting equation.
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Cancer starvation therapy have received continuous attention as an efficient method to fight against wide-spectrum cancer. However, during cancer starvation therapy, the protective autophagy promotes cancer cells survival, compromising the therapeutic effect. Herein, a novel strategy by combination of autophagy-activated fluorescent photosensitizers (PSs) and cancer starvation therapy to realize the controllable and efficient ablation of tumor is conceived. Two dual-emissive self-reporting aggregation-induced emission luminogens (AIEgens), TPAQ and TPAP, with autophagy-activated reactive oxygen species (ROS) generation are prepared to fight against the protective autophagy in cancer starvation therapy. When protective autophagy occurs, a portion of TPAQ and TPAP will translocate from lipid droplets to acidic lysosomes with significant redshift in fluorescence emission and enhanced ROS generation ability. The accumulation of ROS induced by TPAQ-H and TPAP-H causes lysosomal membrane permeabilization (LMP), which further results in cell apoptosis and promotes cell death. In addition, TPAQ and TPAP can enable the real-time self-reporting to cell autophagy and cell death process by observing the change of red-emissive fluorescence signals. Particularly, the efficient ablation of tumor via the combination of cancer starvation therapy and photodynamic therapy (PDT) induced by TPAQ has been successfully confirmed in 3D tumor spheroid chip, suggesting the validation of this strategy.
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Neoplasias , Fotoquimioterapia , Humanos , Fármacos Fotosensibilizantes/farmacología , Fármacos Fotosensibilizantes/uso terapéutico , Especies Reactivas de Oxígeno/metabolismo , Fotoquimioterapia/métodos , Neoplasias/tratamiento farmacológico , AutofagiaRESUMEN
Visualizing Ribonucleic acid (RNA) dynamics inside live cells is crucially important for the research of life science. However, almost all of the reported RNA probes target RNA with cationic groups, and mitochondria with high negative transmembrane potential may bring significant interferences. As a result, precise visualization of RNA in living cells is still a greatly challenging task. To overcome this problem, in this work, we proposed a novel charge-elimination strategy to construct a fluorescent probe (H-SMBT) specific for RNA undisturbed by mitochondria in live cells. Probe H-SMBT was designed to target the negative groove of RNA with a cationic group, and an additional hydroxyl group was modified to overcome the interference from mitochondria. H-SMBT will change from cationic structure to a charge-eliminated state in mitochondria with weak alkalic environment and detach from mitochondria, and therefore, it can exclusively stain RNA in live cells. Using M-SMBT with a methoxy group as a comparative molecule, we confirmed that the phenol group in H-SMBT played a decisive role to achieve the RNA specificity. Furthermore, H-SMBT can fast stain live cells in 5 min with excellent RNA selectivity. The probe can also monitor cellular damage processes, and successfully be applied to live zebrafish imaging due to the good tissue permeability. This work provides a new design strategy for constructing RNA-selective fluorescent probes avoiding the interference from mitochondria, and the designed RNA probe can be widely used for RNA-related life science research.
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Colorantes Fluorescentes , ARN , Animales , Pez Cebra , Mitocondrias , Membrana CelularRESUMEN
Advanced materials with high performance and distinctive function are one of the main driving forces for the development of human society. The selection of appropriate materials and adequately utilizing their features to apply them in a specific area rationally are of great significance but remain challenging. Herein, an aggregation-induced emission (AIE)-active nanocomposite (NC) hydrogel is developed by introducing a pH-responsive AIE luminogen (AIEgen) into a Laponite XLS/polyacrylamide-based NC hydrogel (Laponite is a trademark of the company BYK Additives Ltd.). The AIEgen can protonate to interact with the negatively charged clay through the electrostatic interaction, which results in a drastic fluorescence enhancement due to the restriction of intramolecular motion by the rigid clay to the protonated AIEgen. This behavior facilitates the input of fluorescent information with a high contrast ratio in the hydrogel by acid stimulation. Moreover, by utilizing the excellent resilience of the hydrogel, hierarchically inputting and displaying the information in the original and stretched states of the hydrogel film is realized, which achieves information-storage expansion and dual-encryption via switching between stretching and restoring the film. This work showcases fully and synergistically utilizing the superiorities of various advanced materials to achieve superior applications and should guide the future development of advanced materials in emerging areas.
