Search Results (488)

Background/Objectives: Letermovir is effective for cytomegalovirus (CMV) prophylaxis after kidney transplantation, but cytochrome P450 3A inhibition may alter tacrolimus and everolimus exposure. We evaluated changes in trough concentrations and dose requirements during letermovir initiation and discontinuation in de novo kidney transplant recipients. Methods: This single-center retrospective observational study included 20 living-donor kidney transplant recipients with D+/R− or R+ CMV serostatus receiving everolimus-based reduced-tacrolimus immunosuppression and letermovir prophylaxis. Trough concentrations, daily doses, kidney allograft function, CMV outcomes, and safety were assessed descriptively. Results: After letermovir initiation, mean tacrolimus trough concentrations increased by approximately 40–60% and everolimus trough concentrations by approximately 35–50%, requiring dose reductions of approximately 45% and 30–40%, respectively. By day 7 after discontinuation, mean tacrolimus and everolimus trough concentrations decreased to 57.4% and 65.9% of baseline, respectively, while daily doses increased to 136.6% and 132.5%, respectively. Group mean serum creatinine and eGFR remained within approximately 10% of baseline. No CMV DNAemia or disease, leukopenia, neutropenia, acute T cell-mediated rejection, or antibody-mediated rejection occurred during letermovir prophylaxis; gastrointestinal symptoms occurred in three recipients. Conclusions: Letermovir was associated with rapid, reversible changes in tacrolimus and everolimus trough concentrations and dose requirements. Intensive therapeutic drug monitoring is warranted at both initiation and discontinuation. Full article

The taxonomy of Opsariichthys Bleeker, 1863 in southern China is complicated by conflicting morphological and molecular evidence, particularly the status of O. minutus Nichols, 1926, long treated as a junior synonym of O. bidens Günther, 1873 until its revalidation. To resolve this, we adopted an integrative approach combining D-loop sequencing to anchor specimens to the O. minutus lineage (supplementing scarce GenBank data with vouchers bearing undulating jaw margins), genus-wide cytochrome b (cyt b) phylogenetics to avoid circular reasoning, and detailed comparative morphology. D-loop analyses revealed that all specimens with undulating jaw margins formed two clades: Fujian Province populations clustered with O. bidens, while Guangxi Zhuang Autonomous Region populations formed a distinct monophyletic lineage. Cyt b phylogenetics confirmed Fujian populations shared <2.0% uncorrected p-distances with O. bidens and showed extensive morphological overlap, reaffirming O. minutus as a junior synonym of O. bidens. In contrast, Guangxi populations from the Xijiang River basin (middle and upper Pearl River system, including the Hongshuihe, Guijiang and Liujiang Rivers) formed a distinct monophyletic lineage with 7.4–15.9% uncorrected p-distances from congeners, diagnosed by 49–55 lateral-line scales, two pharyngeal tooth rows, and prominent lower-jaw tubercles arranged in 1–2 rows in breeding males. This lineage is described as a new species of Opsariichthys, with its type locality in the Hongshuihe River. Collectively, our findings not only resolve a long-standing taxonomic puzzle but also underscore the efficacy of integrative taxonomy in uncovering cryptic diversity within the highly biodiverse yet understudied Pearl River basin. Full article

Hydrogen peroxide (H₂O₂) contributes to oxidative stress in Alzheimer’s disease (AD), yet conventional antioxidants that directly scavenge H₂O₂ have shown limited therapeutic efficacy. Here, we investigated K34 (3,3″-dihydroxy-6′-desmethyl terphenyllin), a catechol-bearing para-terphenyl isolated from the marine fungus Penicillium janthinellum 168CLC-17.1, as an indirect H₂O₂-decomposing agent. Complementary cell-free assays showed that K34 promoted peroxidase-dependent H₂O₂ decomposition, including Hb-supported H₂O₂ decomposition, while molecular docking predicted a plausible binding pose near the Hbβ heme pocket. In amyloid-β₄₂ (Aβ₄₂)-stimulated primary astrocytes, K34 reduced intracellular H₂O₂. In APPswe/PSEN1dE9 (APP/PS1) mice subjected to focal GFAP-inducible diphtheria toxin receptor (fGiD), systemic K34 attenuated memory impairment, preserved NeuN-positive neurons, reduced GFAP immunoreactivity, and improved hippocampal spike probability. K34 also exhibited low passive blood–brain barrier (BBB) permeability and limited inhibition of five major cytochrome P450 (CYP) isoforms under the tested conditions. These findings extend Hb-supported H₂O₂ decomposition to a structurally distinct marine natural product and support further evaluation of K34 as an antioxidant scaffold for modulating oxidative stress-associated neurodegenerative phenotypes. Full article

