Journal of Food Measurement and Characterization
·
2024
·
IF 2.9
Meng Wenya,Xue Yuhan,Zhang Jing,Wang Ye,Hou Hu,Li Jing,Dong Ping
Fish sperm extracts have been shown to have various activities, but it has not been effectively developed and utilized at present due to the lack of effective methods for preparing edible grade products. This study proposed a method for preparing edible fish sperm DNA from Alaska pollock (PSD) and evaluated the hepatoprotective effect of the extracted DNA. Firstly, the preparation conditions of fish sperm DNA were optimized by response surface methodology. Under the ideal conditions that the ratio of solid to liquid was 3:1, the pH of extractant was 7.5, the purified solvent was saturated sodium carbonate solution and absolute ethanol was twice the volume of purified solvent, the yield of PSD was (13.657 ± 0.577) µg/mg. To evaluate the liver protective effects of the extracted DNA, the digestion products by three digestion enzymes (pepsin, DNase I and DNase II) compared using an alcohol-induced HepG2 cell model. The DNase I hydrolysis product was more effective in preventing cell alcohol damage, as shown by reducing the content of lipid peroxides. Transcriptomics and metabolomics data revealed that the DNase I hydrolysis product activated the Nrf2/HO-1 pathway through the PI3K/Akt signaling pathway and regulated abnormal lipid synthesis through the steroidogenic pathway. This study will provide an important basis for the evaluation and analysis of the hepatoprotective effect of fish sperm DNA in vivo. Overall, our findings will promote the comprehensive utilization of fish sperm and other aquatic wastes, and provide scientific guidance for the development of functional nucleic acid foods.
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高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2025
·
IF 3.8
Lai Shanshan,Guo Jiahong,Li Xin,Yu Xiaoxiao,Lai Guosong
Fluorescence biosensors hold significant importance for testing antibiotic residues which seriously endanger public health. However, how to adopt appropriate strategies to address the false result disadvantage involved in traditional single-channel biosensors is still a great challenge. Meanwhile, too much attention focused on designing signal amplification strategies of biosensors unavoidably decreases their detection efficiency. Herein, we combined the designed dual DNA recycling amplification strategy with CRISPR/Cas12a-mediated dual-channel signal output mode to successfully develop a novel ratiometric fluorescence biosensor for testing kanamycin (Kana) residues in complex sample matrices. The first recycling was formed from an exonuclease-assisted aptamer recognition reaction, which also triggered another cascade DNA recycling to amplify the release of the Cas12a activator. With the non-discrimination cleavage of Cas12a to cause reverse fluorescence changes of copper nanoclusters and an AMAC-labeled signal DNA, the ratiometric signal transduction strategy was constructed. Under optimal conditions, this biosensor could be applied for ultrasensitive testing of Kana antibiotics in a five-order of magnitude wide linear range with a low detection limit of 17.2 fg mL −1 . Benefiting from the self-correction function of the ratiometric signal transduction mode, it showed promising practicality in lake water and milk samples with the relative error less than 4.9% to the standard ELISA results. Besides CRISPR/Cas12a-based fluorescence output efficiency improvement, this biosensor also excluded the complicated manipulations and expensive instruments required in traditional methods. Therefore, it provides a good choice for expanding the application of fluorescence biosensing technology for practical analysis application. Graphical
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高影响因子
PNAS Nexus
·
2025
·Vol. 4
·期 5
·
IF 3.8
Sun Kai,Shi Zeyu,Dai Lingzhi,Si Youbin,Ma Junchao,Lin Hui,Yu Han-Qing
Endocrine-disrupting chemicals (EDCs), such as 17β-estradiol (E2) and bisphenol A (BPA), can induce DNA damage, leading to genomic instability and cell death. Laccase, an enzyme secreted by diverse organisms, plays a critical role in mitigating the cytotoxicity of these contaminants. Despite its importance, the dynamic evolution and interaction mechanisms of EDCs and DNA in laccase catalysis remain poorly understood. This study investigates the interactions between EDCs and DNA during laccase-induced polymerization transfer at a molecular level. As the DNA concentration was increased from 0 to 7.575 nM, the pseudo-first-order kinetic constants for E2 and BPA decreased by 2.03 and 2.10 times, respectively. DNA-bound EDCs disrupted the catalytic activity and stability of laccase, thereby delaying the polymerization transfer rate of EDCs. E2 and BPA bound to DNA base pairs via groove and intercalative modes, respectively. Laccase-induced polymerization reduced damage to the DNA helix and base stacking caused by EDC binding. Moreover, the resulting DNA–EDC-precipitated polymers, formed through continuous laccase polymerization, exhibited denser and more complex structures compared with spherical EDC-precipitated polymers, confirming DNA encapsulation and/or binding. This work underscores the intramolecular mechanisms of EDC interaction