Dual-mode aptasensor based on P-CeO2NR@Mxene and exonuclease I-assisted target recycling for malachite green detection.

Malachite green (MG) has been illicitly employed in aquaculture as a parasiticide, however, its teratogenic and carcinogenic effects pose a significant human health threat. Herein, a dual-mode colorimetric and electrochemical aptasensor was fabricated for MG detection, capitalizing on the robust catalytic and peroxidase-like activity of P-CeO2NR@Mxene and good capture efficiency of a tetrahedral DNA nanostructure (TDN) designed with multiple aptamers (m-TDN). P-CeO2NR@Mxene-modified complementary DNA (cDNA) served as both colorimetric and electrochemical probe. m-TDN was attached to AuE to capture MG and P-CeO2NR@Mxene/cDNA. The superior aptamer and MG binding to cDNA regulated signals and enabled precise MG quantification. The further introduced Exo I enabled aptamer hydrolysis, releasing MG for further binding rounds, allowing target recycling amplification. Under the optimal conditions, the aptasensor reached an impressively low detection limit 95.4 pM in colorimetric mode and 83.6 fM in electrochemical mode. We believe this dual-mode approach holds promise for veterinary drug residue detection.

Targeting initial tumour-osteoclast spatiotemporal interaction to prevent bone metastasis.

Bone is the most common site of metastasis, and although low proliferation and immunoediting at the early stage make existing treatment modalities less effective, the microenvironment-inducing behaviour could be a target for early intervention. Here we report on a spatiotemporal coupling interaction between tumour cells and osteoclasts, and named the tumour-associated osteoclast 'tumasteoclast'-a subtype of osteoclasts in bone metastases induced by tumour-migrasome-mediated cytoplasmic transfer. We subsequently propose an in situ decoupling-killing strategy in which tetracycline-modified nanoliposomes encapsulating sodium bicarbonate and sodium hydrogen phosphate are designed to specifically release high concentrations of hydrogen phosphate ions triggered by tumasteoclasts, which depletes calcium ions and forms calcium-phosphorus crystals. This can inhibit the formation of migrasomes for decoupling and disrupt cell membrane for killing, thereby achieving early prevention of bone metastasis. This study provides a research model for exploring tumour cell behaviour in detail and a proof-of-concept for behaviour-targeting strategy.

Fe-Co-Based Metal-Organic Frameworks as Peroxidase Mimics for Sensitive Colorimetric Detection and Efficient Degradation of Aflatoxin B1.

Building multifunctional platforms for integrating the detection and control of hazards has great significance in food safety and environment protection. Herein, bimetallic Fe-Co-based metal-organic frameworks (Fe-Co-MOFs) peroxidase mimics are prepared and applied to develop a bifunctional platform for the synergetic sensitive detection and controllable degradation of aflatoxin B1 (AFB1). On the one hand, Fe-Co-MOFs with excellent peroxidase-like activity are combined with target-induced catalyzed hairpin assembly (CHA) to construct a colorimetric aptasensor for the detection of AFB1. Specifically, the binding of aptamer with AFB1 releases the prelocked Trigger to initiate the CHA cycle between hairpin H2-modified Fe-Co-MOFs and hairpin H1-tethered magnetic nanoparticles to form complexes. After magnetic separation, the colorimetric signal of the supernatant in the presence of TMB and H2O2 is inversely proportional to the target contents. Under optimal conditions, this biosensor enables the analysis of AFB1 with a limit of detection of 6.44 pg/mL, and high selectivity and satisfactory recovery in real samples are obtained. On the other hand, Fe-Co-MOFs with remarkable Fenton-like catalytic degradation performance for organic contaminants are further used for the detoxification of AFB1 after colorimetric detection. The AFB1 is almost completely removed within 120 min. Overall, the introduction of CHA improves the sensing sensitivity; efficient postcolorimetric-detection degradation of AFB1 reduces the secondary contamination and risk to the experimental environment and operators. This strategy is expected to provide ideas for designing other multifunctional platforms to integrate the detection and degradation of various hazards.

Detection of acrylamide in food based on MIL-glucose oxidase cascade colorimetric aptasensor.