Oxidative stress is a central and well-established driver of cardiovascular disease, contributing to mitochondrial dysfunction, endothelial injury, inflammatory activation, and progressive myocardial and vascular remodeling. Although the major endogenous sources of cardiovascular reactive oxygen species (ROS), including NADPH oxidases, mitochondrial electron transport chain leakage, and uncoupled nitric oxide synthase, are well characterized, an additional and underappreciated contributor has recently emerged: cytochrome P450 2D6 (CYP2D6), an enzyme classically regarded as a hepatic drug-metabolizing protein. Accumulating evidence indicates that CYP2D6 is expressed extrahepatically in cardiac, vascular, and neural tissue, where uncoupled catalytic cycling is proposed to generate ROS independently of its canonical xenobiotic-metabolizing role, although direct experimental evidence for this pathway in human cardiac and vascular tissue remains limited. CYP2D6-derived oxidative processes may interact with mitochondrial respiratory function, endothelial nitric oxide bioavailability, and redox-sensitive inflammatory pathways, potentially contributing to cardiovascular vulnerability under specific genetic or pathological conditions. Critically, the magnitude of this oxidative contribution is not fixed: it is dynamically shaped by inherited CYP2D6 genetic variation, with poor and ultra-rapid metabolizer phenotypes exhibiting divergent oxidative burden and pharmacokinetic vulnerability, and is further amplified by polypharmacy, multimorbidity, and inflammation-driven phenoconversion, whereby clinically expressed CYP2D6 activity diverges from inherited genotype in ways that intensify redox imbalance. These dynamics are particularly relevant in East Asian populations, where the decreased-function CYP2D6*10 allele is highly prevalent. In this narrative, hypothesis-generating review, we integrate evidence from pharmacogenomics, redox biology, and cardiovascular pharmacology to propose a conceptual framework that reframes CYP2D6 as a genetically and pharmacologically tunable node within cardiovascular redox biology. We further examine emerging redox biomarkers, multi-omics platforms, and AI-assisted modeling as translational strategies for capturing this dynamic oxidative risk in real time. This framework supports a shift from static genotype-guided prescribing toward oxidative-risk-informed, adaptive cardiovascular precision medicine. Importantly, our focus on CYP2D6 should not be interpreted as evidence that it is a major cardiovascular CYP isozyme or an established driver of cardiovascular pathology. Rather, CYP2D6 is examined here as a deliberately hypothesis-generating candidate whose unusually strong pharmacogenetic variability, clinically important cardiovascular drug substrates, dynamic susceptibility to phenoconversion, extrahepatic expression, and mechanistically plausible links to endogenous substrate metabolism and CYP-associated ROS generation provide a convergent rationale for focused investigation. The mechanistic framework proposed in this review has not yet been experimentally and prospectively validated and should not be applied directly to clinical decision-making without supporting clinical data. Full article