with DNA in vitro during the laccase-induced polymerization, offering efficient ways to mitigate the genotoxicity of EDCs.Endocrine-disrupting chemicals (EDCs) are often described as “a ticking time bomb threatening human survival” (1). Globally, the widespread presence of both natural and synthetic EDCs, such as 17β-estradiol (E2) and bisphenol A (BPA), has raised significant concerns about their detrimental effects on wildlife and human health (2, 3). Due to their persistent and bioaccumulative nature, EDCs represent a latent crisis for both environmental and human health. Moreover, these chemicals undergo transformations in the environment to form secondary pollutants, which are often more toxic than their precursors (4). These compounds, entering organisms directly or indirectly, have been linked to reproductive disorders, deformities in sexual organs, sex reversal, and feminization in wildlife species (5). Recent studies also correlate the exposure to EDCs with increasing rates of large-bowel, breast, cervical, and ovarian cancers in women and declining sperm count and motility in men (6, 7).The hazardous effects of EDCs are attributed to their interactions with DNA, disrupting DNA repair, replication, and transcription processes, and thereby causing enhanced DNA damage and cell death (8, 9). Understanding the dynamic processes and intermolecular interactions of EDCs with DNA is crucial for elucidating their mechanisms of genotoxicity.DNA, the carrier and transmitter of hereditary information in nearly all living systems, plays a central role in vital biogenetic processes, such as gene expression, transcription, mutation, and carcinogenesis (10). Composed of pentose sugar, phosphoric acid, and nitrogenous bases, DNA's genetic information is encoded by specific sequences of these bases. The stability of the DNA helix is significantly maintained by the stacking and hydrogen-bonding interactions between nucleobases (11). Thus, the interaction of EDCs with DNA could directly impact its structure and function, leading to DNA damage in exposed cells and altering its existing functions (1). Organic pollutants of biological importance are known to interact with DNA through noncovalent interactions, including electrostatic interactions, groove binding, and intercalative binding (12–14).Laccase (EC 1.10.3.2, oxygen oxidoreductase), a member of the multicopper oxidase superfamily, is naturally secreted by a variety of organisms such as bacteria, fungi, plants, and insects. It has garnered attention for its high activity, low energy requirements, and nontoxicity in degrading harmful phenolic substances (15, 16). Laccase catalyzes the oxidation of EDCs, facilitating the formation of phenoxy radicals, which subsequently polymerize into dimers, trimers, tetramers, oligomers, and polymers (17, 18). Despite its potential, the detailed understanding of the dynamic evolution and molecular mechanisms of interaction between EDCs and DNA during laccase-induced polymerization remains scant.This work aims to elucidate, at the molecular level, the dynamic processes and interaction mechanisms between EDCs and DNA during the laccase-induced polymerization transfer. We explored the reaction kinetics of EDCs under varying DNA concentrations using a pseudo-first-order kinetic model and utilized high-resolution mass spectrometry (HRMS) to identify key oxidation products. Advanced spectroscopic techniques, such as UV-Vis spectroscopy, attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), and circular dichroism (CD) spectroscopy, were employed to reveal the dynamic interactions of DNA with EDCs during polymerization. Atomic force microscopy (AFM) was used to unmask the dynamic variations in DNA morphology on interaction with EDCs and their oligomers from the molecular perspective. Further, the precipitated polymers were characterized using scanning electron microscopy (SEM) equipped with an energy-dispersive X-ray spectrometer (EDS), FTIR, and 13C-nuclear magnetic resonance spectroscopy (13C-NMR). Molecular dynamics simulation (MDS) was conducted to verify the intermolecular interactions between DNA and the oligomers of EDCs. The cytotoxicity of DNA–EDC solutions treated by laccase-induced polymerization was evaluated over time. By uncovering the dynamic evolution and intermolecular interactions of EDCs and DNA in vitro during the laccase-induced polymerization, this study offers insights into molecular strategies for mitigating the genotoxicity of EDCs.Increasing DNA concentrations significantly impeded the laccase-induced single-electron oxidation transfer of EDCs. Figure S1 illustrates that the oxidation rates of E2 and BPA decreased from 97.25 to 80.37% and 71.64 to 54.18%, respectively, after a 3-h reaction, as the DNA concentration was increased from 0 to 7.575 nM. The laccase-mediated oxidation of EDCs adhered to a pseudo-first-order kinetics model within 1 h, with k1 values for E2 and BPA decreasing by factors of 2.03 and 2.10, respectively, as the DNA concentration was increased (R2 = 0.96–0.99). This suggests that laccase activity was inversely related to the DNA concentration during the enzyme-catalyzed oxidation of EDCs (Fig. S2). Linear regression analysis further elucidated the dependency of the k1 value on low DNA