Secreted phospholipase PLA2G2E contributes to regulation of T cell immune response against influenza virus infection.

The involvement of secreted phospholipase A2s in respiratory diseases, such as asthma and respiratory viral infections, is well-established. However, the specific role of secreted phospholipase A2 group IIE (PLA2G2E) during influenza virus infection remains unexplored. Here, we investigated the role of PLA2G2E during H1N1 influenza virus infection using a targeted mouse model lacking Pla2g2e gene (Pla2g2e-/-). Our findings demonstrated that Pla2g2e-/- mice had significantly lower survival rates and higher viral loads in lungs compared to wild-type mice following influenza virus infection. While Pla2g2e-/- mice displayed comparable innate and humoral immune responses to influenza virus challenge, the animals showed impaired influenza-specific cellular immunity and reduced T cell-mediated cytotoxicity. This indicates that PLA2G2E is involved in regulating specific T cell responses during influenza virus infection. Furthermore, transgenic mice expressing the human PLA2G2E gene exhibited resistance to influenza virus infection along with enhanced influenza-specific cellular immunity and T cell-mediated cytotoxicity. Pla2g2e deficiency resulted in perturbation of lipid mediators in the lung and T cells, potentially contributing to its impact on the anti-influenza immune response. Taken together, these findings suggest that targeting PLA2G2E could hold potential as a therapeutic strategy for managing influenza virus infections.IMPORTANCEThe influenza virus is a highly transmissible respiratory pathogen that continues to pose a significant public health concern. It effectively evades humoral immune protection conferred by vaccines and natural infection due to its continuous viral evolution through the genetic processes of antigenic drift and shift. Recognition of conserved non-mutable viral epitopes by T cells may provide broad immunity against influenza virus. In this study, we have demonstrated that phospholipase A2 group IIE (PLA2G2E) plays a crucial role in protecting against influenza virus infection through the regulation of T cell responses, while not affecting innate and humoral immune responses. Targeting PLA2G2E could therefore represent a potential therapeutic strategy for managing influenza virus infection.

Quantification of safety improvements and human-machine tradeoffs in the transition to automated driving.

The assumption of reduced human error-related crashes with increasing levels of automation in pursuing Level 5 automation lacks empirical evidence. As automation levels rise, human error-induced safety hazards are anticipated to decrease, while machine error-induced hazards will increase. However, a quantitative index capturing this tradeoff is absent. Additionally, theoretical modeling of safety improvements during the transition to automated driving remains unexplored, particularly concerning reducing human error-related hazards. These limitations impede the understanding of safety from human and machine perspectives for Automated Vehicle (AV) specialists and manufacturers. This research addresses these gaps by investigating safety performance associations between human and machine factors using the "Human-Machine conflict reduction ratio" (H/M ratio), a novel metric. The study aims to establish safety improvements related to human errors under various automation levels. Sixty participants completed driving tasks on a driving simulator at Levels 0, 4, 3, and 2. Safety performance measures, including conflict frequency and severity, were computed. As a result, Level 4 exhibits the largest decrease (93.3%) compared to manual driving, followed by Level 2 (70.7%) and Level 3 (40.5%). The H/M ratio measures the tradeoff between reducing human and machine error-induced hazards, with Level 2 demonstrating the highest ratio, followed by Levels 4 and 3. Safety performance is evaluated by considering all possible types of human errors at each automation level. Theoretical models from a human factor's perspective are employed to estimate safety improvements at each level. This research contributes to a comprehensive understanding of safety in the "human-machine cooperative driving" phase, offering insights to AV industry practitioners and stakeholders.

Evidence of automated vehicle safety's influence on people's acceptance of the automated driving technology.

Existing studies identified targeted audiences showing increases in Automated Vehicles (AV) acceptance after experiencing automated driving. However, there is still uncertainty regarding the reasons. Although some studies cited safety as the primary reason, there is no objective evidence from safety performance in verifying its impact on AV acceptance. This study contributes to the literature by quantitatively revealing why AV acceptance is changed after experiencing automated driving via a Structural Equation Modeling (SEM) method and objectively validating that safety is the primary factor in determining AV acceptance. Sixty drivers completed driving tasks on a driving simulator under Levels 0, 4, 3, and 2 and survey questions in between. As a result, the safety-related perceptions of AV were identified as reasons for affecting AV acceptance. Particularly, the evaluation of traffic conflicts and conflict severity validates the results from SEM, proving that safety is the primary and significant reason for influencing AV acceptance.