The aryl hydrocarbon receptor (AHR) and the vitamin D receptor (VDR) were long regarded as independent transcription factors governing distinct physiology—xenobiotic sensing and calcium–vitamin D homeostasis, respectively. AHR, a basic helix–loop–helix/PAS protein, heterodimerizes with ARNT and binds xenobiotic response elements (XREs) to drive cytochrome P450 genes such as CYP1A1; VDR, a nuclear receptor activated by 1,25-dihydroxyvitamin D₃, heterodimerizes with RXR and binds vitamin D response elements (VDREs). Although their genes reside on separate chromosomes (AHR, Chr 7; VDR, Chr 12), an integrated view recognizes the two pathways as extensively cross-regulatory. This review synthesizes the molecular, immunological, and tissue-level evidence for VDR–AHR interplay. At the molecular level, the receptors cooperate at composite promoter architectures—most notably an everted-repeat VDRE positioned adjacent to an XRE in the CYP1A1 promoter—while AHR ligands reciprocally enhance CYP24A1-mediated catabolism of active vitamin D. Tryptophan metabolism provides a bidirectional hub: kynurenine and the UVB photoproduct FICZ serve as endogenous AHR ligands whose balance, modulated by VDR, shapes signaling output. The tumor suppressor p53 functions as a shared upstream regulator coupling genotoxic stress to both receptors, with convergence on the CDKN1A (p21) checkpoint. Functionally, AHR and VDR converge on the regulatory T cell (Treg)/Th17 axis to influence immune tolerance: sustained AHR activation by TCDD favors Foxp3⁺ Treg differentiation, transient FICZ-driven activation promotes Th17 responses, and VDR reinforces the tolerogenic arm while independently repressing IL-17. The receptors further cooperate in maintaining intestinal epithelial barrier integrity and NF-κB restraint, with parallel impairment in inflammatory bowel disease, and are co-activated in skin by solar UVB, which simultaneously generates vitamin D₃ and the AHR ligand FICZ within keratinocytes. In cancer, VDR acts as a tumor suppressor, AHR exhibits context-dependent pro- and anti-tumor roles, and a three-way AHR–VDR–p53 interaction—inverted by mutant p53—forms a critical regulatory node. Throughout, the direction and magnitude of cross-talk prove highly dependent on cell type, ligand identity and kinetics, and species—distinctions often underappreciated in the literature. Clarifying these context-specific determinants is essential for translating AHR–VDR cross-regulation into rational therapies in autoimmunity, mucosal inflammation, dermatology, and oncology. Full article

Objective: Saline–alkali stress (composed of neutral and alkaline salts) severely constrains soybean productivity. Although nicotinamide can enhance plant salt tolerance, its most effective concentration and molecular basis in soybean remain unclear. This study aimed to systematically investigate the regulatory effect of exogenous nicotinamide on soybean saline–alkaline tolerance and to identify its candidate genes and putative mechanisms under salt-stressed conditions. Methods: Soybean cultivar ‘Hefeng 25’ seedlings were exposed to 200 mmol/L mixed salt stress and treated with nicotinamide at 0 (CK), 10, 50, and 200 mg/L. Growth parameters, ROS levels, ion contents, photosynthetic pigments, osmoregulatory substances, and antioxidant enzyme activities were measured. Transcriptome sequencing and RT-qPCR were performed to identify differentially expressed genes (DEGs), followed by enrichment analysis. Results: Exogenous nicotinamide significantly improved soybean saline–alkaline tolerance relative to the salt-stressed CK group, with 50 mg/L (SA2) showing the most pronounced physiological improvements among all tested concentrations. SA2 treatment enhanced plant height, biomass, K+ content, and antioxidant enzyme activities while reducing Na+ accumulation and ROS levels compared with CK. Transcriptome analysis identified PM1 as the shared gene across the five non-most effective comparisons. Enrichment analysis implicated DEGs primarily in ko03110 (chaperones and folding catalysts), ko00199 (cytochrome P450), and ko04141 (protein processing in the endoplasmic reticulum). RT-qPCR confirmed that PM1, P450 genes (CYP74A1, CYP83D1), HSPs (HSP23, HSP18), and ROS scavengers (NAC1, GSTU43) were upregulated under non-most effective doses (SA1/SA3) but downregulated to levels comparable to the stressed CK in SA2; conversely, SA2 specifically restored the expression levels of stress-repressed WRKY57 and LBD15. Transcriptomically, SA2 exhibited a profile most similar to the saline–alkali-stressed CK group, with the fewest number of differentially expressed genes among all treatments, whereas non-optimal nicotinamide concentrations were associated with extensive transcriptional changes in ER protein processing, detoxification-related, and oxidative stress-related pathways. The limited transcriptional changes in SA2 indicate greater transcriptional similarity to the stressed control, which paralleled its superior physiological performance. Conclusions: Nicotinamide effectively alleviates soybean saline–alkaline stress at 50 mg/L, with the most significant physiological benefits and the fewest transcriptional changes relative to the stressed control. The non-most effective nicotinamide concentrations show extensive stress-related transcriptional reprogramming in protein folding, detoxification, and stress-response pathways relative to the salt-stressed control, putatively implicating ER stress, detoxification, and oxidative stress-related responses at the transcript level, with PM1 acting as a potential candidate transcriptional marker of dose-dependent transcriptional regulation under saline–alkali stress. Full article