concentration (0–0.7575 nM), revealing a linear relationship (R2 ≥ 0.98) and calculated pseudo-second-order rate coefficients (k2) for E2 and BPA of 1.52 and 0.33 nM−1·h−1, respectively (Fig. 1a and b).High levels of phenoxy radicals, produced by the laccase-induced oxidation of EDCs, facilitated the polymerization transfer. HRMS identified the polymerization products, revealing their molecular architectures through computational analysis (Table S1 and Fig. 1c). Notably, three major E2 oligomers—dimer (m/z = 541.3310), trimer (m/z = 811.4930), and tetramer (m/z = 1,081.6555)—suggested the pattern of n × M − 2(n − 1) for EDC polymerization products, where n is the number of polymerized EDC monomers and M is the molecular mass of the monomer. Additionally, a peak at m/z = 269.1548 indicated the formation of estrone (E1) by deprotonation, accounting for <2.4% of the initial E2 after 1 h of reaction. This pattern was consistent with the laccase-induced polymerization of BPA, suggesting that polymerization transfer is the predominant pathway in EDC oxidation by laccase, regardless of the presence of DNA.UV-Vis absorption spectroscopy confirmed the interaction modes between DNA and EDCs. The UV-Vis absorption spectra of DNA displayed two peaks at 234 and 260 nm (Fig. S3). The addition of E2 resulted in a hyperchromic effect at 260 nm, indicating damage to the DNA helix through a nonintercalative interaction mode. The degree of E2 polymerization transfer correlated negatively with the intensities of the characteristic DNA peaks (Fig. 2a). In contrast, the interaction with BPA showed a hypochromic effect, suggesting an intercalative binding that involves strong π–π aromatic stacking interactions between BPA and the phosphate backbone of the DNA helix (19). The degree of BPA polymerization transfer correlated positively with DNA peak intensities (Fig. 2b). Clearly, laccase-induced polymerization effectively reduced the concentration of EDCs and mitigated their interaction with DNA.The molecular fingerprint of DNA is located in the spectral region of 1,800–700 cm−1 due to nitrogenous bases, phosphate, and deoxyribose stretching vibrations. Binding of EDCs to DNA bases was investigated by analyzing spectral changes at 1,691, 1,648, 1,605, and 1,485 cm−1 that were attributed to guanine, thymine, adenine, and cytosine base vibrations, respectively. FTIR spectra explored the binding sites of DNA and EDCs, with characteristic DNA peaks indicating interactions at thymine and adenine bases, which appeared to be primary binding sites for EDCs (Fig. 2c and d). CD spectroscopy further revealed conformational changes in DNA induced by EDCs, highlighting the distinct interaction modes of E2 and BPA with DNA. The upward shift in the positive peak suggested enhanced base stacking due to groove binding by E2, while the downward shift in the negative peak for BPA indicated intercalative binding (Fig. 2e and f).AFM does not require any additional sample pretreatment to observe samples in liquids, making it exceptionally suitable for DNA imaging and therefore plays an important role in probing the intermolecular interactions of EDCs and DNA during the laccase-induced polymerization. AFM offered direct evidence of DNA binding with EDCs at the molecular level (Fig. 3). No obvious change in the contour length of DNA was observed in the presence of E2, confirming that E2 bound to DNA by groove mode (Fig. 3a). However, the interaction of intercalating BPA with DNA could lead to an increase in the contour length of DNA by bridging two or more DNA molecules (Fig. 3e). Further, the enhanced degree of EDC polymerization boosted DNA encapsulation and/or binding, forming larger spherical particles (Fig. 3b–d and f–h).SEM analysis showed that compared with the spherical E2-precipitated polymers, DNA–E2-precipitated polymers displayed a denser and thicker morphology, indicative of tighter and more complex structural integration (Fig. 4a and b). EDS confirmed a lower carbon-to-oxygen ratio in the DNA–E2-precipitated polymers compared with those without DNA, suggesting more extensive oxygen bonding. A similar phenomenon was also found in the BPA and DNA–BPA-precipitated polymers (Fig. 4e and f). As demonstrated in Fig. 4c and g, the EDC-precipitated polymers displayed the functional characteristics of the aromatic skeleton, phenolic –OH, and ether linkage. By comparison, the intensities and positions of the above functional groups in the DNA–EDC-precipitated polymers changed unmistakably. 