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Sweat biomarkers have the potential to offer valuable clinical insights into an individual's health and disease condition. Current sensors predominantly utilize enzymes and antibodies as biometric components to measure biomarkers present in sweat quantitatively. However, enzymes and antibodies are susceptible to interference by environmental factors, which may affect the performance of the sensor. Herein, we present a wearable microfluidic surface-enhanced Raman scattering (SERS) biosensor that enables the non-invasive and label-free detection of biomarkers in sweat. Concretely, we developed a bimetallic self-assembled anti-opal array structure with uniform hot spots, enhanced the Raman scattering effect, and integrated it into a silk fibroin-based sensing patch. Utilizing a silk fibroin substrate in the wearable SERS sensor imparts desirable properties such as softness, breathability, and biocompatibility, which enables the sensor to establish close contact with the skin without causing chemical or physical irritation. In addition, introducing microfluidic channels enables the controlled and high temporal resolution management of sweat, facilitating more efficient sweat collection. The proposed label-free SERS sensor can offer chemical 'fingerprint' information, enabling the identification of sweat analytes. As an illustration of the feasibility, we have effectively monitored the creatinine and uric acid levels in sweat. This study presents a versatile and highly sensitive approach for the simultaneous detection of biomarkers in human sweat, showcasing significant potential for application in point-of-care monitoring.
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Extracellular vesicles (EVs) have great potential in oncology drug delivery because of their unique biological origin. Apoptotic bodies (ABs), as a member of the EV family, offer distinct advantages in terms of size, availability and membrane properties, but have been neglected for a long time. Here, using ABs and Ti2N nanosheets, we propose a novel drug delivery system (Ti2N-DOX@ABs), which exhibit a homologous targeting ability for dual-strategy tumor therapy with intrinsic biological property. The experimental results demonstrate that such a drug delivery system possesses a drug loading capacity of 496.5% and a near-infrared photothermal conversion efficiency of 38.4%. In addition, the investigation of drug internalization process proved that Ti2N-DOX@ABs featured a supreme biocompatibility. Finally, the dual-strategy response based on photothermal and chemotherapeutic effects was studied under near-infrared laser radiation. This work explores the opportunity of apoptosome membranes in nanomedicine systems, which provides a technical reference for cancer-oriented precision medicine research.
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Doxorrubicina , Terapia Fototérmica , Titânio , Humanos , Terapia Fototérmica/métodos , Titânio/química , Titânio/farmacologia , Doxorrubicina/farmacologia , Doxorrubicina/química , Sistemas de Liberação de Medicamentos/métodos , Nanoestruturas/química , Linhagem Celular Tumoral , Vesículas Extracelulares/química , Antineoplásicos/farmacologia , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Raios Infravermelhos , Animais , Sobrevivência Celular/efeitos dos fármacosRESUMO
The urgent demand for addressing dye contaminants in water necessitates the development of microrobots that exhibit remote navigation, rapid removal, and molecular identification capabilities. The progress of microrobot development is currently hindered by the scarcity of multifunctional materials. In this study, a plasmonic MXene hydrogel (PM-Gel) is synthesized by combining bimetallic nanocubes and Ti3C2Tx MXene through the rapid gelation of degradable alginate. The hydrogel can efficiently adsorb over 60% of dye contaminants within 2 min, ultimately achieving a removal rate of >90%. Meanwhile, the hydrogel exhibits excellent sensitivity in surface enhanced Raman scattering (SERS) detection, with a limit of detection (LOD) as low as 3.76 am. The properties of the plasmonic hydrogel can be further adjusted for various applications. As a proof-of-concept experiment, thermosensitive polymers and superparamagnetic particles are successfully integrated into this hydrogel to construct a versatile, light-responsive microrobot for dye contaminants. With magnetic and optical actuation, the robot can remotely sample, identify, and remove pollutants in maze-like channels. Moreover, light-driven hydrophilic-hydrophobic switch of the microrobots through photothermal effect can further enhance the adsorption capacity and reduced the dye residue by up to 58%. These findings indicate of a broad application potential in complex real-world environments.