Plants are continuously exposed to volatile organic compounds (VOCs) emitted by neighbors. Although the mechanisms governing VOC uptake and metabolism remain unclear, cytochrome P450 monooxygenases (CYP450s) are thought to participate in the detoxification and metabolic conversion of absorbed VOCs. Data from previously conducted cDNA microarray transcriptomic profiling in potato exposed to French marigold essential oil (FM-EO) was here used to filter differentially expressed sequences, and identified 54 unique CYP450 transcripts. Among the 10 most highly expressed sequences, two CYP81D1-like (81D1-1 and 81D1-2) and one CYP81D11-like (81D11-1) transcripts were found. RT-qPCR confirmed their strong induction within 8 h of volatile exposure. Comprehensive bioinformatics identified the most highly induced 81D1-1 gene as a CYP450 containing a predicted N-terminal hydrophobic signal or membrane-anchor region, the conserved heme-binding signature motif, and regulatory elements associated with oxidative stress responses. The other 81D11-1 gene, exhibiting a comparable expression level, was annotated only as a heme-binding protein but possessed seven distinct cis-regulatory elements, suggesting high transcriptional plasticity. Machine learning predictions assigned the highest interaction probability to (Z)-β-ocimene, whereas structure-based docking yielded the most favorable mean score for piperitone. This study provides the first characterization of the potato CYP450 superfamily in the context of volatile-mediated plant–plant interactions and identifies two CYP81D members as strong candidates for the oxidative metabolism of absorbed VOCs. The results support a proposed detoxification pathway in which CYP81-mediated oxidation precedes glutathione conjugation and intracellular sequestration of VOCs. These candidate genes provide a valuable foundation for future functional studies and may facilitate the development of sustainable crop protection strategies based on volatile-mediated plant defense. Full article

The cytochrome P450 superfamily is vital for plant growth and stress responses, yet the CYP90 gene family in Brassica napus remains poorly characterized. In this study, we systematically identified 25 BnCYP90 genes across 16 chromosomes, classifying them into CYP90A, CYP90B, CYP90C, and CYP90D phylogenetic groups. Synteny and evolutionary analyses were consistent with purifying selection acting on this gene family. Promoter analysis identified diverse stress- and hormone-responsive elements, while network analyses predicted BnCYP90_4, BnCYP90_6, and BnCYP90_11 as highly connected nodes in candidate protein–protein interaction and miRNA-mediated regulatory networks. Furthermore, single nucleotide polymorphism (SNP) analysis of a natural population demonstrated that a missense variant (C09:66,297,283) in BnCYP90_25 was significantly associated with seed germination rate and aboveground biomass under salt stress. Transcriptome and RT-qPCR analyses highlighted distinct spatiotemporal expression profiles under abiotic stresses: BnCYP90_25 showed sustained activation under salt stress, BnCYP90_9 and BnCYP90_20 exhibited mid-stage responses to osmotic stress, and BnCYP90_21 and BnCYP90_25 rapidly responded to alkaline stress. Finally, weighted gene co-expression network analysis (WGCNA) identified BnCYP90_13 as a hub gene in the T1 developmental stage. Overall, this study comprehensively characterizes the BnCYP90 family, providing valuable candidate genes for future functional validation in breeding stress-tolerant B. napus. Full article