13C-NMR spectra of the precipitated polymers revealed a predominance of aromatic and phenolic carbon, indicating that DNA was potentially encapsulated and/or bound to the EDC-precipitated polymers during the laccase-induced polymerization and thus facilitated more complex carbon backbone formations (Fig. 4d and h).The dynamics of polymerization and the molecular interactions were extensively analyzed to elucidate the complex behavior of EDCs in the presence of DNA during laccase catalysis. These insights provide a deeper understanding of the intramolecular mechanisms through which laccase-mediated processes may mitigate the genotoxicity of EDCs, highlighting the potential applications of laccase polymerization for the decontamination and detoxification of EDCs at the molecular level.To support the above experimental results, we further explored the stability and position changes of EDCs and their oligomers binding to DNA by computer-aided MDS. Root-mean-square deviation (rmsd) serves as a measure of the stability and interaction between DNA and substrates. Higher rmsd values suggest more unstable interactions. In this study, the interactions of DNA with EDCs and their oligomers (dimer, trimer, and tetramer) were examined over a 100-ns MDS period. As illustrated in Fig. 5a–e, rmsd values for DNA-bound E2 and its oligomers ranged from 0 to 7.0 Å. The average rmsd for DNA-bound E2 was 1.05 Å, indicating more stable interactions compared with DNA-bound E2 dimer (2.68 Å), trimer (1.73 Å), and tetramer (2.10 Å). Hydrogen bonds, crucial for the affinity and selectivity of the DNA-substrate complex, diminished in number from the monomer to the oligomers, underscoring the enhanced stability of DNA-bound E2. Conversely, for BPA, the mean rmsd values increased with oligomerization; DNA-bound BPA monomer exhibited an rmsd of 3.06 Å, greater than its oligomers—dimer (1.74 Å), trimer (2.45 Å), and tetramer (2.44 Å; Fig. 5f–j), correlating with an increase in hydrogen bonds with higher molecular weights, indicating greater stability in DNA-bound BPA oligomers.As mentioned above, hydroxyl substitution facilitated the binding of EDCs and their oligomers to DNA, but the intermolecular binding was relatively unstable (rmsd > 1). Figure 6 shows the position changes of EDCs and their dimers, trimers, and tetramers relative to DNA at different simulation times. As the MDS time was increased, EDCs and their oligomers gradually deviated from the initial binding sites of DNA. Furthermore, these results also disclosed abnormal DNA structures, which were consistent with the CD spectroscopy. Overall, the MDS results confirm that EDCs and their oligomers unstably bound to their corresponding binding sites during the simulation process.The dynamic interactions and mechanisms of DNA with EDCs during the laccase-induced polymerization remain poorly understood. Laccase facilitates the one-electron oxidation of EDCs, predominantly leading to polymerization products, while DNA hampers this transfer by interacting with functional groups of DNA molecules, such as base pairs and phosphate skeletons. This interaction forms complexes of DNA-bound EDCs, which obstruct laccase's catalytic activity and stability, thereby slowing the polymerization rate of EDCs. In contrast, the polymerization products of EDCs, also bound to DNA via hydrogen bonds, form denser and thicker structures compared with those without DNA, suggesting potential encapsulation and/or binding within the DNA–EDC polymers. Understanding these dynamic processes and interactions provides insights into mitigating the genotoxicity and cancer risks associated with EDCs.The EDC polymers precipitated through laccase-induced polymerization play a crucial role in reducing the cytotoxic effects of EDCs (20). The phytotoxicity assessments of DNA–EDC solutions treated through laccase-induced polymerization, illustrated in Fig. S4, demonstrate significant reductions in EDC-induced inhibitory effects on seed germination and root elongation. For instance, the germination index (GI) of wheat seeds treated with E2 increased from 12.85% (severe inhibition) to 87.48% (no inhibition) over a 9-h laccase polymerization period. The addition of DNA maintained a GI of 57.69%, even after 9 h. Furthermore, the cytotoxicity of polymerization products diminished over time, showcasing the potential of laccase-induced polymerization in alleviating the genotoxic effects of EDCs. Significantly, microbial enzymes may degrade DNA, resulting in the release of DNA-bound EDCs. Future studies are required to explore the stability of DNA-bound EDCs and their oligomers, as well as the long-term effects of the complexes in the environment. Furthermore, the in vivo interactions of DNA with EDCs should also be explored to uncover the self-detoxification mechanisms of laccase-producing organisms at a genomic level.In summary, this work provides comprehensive insights into the complex interactions between EDCs and DNA in the context of laccase-mediated reactions. The findings reveal that DNA significantly alters the kinetics and mechanisms of EDC polymerization, potentially offering a novel approach to mitigate the environmental impact and genotoxicity of these harmful compounds. By elucidating the specific binding interactions and the resulting structural changes in DNA and EDC polymers, this research contributes to a deeper understanding of the intramolecular mechanisms of EDC interaction with DNA in vitro during the laccase-induced polymerization and opens new avenues for the development of more effective decontamination and detoxification strategies in vivo.E2 (C18H24O2, ≥98%), BPA (C15H16O2, ≥99%), double-stranded salmon testes DNA (%G-C content: 41.2%; molecular mass: 1.3 × 106 Da; DNA fragment lengths: ∼2,000 bp), and Trametes versicolor laccase (light brown, ≥0.5 U·mg−1) were sourced from Sigma-Aldrich Co., China. Methanol (CH3OH, suitable for high-performance liquid chromatography [HPLC], ≥99.9%) and acetonitrile (CH3CN, suitable for HPLC, ≥99.9%) were procured from Thermo