A Simplified Amplification-Free Strategy with Lyophilized CRISPR-CcrRNA System for Drug-Resistant Salmonella Detection.

Drug resistance in pathogenic bacteria has become a major threat to global health. The misuse of antibiotics has increased the number of resistant bacteria in the absence of rapid, accurate, and cost-effective diagnostic tools. Here, an amplification-free CRISPR-Cas12a time-resolved fluorescence immunochromatographic assay (AFC-TRFIA) is used to detect drug-resistant Salmonella. Multi-locus targeting in combination crRNA (CcrRNA) is 27-fold more sensitive than a standalone crRNA system. The lyophilized CRISPR system further simplifies the operation and enables one-pot detection. Induction of nucleic acid fixation via differentially charged interactions reduced the time and cost required for flowmetric chromatography with enhanced stability. The induction of nucleic acid fixation via differentially charged interactions reduces the time and cost required for flowmetric chromatography with enhanced stability. The platform developed for the detection of drug-resistant Salmonella has an ultra-sensitive detection limit of 84 CFU mL-1 within 30 min, with good linearity in the range of 102 -106 CFU mL-1 . In real-world applications, spiked recoveries range from 76.22% to 145.91%, with a coefficient of variation less than 10.59%. AFC-TRFIA offers a cost-effective, sensitive, and virtually equipment-independent platform for preventing foodborne illnesses, screening for drug-resistant Salmonella, and guiding clinical use.

Recent advances in biosynthesis of mycotoxin-degrading enzymes and their applications in food and feed.

Mycotoxins are secondary metabolites produced by fungi in food and feed, which can cause serious health problems. Bioenzymatic degradation is gaining increasing popularity due to its high specificity, gentle degradation conditions, and environmental friendliness. We reviewed recently reported biosynthetic mycotoxin-degrading enzymes, traditional and novel expression systems, enzyme optimization strategies, food and feed applications, safety evaluation of both degrading enzymes and degradation products, and commercialization potentials. Special emphasis is given to the novel expression systems, advanced optimization strategies, and safety considerations for industrial use. Over ten types of recombinases such as oxidoreductase and hydrolase have been studied in the enzymatic hydrolysis of mycotoxins. Besides traditional expression system of Escherichia coli and yeasts, these enzymes can also be expressed in novel systems such as Bacillus subtilis and lactic acid bacteria. To meet the requirements of industrial applications in terms of degradation efficacy and stability, genetic engineering and computational tools are used to optimize enzymatic expression. Currently, registration and technical difficulties have restricted commercial application of mycotoxin-degrading enzymes. To overcome these obstacles, systematic safety evaluation of both biosynthetic enzymes and their degradation products, in-depth understanding of degradation mechanisms and a comprehensive evaluation of their impact on food and feed quality are urgently needed.

Combined metabolomics and transcriptomics analysis reveals the mechanism of antibiotic resistance of Salmonella enterica serovar Typhimurium after acidic stress.

Drug-resistant Salmonella is widely distributed in the meat production chain, endangering food safety and public health. Acidification of meat products during processing can induce acid stress, which may alter antibiotic resistance. Our study investigated the effects of acid stress on the antibiotic resistance and metabolic profile of Salmonella Typhimurium, and explored the underlying mechanisms using metabolomic and transcriptomic analysis. We found that acid-stressed 14028s was more sensitive to small molecule hydrophobic antibiotics (SMHA) while more resistant to meropenem (MERO). Metabolomic analysis revealed that enhanced sensitivity to SMHA was correlated with increased purine metabolism and tricarboxylic acid cycle. Transcriptomic analysis revealed the downregulation of chemotaxis-related genes, which are also associated with SMHA sensitivity. We also found a significant downregulation of the ompF gene, which encodes a major outer membrane protein OmpF of Salmonella. The decreased expression of OmpF porin hindered the influx of MERO, leading to enhanced resistance of the bacteria to the drug. Our findings contribute to greatly improve the understanding of the relationship between Salmonella metabolism, gene expression, and changes in drug resistance after acid stress, while providing a structural framework for exploring the relationship between bacterial stress responses and antibiotic resistance.

Exploring the impact of fungal spores from agricultural environments on the mice lung microbiome and metabolic profile.

