The pervasive threat of organophosphorus pesticides (OPs) contamination and associated poisoning incidents demands detection strategies that go beyond simple quantification, specifically targeting the identification of unknown analogs within matrices. Herein, we report a novel chemiluminescent (CL) sensing platform driven by rationally designed, enzyme-specific probes (ACh-CL and BCh-CL) that exploit distinct steric and electronic interactions with acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Unlike conventional assays, we constructed the classification models by integrating distinct signal patterns across three concentration gradients with machine learning models, including XGBoost and Random Forest. Based on that, we successfully discriminated OPs from non-OPs and non-pesticide analogs (NPAs) in samples of unknown category and concentration. Achieving high sensitivity (low to 0.1 ng/mL) and rapid response (10 min), the method showed good anti-interference in vegetables, soil, water and serum, with recoveries of 74.21 %–113.8 %. Overall, this CL technique-driven, data-enhanced protocol enables high-throughput screening and risk assessment in food safety and environmental monitoring.

Acute lung injury (ALI) has garnered increasing attention due to its high morbidity and mortality, particularly in light of the recent global COVID-19 pandemic. The use of biomarkers presents a promising avenue for the early evaluation of clinical conditions and the prompt initiation of treatment. Especially, it potentially mitigates disease progression without radiation exposure and the reliance on already present and even severe symptoms. Carboxylesterase (CES) has been demonstrated to be a promising biomarker for ALI due to its crucial role in regulating pulmonary surfactant homeostasis and cholesterol mobilization. Herein, we designed three novel chemiluminescent probes (CES-AT, CES-TA and CES-CA) by tethering an acyl group to the chemiluminescent scaffold, enabling the facile and rapid detection of CES. Among them, CES-AT exhibited the highest signal-to-background ratio and the best stability. We further demonstrated that probe CES-AT was suitable for sensing endogenous CES in cells and mice with good cell viability and sensitivity (able to detect as few as 10 cells), thus for real-time imaging of CES levels, varying with the progression of ALI and its treatment. Overall, probe CES-AT can effectively reflect lung injury status, making it a valuable tool for diagnosing ALI and assessing treatment efficacy by virtue of its simplicity and rapid detection capabilities.

Antibiotic-resistant bacteria, primarily driven by β-lactamases, significantly undermine the effectiveness of β-lactam antibiotics, posing a grave threat to human health. As such, it is crucial to develop sensitive and rapid diagnostic methods to distinguish antibiotic-resistant bacteria from antibiotic-susceptible ones, thereby guiding clinical drug use. In this study, we present a selective "turn-on" chemiluminescent probe (HS-CL) created by attaching a 2-iodobenzoyl group to a phenoxy-dioxetane scaffold. The interaction of antibiotic-resistant bacteria with β-lactams generates hydrogen sulfide molecules, which selectively trigger probe HS-CL, resulting in a noticeable increase in the chemiluminescent signal. This enables point-of-care testing for bacterial resistance. Probe HS-CL demonstrated excellent specificity and a linear response to hydrogen sulfide concentration ranging from 5 to 100 μM (R2 = 0.9822), with a detection limit of 1.02 μM. We tested this method with four different antibiotic-resistant bacterial strains, including Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus, Escherichia coli, and Klebsiella pneumoniae, and two antibiotic-susceptible strains, Staphylococcus aureus and Escherichia coli. This probe can directly indicate the presence of antibiotic resistance in examined β-lactams. Most notably, when we applied this technique to bacteria isolated from patients, including Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa, the chemiluminescence probe successfully distinguished β-lactam-resistant strains from antibiotic-susceptible ones.