Fisher Scientific Inc., USA. Tris-HCl buffer solution (10 mM, pH 7), citrate-phosphate buffer solution (CPBS, 10 mM), and Murashige and Skoog (MS) medium were obtained from Aladdin Chemistry Co., China. Stock solutions of E2 and BPA were prepared in methanol at 1 mM. Laccase stock solution (100 U·mL−1) and DNA stock solution (dissolving 75.75 nmol of DNA in 100 mL Tris-HCl buffer) were prepared in Milli-Q water. UV absorption at 260 nm with a molar extinction coefficient (ε260) of 6,600 L·mol−1·cm−1 was used to determine the DNA concentration in the solution. The DNA purity, indicated by an absorbance ratio at 260 and 280 nm (A260/A280) of >1.83, confirmed high purity. Wheat seeds (Triticum aestivum L.) were supplied by the Anhui Academy of Agriculture Sciences, China. All solutions were prepared using Milli-Q water (18.25 MΩ·cm) and stored at 4 °C prior to use.The reaction kinetics of EDCs catalyzed by laccase in the presence of DNA were conducted in 20 mL of CPBS at 25 ± 0.5 °C and pH 5 under dark conditions. A concentration of 0.5 U·mL−1 laccase was added to solutions containing 5 μM of either E2 or BPA and varying concentrations of DNA (0–7.575 nM). Periodically, 0.5 mL samples were extracted from the reaction mixture, quenched immediately with an equivalent volume of methanol, and filtered through a 0.22-μm membrane for analysis. Experiments were replicated in triplicate, and the oxidation kinetics of EDCs were modeled by pseudo-first-order kinetics:where k1 is the pseudo-first-order rate constant and [EDCs] is the residual concentration of free EDCs in the reaction system at a given time.Details on the determination of laccase activity are provided in the Supporting Information.Quantitative and qualitative analyses, including HPLC and HRMS, are described in the Supporting Information.The interaction mode between DNA and EDCs during enzymatic catalysis was examined using a UV-Vis spectrophotometer (UV-2550, Shimadzu Co., Japan) equipped with 1 cm quartz cuvettes. Samples were analyzed at preset intervals (0, 5, 10, 15, 30, 45, 60, 90, 120, 150, and 180 min) for UV absorption spectrum (220–300 nm) in 1-nm increments at 25 ± 0.5 °C. ATR-FTIR spectroscopy (NEXUS-870, Thermo Nicolet Co., USA) and CD spectroscopy (Chirascan, APL, UK) were also employed to further explore the binding mechanism between DNA and EDCs.The morphological changes of DNA binding to EDCs and their oligomers were directly observed by AFM. DNA (7.575 nM) and EDCs (5 μM) were incubated at 25 ± 0.5 ° C and pH 5.0 to ensure that EDCs were adequately bound to DNA. Next, the enzymatic reaction was initiated by adding 0.5 U·mL−1 laccase to the mixture. By adjusting the pH of the mixture to 1.5, the enzymatic reaction was periodically terminated. To prepare the sample for AFM, 50 μL of the mixed solution was pipetted onto freshly cracked mica sheets, gently washed with ultrapure water and air-dried, and then imaged by AFM (Dimension Icon, Bruker Co., USA). AFM images were recorded at a resolution of 500 × 500 pixels and a scanning line speed of 0.600–0.998 Hz.Precipitated polymers collected from the laccase-induced polymerization transfer of EDCs were freeze-dried, ground, and sieved. Their microstructures, functional groups, and chemical compositions were characterized using SEM (Regulus 8100, Hitachi, Japan) equipped with an EDS (Bruker Co., Germany), FTIR, and 13C-NMR (Avance III, Bruker Co., Germany).MDS was performed to model the interaction of DNA with EDCs and their oligomers (dimer, trimer, and tetramer). The 3D structure of B-DNA dodecamer d(CGCGAATTCGCG)2 was sourced from the Protein Data Bank (PDB ID: 2B0 K). The structures of EDCs and their oligomers were built using ChemBioDraw Ultra 14.0, optimized with ChemBio3D Ultra 14.0, with nonpolar hydrogen atoms merged and Kollman charges added using MGLTools 1.5.6. Simulations were conducted using GROMACS 2016.4 with the AMBER99SB force field. Rmsd was calculated to assess the stability of each complex:where Natm is the number of atoms, rA is the position of atom A, and rAref is the reference position of atom A.The phytotoxicity of DNA and EDC solutions treated by laccase-induced polymerization was evaluated using a GI. Wheat seeds were monitored over a 3-day cultivation period in MS medium under controlled conditions. The GI was calculated using the formula:where NCRS and NMW are the number of seeds germinated in the reaction solution and Milli-Q water, respectively, and RCRS and RMW are the root lengths in the reaction solution and Milli-Q water, respectively.Supplementary material is available at PNAS Nexus online.This work was financially supported by the National Natural Science Foundation of China (42277019 and 51821006) and the Natural Science Foundation-Excellent Youth Fund Project of Anhui Province, China (2308085Y28).K.S., Z.S., and J.M. were involved in data curation and formal analysis. L.D. was involved in formal analysis. Y.S. was involved in software, validation, and writing—review and editing. H.L. was involved in software and validation. H.-Q.Y. was involved in conceptualization, resources, supervision, funding acquisition, writing—original draft, project administration, and writing—review and editing.All data are included in the article and Supporting Information.