Determination, Quality, and Health Assessment of Pesticide Residues in Kumquat in China.

Pesticide residues in kumquat fruits from China, and the quality and chronic/acute intake risks in Chinese consumers, were assessed using the QuEChERS procedure and UHPLC-MS/MS and GC-MS/MS methods. Our 5-year monitoring and survey showed 90% of the 573 samples of kumquat fruits collected from two main production areas contained one or multiple residual pesticides. Overall, 30 pesticides were detected, including 16 insecticides, 7 fungicides, 5 acaricides, and 2 plant growth modulators, of which 2 pesticides were already banned. Two or more residual pesticides were discovered in 81% of the samples, and pesticide residues in 9.4% of the samples surpassed the MRLs, such as profenofos, bifenthrin, triazophos, avermectin, spirodiclofen, difenoconazole, and methidathion. The major risk factors on the safety of kumquat fruits before 2019 were profenofos, bifenthrin, and triazophos, but their over-standard frequencies significantly declined after 2019, which was credited to the stricter supervision and management policies by local governments. Despite the high detection rates and multi-residue occurrence of pesticides in kumquat fruits, about 81% of the samples were assessed as qualified. Moreover, the accumulative chronic diet risk determined from ADI is very low. To better protect the health of customers, we shall formulate stricter organic phosphorus pesticide control measures and stricter use guidelines, especially for methidathion, triazophos, chlorpyrifos, and profenofos. This study provides potential data for the design of kumquat fruit quality and safety control guidelines and for the reduction in health risks to humans.

Dissipation and dietary exposure risk assessment of spinosad, thiocyclam, and its metabolite nereistoxin in cucumber and groundwater for different population groups.

A QuEChERS (quick, easy, cheap, effective, rugged, and safe) technique using ultrahigh-performance liquid chromatography with tandem mass spectrometry for the analysis of spinosad (spinosyn A + spinosyn D), thiocyclam, and nereistoxin in cucumber was developed with mean recoveries of 93-104%, relative standard deviations of ≤9%, and limits of quantification of 0.01 mg/kg. Field trials of spinosad and thiocyclam were performed in 12 representative cultivating areas in China. Field trial results indicate that spinosyn A and spinosyn D easily dissipated in cucumber with half-lives of 2.48-6.24 and <3 days, respectively. Nereistoxin was produced after thiocyclam application and was more persistent than its parent. The terminal residues of spinosad were all below the maximum residue limits (0.2 mg/kg) in China, whereas the terminal concentration of nereistoxin (calculated as the stoichiometric equivalent of thiocyclam), which was much higher than that of thiocyclam, was far beyond the maximum residue limits of thiocyclam in cucumber (0.01 mg/kg) established by the European Union. The predicted no-effect concentrations of spinosyn A, spinosyn D, thiocyclam, and nereistoxin leaching into groundwater were estimated using China-PEARL (Pesticide Emission Assessment at Regional and Local scales) models after application. However, the dietary (food and water) exposure risk quotient for different populations was below 1 with a preharvest interval set at 5 days after the last application, indicating that the application of spinosad and thiocyclam in cucumber was unlikely to pose unacceptable risk for human health. This study provides data for the safe use of spinosad and thiocyclam in cucumber ecosystem.

Ascorbic acid-mediated in situ growth of gold nanostars for photothermal immunoassay of ochratoxin A.

Currently, the development of efficient mycotoxins detection methods, particularly using portable devices as readout devices, remains a great challenge. Herein, a photothermal enzyme-linked immunosorbent assay (ELISA) based on gold nanostars (AuNSs) for the detection of ochratoxin A (OTA) using a "thermometer" was proposed for the first time. AuNSs with photothermal conversion capacity were parepared using an ascorbic acid (AA)-mediated in situ growth methd. Quantification was based on the alkaline phosphatase catalyzing the dephosphorylation of ascorbic acid 2-phosphoate to AA, thereby converting OTA concentration to the amount of in situ synthesized AuNSs, thus achieving straightforward readout by temperature. Benefiting from the classical tyramine signal amplification strategy, a detection limit of 0.39 ng mL-1 was obtained. The recoveries of grape juice and maize samples spiked with 10 ng mL-1 and 30 ng mL-1 OTA ranged from 86.53% to 116.9%. Our method has great potential in on-site OTA detection for food safety.