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Janus structure plays a crucial role in achieving chemically driven nanomotors with exceptional motion performance. However, Janus-structured chemically driven nanomotors with magnetic responsiveness are commonly fabricated by sputtering metal films. In the study, a self-assembly technique is employed to asymmetrically modify the surfaces of magnetic silica (SiO2@Fe3O4) nanoparticles with platinum nanoparticles, resulting in the formation of this kind nanomotors. Compared to platinum film, platinum nanoparticles exhibit a larger surface area and a higher catalytic activity. Hence, the nanomotors demonstrate improved diffusion capabilities at a significantly lower concentration (0.05%) of hydrogen peroxide (H2O2). Meanwhile, exosomes have gained attention as a potential tool for the efficient delivery of biological therapeutic drugs due to their biocompatibility. However, the clinical applications of exosomes are limited by their restricted tropism. The previously obtained nanomotors are utilized to deliver exosomes, greatly enhancing its targetability. The drug doxorubicin (DOX) is subsequently encapsulated within exosomes, acting as a representative drug model. Under the conditions of H2O2 concentration at the tumor site, the exosomes exhibited a significantly enhanced rate of entry into the breast cancer cells. The utilization of the nanomotors for exosomes presents a novel approach in the development of hybrid chemically and magnetically responsive nanomotors.
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Exosomes contain a wealth of proteomic information, presenting promising biomarkers for the noninvasive early diagnosis of diseases, especially cancer. However, it remains a great challenge to accurately and reliably distinguish exosomes secreted from different types of cell lines. Fluorescence immunoassay is frequently used for exosome detection. Nonspecific adsorption in immunoassays is unavoidable and affects the reliability of assay results. Despite the fact that various methods have been proposed to reduce nonspecific adsorption, a more effective method that can eliminate the influence of nonspecific adsorption is still lacking. Here, we report a more convenient way (named SR-TFC) to remove the artifacts caused by nonspecific adsorption, which combines tricolor fluorescence labeling of target exosomes, tricolor super-resolution imaging, and pixel counting. The pixel counting method (named CFPP) is realized by MATLAB and can eliminate nonspecific binding sites at the single-pixel level, which has never been achieved before and could improve the reliability of detection to the maximum extent. Furthermore, as a proof-of-concept, profiling of exosomal membrane proteins and identification of breast cancer subpopulations are demonstrated. To enable multiplex breast cancer phenotypic analysis, three kinds of specific proteins are labeled to obtain the 3D phenotypic information on various exosomes. Breast cancer subtypes can be accurately identified according to the super-resolution images of some clinically relevant exosomal proteins. Worth mentioning is that, by selecting other biomarkers, classification of other cancers could also be realized using SR-TFC. Hence, the present work holds great potential in clinical cancer diagnosis and precision medicine.
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Neoplasias da Mama , Exossomos , Humanos , Feminino , Exossomos/metabolismo , Proteômica , Reprodutibilidade dos Testes , Biomarcadores/análise , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/metabolismo , Fenótipo , Proteínas de Membrana/metabolismoRESUMO
Selective ion transport underpins fundamental biological processes for efficient energy conversion and signal propagation. Mimicking these 'ionics' in synthetic nanofluidic channels has been increasingly promising for realizing self-sustained systems by harvesting clean energy from diverse environments, such as light, moisture, salinity gradient, etc. Here, we report a spatially nanoconfined ion separation strategy that enables harvesting electricity from CO2 adsorption. This breakthrough relies on the development of Nanosheet-Agarose Hydrogel (NAH) composite-based generators, wherein the oppositely charged ions are released in water-filled hydrogel channels upon adsorbing CO2. By tuning the ion size and ion-channel interactions, the released cations at the hundred-nanometer scale are spatially confined within the hydrogel network, while ångström-scale anions pass through unhindered. This leads to near-perfect anion/cation separation across the generator with a selectivity (D-/D+) of up to 1.8 × 106, allowing conversion into external electricity. With amplification by connecting multiple as-designed generators, the ion separation-induced electricity reaching 5 V is used to power electronic devices. This study introduces an effective spatial nanoconfinement strategy for widely demanded high-precision ion separation, encouraging a carbon-negative technique with simultaneous CO2 adsorption and energy generation.
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The electrochemical extraction of lithium (Li) from aqueous sources using electrochemical means is a promising direct Li extraction technology. However, to this date, most electrochemical Li extraction studies are confined to Li-rich brine, neglecting the practical and existing Li-lean resources, with their overall extraction behaviors currently not fully understood. More still, the effect of elevated sodium (Na) concentrations typically found in most Li-lean water sources on Li extraction is unclear. Hence, in this work, we first understand the electrochemical Li extraction behaviors from ultradilute solutions using spinel lithium manganese oxide as the model electrode. We discovered that Li extraction depends highly on the Li concentration and cell operation current density. Then, we switched our focus on low Li to Na ratio solutions, revealing that Na can dominate the electrostatic screening layer, reducing Li ion concentration. Based on these understandings, we rationally employed pulsed electrochemical operation to restructure the electrode surface and distribute the surface-adsorbed species, which efficiently achieves a high Li selectivity even in extremely low initial Li/Na concentrations of up to 1:20,000.