Exposure to particulate matter (PM) from agricultural environments has been extensively reported to cause respiratory health concerns in both animals and agricultural workers. Furthermore, PM from agricultural environments, containing fungal spores, has emerged as a significant threat to public health and the environment. Despite its potential toxicity, the impact of fungal spores present in PM from agricultural environments on the lung microbiome and metabolic profile is not well understood. To address this gap in knowledge, we developed a mice model of immunodeficiency using cyclophosphamide and subsequently exposed the mice to fungal spores via the trachea. By utilizing metabolomics techniques and 16 S rRNA sequencing, we conducted a comprehensive investigation into the alterations in the lung microbiome and metabolic profile of mice exposed to fungal spores. Our study uncovered significant modifications in both the lung microbiome and metabolic profile post-exposure to fungal spores. Additionally, fungal spore exposure elicited noticeable changes in α and ß diversity, with these microorganisms being closely associated with inflammatory factors. Employing non-targeted metabolomics analysis via GC-TOF-MS, a total of 215 metabolites were identified, among which 42 exhibited significant differences. These metabolites are linked to various metabolic pathways, with amino sugar and nucleotide sugar metabolism, as well as galactose metabolism, standing out as the most notable pathways. Cysteine and methionine metabolism, along with glycine, serine and threonine metabolism, emerged as particularly crucial pathways. Moreover, these metabolites demonstrated a strong correlation with inflammatory factors and exhibited significant associations with microbial production. Overall, our findings suggest that disruptions to the microbiome and metabolome may hold substantial relevance in the mechanism underlying fungal spore-induced lung damage in mice.

Recent Advances in g-C3 N4 -Based Photocatalysts for Pollutant Degradation and Bacterial Disinfection: Design Strategies, Mechanisms, and Applications.

Emerging photocatalytic technology promises to provide an effective solution to the global energy crisis and environmental pollution. Graphite carbon nitride (g-C3 N4 ) has gained extensive attention in the scientific community due to its excellent physical and chemical properties, attractive electronic band structure, and low cost. In this paper, research progress in design strategies for g-C3 N4 -based photocatalysts in the past five years is reviewed from the perspectives of nanostructure construction, element doping, and heterostructure construction. To clarify the relationship between application requirements and structural design, variations in the morphology, electronic energy band structure, light absorption capacity, as well as interfacial charge transfer caused by various modification strategies are discussed in detail. The recent applications of g-C3 N4 -based photocatalysts for pollutant degradation and bacterial disinfection are reviewed, as well as the antimicrobial activity and degradation mechanisms. Finally, current challenges and future development directions for the practical application of g-C3 N4 -based photocatalysts are tentatively discussed.

Advances in the study of liver microsomes in the in vitro metabolism and toxicity evaluation of foodborne contaminants.

Foodborne contaminants are closely related to anthropologic activities and represent an important food safety hazard. The study of metabolic transformation and toxic side effects of foodborne contaminants in the body is important for their safety assessment. Liver microsomes contain a variety of enzymes related to substance metabolism and biotransformation. An in vitro model simulating liver metabolic transformation is associated with a significant advantage in the study of the metabolic transformation mechanisms of contaminants. This review summarizes the recent progress in the application of liver microsomes in metabolic transformation and toxicity evaluation of various foodborne pollutants based on metabolic kinetics, molecular docking and enzyme inhibition studies. The purpose of this review is to distinguish the existing studies involving liver microsomes and provide strategies for their application in the future. Finally, the prospects and challenges of the liver microsomal model are discussed.

Occurrence, transformation, and toxicity of fumonisins and their covert products during food processing.

Fumonisins comprise structurally related metabolites mainly produced by Fusarium verticillioides and Fusarium proliferatum. Contamination with fumonisins causes incalculable damage to the economy and poses a great risk to animal and human health. Fumonisins and their covert products are found in cereals and cereal products. Food processing significantly affects the degradation of toxins and the formation of covert toxins. However, studies on fumonisins and their covert mycotoxins remain inadequate. This review aims to summarize changes in fumonisins and the generation of covert fumonisins during processing. It also investigates the toxicity and determination methods of fumonisins and covert fumonisins, and elucidates the factors affecting fumonisins and their covert forms during processing. In addition to the metabolic production by plants and fungi, covert fumonisins are mainly produced by covalent or noncovalent binding, complexation, or physical entrapment of fumonisins with other substances. The toxicity of covert fumonisins is similar to that of free fumonisins and is a non-negligible hazard. Covert fumonisins are commonly found in food matrices, and methods to analyze them have yet to be improved. Food processing significantly affects the conversion of fumonisins to their covert toxins.