Vivipary adversely affects the production process of the cucumber seed industry and greatly limits the popularization of cucumber varieties. Identification of the cucumber seed vivipary phenotype and screening of vivipary-associated genes will provide important theoretical value and practical significance for solving this problem in agricultural production. In this study, cucumber near-isogenic lines with significant differences in vivipary traits (viviparous line F and non-viviparous line BF) were successfully screened and used to construct genetic populations. Bulk segregant analysis (BSA) and QTL-seq were performed to fine-map the major-effect quantitative trait locus associated with vivipary variation. Based on QTL and BSA analyses, CsaV3_3G044640 (designated CsCYP82D47), which encodes a cytochrome P450 family protein, was identified as a candidate gene associated with cucumber vivipary. CsCYP82D47 exhibits obvious tissue specificity and is highly expressed in leaves, sprouts, and seeds. However, no significant difference in CsCYP82D47 expression was detected between viviparous and non-viviparous cucumber materials. Further sequence analysis revealed multiple mutation sites in this gene between different genotypes. Specifically, the CYP82D47 protein in viviparous materials harbours one amino acid insertion (L63) and two missense mutations (M71L and S124L). In addition, the altered leucine residue distribution in viviparous cucumber may enlarge the substrate channel and enhance substrate catalytic efficiency, which may contribute to the vivipary phenotype. In summary, this study identifies a promising candidate gene potentially related to cucumber vivipary, which lays a foundation for further exploration of the molecular mechanism underlying cucumber vivipary and provides a potential genetic resource for cucumber molecular breeding. Full article

In drug-induced liver injury (DILI) prediction field, animal models and in vitro cell models are most commonly used. However, animal models require long experimental timelines and may exhibit species-specific differences compared with humans, whereas conventional two-dimensional (2D) cell culture models lack cell-to-cell and cell-to-extracellular matrix (ECM) interaction. Liver organoid models and liver organ-on-a-chip can better simulate the human liver microenvironment; however, the construction of liver organoids requires a long cycle and high costs, while liver organ-on-a-chip systems demand specialized equipment and professional technicians. Herein, we selected the human C3A cell line, characterized by its low cost and facile culture conditions to establish an in vitro three-dimensional (3D) liver model. Briefly, C3A cells were embedded in Matrigel and cultured for 7 days to allow model maturation. Compared with their 2D-cultured cell model, the established 3D model exhibited elevated mRNA expression levels of drug-metabolizing cytochrome P450 enzymes (CYPs). Moreover, the model displayed robust expression of key hepatic biomarkers, as well as bile duct biomarkers. To evaluate the model’s applicability for DILI prediction, we performed toxicity assessments using a panel of six well-characterized hepatotoxicants and three non-hepatotoxic compounds. Notably, the 3D C3A model achieved a sensitivity of 83.3%, specificity of 100%, and overall accuracy of 88.9%. Furthermore, treatment of this model with chlorpromazine, a well-characterized cholangiotoxic agent, resulted in suppressed expression of the bile duct biomarker cytokeratin 19 (CK19) and bile salt export pump (BSEP), accompanied by impaired bile acid transport capacity. Taken together, this study provided a simple, low-cost, easy to culture, and more readily scalable 3D hepatic model in comparison with conventional 2D primary human hepatocyte (PHHs) models and other advanced 3D liver models. Notably, the model displayed dual hepatic and biliary characteristics, supporting predictions of both DILI and drug-induced bile duct injury. It provided a promising in vitro platform for assessing drug-induced hepatobiliary toxicity, with potential to reduce reliance on animal experiments and accelerate early-stage screening of novel pharmaceutical candidates. Full article

VERO cells, derived from the kidney epithelium of the African green monkey, are widely used in virology, but their ability to metabolize xenobiotics is not fully understood. Since cytochrome P450 (CYP) enzymes participate in xenobiotic metabolism, we investigated which CYP genes are expressed in VERO-E6 cells. Reverse transcription–quantitative polymerase chain reaction (RT-qPCR) showed that VERO-E6 cells express CYP3A4, CYP3A5, and CYP3A7. In contrast, CYP1A1, CYP1A2, CYP1B1, CYP2E1, CYP2D6, and CYP2C9 transcripts were either not detected or at a low detection level. To determine whether the encoded enzymes have the potential to activate aflatoxin B1 (AFB1), we used artificial intelligence (AI)-based structural modeling along with molecular docking. AI modeling suggested that CYP3A enzymes can position AFB1 in an orientation compatible with the formation of the reactive intermediate, and CYP3A4 showed the most favorable predicted interaction (docking score: −16.3 kcal/mol). To demonstrate AFB1 bioactivation, we exposed VERO-E6 cells to 200 nmol/L AFB1. After 10 days, we observed about 40% cell death. Liquid chromatography–tandem mass spectroscopy (LC–MS/MS) analysis confirmed the presence of AFB1-derived DNA adducts, indicating that metabolic activation occurred in these cells. These findings support the presence of CYP-dependent AFB1 bioactivation in VERO-E6 cells. Thus, combining computational and experimental approaches elucidates xenobiotic metabolism in cells where biochemical data are limited. Full article