Various supported metal catalysts have been developed to immobilize and stabilize metal species with small sizes and high dispersity. A hierarchical branch-like metal-organic frameworks (MOF) with nanosized branches and empty space between its building units facilitates the accession of substrates to metal active sites and the loading of nanoparticles, which make it an ideal carrier for developing supported metal nanocatalysts due to its high catalytic activity and load capacity. Herein, a Ag nanoparticle-loaded hierarchical CoZn-MOF (AgCZM) with enhanced peroxidase (POD)-like activity was prepared for the sensitive chemiluminescence (CL) assay. To prepare AgCZM, a pine-needle-cluster-like CoZn-MOF with POD-like activity was synthesized first via a one-pot solvothermal method. Then, the high-loaded CoZn-MOF-supported Ag nanoparticle catalyst for boosting its activity was synthesized by simply mixing CoZn-MOF and AgNO3 in polyvinylpyrrolidone (PVP) aqueous solution without using strong reducing agents, thereby enabling CoZn-MOF to retain its hierarchical structure. Further, the reactive oxygen species (ROS) generated during AgCZM-based and CoZn-MOF-based catalytic decomposition of H2O2 were verified and compared via scavenger experiments and electron paramagnetic resonance (EPR) tests, and the possible AgCZM-based luminol CL mechanism was explored. To validate the feasibility of the AgCZM-based CL assay, a CL system consisting of luminol, H2O2, and AgCZM was fabricated for dopamine (DA) detection with a negative response signal. The proposed AgCZM-based CL assay exhibited good linearities at low (5-125 nM) and high (0.625-1.75 mu M) DA concentrations with detection limits of 3.91 and 73.87 nM, respectively.

Mycoplasma pneumoniae (MP) is a leading pathogen responsible for community-acquired pneumonia in the whole population, posing a life-threatening risk in severe disease. Current diagnostic methods, including culture, serology, and PCR, are limited by low sensitivity, delayed results, or operational complexity, highlighting the urgent need for rapid and reliable alternatives. Critically, MP metabolism produces hydrogen peroxide (H2O2), a key virulence factor, making it a promising biomarker for MP infection. Compared to other optical approaches, chemiluminescence-based one offers a powerful alternative for pathogen detection due to their high sensitivity, low cost, and rapid response. Herein, we report the first application of a chemiluminescent probe (MPCL) based on the established phenoxy-dioxetane scaffold with a 4-nitro-α-ketoamide trigger for direct MP detection. Our study demonstrated the practical value of this probe in distinguishing MP from other respiratory pathogens through quantitative H2O2 detection. MPCL enabled sensitive quantification and real-time imaging of H2O2 in MP-infected mice, and, when applied to clinical respiratory samples, achieved excellent diagnostic performance, with ROC analysis showing an area under the curve of 0.992 (95% CI: 0.974-1.000), 100% sensitivity, and 96.5% specificity. These results establish MPCL as a practical chemiluminescent tool with strong translational potential for rapid, low-cost, and reliable diagnosis of MP infections in both clinical and preclinical settings.

Imaging detection of interlinked dual proteases is imperative for precise tumor imaging, which remains challenging due to limited modification position of specific substrate and possible steric hindrance. Herein, we have developed a unimolecular chemiluminescent probe (LGP-CL) tandemly activated by two proteases interlinked with liver cancer to achieve precise tumor imaging. Probe LGP-CL consists of a phenoxy-dioxetane scaffold caged by a tripeptide substrate (LGP, leucine-glycine-proline) as the sensing layer, which can be cleaved sequentially by aminopeptidase N (APN) and dipeptidyl peptidase IV (DPPIV) to turn on a strong chemiluminescent signal, and silenced by specific inhibitor of each enzyme, which accounts for an integrated logic gate (AND, OR and INHIBIT). The successful cleavage of dual proteases on the metabolic site depends on the proper structure of the tripeptide substrate, as confirmed by two probes design. Probe LGP-CL (LGP as the substrate) enables the excellent “dual-lock-dual-key” fit with a 382-fold enhancement of chemiluminescent emission while no obvious signal is observed by using GPL-CL (GPL as the substrate). By virtue of its rapid response (several minutes), high sensitivity and good cell viability, probe LGP-CL has been utilized to evaluate upregulated levels of proteases in vitro and in living systems, especially to distinguish liver tumor cells (HepG2) from others (LO2, MCF-7, MCF-10a and RAW264.7). Overall, the newly developed CL probe may facilitate rapid investigation into the role played by proteases in liver diseases, enabling timely selection appropriate treatment. Therefore, our work not only sheds light on the rational design of optical probes for dual protease imaging, but provides a promising tool for clinical diagnosis and even drug discovery.

