Tau class glutathione S-transferases (GSTs) have been identified as the main degradation route for thiram in fruits. However, the degradation rates of thiram under natural conditions are insufficient to guarantee its absence upon delivery to consumers. Here, the application of 2 mM kojic acid (KA), a non-toxic metabolite widely utilized in cosmetic industry, on thiram-containing papaya fruit enhanced H2O2 content by 58.0 % following treatment. This effect resulted in an antioxidant response over the long term (1, 3, and 5 days after treatment). Catalase and superoxide dismutase activities increased by 341.9 % and 201.3 %, respectively, on day 3 post-treatment. Tau class GSTs were upregulated, including the GST identified by GenBank number XP_021896923.1, which exhibited a 14.1-fold increase after treatment with 2 mM KA. Thiram degradation in papaya peel reached 89.7 % after 5 days without KA. However, applying 1 and 2 mM KA led to degradation rates of 98.4 % and 98.9 %, respectively. KA exhibited effects similar to those of 0.1 % v/v H2O2. The application of KA with iron(II) had no apparent effects on H2O2 content and peroxidase activity, indicating that KA may activate the antioxidant response by lowering free iron(II) content in papaya, which in turn leads to a reduction in peroxidase activity. On day 5 post-application, 2 mM KA enhanced the degradation of azoxystrobin, chlorothalonil, difenoconazole, fluxapyroxad, pyraclostrobin, thiabendazole, tebuconazole, trifloxystrobin, and triflumizole by 95.2 %, 61.3 %, 40.9 %, 68.5 %, 59.3 %, 41.2 %, 59.9 %, 47.7 %, and 67.0 %, respectively, in a GST-dependent manner. These findings may have significant implications for managing toxic residues in postharvest products.

Protein-protein interactions (PPIs) monitoring is critical to reveal cellular fundamental mechanism and facilitate drugs discovery. Nevertheless, the facial and robust protein 53 (p53)-murine double minute2 (MDM2) interaction detection methods remain challenging. Herein, we developed a PPIs detection platform based on CRISPR-Cas12a sensing system for the first time for p53-MDM2 interaction monitoring and inhibitors potency evaluation, designated Cas-PPIor (Cas12a-mediated PPIs detector). Inspired by the conformational-dependent consensus DNA binding ability of p53, a double stranded DNA (dsDNA) probe was designed elaborately through incorporating Cas12a activation DNA sequence into the specific p53 DNA-binding sequence. Thus, the PPI event can then be transduced into the collateral cleavage activity of Cas12a. As both the Cas12a/CRISPR RNA (crRNA) complex and p53 can bind to the same dsDNA probe in a competitive manner, the activated Cas12a mediated fluorescent readout scaled negatively with the p53 binding. Accordingly, we monitored the p53-MDM2 interaction process as well as its inhibition by a small-molecule antagonist. Based on the sequence specific binding of wild p53, our designed Cas-PPIor achieved high accuracy for differentiation of wild-type p53 cells from p53 mutation or deletion cells. Through discriminating p53 levels from cellular matrix treated with enantiomers of Nutlin-3, the assay also demonstrated reliable capability to evaluate drugs antiproliferative potency. By integrating the DNA binding ability of proteins with intrinsic flexible programmability and outstanding sensitivity of Cas12a, the proposed Cas-PPIor methodology hold great potential to enable accurate and facial PPIs monitoring and inhibitors potency evaluations.

Protein-protein interaction s (PPIs) monitoring is critical to reveal cellular fundamental mechanism and facilitate drugs discovery. Nevertheless, the facial and robust protein 53 (p53)-murine double minute2 (MDM2) interaction detection methods remain challenging. Herein, we developed a PPIs detection platform based on CRISPR-Cas12a sensing system for the first time for p53-MDM2 interaction monitoring and inhibitors potency evaluation, designated Cas-PPIor ( Cas 12a-mediated PPI s detect or ). Inspired by the conformational-dependent consensus DNA binding ability of p53, a double stranded DNA (dsDNA) probe was designed elaborately through incorporating Cas12a activation domain into the specific p53 DNA-binding sequence. Thus, the PPI event can then be transduced into the collateral cleavage activity of Cas12a. As both the Cas12a/single-guide RNA (sgRNA) complex and p53 can bind to the same dsDNA probe in a competitive manner, the activated Cas12a mediated fluorescent readout scaled negatively with the p53 binding. Accordingly, we monitored the p53-MDM2 interaction process as well as its inhibition by a small-molecule antagonist. Based on the sequence specific binding of wild p53, our designed Cas-PPIor achieved high accuracy for differentiation of wild-type p53 cells from p53 mutation or deletion cells. Through discriminating p53 levels from cellular matrix treated with enantiomers of Nutlin-3, the assay also demonstrated reliable capability to evaluate drugs antiproliferative potency. By integrating the DNA binding ability of proteins with intrinsic flexible programmability and outstanding sensitivity of Cas12a, the proposed Cas-PPIor methodology hold great potential to enable accurate and facial PPIs monitoring and inhibitors potency evaluations.

On-site monitoring of trace organic pollutants with facile methods is critical to environmental pollutant prevention and control. Herein, we proposed a CRISPR-Cas12a-based aptasensor platform (named as MC-LR-Casor) for on-site and sensitive detection of microcystin-LR (MC-LR). After hybridization with blocker DNA, the MC-LR aptamers were conjugated to magnetic beads (MBs) to get the MB aptasensor. In the presence of MC-LR, their interactions with aptamers were triggered and the specific binding caused the release of blocker DNA. Using the programmability of the CRISPR-Cas system, the released blocker DNA was designed to activate a Cas12a-crRNA complex. Single strand DNA reporters were rapidly cleaved by the complex. Signal readout could be achieved by fluorometer or lateral flow strips, which were positively correlated to MC-LR concentration. Benefiting from the CRISPR-Cas12a amplification system, the proposed sensing platform exhibited high sensitivity and reached the limit of detection of ∼3 × 10-6 μg/L (fluorescence method) or 1 × 10-3 μg/L (lateral flow assay). In addition, the MC-LR-Casor showed excellent selectivity and good recovery rates, demonstrating their good applicability for real water sample analysis. During the whole assay, only two steps of incubation at a constant temperature were required and the results could be visualized when employing flow strips. Therefore, the proposed assay offered a simple and convenient alternative for in situ MC-LR monitoring, which may hold great promise for future environmental surveillance.

中华文明上下五千年的历史进程中,无数清官名臣的光辉事迹被载入史册,其中蕴含着尊老孝亲、治国安邦、作风俭廉、忧国忧民、敢于担当等丰富内涵和精神特质的人格力量也随之流芳千古、熠熠生辉.当前我国正处于推进国家治理体系和治理能力现代化建设的关键时期,应深刻总结古代清官人格力量的形成机制,探寻其向现代转换的实现路径,建构新时代的政德文化:弘扬古代清官"修身正己"的人格力量,提高领导干部自持力;弘扬古代清官"勤政恤民"的人格力量,增强领导干部为民意识;弘扬古代清官"为政俭廉"的人格力量,引导领导干部廉洁从政.

文章首先以"在绵延中直觉主体的生存自由"为切入点,对柏格森的"绵延、物象、直觉、生命冲动和自由"等概念进行了逻辑上的展开与把握;其次,文章阐述了"柏格森生命哲学的中国式流变";最后,文章以柏格森的生命哲学为基础,以冯契先生实践着的"转识成智"为方法,发挥主体的"原始冲力",从而促成"个体性自由与开放社会的生成".