Background: Pharmacogenetic (PGx) testing could identify actionable drug–gene interactions, reducing the risks of inappropriate prescribing of certain medications in some patients. An area of growing public health concern is rising global rates of depression and antidepressant use over the last two decades. Prior research has elucidated perspectives of healthcare providers who prescribe antidepressants regarding the clinical utility of genetic information, including PGx testing, but there is a gap in understanding how individual perspectives and systemic contextual factors may combine to influence PGx testing adoption. Objective: The objective of this study was to elucidate combinations of individual and contextual conditions associated with willingness to adopt PGx testing for Cytochrome P450 Subfamily IID, Polypeptide 6 (CYP2D6) and Subfamily IIC, Polypeptide 19 (CYP2C19) among antidepressant prescribers. Methods: We conducted a cross-sectional, mixed-methods study using structured questionnaires and semi-structured interviews with healthcare providers who prescribe antidepressants within their scope of practice across four healthcare systems in the United States. We collected data on implementation science concepts from the Theoretical Domains Framework, the Consolidated Framework for Implementation Research (CFIR), and the Implementation Outcomes Framework. Coincidence analysis (CNA), a case-based, Boolean logic-based method that identifies minimally sufficient combinations of conditions that lead to a particular outcome, was used to identify combinations of conditions for PGx test adoption among antidepressant prescribers. Interviews were also conducted with 10 patients who received pharmacogenetic testing within these healthcare systems to contextualize findings with patient perspectives. Results: Prescribers adopted PGx testing when they believed it would be beneficial to patients and were not deterred by cost-related concerns; the combination of these conditions led to PGx adoption in the most highly supported CNA model. Patient perspectives were also consistent with the selected model, with data suggesting they may have greater willingness to tolerate costs when they perceived or experienced benefits from testing. Conclusions: Insights from this study may be used by health system administrators and public health policymakers to inform future PGx implementation strategies that enhance uptake and awareness of existing evidence for clinical benefits of PGx testing and mitigate cost-related barriers to adoption. Full article

Wheat Fusarium head blight (FHB) is a globally prevalent and destructive fungal disease predominantly caused by Fusarium graminearum. Pydiflumetofen, a novel succinate dehydrogenase inhibitor (SDHI) fungicide, exhibits strong inhibitory activity against F. graminearum; however, the molecular regulatory mechanisms underlying the field-developed resistance in this pathogen remain poorly defined. In the present study, a field-evolved resistant isolate W24-039 and a sensitive isolate W24-016 were subjected to multi-omics analysis. The sequencing results identified compound mutations C89S/A93V in SdhC2, and A21T/S30F in SdhD of the resistant strain, which confer stable fungicide resistance without any detectable fitness costs. Physiological tests revealed that these target mutations sustain the homeostasis of succinate dehydrogenase (SDH) activity and intracellular ATP production. Following pydiflumetofen treatment, the sensitive isolate displayed remarkable declines in SDH activity, intracellular ATP content and deoxynivalenol (DON) biosynthesis, accompanied by markedly elevated cell membrane permeability. Transcriptomic sequencing uncovered 2221 differentially expressed genes (DEGs) in the sensitive strain under fungicide stress, and 2566 DEGs in the resistant isolate under the same conditions. The genes associated with detoxification and drug efflux, including cytochrome P450, glutathione S-transferase (GST), ABC and MFS transporters, were significantly upregulated in the resistant isolate. Metabolomic analysis indicated that differential metabolites were mainly enriched in the tricarboxylic acid (TCA) cycle, amino acid metabolism and membrane lipid biosynthesis pathways. The resistant strain maintained intact TCA cycle operation and accumulated high levels of pivotal metabolites such as phosphatidylcholine, unsaturated fatty acids and reduced glutathione. Integrated multi-omics analysis verified that the ABC transporter and glutathione metabolism pathways serve as core regulatory modules governing fungicide resistance. Collectively, F. graminearum develops resistance via the synergistic effects of SDH compound mutations, enhanced detoxification and efflux, and global metabolic remodeling, demonstrating that target-site mutation alone is not the sole driver of resistance, which is instead controlled by an intricate regulatory network involving multiple coordinated pathways. This study systematically characterizes the resistance regulatory network of F. graminearum against pydiflumetofen, and provides theoretical guidance for the rational application and sustainable field resistance management of this fungicide. Full article