Advances on the rapid and multiplex detection methods of food allergens.

With the gradually increasing prevalence of food allergy in recent years, food allergy has become a major public health problem worldwide. The clinical symptoms caused by food allergy seriously affect people's quality of life; there are unknown allergen components in novel food and hidden allergens caused by cross contamination in food processing, which pose a serious risk to allergy sufferers. Thus, rapid and multiplex detection methods are required to achieve on-site detection or examination of allergic components, so as to identify the risk of allergy in time. This paper reviews the progress of high-efficiency detection of food allergens, including enhanced traditional detection techniques and emerging detection techniques with the ability high-throughput detection or screening potential food allergen, such as xMAP, biosensors, biochips, etc. focusing on their sensitivity, applicability of each method in food, along with their pretreatment, advantages, limitation in the application of food analysis. This paper also introduces the challenges faced by these high-efficiency detection technologies, as well as the potential of customized allergen screening methods and rapid on-site detection technology as future research directions.

Microbial detoxification of mycotoxins in food and feed.

Mycotoxins are metabolites produced by fungi growing in food or feed, which can produce toxic effects and seriously threaten the health of humans and animals. Mycotoxins are commonly found in food and feed, and are of significant concern due to their hepatotoxicity, nephrotoxicity, carcinogenicity, mutagenicity, and ability to damage the immune and reproductive systems. Traditional physical and chemical detoxification methods to treat mycotoxins in food and feed products have limitations, such as loss of nutrients, reagent residues, and secondary pollution to the environment. Thus, there is an urgent need for new detoxification methods to effectively control mycotoxins and treat mycotoxin pollution. In recent years, microbial detoxification technology has been widely used for the degradation of mycotoxins in food and feed because this approach offers the potential for treatment with high efficiency, low toxicity, and strong specificity, without damage to nutrients. This article reviews the application of microbial detoxification technology for removal of common mycotoxins such as Aflatoxin, Ochratoxin, Zearalenone, Deoxynivalenol, and Fumonisins, and discusses the development trend of this important technology.

Highly sensitive real-time detection of intracellular oxidative stress and application in mycotoxin toxicity evaluation based on living single-cell electrochemical sensors.

Single-cell electrochemical sensor is widely used in the local selective detection of single living cells because of its high spatial-temporal resolution and sensitivity, as well as its ability to obtain comprehensive cellular physiological states and processes with increased accuracy. Functionalized nanoprobes can detect the oxidative stress response of cells in single-cell electrochemical sensors. Moreover, the T-2 toxin is one of the most toxic mycotoxins and widely occurs in field crops. T-2 toxin can cause mitochondrial damage in cells and increase intracellular reactive oxygen species (ROS) in various cells. As the most representative free radical of intracellular ROS, H2O2 can effectively reflect the toxic effects of intracellular T-2 toxin. In this study, a functionalized gold nanoprobe was used to dynamically monitor the production of H2O2 in a single live human hepatoma cell HepG2 stimulated by mycotoxin T-2. The concentration of H2O2 produced by HepG2 cells stimulated by T-2 toxin at 1 ppb-1 ppm was linearly correlated, R2 = 0.99055, and LOD = 0.13807 ng mL-1. Sample spiking experiments were conducted, and the recovery rate of spiking was 81.19%-130.17%. A comparative analysis of differences in the current produced by multiple toxins, HT-29 cells, as well as single cells in cell populations, was performed. This method can be applied in real-time monitoring of mycotoxin toxicity during food processing in living cells and provides a novel idea for enhancing food quality and safety in a nanoenvironment.

One-step time-resolved fluorescence microsphere immunochromatographic test strip for quantitative and simultaneous detection of DON and ZEN.

Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins that contaminate a wide range of grains and crops. In this study, a one-step time-resolved single-channel immunochromatographic test strip based on europium ion polystyrene fluorescence microspheres was first developed for sensitive and quantitative detection of DON and ZEN. The concentration of the artificial antigen and the mass ratio of the monoclonal antibody to fluorescent microspheres for conjugation were optimized to simplify the sample addition process during immunochromatographic assay and improve the on-site detection efficiency. The limits of detection (LOD) of the single-channel immunochromatographic test strip for DON and ZEN detection were 0.17 and 0.54 µg/L, respectively. Meanwhile, the dual-channel immunochromatographic test strip was designed to simultaneously detect DON and ZEN, with LODs of 0.24 and 0.69 µg/L achieved for DON and ZEN, respectively. The developed test strips also yielded recovery results consistent with that obtained by LC-MS/MS for DON and ZEN detection in real samples of wheat and corn flour, confirming the practicability and reliability of the test strip. The developed immunochromatographic test strips realize quick and sensitive detection of DON and ZEN, exhibiting potential for broad applications in the point-of-care testing platform of multiple mycotoxins in agricultural products. Graphic abstract.