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Lítio , Sódio , Lítio/química , Eletrodos , Íons , Sódio/química , ÁguaRESUMO
Extravasation, as one of the key steps in cancer metastasis, refers to the process where tumor cells escape the bloodstream by crossing the vascular endothelium and invade the targeted tissue, which accounts for the low five-year survival rate of cancer patients. Understanding the mechanism of cancer metastasis and inhibiting extravasation are crucial to improve patient prognosis. Here, a 3D organotypic microï¬uidic chip combined with SERS-based protein imprinted nanomaterials (SPINs) was proposed to study the extravasation process in vitro. The chip consists of a collagen gel channel and a vascular channel where human vein endothelial cells (HUVECs) and breast cancer cells are injected sequentially to induce extravasation. By comparing two subtypes of breast cancer cells (MCF-7 and MDA-MB-231), we successfully observed the difference in extravasation capabilities between two kinds of cells through fluorescence imaging. Meanwhile, thanks to the high specificity of molecular imprinting technology and the high sensitivity of surface enhanced Raman scattering (SERS), SPINs were utilized to analyze the concentration of several cancer secretions (interleukin-6 and interleukin-8) in complex biological fluid in real-time. Further, our model showed that downregulation of secretions by therapeutic drugs can inhibit the extravasation of breast cancers. This microfluidic model may pave the way for the fundamental research of the cancer metastasis and evaluating the therapeutic efficacy of potential drugs.
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Neoplasias da Mama , Nanoestruturas , Humanos , Feminino , Microfluídica/métodos , Neoplasias da Mama/patologia , Células Endoteliais , Colágeno , Análise Espectral Raman/métodosRESUMO
Matrix metalloproteinase 2 (MMP-2) has been considered a promising molecular biomarker for cancer diagnosis due to its related dysregulation. In this work, a core-satellite structure-powered ratiometric surface-enhanced Raman scattering (SERS) nanosensor with high sensitivity and specificity to MMP-2 was developed. The SERS nanosensor was composed of a magnetic bead encapsulated within a 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB)-labeled gold shell as the capture core and a 4-mercaptobenzonitrile (MBN)-encoded silver nanoparticle as the signal satellite, which were connected through a peptide substrate of MMP-2. MMP-2-triggered cleavage of peptides from the core surface resulted in a decrease of the SERS intensity of MBN. Since the SERS intensity of DTNB was used as an internal standard, the reliable and sensitive quantification of MMP-2 activity would be realized by the ratiometric SERS signal, with a limit of detection as low as 2.067 ng/mL and a dynamic range from 5 to 100 ng/mL. Importantly, the nanosensor enabled a precise determination of MMP-2 activity in tumor cell secretions, which may provide an avenue for early diagnosis and classification of malignant tumors.
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Nanopartículas Metálicas , Nanopartículas Metálicas/química , Metaloproteinase 2 da Matriz , Análise Espectral Raman/métodos , Ácido Ditionitrobenzoico , Prata/química , Ouro/químicaRESUMO
BACKGROUND: To date, the classification of mesiodens has been based on the location, crown orientation, and morphology; however, there is no assistance aid focusing on choosing surgical approach. PURPOSE: This study aimed to introduce and evaluate a new surgical assistance aid for mesiodens extraction based on surgical approach. STUDY DESIGN, SETTING, SAMPLE: For the retrospective trial part of this study, case data from mesiodens patients who had surgery at the Affiliated Stomatological Hospital was collected, and a new surgical assistance aid was developed. A prospective randomized controlled trial was conducted on mesiodens patients who were seen in our department (patients with one mesiodens were included). PREDICTOR VARIABLE: The predictor variable was surgical approach either with or without the surgical assistance aid. Subjects were randomized to one of the two study groups. For subjects assigned to the group using the surgical assistance guide, the approach was selected according to the aid detailed in this study. For subjects assigned to the group without the surgical assistant aid, 2 residents chose an approach based on their judgment and review of relevant imaging and physical examination. MAIN OUTCOME VARIABLES: The preoperative evaluation time, operative time, and complications associated with surgery were recorded separately for the two groups. COVARIATES: The age and sex were also recorded. ANALYSES: Variables were analyzed using the independent t-test and χ2 test. The level of statistical significance is P < .05. RESULTS: In the retrospective trial part, a new surgical assistance aid for mesiodens extraction was developed based on the ideal surgical approach. In the prospective randomized controlled trial, the experimental group (n = 50) was statistically significant in preoperative evaluation time (4.51 ± 0.34 mins vs 5.43 ± 0.34 mins) and operative time (31.87 ± 5.57 mins vs 36.32 ± 5.28 mins) compared to the control group (n = 50) (P < .001). There was no significant intergroup difference in complications associated with surgery (P > .05). CONCLUSION AND RELEVANCE: The new surgical assistance aid developed in this study guides surgeons to ease the selection of surgical approaches and shorten the operative time.