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高影响因子
Alexandria Engineering Journal
·
2024
·
IF 6.2
Jia Wang,Juelan Ye,Gang Li
Human papillomavirus (HPV) is the primary cause of cervical cancer, a major global health concern affecting women worldwide. Early detection of high-risk HPV genotypes is crucial for timely intervention and reducing disease burden. However, current detection methods often lack sensitivity, speed, or cost-effectiveness, especially in resource-limited settings. This work addresses this critical need by developing an ultrasensitive electrochemical biosensing platform based on nitrogen-doped graphene paper electrodes for quantifying high-risk HPV16 DNA sequences. The N-graphene nanohybrid, fabricated via simple solvothermal treatment of biomass-derived graphene oxide, demonstrated significantly enhanced electrical and electrocatalytic properties compared to undoped graphene. DNA detection was achieved through characteristic oxidation signals of guanine bases, which displayed a wide linear dynamic range (0.5–100 μg/mL), low detection limit (75 pg/mL), and excellent reproducibility (relative standard deviation <3.5 %). Detailed spectroscopic and electrochemical analyses revealed the key roles of pyridinic nitrogen defects in improving interfacial charge transfer kinetics and mitigating biofouling issues. This synergistic combination of high electrical conductivity and versatile surface functionality paves the way for equipment-free, point-of-care genosensor devices tailored for quantitative biomarker screening in resource-constrained environments.
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高影响因子
TOXICOLOGY AND APPLIED PHARMACOLOGY
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2025
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IF 3.4
Yan Jiang,Jiayu Zhou,Shusen Fang,Yufei Liu,Yueting Shao,Yiguo Jiang
Smoking, a pivotal environmental risk factor, drives diseases including lung cancer through genetic and epigenetic alterations. While 2-Naphthylamine (2-NA), a tobacco-derived carcinogen, is established as a bladder carcinogen via DNA damage, its role in lung carcinogenesis remains mechanistically uncharacterized despite epidemiological associations. This study identifies bronchial epithelium as a direct target of 2-NA, demonstrating dose-dependent DNA damage in 16HBE cells (peak at 250 μM, 12 h; P < 0.05), accompanied by S-phase arrest, apoptosis, reduced proliferation, and reactive oxygen species (ROS) generation. 2-NA exposure upregulated the expression of circSHOC1, which is a circular RNA derived from the SHOC1 gene, in a dose-dependent manner. Functional assays revealed that circSHOC1 overexpression exacerbated 2-NA-induced DNA damage by enhancing ROS production and 8-hydroxy-2′-deoxyguanosine (8-OHdG) accumulation, whereas knockdown attenuated these effects. Mechanistically, circSHOC1 interacted with mitochondrial protein SLC25A3 (Solute Carrier Family 25 Member 3), a key oxidative stress regulator; SLC25A3 knockdown mitigated DNA damage ( P < 0.01), and co-transfection experiments confirmed SLC25A3 as a critical mediator of circSHOC1-driven genotoxicity. Collectively, this work provides the first experimental evidence that 2-NA induces bronchial epithelial DNA damage via a circSHOC1-SLC25A3-ROS axis, supporting a novel mechanism for tobacco-associated lung carcinogenesis and highlighting 2-NA as a potential pulmonary carcinogen.
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高影响因子
INTERNATIONAL IMMUNOPHARMACOLOGY
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2025
·
IF 4.7
Liyun Jie,He Huang,LuYing Sha,Lidan Zhang,Ye Wang,Yu Cao
Cutaneous squamous cell carcinoma (cSCC) is a common type of skin tumor with high incidence and significant disease burden. Current clinical treatments face challenges such as high recurrence rates and drug resistance, making the discovery of novel drugs and therapeutic targets critically important. Previous studies have confirmed that Mangiferin(MF) can inhibit the proliferation of various tumors, promote apoptosis, and regulate cellular oxidative stress. This study investigated the therapeutic effects and mechanisms of MF in cutaneous squamous cell carcinoma (cSCC). CCK-8 and EDU assays demonstrated that MF significantly inhibited A431 cell proliferation, induced apoptosis, and caused G0/G1 phase arrest. Network pharmacology, molecular docking, and molecular dynamics simulations identified HSP90α as a direct target of MF, which was highly expressed in clinical cSCC tissues and validated through CETSA and DARTS experiments. Genetic manipulation experiments revealed that HSP90α knockdown promoted apoptosis, while its overexpression accelerated proliferation and attenuated MF's effects. Mechanistically, MF induced ROS generation, leading to mitochondrial and DNA damage, which could be reversed by the antioxidant NAC. Furthermore, ROS-mediated activation of the MAPK pathway contributed to apoptosis and DNA damage. In vivo, MF effectively suppressed cSCC growth without causing organ toxicity. In conclusion, MF inhibits cSCC progression both in vitro and in vivo with minimal toxicity, primarily through targeting HSP90α and activating the ROS/MAPK pathway.