A novel concentration gradient microfluidic chip for high-throughput antibiotic susceptibility testing of bacteria.

Antibiotic resistance has become a serious threat to food safety and public health globally. Therefore, the development of a sensitive, quick, and simple method for antibiotic susceptibility testing is an urgent and crucial need. A novel concentration gradient microfluidic chip was designed in this work to generate antibiotic concentration gradient, culture bacteria, and produce fluorescence emission. An in-house-assembled fluorescence detection platform was constructed, and experiments were conducted to verify the linearity of the generated concentration gradient, explore the appropriate incubation time and flow rate for the microfluidic chip, and study the effect of long-term acid-based food processing on antibiotic susceptibility testing. Experimental results show that the concentration gradient generated by the microfluidic chip exhibited good linearity, stability, and controllability. The appropriate flow rate and incubation time for the microfluidic chip were 2 µL/min and 5 h, respectively. The use of this microfluidic chip for testing antibiotic resistance of Salmonella to ofloxacin and ampicillin generated results that were completely consistent with test results obtained using the gold-standard method. Furthermore, Salmonella showed greater sensitivity to antibiotics under strong acid conditions, confirming the potential influence of acid-based food processing on antibiotic susceptibility testing of real samples. The designed microfluidic chip provides a high-throughput, sensitive, and rapid antibiotic susceptibility testing method that combines the microfluidic chip and the fluorescence detection platform. The application of this method would facilitate determination of antibiotic-resistant bacterial strains for clinicians and researchers, and enable monitoring of changes in bacterial resistance during food processing.

Opposite estrogen effects of estrone and 2-hydroxyestrone on MCF-7 sensitivity to the cytotoxic action of cell growth, oxidative stress and inflammation activity.

There are many kinds of estrogens, and endogenous estrogens produce a variety of estrogen metabolites with similar structure but with different physiological effects after metabolism in vivo. Studies have shown that estrone (E1) widely occurs in the environment and animal-derived food. Because of its estrogen effect, E1 can have adverse effects on the human body as an endocrine disruptor. In this study, we found that E1 and 2-hydroxyestrone (2-OH-E1), the hydroxylation metabolite of estrogen, have opposite proliferative effects on breast cancer cells (MCF-7) through cell proliferation experiments and comparison of their effects by molecular docking and detection of ROS, Ca2+, and cell pathway proteins. The effects of 2-methoxyestrone (2-MeO-E1) and 16α-hydroxyestrone (16α-OH-E1) on the biochemical and protein levels of MCF-7 were further studied to compare the effects of metabolic sites and modes on estrogen effects. Hydroxylation of E1 at the C2 site weakened the estrogen effect, down-regulated the expression of the mammalian target of rapamycin (mTOR) and protein kinase B (Akt) pathway proteins, inhibited the proliferation of cancer cells, and enhanced anti-oxidative stress and anti-inflammation. Methoxylation at the C2 position also inhibited the expression of inflammatory and oxidative stress pathway proteins but did not greatly affect the estrogen effects. However, hydroxylation on C16 had no significant effect on the biological effects of estrogen. Therefore, the structural changes of estrogen on C2 are important reasons for the different physiological effects of estrogen and its metabolites. Thus, by regulating the gene Cytochrome P450 1B1(CYP1B1), which affects the hydroxylation metabolism of estrogen, and promoting the hydroxylation of estrone at the C2 position, the estrogen effect of estrone can be effectively reduced, thus reducing the harm its poses in food and the environment.

A novel fluorescent molecularly imprinted polymer SiO2 @CdTe QDs@MIP for paraquat detection and adsorption.