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Dente Supranumerário , Humanos , Estudos Retrospectivos , Estudos Prospectivos , Dente Supranumerário/cirurgia , Projetos de Pesquisa , Cuidados Pré-OperatóriosRESUMO
For the long-time tracking of biological events, maintaining the bioactivity of the analytes during the detection process is essential. Here, we show a versatile surface-enhanced Raman Scattering (SERS) platform, termed a superwettable-omniphobic lubricous porous SERS (SOLP-SERS) substrate. The SOLP-SERS substrate could generate a three-dimensional liquid "hotspots" matrix with an ultra-long lifetime (tens of days) by confining tiny amounts of liquids within the gaps between nanoparticles. Then, the analytes are trapped in the uniform liquid "hotspots", whose bioactivity can be well maintained over a long period of time during SERS detection. Limits of detection down to femtomolar levels were achieved for various molecules. More importantly, SERS signals were uniform within the substrate and remained stable for more than 30 days. As a proof-of-concept experiment, the dynamic detection of the polymerization of Aß peptides into amyloids was monitored by the SOLP-SERS substrate within 48 h. Moreover, the exosomes secreted by breast cancer cells, an important biomarker of cancer, were also measured. These results demonstrate that the SOLP-SERS platform will provide new insights into the development of real-time biochemical sensors with ultrahigh sensitivity.
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Exossomos , Nanopartículas , Tetranitrato de Pentaeritritol , Transporte Biológico , PolimerizaçãoRESUMO
Covalent modification is commonly used to tune the channel size and functionality of 2D membranes. However, common synthesis strategies used to produce such modifications are known to disrupt the structure of the membranes. Herein, we report less intrusive yet equally effective non-covalent modifications on Ti3C2Tx MXene membranes by a solvent treatment, where the channels are robustly decorated by protic solvents via hydrogen bond network. The densely functionalized (-O, -F, -OH) Ti3C2Tx channel allows multiple hydrogen bond establishment and its sub-1-nm size induces a nanoconfinement effect to greatly strengthen these interactions by maintaining solvent-MXene distance and solvent orientation. In sub-1-nm ion sieving and separation, as-decorated membranes exhibit stable ion rejection, and proton-cation (H+/Mn+) selectivity that is up to 50 times and 30 times, respectively, higher than that of pristine membranes. It demonstrates the feasibility of non-covalent methods as a broad modification alternative for nanochannels integrated in energy-, resource- and environment-related applications.
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Point-of-care analysis of neurotransmitters in body fluids plays a significant role in healthcare improvement. Conventional approaches are limited by time-consuming procedures and usually require laboratory instruments for sample preparation. Herein, we developed a surface enhanced Raman spectroscopy (SERS) composite hydrogel device for the rapid analysis of neurotransmitters in whole blood samples. The PEGDA/SA composite hydrogel enabled fast separation of small molecules from the complex blood matrix, while the plasmonic SERS substrate allowed for the sensitive detection of target molecules. 3D printing was employed to integrate the hydrogel membrane and the SERS substrate into a systematic device. The sensor achieved highly sensitive detection of dopamine in whole blood samples with a limit of detection down to 1 nM. The whole detection process from sample preparation to SERS readout can be finished within 5 min. Due to the simple operation and rapid response, the device shows great potential in point-of-care diagnosis and the monitoring of neurological and cardiovascular diseases and disorders.