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高影响因子
Insects
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2025
·Vol. 16
·期 11
·1147
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IF 2.9
Feiyang Liang,Wei Zeng,Sheng Li,Xingyue Liu
Bark lice are common in terrestrial ecosystems, yet the poor taxonomic study of them renders species identification difficult. In this study, we investigated the species identification of the bark louse genus Neostenopsocus using DNA barcodes (675 bp fragment of the mitochondrial COI gene). We examined newly collected specimens and the type materials of some valid species of Neostenopsocus , redescribing 13 valid species and proposing 39 new synonyms. A total of 110 sequences of Neostenopsocus species were obtained from newly collected specimens and public data. The results show that the COI barcode is sufficient to distinguish most Neostenopsocus species, whereas more molecular marks and distribution data are necessary to identify the closely similar species. The family Stenopsocidae exhibits a rich biodiversity in China, yet species identification based on morphological characters remains problematic due to insufficient study and absent molecular data. DNA barcoding has been widely employed for rapid species identification in insects, particularly through standardized COI gene sequencing. In this study, we conducted species delimitation for Neostenopsocus in China based on the morphological characters and DNA barcodes, comprising 20 morphological species with 110 barcodes. We redescribed 13 species from China and proposed 39 new synonyms. This study supports the applicability of DNA barcoding for species identification in Neostenopsocus . The K2P distance between the morphological species of Neostenopsocus ranges from 0.0051 to 0.2040, with most exceeding 0.10. The maximum intraspecific divergence reaches 0.0778 in N. nepalensis . The external morphology of Neostenopsocus shows considerable structural uniformity, and the genitalia provide limited diagnostic value for species identification. Instead, the marking patterns of the head and forewing are considered to be useful for species identification. Therefore, delimiting closely related species demands an integrated taxonomy approach that combines extensive geographical sampling, detailed morphological analysis, and nuclear DNA data.
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高影响因子
BIOMATERIALS
·
2025
·
IF 12.9
Bing Ye,Huiling Lei,Xirui Jing,Qiuyue Ding,Sheng Yao,Hong Wang,Chunqing Meng,Xiaodong Guo,Bin Wu,Yuzhou Wu,Tingfang Sun
Vascularization-ossification coupling is pivotal for bone defect repair. Neovascularization guides the migration of osteogenic precursor cells, and osteoblasts in the perivascular area support vascular regeneration through the secretion of cytokines. Delivery systems are being engineered to orchestrate the coupling of vascularization and ossification precisely. However, most delivery systems lack sensitivity to the inherent repair environment, hampering timely bone formation. This study developed a vascular environment-responsive biomaterial: a true bone ceramic/small intestine submucosa (TBC/SIS) scaffold loaded with nanoswitch and Apt19s-functionalized nanoparticles. This scaffold is designed to release the functionalized nanoparticles via an "unfolding–refolding" nanoswitch based on base pairing and protein-nucleic acid recognition. The TBC/SIS scaffold initiates neovascularization, prompting endothelial cells to secrete nucleolin. The nanoswitches detect nucleolin and trigger the release of nanoparticles, thereby creating a responsive vascular environment. Moreover, when modified with aptamer-19s, the nanoparticles can attract mesenchymal stem cells to perivascular areas, facilitating spatial coupling with blood vessels. Encapsulating miRNA-26a in nanoparticles enhances osteogenic activity and stimulates the secretion of vascular endothelial growth factor. As vascular endothelial cells infiltrate, nanoparticles are continuously released from the scaffold, establishing a positive feedback loop that promotes vascularization-ossification coupling and ultimately enhances bone regeneration. Our findings provide valuable insights for designing intelligent materials that utilize endogenous stimuli to enhance spatiotemporal interactions between vascularization and ossification. Further in-depth studies on the expression of nucleolin in vivo will continue to refine this promising strategy.
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高影响因子
BIOMEDICINE & PHARMACOTHERAPY
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2021
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IF 6.53
Qi Liu,Xiaoli Wang,Jun Li,Jiaojiao Wang,Guohui Sun,Na Zhang,Ting Ren,Lijiao Zhao,Rugang Zhong
Chloroethylnitrosoureas (CENUs) are an important family of chemotherapies in clinical treatment of cancers, which exert antitumor activity by inducing the formation of DNA interstrand crosslinks (dG-dC ICLs). However, the drug resistance mediated by O 6 -alkylguanine-DNA alkyltransferase (AGT) and absence of tumor-targeting ability largely decrease the antitumor efficacy of CENUs. In this study, we synthesized an azobenzene-based hypoxia-activated combi-nitrosourea prodrug, AzoBGNU, and evaluated its hypoxic selectivity and antitumor activity. The prodrug was composed of a CENU pharmacophore and an O 6 -benzylguanine ( O 6 -BG) analog moiety masked by a N , N -dimethyl-4-(phenyldiazenyl)aniline segment as a hypoxia-activated trigger, which was designed to be selectively reduced via azo bond break in hypoxic tumor microenvironment, accompanied with releasing of an O 6 -BG analog to inhibit AGT and a chloroethylating agent to induce dG-dC ICLs. AzoBGNU exhibited significantly increased cytotoxicity and apoptosis-inducing ability toward DU145 cells under hypoxia compared with normoxia, indicating the hypoxia-responsiveness as expected. Predominant higher cytotoxicity was observed in the cells treated by AzoBGNU than those by traditional CENU chemotherapy ACNU and its combination with O 6 -BG. The levels of dG-dC ICLs in DU145 cells induced by AzoBGNU was remarkably enhanced under hypoxia, which was approximately 6-fold higher than those in the AzoBGNU-treated groups under normoxia and those in the ACNU-treated groups. The results demonstrated that azobenzene-based combi-nitrosourea prodrug possessed desirable tumor-hypoxia targeting ability and eliminated chemoresistance compared with the conventional CENUs.