Being a class of highly ordered porous materials, hydrogen-bonded organic frameworks (HOFs) hold great potential for developing promising enzyme mimics in sensing applications, yet remain underutilized in this field. Herein, a robust HOF@Co/Heme with stable peroxidase-like activity was synthesized by loading Heme and Co2+ to HOF for establishing a sensitive chemiluminescence (CL) assay of glucose. The scavenger experiment and electron paramagnetic resonance test revealed that reactive oxygen species (ROS) including 1O2, OH center dot and O2 center dot- were generated in HOF@Co/Heme-based catalytic decomposition of H2O2, and thereafter notably enhanced the CL signal of luminol-H2O2 CL system. Consequently, a CL biosensor based on glucose oxidase (GOx) and HOF@Co/Heme tandem catalysis was developed for sensitive detection of glucose, where GOx could catalyze glucose oxidation and offer H2O2 to HOF@Co/Heme for generating ROS and enhancing CL signal. Under optimized conditions, the GOx-HOF@Co/Heme-based biosensor exhibited a linear detection range from 0.06 to 6 mu M with an impressively low detection limit of 54 nM. Furthermore, its practicality was confirmed by the successful application in quantifying glucose in human serum samples.

Optical analysis is non-destructive, real-time with a specific spatial resolution, which has been developed as an essential technology to study the occurrence, development, diagnosis, and treatment of diseases. It contains fluorescent (FL), bioluminescent (BL) and chemiluminescent (CL) methods. Among them, CL probes with an adamantane-dioxetane chemiluminescence (AD-CL) scaffold attracted much attention. Recently, significant improvement on these probes has been achieved with the elimination of external light source, low phototoxicity, high sensitivity, and a facile system without additional reagents, such as oxidants. Until now, the CL probes were further developed with special modifications and new synthesis routes based on the AD-CL scaffold, realizing the detection and optical imaging of various biomolecules in living systems with enhanced properties. Herein, the recent research progress on AD-CL probes has been reviewed. The review is divided into two parts. The first part will mainly introduce the molecular modification strategy of AD-CL probes and the second part will focus on their application in several cases.

Aminopeptidase N, as a target for drug discovery, shows marked relationships with many diseases, especially liver injury and cancer. Here, we explored a chemiluminescence (CL) probe for sensing APN by tethering the APN-specific substrate group to the ortho-acrylated phenoxy-dioxetane scaffold. In this way, two CL probes (APN-CL and BAPN-CL) were designed with noncapped leucine and butoxy-carbonyl capped leucine as the protecting group to preserve the chemiexcitation energy. The uncovered leucine was demonstrated to be essential for detection of APN activity by comparing the CL intensity of two CL probes. Probe APN-CL was turned on upon APN cleavage, resulting in a high chemiluminescent emission, whereas the chemiexcitation energy of probe BAPN-CL was still restrained even with the high-level APN. The result was further elucidated by molecular docking simulations. Probe APN-CL exhibited a fast response and high sensitivity with a detection limit of 0.068 U/L, and an excellent specificity for the discrimination of APN from biological ions, small molecules, and other proteases commonly found in living system. By virtue of good stability and cell viability, probe APN-CL imaged abnormal levels of APN in tumour cells and tumour-bearing mice. Moreover, this probe APN-CL could be easily used to evaluate APN inhibitors and APN levels in plasma samples from 20 patients. Overall, as a facile and cost-effective probe, APN-CL will be a promising alternative in the early diagnosis of pathologies and for cost-effective screening of inhibitors.