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Nanopartículas Metálicas , Sistemas Automatizados de Assistência Junto ao Leito , Hidrogéis/química , Análise Espectral Raman/métodos , Neurotransmissores , Dopamina , Nanopartículas Metálicas/químicaRESUMO
The variation of tumor-associated metabolites in extracellular microenvironment timely reflects the development, the progression and the treatment of cancers. Conventional methods for metabolite detection lack the efficiency to grasp the dynamic metabolic alterations. Herein, we developed a SERS bionic taster which enabled real-time analysis of extracellular metabolites. The instant information of cell metabolism was provided by the responsive Raman reporters, which experienced SERS spectral changes upon metabolite activation. Such a SERS sensor was integrated into a 3D-printed fixture which fits the commercial-standard cell culture dishes, allowing in-situ acquisition of the vibrational spectrum. The SERS taster can not only accomplish simultaneous and quantitative analysis of multiple tumor-associated metabolites, but also fulfill the dynamic monitoring of cellular metabolic reprogramming, which is expected to become a promising tool for investigating cancer biology and therapeutics.
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Nanopartículas Metálicas , Biônica , Análise Espectral Raman/métodos , Impressão TridimensionalRESUMO
Tumor cell exosomes play a very important role in the process of tumor cell proliferation and metastasis. However, due to the nanoscale size and high heterogeneity of exosomes, in-depth understanding of their appearance and biological characteristics is still lacking. Expansion microscopy (ExM) is a method that embeds biological samples in a swellable gel to physically magnify the samples to improve the imaging resolution. Before the emergence of ExM, scientists had invented several super-resolution imaging techniques that could break the diffraction limit. Among them, single molecule localization microscopy (SMLM) usually has the best spatial resolution (20-50 nm). However, considering the small size of exosomes (30-150 nm), the resolution of SMLM is still not high enough for detailed imaging of exosomes. Hence, we propose a tumor cell exosomes imaging method that combines ExM and SMLM (i.e. Expansion SMLM, denoted as ExSMLM), which can realize the expansion and super-resolution imaging of tumor cell exosomes. In this technique, immunofluorescence was first performed to fluorescently label the protein markers on the exosomes, then the exosomes were polymerized into a swellable polyelectrolyte gel. The electrolytic nature of the gel made the fluorescently labeled exosomes undergo isotropic linear physical expansion. The expansion factor obtained in the experiment was about 4.6. Finally, SMLM imaging of the expanded exosomes was performed. Owing to the improved resolution of ExSMLM, nanoscale substructures of closely packed proteins were observed on single exosomes, which has never been achieved before. With such a high resolution, ExSMLM would have a great potential in detailed investigation of exosomes and exosome-related biological processes.
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Exossomos , Neoplasias , Humanos , Microscopia/métodos , Neoplasias/diagnóstico por imagem , ProteínasRESUMO
In recent years, nanoparticles camouflaged by red blood cell membrane (RBCM) have become a potential nano-drug delivery platform due to their good biocompatibility and immune evasion capability. Here, a multifunctional drug nanocarrier based on RBCM camouflaged mesoporous silica nanorods (MSNR) is presented, which can be used in pH and near-infrared (NIR) light triggered synergistic chemo-photothermal killing of cancer cells. To fabricate such a nanocarrier, MSNR and RBCM were prepared by the sol-gel method and modified hypotonic lysis method, respectively. Drugs were loaded into the pores of MSNR. Finally, RBCM was coated on the surface of MSNR by extrusion through a polycarbonate membrane. The advantages of the nanocarrier include: 1) MSNR can induce more cellular uptake than sphere shaped mesoporous silica nanoparticles. 2) The RBCM can reduce drug leakage and prevent clearance of the nanocarriers by macrophages. 3) By simultaneous loading doxorubicin (DOX) and indocyanine green (ICG), pH and NIR triggered synergistic chemo-photothermal therapy can be realized. In the experiment, we studied the drug releasing and cellular uptake of the nanocarriers in a breast cancer cell line (SKBR3 cells), in which a sufficient killing effect was observed. Such a multifunctional drug nanocarrier holds a broad application prospect in cancer treatment.