BACKGROUND: Maillard reaction involves the polymerization, condensation, and other reactions between compounds containing free amino groups and reducing sugars or carbonyl compounds during heat processing. This process endows unique flavors and colors to food, while it can also produce numerous hazards. Acrylamide (AAm) is one of Maillard's hazards with neurotoxicity and carcinogenicity, these effects can trigger mutations and alternations in gene expression in human cells and accelerate organ aging. An accurate and reliable acrylamide detection method with high sensitivity and specificity for future regulatory activities is urgently needed. RESULTS: Herein, we constructed a colorimetric aptasensor with the hybridization of MIL-glucose oxidase (MGzyme)-cDNA and magnetic nanoparticle-aptamer (MNP-Apt) to specifically detect AAm. The incorporation of MB-Apt and AAm released MGzyme-cDNA in the supernatant, took the supernatant out, with the addition of glucose and TMB, MGzyme would oxidize glucose, the resulting •OH facilitated the oxidation of colorless TMB to blue ox-TMB. The absorbance value at 652 nm, which indicates the characteristic absorption peak of ox-TMB, exhibited a proportion to the concentration of AAm. MGzyme avoided the addition of harmful intermediate H2O2 and created an acid microenvironment for the catalytic reaction. MNP-Apt possessed the advantages of high specificity and simplified separation. Under optimal conditions, this method displayed a linear range of 0.01-100 µM with the limit of detection of 1.53 nM. With the spiked analysis data cross-verified by ELISA kit, this aptasensor was proven to specifically detect AAm at low concentrations. SIGNIFICANCE: This colorimetric aptasensor was the integration of aptamer and the enzyme-cascade system, which could broaden the applicable range of enzyme-cascade system, break the limits of specific detection of substrates, eliminate the need for harmful intermediates, improve the reaction efficiency, implement the specific detection, whilst enabling the accurate detection of AAm. Given these remarkable performances, this method has shown significant potential in the field of food safety inspection.

Deoxynivalenol (DON)-Triggered Dual-Color Composite Probe Based on Gold Nanoclusters for Simultaneous Imaging of DON and miR-34a in Living Cells.

Deoxynivalenol (DON) is a mycotoxin secreted by Fusarium species, posing great harm to food safety and human health. Therefore, it is of great significance to study its toxic effects and mechanism. miR-34a is a representative biomarker during the process of DON-induced apoptosis. Herein, a DON-triggered dual-color composite probe was constructed for simultaneous imaging of DON and miR-34a in living cells. The aptamer blocks the recognition sequence of miR-34a to realize DON-triggered cell imaging. The specific binding of DON with its aptamer and HCR induced by miR-34a resulted in the recovery of fluorescence of the dual-color Au NCs. Under the optimal conditions, the correlation between the relative fluorescence intensities of dual-color Au NCs showed good linear relationships with the logarithm of DON and miR-34a concentration, respectively. With the increase in DON concentration (0-20 µg/mL) and stimulation time (0-12 h), the fluorescence of dual-color Au NCs gradually recovered. This dual-color Au NCs composite probe can realize simultaneous detection of DON and miR-34a induced by DON, which is significant for verifying the cytotoxic mechanism of DON.

Neutralization of Intracellular pH Homeostasis to Inhibit Osteoclasts Based on a Spatiotemporally Selective Delivery System.

Osteoporosis is a global disease caused by abnormal overactivation of osteoclasts. The acidic environment in sealing zone of osteoclasts with H+ pumped from cytoplasm is critical to the maturation of osteoclasts. Therefore, reducing the intracellular H+ concentration can reduce the H+ secretion of osteoclasts from the source. In our study, we developed a novel nanovesicle which encapsulates Na2HPO4 with a liposome hybridizes with preosteoclast membrane (Na2HPO4@Lipo-pOCm). These nanovesicles release Na2HPO4 into the preosteoclast by targeting preosteoclasts and membrane fusion, reducing the intracellular H+ concentration, and achieve biological cascade regulation of osteoclasts through simple pH regulation. In vitro and in vivo experiments confirmed that these nanovesicles reduce mitochondrial membrane potential by decreasing intracellular H+ concentration, thereby reducing the ROS in osteoclasts as well as the expression of the upstream transcription factor FOXM1 of Acp5. In short, this nanovesicle can significantly inhibit the osteoclasts and ameliorate osteoporosis caused by OVX.