Influenza Virus Carrying a Codon-Reprogrammed Neuraminidase Gene as a Strategy for Live Attenuated Vaccine.

Live attenuated influenza vaccines offer broader and longer-lasting protection in comparison to inactivated influenza vaccines. The neuraminidase (NA) surface glycoprotein of influenza A virus is essential for the release and spread of progeny viral particles from infected cells. In this study, we de novo synthesized the NA gene, in which 62% of codons were synonymously changed based on mammalian codon bias usage. The codon-reprogrammed NA (repNA) gene failed to be packaged into the viral genome, which was achievable with partial restoration of wild-type NA sequence nucleotides at the 3' and 5' termini. Among a series of rescued recombinant viruses, we selected 20/13repNA, which contained 20 and 13 nucleotides of wild-type NA at the 3' and 5' termini of repNA, respectively, and evaluated its potential as a live attenuated influenza vaccine. The 20/13repNA is highly attenuated in mice, and the calculated LD50 was about 10,000-fold higher than that of the wild-type (WT) virus. Intranasal inoculation of the 20/13repNA virus in mice induced viral-specific humoral, cell-mediated, and mucosal immune responses. Mice vaccinated with the 20/13repNA virus were protected from the lethal challenge of both homologous and heterologous viruses. This strategy may provide a new method for the development of live, attenuated influenza vaccines for a better and more rapid response to influenza threats.

Synthesis of Chainlike ZSM-5 with a Polyelectrolyte as a Second Template for Oleic Acid and Ethanol Cracking into Light Olefins.

Chainlike ZSM-5 was synthesized in a tetrapropylammonium hydroxide (TPAOH) and poly(diallydimethylammonium chloride) (PDDA) dual-template system. The synthesis parameters and formation mechanism of chainlike zeolites were investigated. The optimized composition of the synthesis mixture was as follows: the PDDA/SiO2, TPAOH/SiO2, SiO2/Al2O3, and H2O/SiO2 molar ratios are, respectively, 0.16, 0.4, 50, and 40, with tetraethyl orthosilicate and aluminum nitrate as silicon/aluminum sources. The resultant ZSM-5 showed a cross-linked chainlike morphology, mesopore-dominated pore structure, and mild acidity. The formation of the chainlike zeolite was attributed to synergistic actions between PDDA and TPAOH. TPAOH acted as an alkali source and helped to induce nucleation and control the crystal size. PDDA acted as a soft template to promote crystal nucleation, and a hard template to form a three-dimensional mesoporous structure. Light olefin (C2-4 =) selectivities from cracking of ethanol and oleic acid over the present chainlike ZSM-5 at 400 °C reached 90 and 75.7%, respectively, which were much higher than those from commercial ZSM-5 (75 and 52.3%, respectively), demonstrating the excellent hydrothermal stability and catalytic performance of the synthesized chainlike zeolite.

Preparation of sodium silicate/red mud-based ZSM-5 with glucose as a second template for catalytic cracking of waste plastics into useful chemicals.

ZSM-5 was economically synthesized with red mud (RM) and industrial sodium silicate (ISS) in a tetrapropylammonium bromide (TPABr)-glucose dual-template system. The roles of glucose, Fe and Ca in RM on the formation of ZSM-5 were investigated. The catalytic performances of the resultant ZSM-5 were tested by cracking waste plastics. It was found that the formation of ZSM-5 was attributed to a synergistic effect between TPABr and glucose. The addition of glucose decreased the pH value in the crystallization solution and thus promoted the crystallization effect. Glucose acted as a hard template to generate mesopores. Fe atoms were partly distributed in the framework and partly adsorbed in the pores of ZSM-5, and helped to generate more Lewis acid sites. Ca atoms were mainly adsorbed in the pores of ZSM-5, and showed an inhibitory effect on the formation of zeolites. The synthesized ZSM-5 showed a weakly acidic and mesoporous structure and achieved an enhanced effect on producing gaseous products (gas yield: 85.3%), especially light olefins (C[double bond, length as m-dash] 2-4) (selectivity: 77.1%) from cracking of low density polyethylene at 500 °C. The long-term cracking experiment showed that the synthesized ZSM-5 is superior in converting waste plastics to light olefins (ethylene and propene) than the commercial ZSM-5.