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Organic radicals are unstable and stable radicals usually display non-luminescent properties. Luminescent radicals possess the all-in-one properties of optoelectronics, electronics, and magnetics. To date, the reported structures of luminescent radicals are limited to triphenylmethyl radical derivatives and their analogues, which are stabilized with extended π-conjugation. Here, we demonstrate the first example of a nonconjugated luminescent radical. In spite of the lack of delocalized π-stabilization, the radical polymer readily emits red luminescence in the solid state. A traditional luminescent quencher, 2,2,6,6-tetramethylpiperidin-1-yl turned into a red chromophore when grafted onto a polymer backbone. Experimental data confirm that the emission is associated with the nitroxide radicals and is also affected by the packing of the polymer. This work discloses a novel class of luminescent radicals and a distinctive pathway for luminescence from open-shell materials.
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Luminiscencia , Polímeros , Polímeros/químicaRESUMEN
Microviscosity is a fundamental parameter in the biophysics of life science and governs numerous cellular processes. Thus, the development of real-time quantitative monitoring of microviscosity inside cells is important. The traditional probes for detecting microviscosity via time-resolved luminescence imaging (TRLI) are generally disturbed by autofluorescence or surrounding oxygen in cells. Herein, we developed loose packing nanoaggregates with aggregation-induced delayed fluorescence (FKP-POA and FKP-PTA) and free from the effect of oxygen and autofluorescence for viscosity mapping via TRLI. The feasibility of FKP-PTA nanoparticles (NPs) for microviscosity mapping through TRLI was demonstrated by monitoring the variation of microviscosity inside HepG2 cancer cells, which demonstrated a value change from 14.9 cP to 216.9 cP during the apoptosis. This indicates that FKP-PTA NP can be used as a probe for cellular microviscosity mapping to help people to understand the physiologically dynamic microenvironment. The present results are expected to promote the advancement of diagnostic and therapeutic methods to cope with related diseases.
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Oxígeno , Humanos , Viscosidad , Polarización de Fluorescencia , BiofisicaRESUMEN
The integration of aggregation-induced emission luminogens (AIEgens) and inorganic constituents to generate multifunctional nanocomposites has attracted much attention because it couples the bright aggregate-state fluorescence of AIEgens with the diverse imaging modalities of inorganic constituents. Herein, a facile and universal strategy to prepare metal-phenolic-network (MPN)-coated AIE dots in a high encapsulation efficiency is reported. Through precise control on the nucleation of AIEgens and deposition of MPNs in tetrahydrofuran/water mixtures, termed as coacervation, core-shell MPN-coated AIE dots with bright emission are assembled in a one-pot fashion. The optical properties of MPN-coated AIE dots can be readily tuned by varying the incorporated AIEgens. Different metal ions, such as Fe3+ , Ti4+ , Cu2+ , Ni2+ , can be introduced to the nanoparticles. The MPN-coated AIE dots with a red-emissive AIEgen core are successfully used to perform magnetic resonance/fluorescence dual-modality imaging in a tumor-bearing mouse model and blood flow visualization in a zebrafish larva. It is believed that the present study provides a tailor-made nanoplatform to meet the individual needs of in vivo bioimaging.
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Nanopartículas , Imagen Óptica , Animales , Colorantes Fluorescentes , Metales , Ratones , Imagen Óptica/métodos , Pez CebraRESUMEN
Peroxynitrite (ONOO-) is a potent reactive nitrogen species that plays a role as a critical mediator in liver injury elicited by drugs such as acetaminophen (APAP). At a therapeutic dosage, most APAP is metabolized by liver cells and then excreted in the urine. However, excessive APAP intake can cause an acute production of ONOO-, which induces mitochondrial oxidative stress and necrosis of the liver cells. Therefore, the ONOO- levels in hepatocytes have been considered as an early sign of hepatotoxicity associated with drug overdosage. Herein, a ratiometric theranostic system based on aggregation-induced emission luminogens (AIEgens) for the visualization of ONOO- and reduction of drug-induced hepatotoxicity is developed. The AIEgen ATV-PPB shows a ratiometric fluorescence response from red to green upon cleavage of arylboronic ester moieties by ONOO- with high sensitivity and selectivity. Meanwhile, experiments reveal that ATV-PPB not only acts as a fluorescent probe for ONOO- but also as an intracellular ONOO- scavenger to reduce the hepatotoxicity under overdose APAP treatment.