A fluorescence and surface-enhanced Raman scattering dual-mode aptasensor for sensitive detection of deoxynivalenol based on gold nanoclusters and silver nanoparticles modified metal-polydopamine framework.

Deoxynivalenol (DON), a common mycotoxin produced by Fusarium species, poses a great threat to human and animal body. Hence, it is of significance to develop an ultrasensitive and reliable method for DON detection. Herein, a fluorescence and surface-enhanced Raman scattering (FL-SERS) dual-mode aptasensor was designed for the detection of DON based on gold nanoclusters (Au NCs) and silver nanoparticles modified metal-polydopamine framework (Ag NPs/MPDA). In this aptasensor, complementary DNA modified Au NCs (cDNA-Au NCs) was selected as fluorescence probe, and 6-carboxytetramethylrhodamine (TAMRA)-labeled aptamer modified Ag NPs/MPDA (Ag NPs/MPDA-Apt-TAMRA) was employed as SERS probe, in which Ag NPs/MPDA acted as SERS substance and fluorescence quencher, and TAMRA acted as Raman label. The superior binding affinity of the aptamer with DON to cDNA can regulate the fluorescence and Raman signal intensities and realize the quantitative determination of DON. Under the optimal conditions, the aptasensor exhibited a low detection limit of 0.08 ng mL-1 (0.1-100 ng mL-1) in FL mode and 0.06 ng mL-1 (0.1-100 ng mL-1) in SERS mode. In addition, it was successfully applied for DON detection in wheat flour. We believe that the proposed FL-SERS strategy has a promising application in the detection of mycotoxins.

A sensitive fluorescent assay based on gold-nanoclusters coated on molecularly imprinted covalent organic frameworks and its application in malachite green detection.

Malachite green (MG), as a parasiticide, is widely used in aquaculture to increase the production of the fishery industry. It poses a great danger to both the food system and the human body. In this study, a one-pot reverse microemulsion polymerization was employed to combine the gold nanoclusters (AuNCs) with molecularly imprinted polymers (MIPs) and covalent organic frameworks (COFs) to synthesize an efficient fluorescent hybrid probe (AuNCs@COFs@MIPs) for selective detection of MG. The specific recognition of AuNCs@COFs@MIPs towards MG triggers the fluorescence quenching of AuNCs. The fluorescent response was linearly related to the concentration over the range of 10-150 nmol/L with a limit of detection of 2.78 nmol/L. In addition, the proposed probe was further applied to fish and water samples. A favorable recovery ranged from 97.34 to 101.51 % toward trace amounts of MG indicating its promising application for detecting residue of veterinary drugs.

Hydrogel-integrated sensors for food safety and quality monitoring: Fabrication strategies and emerging applications.

Food safety is a global issue in public hygiene. The accurate, sensitive, and on-site detection of various food contaminants performs significant implications. However, traditional methods suffer from the time-consuming and professional operation, restricting their on-site application. Hydrogels with the merits of highly porous structure, high biocompatibility, good shape-adaptability, and stimuli-responsiveness offer a promising biomaterial to design sensors for ensuring food safety. This review describes the emerging applications of hydrogel-based sensors in food safety inspection in recent years. In particular, this study elaborates on their fabrication strategies and unique sensing mechanisms depending on whether the hydrogel is stimuli-responsive or not. Stimuli-responsive hydrogels can be integrated with various functional ligands for sensitive and convenient detection via signal amplification and transduction; while non-stimuli-responsive hydrogels are mainly used as solid-state encapsulating carriers for signal probe, nanomaterial, or cell and as conductive media. In addition, their existing challenges, future perspectives, and application prospects are discussed. These practices greatly enrich the application scenarios and improve the detection performance of hydrogel-based sensors in food safety detection.

Obacunone targets macrophage migration inhibitory factor (MIF) to impede osteoclastogenesis and alleviate ovariectomy-induced bone loss.