Paraquat (PQ) residue is harmful for human health, agriculture, and the aquatic environment. This paper proposes a novel fluorescent molecularly imprinted polymer (MIP), SiO2 @CdTe QDs@MIP, for PQ detection and adsorption. The MIP was synthesized using 3-aminopropyltriethoxysilane as the functional monomer, 4,4'-bipyridyl as the template molecule, and tetraethoxysilane as the cross-linker. In addition, CdTe quantum dots featuring unique optical characteristics and excellent photochemical stability were combined as signal reporter. The synthesized MIP had a Brunauer-Emmett-Teller surface area of 68.2 m2 /g, pore volume of 0.42 cm3 /g and pore size of 6.9 nm, demonstrating the potential for both PQ detection and adsorption. For PQ detection, the MIP could achieve three orders of magnitude better than the limit of detection, and one order of magnitude wider detection range than existing methods. The PQ recovery values for real samples of water and corn were 96.4-102.1% and 93.9-97.3%, respectively. The amount of PQ detected by the MIP was within 98.05% on average of that using high-performance liquid chromatography. For PQ adsorption, the MIP had an adsorption capacity of 3.36 mg/g, and followed a pseudo-second-order kinetic model with excellent toxicological characteristics. Overall, the novel SiO2 @CdTe QDs@MIP proposed in this paper could facilitate an efficient and convenient method for PQ detection and adsorption.

Loop-mediated isothermal amplification-based microfluidic chip for pathogen detection.

Due to the significant growth of food production, the potential likelihood of food contamination is increasing. Foodborne illness caused by bacterial pathogens has considerably increased over the past decades, while at the same time, the species of harmful microorganisms also varied. Conventional bacterial culturing methods have been unable to satisfy the growing requirement for food safety inspections and food quality assurance. Therefore, rapid and simple detection methods are urgently needed. The loop-mediated isothermal amplification (LAMP) technology is a highly promising approach for the rapid and sensitive detection of pathogens, which allows nucleic acid amplification under isothermal conditions. The integration of the LAMP assay onto a microfluidic chip is highly compatible with point-of-care or resource-limited settings, as it offers the capability to perform experiments in combination with high screening efficiency. Here, we provide an overview of recent advances in LAMP-based microfluidic chip technology for detecting pathogens, based on real-time or endpoint determination mechanisms. We also discuss the promoting aspects of using the LAMP technique in a microfluidic platform, to supply a guideline for further molecular diagnosis and genetic analysis.

Ultrasensitive Fluorometric Angling Determination of Staphylococcus aureus in Vitro and Fluorescence Imaging in Vivo Using Carbon Dots with Full-Color Emission.

Rapid, accurate, and safe screening of foodborne pathogenic bacteria is essential to effectively control and prevent outbreaks of foodborne illness. Fluorescent sensors constructed from carbon dots (CDs) and nanomaterial-based quenchers have provided an innovative method for screening of pathogenic bacteria. Herein, an ultrasensitive magnetic fluorescence aptasensor was designed for separation and detection of Staphylococcus aureus (S. aureus). Multicolor fluorescent CDs with a long fluorescent lifetime (6.73 ns) and high fluorescence stability were synthesized using a facile hydrothermal approach and modified cDNA as a highly sensitive fluorescent probe. CD fluorescence was quenched by Fe3O4 + aptamer via fluorescence resonance energy transfer (FRET). Under optimal conditions, the FRET-based aptasensor can detect S. aureus accompanied by a wide linear range of 50-107 CFU·mL-1 and a detection limit of 8 CFU·mL-1. Compared with other standard methods, this method was faster and more convenient, and the entire test was finished within 30 min. The capability of the aptasensor was simultaneously investigated on food samples. Additionally, the developed CDs exhibited excellent biocompatibility and were thus applied as fluorescent probes for bioimaging both in vitro and in vivo. This new platform provided an excellent application of the CDs for detecting and bioimaging pathogenic bacteria.

Ultrasensitive fluorometric determination of iron(iii) and inositol hexaphosphate in cancerous and bacterial cells by using carbon dots with bright yellow fluorescence.

An ON-OFF-ON dual-function fluorescent nanoprobe is described for the trace detection of ferric ions and inositol hexaphosphate (IP6) in living cells. It is based on the use of yellow-fluorescent nitrogen-doped carbon dots (YN-CDs). Highly fluorescent YN-CDs were synthesized by a hydrothermal process. They have an absolute quantum yield of 2.15% and excitation/emission peaks at 420/575 nm. Fluorescence is quenched by Fe3+via photo-induced electron transfer. The quenchometric assay has a 34 nM detection limit for Fe(iii). On addition of IP6 which has a high affinity for Fe3+ due to the formation of Fe-O-P bonds, fluorescence becomes gradually restored. The resulting ON-OFF-ON assays for Fe(iii) and IP6 are reliable and sensitive. IP6 can be detected at concentrations as low as 2 nM. The nanoprobe was then applied to the determination of Fe3+ and IP6 in living cells in a food matrix. Furthermore, YN-CDs exhibited excellent biocompatibility. Hence, the probe can be applied as a fluorescent ink for bioimaging, both in vitro (cancer cells and bacteria) and in vivo (nematodes and mice).