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Hipertermia Induzida , Nanopartículas , Nanotubos , Dióxido de Silício , Terapia Fototérmica , Fototerapia , Doxorrubicina/farmacologia , Eritrócitos , Linhagem Celular TumoralRESUMO
All-inorganic perovskite nanocrystals (CsPbX3 NCs, X = Cl, Br, I) are promising fluorescence materials for biological detection due to their excellent optical properties. However, there is still a challenge to obtain stable CsPbX3 NCs with more biofunctions. Here, we proposed a distinct strategy by absorbing the functionalized metal nanoprobes onto the phospholipid encapsulated CsPbX3 NCs to achieve CsPbX3-metal hybrids as probes for the detection of tumor-derived exosomes. Here, the metal nanoprobes have two functions: first, it endows phospholipid encapsulated CsPbX3 NCs with recognition ability; second, it avoids the fluorescence quenching of CsPbX3 NCs during the biological modification process by using metal nanoparticles as a bridge to connect with CsPbX3 NCs and various biomolecules. The obtained CPXD-AD exhibited a bright fluorescence signal, narrow full width at half-maximum (FWHM), and high specificity. Under optimal conditions, the CPXD-AD-based fluorescence-linked immunosorbent assay (FLISA) was successfully established and used for both qualitative and quantitative detection of tumor-derived exosomes.
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Exossomos , Nanopartículas Metálicas , Neoplasias , Humanos , Imunoadsorventes , Metais , FosfolipídeosRESUMO
Although two-dimensional (2D) materials have grown into an extended family that accommodates hundreds of members and have demonstrated promising advantages in many fields, their practical applications are still hindered by the lack of scalable high-yield production of monolayer products. Here, we show that scalable production of monolayer nanosheets can be achieved by a facile ball-milling exfoliation method with the assistance of viscous polyethyleneimine (PEI) liquid. As a demonstration, graphite is effectively exfoliated into graphene nanosheets, achieving a high monolayer percentage of 97.9% at a yield of 78.3%. The universality of this technique is also proven by successfully exfoliating other types of representative layered materials with different structures, such as carbon nitride, covalent organic framework, zeolitic imidazolate framework and hexagonal boron nitride. This scalable exfoliation technique for monolayer nanosheets could catalyze the synthesis and industrialization of 2D nanosheet materials.
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Ag+ ions are widely used in various fields of human life due to their unique properties and they threaten the environment and human health. The traditional methods for Ag+ detection commonly suffer from disadvantages including limited sensitivity, expensive equipment and complicated operating steps. Herein, we developed a highly specific dual-color fluorescence co-localization (DFC) strategy based on the C-Ag+-C structure for Ag+ detection. In this strategy, Ag+ ions can be captured to form C-Ag+-C base pairs, and these ions enable single-stranded DNAs to form double strands. The DFC strategy can exclude nonspecific interaction sites and greatly improve the sensitivity and specificity. By DFC of the QDs and Cy5 linked to the DNA strands, highly sensitive Ag+ detection was achieved in the concentration range from 0.14 pM to 200 nM, with a limit of detection (LOD) of 0.14 pM. Moreover, this method has been applied for the detection of Ag+ ions in real environmental samples with satisfactory recoveries. We believe that the DFC strategy is promising for Ag+ detection.
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DNA de Cadeia Simples , Prata , Humanos , Prata/química , DNA/química , Limite de Detecção , ÍonsRESUMO
Tumor cell-derived extracellular vesicles and their cargo of bioactive substances have gradually been recognized as novel biomarkers for cancer diagnosis. Meanwhile, the PD-L1 (Programmed Death-Ligand 1) protein, as an immune checkpoint molecule, is highly expressed on certain tumor cells and holds significant potential in immune therapy. In comparison to PD-L1 monoclonal antibodies, the inhibitory effect of PD-L1 siRNA (small interfering RNA) is more advantageous. In this article, we introduced a microfluidic chip integrating cell cultivation and exosome detection modules, which were intended for the investigation of the gene silencing effect of PD-L1 siRNA. Basically, cells were first cultured with PD-L1 siRNA in the chip. Then, the secreted exosomes were detected via super-resolution imaging, to validate the inhibitory effect of siRNA on PD-L1 expression. To be specific, a "sandwich" immunological structure was employed to detect exosomes secreted from HeLa cells. Immunofluorescence staining and DNA-PAINT (DNA Point Accumulation for Imaging in Nanoscale Topography) techniques were utilized to quantitatively analyze the PD-L1 proteins on HeLa exosomes, which enabled precise structural and content analysis of the exosomes. Compared with other existing PD-L1 detection methods, the advantages of our work include, first, the integration of microfluidic chips greatly simplifying the cell culture, gene silencing, and PD-L1 detection procedures. Second, the utilization of DNA-PAINT can provide an ultra-high spatial resolution, which is beneficial for exosomes due to their small sizes. Third, qPAINT could allow quantitative detection of PD-L1 with better precision. Hence, the combination of the microfluidic chip with DNA-PAINT could provide a more powerful integrated platform for the study of PD-L1-related tumor immunotherapy.