CO2-elevated cell-free protein synthesis.

Gases are the vital nutrition of all organisms as the precursor of metabolism pathways. As a potential biological process, protein synthesis is inevitably regulated by gas transport and utilization. However, the effect of carbon dioxide (CO2) present in many metabolic pathways on protein synthesis has not been studied well. In this work, carbon dioxide combined with oxygen was employed for cell-free protein synthesis (CFPS) in the tube-in-tube reactor with precise control of gas concentration. In this in vitro system, gases could directly affect the protein synthesis process without transmembrane transport. Varied concentrations of carbon dioxide (0-1%) and constant oxygen concentration (21%) were employed for CFPS to assess the effects. The cell-free reactions with 0.3% CO2 and 21% O2 showed the highest protein yields. The combined effect of CO2 and O2 also resulted in relatively high protein expression under high oxygen conditions (0.3% CO2 and 100% O2). Moreover, metabolomics assays were performed to gain insight into metabolic changes, which showed that CO2 slightly improved energy metabolism and redox balance. In particular, the extra supplied CO2 activated the decarboxylating reactions and removed toxic metabolites to recover the protein synthesis activity. The exploration of CO2 on protein synthesis could provide guiding implications for basic studies and biomanufacturing.

CRISPR-Cas12a based fluorescence assay for organophosphorus pesticides in agricultural products.

Herein, we propose a sensitive fluorescent assay for organophosphorus pesticides (OPs) detection based on a novel strategy of activating the CRISPR-Cas12a system. Specifically, acetylcholinesterase (AChE) hydrolyzes acetylthiocholine into thiocholine (TCh). Subsequently, TCh induces the degradation of MnO2 nanosheets and generates sufficient Mn2+ ions to activate the Mn2+-dependent DNAzyme. Then, as the catalytic product of activated DNAzyme, the short DNA strand activates the CRISPR-Cas12a system to cleave the fluorophore-quencher-labeled DNA reporter (FQ) probe effectively; thus, increasing the fluorescence intensity (FI) in the solution. However, in the presence of OPs, the activity of AChE is suppressed, resulting in a decrease in FI. Under optimized conditions, the limits of detection for paraoxon, dichlorvos, and demeton were 270, 406, and 218 pg/mL, respectively. Benefiting from the outstanding MnO2 nanosheets properties and three rounds of enzymatic signal amplification, the proposed fluorescence assay holds great potential for the detection of OPs in agricultural products.

Potential of PKM2 as a drug target in mouse models with type 1 diabetes mellitus.

BACKGROUND: This study aimed to determine the effect of PKM2 knockout in STZ induced type 1 diabetes mellitus (T1D) mouse models and to explore the possible mechanism. METHOD: PKM2fl/fl C57BL/6 mouse was backcrossed with Ins-1cre C57BL/6 mouse to generate ß-cell-specific PKM2 knockout mouse after tamoxifen administration. The expression level of PKM2 in pancreas tissues was detected by quantitative reverse-transcription polymerase chain reaction and western blot analysis. The blood glucose levels in STZ induced T1D mouse models were measured to validate the establishment of T1D models. The pathological changes of T1D mouse were examined by hematoxylin and eosin. The oxidative stress (OS) and inflammatory response in T1D mouse were determined by measuring the expression levels of malondialdehyde, superoxide dismutase, and 8-OHdG in pancreatic tissues and the serum levels of interleukin-6 and tumor necrosis factor-α. The ability to catabolize glucose was assessed through intraperitoneal glucose tolerance test and insulin tolerance test. RESULTS: ß-cell-specific PKM2 knockout was successfully achieved in PKM2fl/flcre+ mouse. T1D mouse with PKM2 knockdown had decreased blood glucose level and suppressed cell apoptosis. PKM2 knockout in T1D mouse attenuated ß cell injury. OS and inflammatory response in T1D mouse with PKM2 knockout were also suppressed compared with T1D mouse without PKM2 knockout. CONCLUSION: PKM2 knockout in T1D mouse can attenuate OS and inflammatory response as well as decrease blood glucose level, suggesting the potential of PKM2 as a drug target for T1D treatment.

Enantioselective determination of phenthoate enantiomers in plant-origin matrices using reversed-phase high-performance liquid chromatography-tandem mass spectrometry.