DNA-based logic gates promote the development of molecular computing and show enormous potential in the fields of nanotechnology and biotechnology. Dumbbell oligonucleotides (DNA) with poly-thymine (poly-T) loops and a nicked random double strand have been demonstrated to be an efficient template for the formation of fluorescent copper nanoclusters (CuNCs) in our previous work. Herein, a new platform technology is presented with which to construct molecular logic gates by employing CuNCs probe as a basic output generator, coupling of functional nucleases as the inputs. Two dumbbell DNAs are used with the difference in stem length (8 bp and 16 bp, respectively). The degradation of DNA templates can be tuned by various nucleic acid enzymes, single-stranded nuclease (S1), double-stranded specific nuclease (DSN), E. coli DNA ligase, exonucleases I and III. Briefly, S1 can digest both DNA templates, while the cleavage ability of DSN will be resistant by the short stem of SS-DNA (short-stem DNA). Exonuclease I and III can degrade these two nicked DNA templates, which are inhibited due to the ligation of E. coli DNA ligase. With this novel strategy, a set of logic gates is successfully constructed at the molecular level, including "YES", "PASS 0", "OR", "INHIBIT", which take the advantages of no label, easy operation, fast speed, high efficiency and low cost. Furthermore, S1 nuclease, as the biomarker of numerous carcinogens, is selectively detected in the range of 0.05-50 U/mL with the detection limit of 0.005 U/mL (1 x10(-6) U) based on this platform. (C) 2021 Published by Elsevier B.V. on behalf of Chinese Chemical Society and Institute of Materia Medica, Chinese Academy of Medical Sciences.

Fibroblast activation protein-alpha (FAPα) is a key modulator of the microenvironment in multiple pathologies and is becoming the next pan-cancer target for cancer diagnostics and therapeutics. Chemiluminescence (CL) luminophores are considered as one of the most sensitive families of probes for detection and imaging applications due to their high signal-to-noise ratio. Until now, however, no such effective CL probe was reported for FAPα detection. Herein, we developed a novel CL probe for the detection of endogenous FAPα activity by incorporating FAPα-specific dipeptide substrates (glycine-proline) to the improved Schaap's adamantylidene-dioxetane. In this manner, we designed three CL probes (CFCL, BFCL, and QFCL) with the dipeptide substrate blocked by N-terminal benzyloxycarbonyl, N-tert-butoxycarbonyl or N-quinoline-4-carboxylic acid, respectively, which was used as the masking group to restrain the chemiexcitation energy. Probe CFCL exhibited the optimal specificity for the discrimination of FAPα from dipeptidase IV and prolyl oligopeptidase, which was elucidated by molecular docking simulation. Upon FAPα cleavage, CFCL was turned on for the highly selective and sensitive detection of FAPα with a limit of detection of 0.785 ng/mL. Furthermore, the ability of CFCL to image FAPα was effectively demonstrated in vitro, including various biological samples (plasma and tissue preparations), and in living systems (tumor cells and tumor-bearing mice). Furthermore, this newly established probe could be easily extended to evaluate FAPα inhibitors. Overall, we anticipate that probe CFCL will offer a facile and cost-effective alternative in the early detection of pathologies, individual tailoring of drug therapy, and drug screening.