Alzheimer’s disease (AD) is a complex neurodegenerative condition marked by a gradual decline in cognitive abilities, a reduction in acetylcholine (ACh) levels, and the accumulation of β-amyloid (Aβ) plaques. In a healthy brain, approximately 80% of ACh is broken down by acetylcholinesterase (AChE). Meanwhile, butyrylcholinesterase (BChE) serves a supportive function, gaining significance as AChE activity diminishes during the progression of Alzheimer’s disease. Modern therapeutic approaches focus on creating dual inhibitors of AChE and BChE that also aim to diminish Aβ-amyloidogenesis through interactions with the peripheral anionic site (PAS). This study combined fragment-based molecular design (CReM), AI-assisted retrosynthetic feasibility assessment, and in silico evaluation (docking, molecular dynamics, and ADMET profiling) to identify novel Donepezil derivatives as potential dual AChE/BChE inhibitors. A series of 10,000 derivatives were developed through computational methods and carefully assessed based on stringent drug-likeness, synthetic accessibility, and medicinal chemistry standards. This was succeeded by comprehensive ADMET profiling. Nine candidates were identified with predicted CNS pharmacokinetics, adequate toxicological profiles and reduced cytochrome P450 liabilities. Molecular docking yielded improved predicted binding affinities relative to Donepezil. Several derivatives, particularly D4 and D5, showed dual-site binding poses spanning both the catalytic gorge and the PAS. MD simulations indicated the stability of these poses over 100 ns. These computational results suggest that the proposed derivatives may preserve or improve upon Donepezil’s pharmacokinetic profile while offering potentially balanced AChE/BChE inhibition and anti-amyloidogenic activity, pending experimental validation. Full article

Dociostaurus maroccanus is a major migratory pest first recorded in Xinjiang, China, in 2025 in the Ili River Valley and Tacheng City. This event represents a new situation of concern in the field of biological security along China’s northwestern border that warrants attention. This study investigated the genetic diversity and population structure of four populations from the China–Kazakhstan border region using combined mitochondrial cytochrome C oxidase subunit I (COI) and cytochrome B (Cytb) gene markers. A total of 41 haplotypes were identified from 74 individuals, with the shared haplotypes H3 and H6 being the most frequent. Haplotype network analysis revealed no clear geographic population structuring. Haplotype diversity was high (Hd = 0.957), whereas nucleotide diversity was low (π = 0.00212), indicating low overall genetic diversity. Frequent gene flow (Nm = 12.31–43.08) and minimal genetic differentiation (Fst = −0.0255 to 0.0199) were detected among populations. AMOVA indicated that genetic variation mainly occurred within populations (100.12%), with no significant differentiation among populations (−0.59%). Neutrality tests (Tajima’s D = −2.45136, p < 0.05; Fu’s Fs = −44.037, p < 0.001), the unimodal mismatch distribution, and goodness-of-fit test results for KZ1 population (SSD = 0.0028, p = 0.91; raggedness = 0.0225, p = 0.86), KZ2 population (SSD = 0.00846, p = 0.25; raggedness = 0.0650, p = 0.13) and YN population further support the expansion phenomenon. Mantel test (r = −0.5948, p = 0.9583) preliminarily indicated that population distribution was not correlated with geographic distance. Therefore, in the region covered by this study, the overall characteristics show recent population expansion features. The preliminary results of this study provide basic materials for understanding the genetic background of this species in China’s border regions and can serve as a reference for subsequent population dynamic monitoring and control work. Full article