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高影响因子
BIOORGANIC CHEMISTRY
·
2020
·
IF 4.831
Qiong Wu,Yue Song,Ruotong Liu,Rui Wang,Wenjie Mei,Weiming Chen,Huanglan Yang,Xicheng Wang
Phenanthroimidazole derivatives containing phenanthroline and imidazole heterocyclic aromatic rings are effective agents to inhibit tumor cell growth. Herein, halogen element-modified imidazo[4,5 f ][1,10]phenanthroline derivatives 1–6 ( 1 , 4-fluorophenyl; 2 , 4-chlorophenyl; 3 , 4-bromobenyl; 4 , 2,3-dichlorophenyl; 5 , 3,4-dichlorophenyl; and 6 , 2,4-dichlorophenyl) were synthesized, and their antitumor activities were investigated. All of the compounds, especially 4 , exhibited an excellent inhibitory effect against nasopharyngeal carcinoma CNE-1 cells. This effect was better than that of doxorubicin. Compound 4 also markedly blocked the proliferation of the CNE-1 cells in a zebrafish xenograft model. The antitumor mechanisms might be attributed to apoptosis induction, which triggered ROS-mediated DNA damage and generated mitochondrial dysfunction by stabilizing c-myc G-quadruplex DNA structure. Results indicated that phenanthroimidazole derivatives could act as promising anticancer agents.
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高影响因子
Nanoscale
·
2018
·Vol. 10
·期 14
·6611-6619
·
IF 7.233
Jinyong Hu,Meiyan Wu,Li Jiang,Zhensheng Zhong,Zhangkai Zhou,Thitima Rujiralai,Jie Ma
The association of a plasmonic nano-antenna with single-molecule FRET technique presents new prospects to investigate the dynamics of biological molecules. However, the presence of a plasmonic nano-antenna significantly modifies the FRET rate and efficiency; this makes its applicability to the prevalent single-molecule FRET experiments unclear. Herein, using gold nanoparticle antennas of different sizes and DNA hairpins labelled with FRET pairs (Cy3 and Cy5) as the model system, we performed experiments to study the folding dynamics of single DNA hairpins at various salt concentrations. Our results indicate that gold nanoparticle antennas can enhance single-molecule fluorescence of Cy3 and Cy5 up to 3–5 folds, substantially reduce the FRET efficiency, and alter the obtained FRET efficiency histograms. However, the folding dynamics of DNA hairpins remains unaffected, and the correct kinetic and dynamic information can still be extracted from the seriously modified FRET efficiencies. Therefore, our experiments demonstrate the feasibility and compatibility for applying plasmonic nano-antennas to the mostly used single-molecule FRET assays, which provide a broad range of possibilities for the future applications of these nano-antennas in studying various essential biological processes.
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高影响因子
ANALYTICA CHIMICA ACTA
·
2021
·
IF 6.558
Bingzhi Li,Peng Zhang,Bin Zhou,Siying Xie,Anqi Xia,Tiying Suo,Shuang Feng,Xing Zhang
Flap endonuclease 1 (FEN1), a ubiquitous enzyme involved in DNA repair and replication, is overexpressed in highly proliferative cancer cells. FEN1 has been recognized as a promising diagnostic marker of cancers; however, very few analytical techniques have been developed for the convenient detection of FEN1. To realize the simplified quantification of FEN1, we developed a FEN1-responsive fluorescent nanoprobe based on DNA-silver nanoclusters (DNA-AgNCs). The nanoprobe was rationally designed with a double-flapped dumbbell conformation, where its 5′ flap was produced with DNA-AgNCs, and the 3′ flap was elongated by a guanine-rich enhancer sequence (GRS). Rigidified by the DNA scaffold, DNA-AgNCs and the GRS are in close proximity, resulting in high fluorescence because of the GRS-induced activation of DNA-AgNCs. Upon the addition of FEN1, the 5’ flap of the nanoprobe is cleaved due to the structure-specific endonuclease activity of FEN1. This cleavage released the DNA-AgNCs from the nanoprobe, broke the proximity between DNA-AgNCs and the GRS, and caused decreased fluorescence. This nanoprobe can be applied in the sensitive detection of FEN1 with a detection limit of 40 fM, and it showed high specificity for the monitoring of FEN1 in clinical samples. As the first attempt to develop biosensors targeting FEN1 based on DNA-AgNCs, this work provided a potent platform for monitoring FEN1 and screening FEN1 inhibitors.
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高影响因子
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