A novel magnetic fluorescent biosensor based on graphene quantum dots for rapid, efficient, and sensitive separation and detection of circulating tumor cells.

We describe a "turn-on" magnetic fluorescent biosensor based on graphene quantum dots (GQDs), Fe3O4, and molybdenum disulfide (MoS2) nanosheets. It is used for rapid, efficient, and sensitive separation and detection of circulating tumor cells (CTCs). A facile approach (electrochemical synthesis method) for the preparation of photoluminescent GQDs functionalized with an aptamer [epithelial cell adhesion molecule (EpCAM) receptors] and a magnetic agent for one-step bioimaging and enrichment of CTCs is described. MoS2 nanosheets, as a fluorescence quencher, and the aforementioned aptamer@Fe3O4@GQD complex were assembled to construct "turn-on" biosensing magnetic fluorescent nanocomposites (MFNs). This system exhibits low cytotoxicity and an average capture efficiency of 90%, which is higher than that of other magnetic nanoparticles on account of the one-step CTC separation method. In addition, the MFNs could quickly identify and label CTCs within 15 min, surpassing other one-step and two-step marker detection methods. Furthermore, because of the presence of aptamers, the MFNs have specific capability to capture CTCs (both low- and high-EpCAM-expressing cells). In addition, high-sensitivity detection of up to ten tumor cells in whole blood was achieved. Therefore, the MFNs have great potential to be used as universal biosensing nanocomposites for fluorescence-guided tumor cell enrichment and bioimaging. Graphical abstract ᅟ.

Recent Developments of High-Resolution Chemical Imaging Systems Based on Light-Addressable Potentiometric Sensors (LAPSs).

A light-addressable potentiometric sensor (LAPS) is a semiconductor electrochemical sensor based on the field-effect which detects the variation of the Nernst potential on the sensor surface, and the measurement area is defined by illumination. Thanks to its light-addressability feature, an LAPS-based chemical imaging sensor system can be developed, which can visualize the two-dimensional distribution of chemical species on the sensor surface. This sensor system has been used for the analysis of reactions and diffusions in various biochemical samples. In this review, the LAPS system set-up, including the sensor construction, sensing and substrate materials, modulated light and various measurement modes of the sensor systems are described. The recently developed technologies and the affecting factors, especially regarding the spatial resolution and temporal resolution are discussed and summarized, and the advantages and limitations of these technologies are illustrated. Finally, the further applications of LAPS-based chemical imaging sensors are discussed, where the combination with microfluidic devices is promising.

Ultrasensitive "FRET-SEF" Probe for Sensing and Imaging MicroRNAs in Living Cells Based on Gold Nanoconjugates.

MicroRNAs (miRNAs), a kind of single-stranded small RNA molecule, play significant roles in the physiological and pathological processes of human beings. Currently, miRNAs have been demonstrated as important biomarkers critically related to many diseases and life nature, including several cancers and cell senescence. It is valuable to establish sensitive assays for monitoring the levels of intracellular up-regulated/down-regulated miRNA expression, which would contribute to the early prediction of the tumor risk and cardiovascular disease. Here, an oriented gold nanocross (AuNC)-decorated gold nanorod (AuNR) probe with "OFF-enhanced ON" fluorescence switching was developed based on fluorescence resonance energy transfer and surface enhanced fluorescence (FRET-SEF) principle. The nanoprobe was used to specifically detect miRNA in vitro, which gave two linear responses represented by the equation F = 1830.32 log C + 6349.27, R2 = 0.9901, and F = 244.41 log C + 1916.10, R2 = 0.9984, respectively, along with a detection limit of 0.5 aM and 0.03 fM, respectively. Furthermore, our nanoprobe was used to dynamically monitor the expression of intracellular up-regulated miRNA-34a from the HepG2 and H9C2 cells stimulated by AFB1 and TGF-ß1, and the experimental results showed that the new probe not only could be used to quantitively evaluate miRNA oncogene in vitro, but also enabled tracking and imaging of miRNAs in living cells.