INTRODUCTION: Osteoporosis is the most common bone disorder where the hyperactive osteoclasts represent the leading role during the pathogenesis. Targeting hyperactive osteoclasts is currently the primary therapeutic strategy. However, concerns about the long-term efficacy and side effects of current frontline treatments persist. Alternative therapeutic agents are still needed. OBJECTIVES: Obacunone (OB) is a small molecule with a broad spectrum of biological activities, particularly antioxidant and anti-inflammatory effects. This study aims to examine OB's therapeutic potential on osteoporosis and explore the rudimentary mechanisms. METHODS: Osteoclast formation and osteoclastic resorption assays were carried out to examine OB's inhibitory effects in vitro, followed by the in-vivo studies of OB's therapeutic effects on ovariectomy-induced osteoporotic preclinical model. To further study the underlying mechanisms, mRNA sequencing and analysis were used to investigate the changes of downstream pathways. The molecular targets of OB were predicted, and in-silico docking analysis was performed. Ligand-target binding was verified by surface plasmon resonance (SPR) assay and Western Blotting assay. RESULTS: The results indicated that OB suppressed the formation of osteoclast and its resorptive function in vitro. Mechanistically, OB interacts with macrophage migration inhibitory factor (MIF) which attenuates receptor activator of nuclear factor kappa B (NF-κB) ligand (RANKL)-induced signaling pathways, including reactive oxygen species (ROS), NF-κB pathway, and mitogen-activated protein kinases (MAPKs). These effects eventually caused the diminished expression level of the master transcriptional factor of osteoclastogenesis, nuclear factor of activated T cells 1 (NFATc1), and its downstream osteoclast-specific proteins. Furthermore, our data revealed that OB alleviated estrogen deficiency-induced osteoporosis by targeting MIF and thus inhibiting hyperactive osteoclasts in vivo. CONCLUSION: These results together implicated that OB may represent as a therapeutic candidate for bone disorders caused by osteoclasts, such as osteoporosis.

A portable paper-based aptasensor for simultaneous visual detection of two mycotoxins in corn flour using dual-color upconversion nanoparticles and Cu-TCPP nanosheets.

A portable paper-based microfluidic aptasensor is established to simultaneously and visually detect zearalenone (ZEN) and ochratoxin A (OTA). The targets at the sample zone can migrate to two detection zones through dual-channels and result in green and blue fluorescence recovery. This is due to the specific recognition by a respective aptamer that destroys fluorescence resonance energy transfer (FRET) from dual-color upconversion nanoparticles (UCNPs) to Cu-TCPP nanosheets. By capturing fluorescent images and analyzing the corresponding RGB value via a smartphone, ZEN and OTA can be analyzed with limits of detection down to 0.44 ng/mL and 0.098 ng/mL in the linear ranges of 0.5-100 ng/mL and 0.1-50 ng/mL, respectively. Satisfactory recoveries are also obtained for ZEN (94.5-103.7 %) and OTA (92.2-106.8 %) in corn flour. With the advantages of simple operation, low sample consumption, and broad adaptability, this promising platform allows for the on-site detection of multiple hazards in food.

A plant-derived natural photosynthetic system for improving cell anabolism.

Insufficient intracellular anabolism is a crucial factor involved in many pathological processes in the body1,2. The anabolism of intracellular substances requires the consumption of sufficient intracellular energy and the production of reducing equivalents. ATP acts as an 'energy currency' for biological processes in cells3,4, and the reduced form of NADPH is a key electron donor that provides reducing power for anabolism5. Under pathological conditions, it is difficult to correct impaired anabolism and to increase insufficient levels of ATP and NADPH to optimum concentrations1,4,6-8. Here we develop an independent and controllable nanosized plant-derived photosynthetic system based on nanothylakoid units (NTUs). To enable cross-species applications, we use a specific mature cell membrane (the chondrocyte membrane (CM)) for camouflage encapsulation. As proof of concept, we demonstrate that these CM-NTUs enter chondrocytes through membrane fusion, avoid lysosome degradation and achieve rapid penetration. Moreover, the CM-NTUs increase intracellular ATP and NADPH levels in situ following exposure to light and improve anabolism in degenerated chondrocytes. They can also systemically correct energy imbalance and restore cellular metabolism to improve cartilage homeostasis and protect against pathological progression of osteoarthritis. Our therapeutic strategy for degenerative diseases is based on a natural photosynthetic system that can controllably enhance cell anabolism by independently providing key energy and metabolic carriers. This study also provides an enhanced understanding of the preparation and application of bioorganisms and composite biomaterials for the treatment of disease.