Quantification of biothiols in living systems is essential to understand their biological applications. Here, we developed two activatable chemiluminescence probes (SHCL and NCCL) and investigated their utility in the bioimaging of intracellular biothiols by directly tethering 2,4-dinitrobenzenesulfonyl to the hydroxyl group of phenoxy-dioxetane. The design of these two probes differed in substituents of phenol-dioxetane, i.e., SHCL contained the ortho chlorine, whereas NCCL had the para hydroxymethyl. Upon glutathione (GSH) cleavage, both probes emitted significantly "turn-on" chemiluminescent signals. However, the chemiluminescence intensity based on NCCL declined with increasing GSH level above 5 mM, while SHCL exhibited much higher chemiluminescent intensity and a wider concentration range (0.5 μM-50 mM), which was much more suitable for sensing endogenous biothiols. We further demonstrated that chlorine substitution in SHCL played an important role in bioimaging owing to the halogen effect, providing a lower pKa value and significant enhancement of the chemiluminescent emission. SHCL imaged the biothiols effectively in tumor cells and tumor-bearing mice. Additionally, this novel chemiluminescence probe can be easily used to evaluate the in vitro activity of acetylcholinesterase. Overall, we anticipate that SHCL may provide a facile and intuitive tool for studying the role of biothiols in diseases.

Aminoglycosides (AGs) are broad-spectrum antibiotics used in both human infection and animal medicine. The overuse of AGs causes undesirable residues in food, leading to serious health problems due to food chain accumulation. In recent years, various methods have been developed to determine AGs in food. Among these methods, fluorescent (FL), colorimetric and chemiluminescent (CL) optical methods possess advantages such as their simple instrumentation, low cost, simple operation, feasibility of realizing visualization, and smartphone imaging. This mini-review summarizes optical assays for the detection of AGs in food developed in recent years. The detection principles for different categories are discussed. Then, the amplification techniques for the ultrasensitive detection of AGs are introduced. We also discuss multiplex methods for the simultaneous detection of AGs. Finally, the challenges and future prospects are discussed in the Conclusions and Perspectives section.

药物分析学科是高等院校药学类专业的重要组成部分,为药品质量检验、药品监管以及制药企业的质量控制部门输送专业人员,一定程度上为人民用药安全提供了保障.针对目前药物分析教学中存在的一些问题,我们提出相应的解决措施,如增加多种教学模式、设计更为丰富的实验课以及提供学生接触该学科的多种方式,从而形成多元化、多层次的药物分析教学新体系,锻炼学生主动学习意识和独立思考能力,增强学生动手能力和提高综合素质.

Bacteria determination, emerging as a critical step in the understanding of the increasingly serious bacterial contaminations, remains a major challenge. Herein, a novel chemiluminescence biosensor was exploited for the ultra-sensitive determination of nuclease activity and bacteria. In which, hemin, the chemiluminescent (CL) tag molecule was encapsulated into ordered mesopores of mesoporous silica nanoparticles with a specific DNA gate. The capped DNA could be specifically switched upon exposure to the DNA nuclease or bacterial lysate and allowed for an increased release of the encapsulated hemin, which therefore resulted in an obviously enhanced CL signal for the luminol-H2O2 system. Attributed to this unique behavior with the linear/sigmoidal relationship between CL intensity and DNA nuclease/bacteria concentration, the as-prepared CL biosensor could detect S1 nuclease activity in the concentration of 0.01-10.0 U with the detection limit of 0.1 mU, and Escherichia coli O157:H7 (E. coli) or Stahylococcus aureus (S. aureus) concentration ranges of 101-109 CFU mL-1. The detection limit of E. coli and S. aureus was calculated to be 3.0 and 2.5 CFU mL-1 respectively, which was comparable or even better than that of previous works. Thus, this detection method could reach detectable levels without cell enrichment overnight. Moreover, the proposed biosensing system could be conducted in the homogeneous solution without separation and washing, greatly improving the reaction efficiency and simplifying the procedure. As expected, the novel CL biosensor promised a great potential for simple and convenient detection of nuclease and bacteria in fields such as food bacterial contamination, pharmaceuticals and clinical analysis.

A self-floating photothermal membrane with simultaneous mechanical stability and antibacterial activity is facilely prepared for efficient solar-driven interfacial water evaporation.