Phenthoate is a chiral organophosphate pesticide with a pair of enantiomers which differ in toxicity, behavior and insecticidal activity, and its acute toxicity on human health owing to the inhibition of acetylcholinesterase highlights the need for enantioselective detection of enantiomers. Therefore, this study aimed to establish a simple rapid method for separation and detection of phenthoate enantiomers in fruits, vegetables and grains. The enantiomers were separated using reversed-phase high-performance liquid chromatography-tandem mass spectrometry for the first time. Rapid chiral separation (within 9 min) of the target compound was achieved on a chiral OJ-RH column with the mobile phase of methanol-water = 85:15(v/v), at a flow rate of 1 ml/min and a column temperature of 30°C. Acetonitrile and graphitized carbon black were used as the extractant and sorbent for pretreatment, respectively. This method provides excellent linearity (correlation coefficient ≥0.9986), high sensitivity (limit of quantification 5 µg/kg and limit of detection <0.25 µg/kg), satisfactory mean recoveries (76.2-91.0%) and relative standard deviation (intra-day RSDs ranged from 2.0 to 7.9% and inter-day RSDs ranged from 2.4 to 8.4%). In addition, a field trial to explore the stereoselective degradation of phenthoate enantiomers in citrus showed that (-)-phenthoate degraded faster than its antipode, resulting in the relative accumulation of (+)-phenthoate.

Supramolecular protein assembly in cell-free protein synthesis system.

Protein-based biomaterials have the characteristics of stability and biocompatibility. Based on these advantages, various bionic materials have been manufactured and used in different fields. However, current protein-based biomaterials generally need to form monomers in cells and be purified before being assembled in vitro. The preparation process takes a long time, and the complex cellular environment is challenging to be optimized for producing the target protein product. Here this study proposed technology for in situ synthesis and assembly of the target protein, namely the cell-free protein synthesis (CFPS), which allowed to shorten the synthesis time and increase the flexibility of adding or removing natural or synthetic components. In this study, successful expression and self-assembly of the dihedral symmetric proteins proved the applicability of the CFPS system for biomaterials production. Furthermore, the fusion of different functional proteins to these six scaffold proteins could form active polymers in the CFPS system. Given the flexibility, CFPS is expected to become a powerful tool as the prototyping and manufacturing technology for protein-based biomaterials in the future.

Determination of five pesticides in kumquat: Dissipation behaviors, residues and their health risk assessment under field conditions.

The present study was carried out to profile the dissipation patterns and residues of five pesticides (triazophos, profenofos, chlorpyrifos, etoxazole and bifenthrin) on kumquat using QuEChERS method coupled with HPLC-MS/MS. The corresponding dietary health risks were also estimated. In the method validation, satisfactory results of good linearity (r2 ≥ 0.9956), sensitivity (limits of quantification ≤0.01 mg/kg), recoveries (71.0-95.7%) with relative standard deviations (0.70-9.4%) were obtained. The half-lives of the five pesticides in kumquat were 13.6-38.5 d under field conditions according to first-order kinetics. Based on the final residue experiment, dietary exposure risks of profenofos, chlorpyrifos, etoxazole and bifenthrin were all acceptably low, with RQc and RQa values of 0.00199-0.122 and 0.00145-0.200, respectively. However, exposure intake of triazophos posed unacceptable acute and chronic health risks for Chinese residents, especially for children with RQa and RQc up to 4.25 and 2.19. Forbidden use suggestion of triazophos and recommended MRLs of profenofos and bifenthrin were put forward in kumquat for safe production and consumption. This work was significant in providing guidance on appropriate application and MRL establishment of pesticides in kumquat.

Strategy of Extra Zr Doping on the Enhancement of Thermoelectric Performance for TiZrxNiSn Synthesized by a Modified Solid-State Reaction.