ZIF-8 base-aptamer "gate-lock" probes enable the visualization of a cascade response between deoxynivalenol and cytochrome c inside living cells.

Zeolitic imidazolate framework (ZIF-8) base-aptamer "gate-lock" biomaterial probes have been synthesized for monitoring intracellular deoxynivalenol (DON) and cytochrome c (cyt c) levels. The aptamer and organic fluorescent dye were regarded as a recognition element and a sensing element, respectively. In the presence of DON, the aptamers of DON and cyt c were specifically bound with the DON and induced cyt c, leading to the dissociation of aptamers from the porous surface of the probes. The gate was subsequently opened to release methylene blue (MB) and Rhodamine 6G (Rh6G), and their fluorescence (emission of MB at 700 nm and Rh6G at 550 nm) significantly recovered within 6 h. Cell imaging successfully monitored the exposure of DON and the biological process of cyt c discharge triggered by the activation of the DON-induced apoptosis pathway. In addition, the response between DON and cyt c was observed during the apoptosis process, which is of high significance for the comprehensive and systematic development of mycotoxins cytotoxicity.

Laser-Printed Paper-Based Microfluidic Chip Based on a Multicolor Fluorescence Carbon Dot Biosensor for Visual Determination of Multiantibiotics in Aquatic Products.

Excessive use of antibiotics in aquaculture severely endangers human health and ecosystems, which has raised significant concerns in recent years. However, conventional laboratory-based approaches regularly required time or skilled manpower. Herein, we propose a point-of-care-testing (POCT) biosensor detection device for the simultaneous determination of multiantibiotics without complex equipment or professional operators. A laser-printed paper-based microfluidic chip loaded with multicolor fluorescence nanoprobes (mCD-µPAD) was developed to rapidly detect sulfamethazine (SMZ), oxytetracycline (OTC), and chloramphenicol (CAP) on-site. These "fluorescence off" detection probes composed of carbon dots (CDs) conjugated with aptamers (donor) and MoS2 nanosheets (acceptor) (CD-apt-MoS2) were based on Förster resonance energy transfer. Upon the addition of target antibiotics, the significantly recovered fluorescence signal on the µPAD can be sensitively perceived by employing a 3D-printed portable detection box through a smartphone. Under optimal conditions, this µPAD allowed for a rapid response of 15 min toward SMZ, OTC, and CAP with considerable sensitivities of 0.47, 0.48, and 0.34 ng/mL, respectively. In shrimp samples, the recoveries were 95.2-101.2, 96.4-105, and 96.7-106.1% with RSD below 6%. This paper-based sensor opens an avenue for on-site, high-throughput, and rapid detection methods and can be widely used in POCT in food safety.

Demonstration of constant-cladding tapered-core Yb-doped fiber for mitigating thermally-induced mode instability in high-power monolithic fiber amplifiers.

In this work, a large-mode-area (LMA) step-index constant-cladding tapered-core (CCTC) Yb-doped fiber with a cladding diameter of ∼600 µm is successfully fabricated. The CCTC fiber has a small-core region (diameter of ∼20 µm) at both ends and a large-core region (diameter of ∼36 µm) in the middle. To prove the laser performance of the CCTC fiber, a detailed comparison experiment with conventional uniform fiber with the same effective core diameter is carried out in a multi-kW all-fiber MOPA configuration. The experimental results show that employing the CCTC fiber can effectively mitigate the thermally-induced transverse mode instability (TMI) in both co-pump and counter-pump schemes, and realize high slope efficiency and single-mode beam quality (M2∼1.30). Under the counter-pump scheme, the TMI threshold of the CCTC fiber is observed at ∼2.49 kW with a slope efficiency of 86.2%, while the uniform fiber amplifier exhibits a TMI threshold of ∼2.05 kW. The theoretical analysis based on a semi-analytical model indicates this CCTC fiber can effectively improve the TMI threshold owing to a stronger gain saturation. Our results verify the great potential of such an LMA CCTC fiber to mitigate thermal-induced TMI effect and achieve single-mode operation without sacrifice of laser efficiency in high power monolithic fiber lasers, and the further power scaling is expected by optimizing the fiber design.