Objective To evaluate health literacy among parents of primary school students in Shanghai.Methods A total of 1 912 pupils from 4 primary schools in Shanghai urban area and 1 371 pupils from 2 primary schools in suburb area were selected by convenient sampling method.The Chinese Citizens Health Literacy Questionnaire was implemented among parents among all participants during September and October in 2015.Results Among all the parents surveyed,21.6％ of them possessed health literacy.Parents with higher education levels possessed higher health literacy.35％ parents with master education level possessed health literacy,and the figure was only 11％ among parents with "junior middle school or below".About 24.56％ local Shanghai and foreign national parents possessed higher health literacy,compared with parents whose children registered out of Shanghai(16.69％).In addition,only 0.27％ parents could identify all safety signs properly;only 7.58％ parents were aware of diseases infected through mosquitoes,flies,mice and cockroaches properly and 10.94％ parents were able to avoid sharing towels with others.Conclusion It's necessary to promote health literacy among parents with low education background and migrant workers.

A novel assay for histidine and cysteine has been constructed based on modulation of fluorescent copper nanoclusters (CuNCs) by molecular switches. In our previous work, a dumbbell DNA template with a poly-T (thymine) loop has been developed as an excellent template for the formation of strongly fluorescent CuNCs. Herein, for the first time, we established this biosensor for sensing two amino acids by using dumbbell DNA-templated CuNCs as the single probe. Among 20 natural amino acids, only histidine and cysteine can selectively quench fluorescence emission of CuNCs, because of the specific interaction of these compounds with copper ions. Furthermore, by using nickel ions (Ni2+) and N-ethylmaleimide as the masking agents for histidine and cysteine respectively, an integrated logic gate system was designed by coupling with the fluorescent CuNCs and demonstrated selective and sensitive detection of cysteine and histidine. Under optimal conditions, cysteine can be detected in the concentration ranges of 0.01–10.0 μM with the detection limit (DL) of as low as 98 pM, while histidine can be detected in the ranges of 0.05–40.0 μM with DL of 1.6 nM. In addition, histidine and cysteine can be observed with the naked eye under a hand-held UV lamp (DL, 50 nM), which can be easily adapted to automated high-throughput screening. Finally, the strategy has been successfully utilized for biological fluids. The proposed system can be conducted in homogeneous solution, eliminating the need for organic cosolvents, separation processes of nanomaterials, or any chemical modifications. Overall, the assay provides an alternative method for simultaneous detection of cysteine and histidine by taking the advantages of high speed, no label and enzyme requirement, and good sensitivity and specificity, and will satisfy the great demand for determination of amino acids in fields such as food processing, biochemistry, pharmaceuticals, and clinical analysis.

Disease-related biomarkers are objectively measurable molecular signatures of physiological status that can serve as disease indicators or drug targets in clinical diagnosis and therapy, thus acting as a tool in support of personalized medicine. For example, the prostate-specific antigen (PSA) biomarker is now widely used to screen patients for prostate cancer. However, few such biomarkers are currently available, and the process of biomarker identification and validation is prolonged and complicated by inefficient methods of discovery and few reliable analytical platforms. Therefore, in this Perspective, we look at the advanced chemistry of aptamer molecules and their significant role as molecular probes in biomarker studies. As a special class of functional nucleic acids evolved from an iterative technology termed Systematic Evolution of Ligands by Exponential Enrichment (SELEX), these single-stranded oligonucleotides can recognize their respective targets with selectivity and affinity comparable to those of protein antibodies. Because of their fast turnaround time and exceptional chemical properties, aptamer probes can serve as novel molecular tools for biomarker investigations, particularly in assisting identification of new disease-related biomarkers. More importantly, aptamers are able to recognize biomarkers from complex biological environments such as blood serum and cell surfaces, which can provide direct evidence for further clinical applications. This Perspective highlights several major advancements of aptamer-based biomarker discovery strategies and their potential contribution to the practice of precision medicine.