Half-Heusler alloys, which possess the advantages of high thermal stability, a large power factor, and good mechanical property, have been attracting increasing interest in mid-temperature thermoelectric applications. In this work, extra Zr-doped TiZrxNiSn samples were successfully prepared by a modified solid-state reaction followed by spark plasma sintering. It demonstrates that extra Zr doping could not only improve the power factor on account of an increase in the Seebeck coefficient but also suppress the lattice thermal conductivity originated from the strengthened phonon scattering by the superlattice nanodomains and the secondary nanoparticles. As a consequence, an increased power factor of 3.29 mW m-1 K-2 and a decreased lattice thermal conductivity of 1.74 W m-1 K-1 are achieved in TiZr0.015NiSn, leading to a peak ZT as high as 0.88 at 773 K and an average ZT value up to 0.62 in the temperature range of 373-773 K. This work gives guidance for optimizing the thermoelectric performance of TiNiSn-based alloys by modulating the microstructures on the secondary nanophases and superlattice nanodomains.

Comparative Studies of the Structural and Transport Properties of Molten Salt FLiNaK Using the Machine-Learned Neural Network and Reparametrized Classical Forcefields.

Despite surging interest in molten salt reactors and thermal storage systems, knowledge of the physicochemical properties of molten salts are still inadequate due to demanding experiments that require high temperature, impurity control, and corrosion mitigation. Therefore, the ability to predict these properties for molten salts from first-principles computations is urgently needed. Herein, we developed and compared a machine-learned neural network force field (NNFF) and a reparametrized rigid ion model (RIM) for a prototypical molten salt LiF-NaF-KF (FLiNaK). We found that NNFF was able to reproduce both the structural and transport properties of the molten salt with first-principles accuracy and classical-MD computational efficiency. Furthermore, the correlation between the local atomic structures and the dynamics was identified by comparing with RIMs, suggesting the significance of polarization of anions implicitly embedded in the NNFF. This work demonstrated a computational framework that can facilitate the screening of molten salts with different chemical compositions, impurities, and additives, and at different thermodynamic conditions suitable for the next-generation nuclear reactors and thermal energy storage facilities.

Dissipation of imidacloprid and its metabolites in Chinese prickly ash (Zanthoxylum) and their dietary risk assessment.

Dissipation of imidacloprid (IMI) and its metabolites (urea, olefin, 5-hydroxy, guanidine, 6-chloronicotinic acid) in Chinese prickly ash (CPA) was investigated using QuEChERS combined with UPLC-MS/MS. Good linearity (r2 ≥0.9963), accuracy (recoveries of 71.8-104.3%), precision (relative standard deviations of 0.9-9.4%), and sensitivity (limit of quantification ≤0.05 mg kg-1) were obtained. After application of IMI at dosage of 467 mg a.i. L-1 for three times with interval of 7 d, the dissipation dynamics of IMI in CPA followed first-order kinetics, with half-life of 6.48-7.29 d. IMI was the main compound in CPA, followed by urea and guanidine with small amounts of olefin, 5-hydroxy, and 6-chloronicotinic acid. The terminal residues of total IMI and its metabolites at PHI of 14-21 d were 0.16-7.80 mg kg-1 in fresh CPA and 0.41-10.44 mg kg-1 in dried CPA, with the median processing factor of 3.62. Risk assessment showed the acute (RQa) and chronic dietary risk quotients (RQc) of IMI in CPA were 0.020-0.083% and 0.052-0.334%, respectively. Based on the dietary structures of different genders and ages of Chinese people, the whole dietary risk assessment indicated that RQc was less than 100% for the general population except for 2- to 7-year-old children (RQc of 109.9%), implying the long-term risks of IMI were acceptable to common consumers except for children.

A sensitive and practical ELISA for analyzing naringenin in pummelo and herb samples.

Naringenin, a flavonoid compound found in pummelo, is a key biological active compound in some traditional Chinese medicines, including Citri reticulatae pericarpium, Citri reticulatae pericarpium viride, Aurantii fructus immaturus, and Aurantii fructus. These Chinese medicinal preparations are the peels or immature fruits of certain citrus species. Aiming at detecting naringenin in complex matrices such as pummelo and traditional Chinese medicines, we put forward a sensitive and practical indirect competitive enzyme-linked immunosorbent assay (icELISA) based on anti-naringenin monoclonal antibodies (anti-Nar-mAbs). The median inhibitory concentration (IC50) was 4.43 ng/mL, and the working range was 1.15-15.81 ng/mL. The findings of the icELISA for the analysis of naringenin in pummelo and herb samples had a good correlation with the ultra performance liquid chromatography (UPLC) methodology and showed good accuracy and reproducibility. These data demonstrated that the developed icELISA is reliable, accurate, and suitable for detecting naringenin in pummelo and traditional Chinese medicines.