Low-numerical aperture confined-doped long-tapered Yb-doped silica fiber for a single-mode high-power fiber amplifier.

A low-numerical aperture (NA) confined-doped long-tapered (LCT) Yb-doped fiber is proposed and fabricated by modified chemical vapor deposition combined with solution doping technique. The LCT fiber owns the core NA of ∼0.05 and the gain dopant doping diameter ratio of ∼77%, with a core/cladding diameter of 25/400 µm at both ends and 37.5/600 µm in the middle. The laser performance is demonstrated by a bidirectional pumping all-fiber amplifier, of which a 4.18-kW single-mode (M2 factor ∼1.3) laser output is achieved with a slope efficiency of ∼82.8%. Compared with the conventional fiber, the co-pumped and counter-pumped transverse mode instability thresholds and beam quality of the LCT fiber are remarkably enhanced. Throughout the continuous operation, the LCT fiber amplifier presents high power stability with fluctuation of < 1%. These results indicate that LCT fiber has great potential in power scaling remaining excellent beam quality.

Gasdermin D maintains bone mass by rewiring the endo-lysosomal pathway of osteoclastic bone resorption.

Gasdermin D (GSDMD)-mediated pyroptosis induces immunogenic cell death and promotes inflammation. However, the functions of GSDMD in tissue homeostasis remain unclear. Here, we identify a physiological function of GSDMD in osteoclasts via a non-lytic p20-generated protein, which prevents bone loss to maintain bone homeostasis. In the late stage of RANKL-induced osteoclastogenesis, GSDMD underwent cleavage, which is dependent on RIPK1 and caspase-8/-3, to yield this p20 product. Gsdmd-deficient osteoclasts showed normal differentiation but enhanced bone resorption with excessive lysosomal activity. Mice with complete or myeloid-specific Gsdmd deletion exhibited increased trabecular bone loss and more severe aging/ovariectomy-induced osteoporosis. GSDMD p20 was preferentially localized to early endosomes and limited endo-lysosomal trafficking and maturation, relying on its oligomerization and control of phosphoinositide conversion by binding to phosphatidylinositol 3-phosphate (PI(3)P). We have thus identified an anti-osteoclastic function of GSDMD as a checkpoint for lysosomal maturation and secretion and linked this to bone homeostasis and endosome-lysosome biology.

Extracellular Vesicle Mimetics: Preparation from Top-Down Approaches and Biological Functions.

Extracellular vesicles (EVs) have attracted attention as delivery vehicles due to their structure, composition, and unique properties in regeneration and immunomodulation. However, difficulties during production and isolation processes of EVs limit their large-scale clinical applications. EV mimetics (EVMs), prepared via top-down strategies that improve the yield of nanoparticles while retaining biological properties similar to those of EVs have been used to address these limitations. Herein, the preparation of EVMs is reviewed and their characteristics in terms of structure, composition, targeting ability, cellular uptake mechanism, and immunogenicity, as well as their strengths, limitations, and future clinical application prospects as EV alternatives are summarized.

Suppression of osteoclast multinucleation via a posttranscriptional regulation-based spatiotemporally selective delivery system.

Redundancy of multinucleated mature osteoclasts, which results from the excessive fusion of mononucleated preosteoclasts (pOCs), leads to osteolytic diseases such as osteoporosis. Unfortunately, the currently available clinical drugs completely inhibit osteoclasts, thus interfering with normal physiological bone turnover. pOC-specific regulation may be more suitable for maintaining bone homeostasis. Here, circBBS9, a previously unidentified circular RNA, was found to exert regulatory effects via the circBBS9/miR-423-3p/Traf6 axis in pOCs. To overcome the long-standing challenge of spatiotemporal RNA delivery to cells, we constructed biomimetic nanoparticles to achieve the pOC-specific targeted delivery of circBBS9. pOC membranes (POCMs) were extracted to camouflage cationic polymer for RNA interference with circBBS9 (POCM-NPs@siRNA/shRNAcircBBS9). POCM-NPs endowed the nanocarriers with improved stability, accurate pOC targeting, fusogenic uptake, and reactive oxygen species-responsive release. In summary, our findings may provide an alternative strategy for multinucleated cell-related diseases that involves restriction of mononucleated cell multinucleation through a spatiotemporally selective delivery system.