Search Results (492)

The Blattella germanica L. (Blattodea: Ectobiidae) is a major urban pest with widespread insecticide resistance. This study evaluated the synergistic effect of combining Metarhizium anisopliae with beta-cypermethrin (β-CYP) and investigated its mechanisms. Compatibility assays showed that β-CYP (1–10 μg/mL) was highly compatible with M. anisopliae (1 × 10⁷–1 × 10⁹ cfu/mL), with enhanced compatibility at lower concentrations. Bioassays confirmed significant synergy, increasing mortality and shortening median lethal time. Mechanistically, the combination accelerated histopathological damage to the midgut, hindgut, Malpighian tubules, and fat body, while promoting fungal proliferation in hemolymph and gut tissues. 16S rRNA sequencing revealed gut microbiota dysbiosis: Firmicutes replaced Bacteroidetes as the dominant phylum, opportunistic pathogens (Weissella, Alistipes) increased, and beneficial bacteria (Enterococcus, Fusobacterium) decreased. Weissella recolonization experiments confirmed its mortality-enhancing role. Additionally, the combination suppressed carboxylesterase activity and modulated immune gene expression (CYP4G19, BgPo), compromising host defense. Collectively, these findings demonstrate that β-CYP and M. anisopliae exert synergistic effects through multi-target mechanisms, offering a promising integrated pest management strategy that reduces chemical usage while enhancing biocontrol efficacy. 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

Bietti crystalline dystrophy like retinal degeneration (BCD) is an inherited retinal degenerative disease caused by defects in the CYP4V2 gene. Recombinant AAV-based gene replacement therapy holds promise for this disease, yet reliable enzymatic activity assays for product potency evaluation remain urgently needed. This study established and validated a method for determining the enzymatic activity of CYP4V2 based on the bioluminescent substrate Luciferin-MultiCYP.HEK293-AAVR cells were seeded at 1 × 10⁴ cells/well and transduced with rAAV-hCYP4V2 across an MOI gradient (1.17 × 10³–2.4 × 10⁶). After incubating at 37 °C for 72 h, medium was replaced with Opti-MEM containing 20 µM Luciferin-MultiCYP and incubated for 90 min ± 30 min. Next, cell supernatants were mixed with luciferase reagent, and bioluminescence was measured. Methodological validation results revealed that the method has good specificity, with an accuracy recovery rate of 100.94% ± 2.84% (RSD = 2.81%). Repeatability (GCV%) was 4.21%, and intermediate precision (RSD) was 6.16%. Linearity was good within the MOI range of 1.17 × 10³ to 2.4 × 10⁶ (R² = 0.9902 ± 0.0061, n = 9). Lastly, the method was successfully applied to activity testing the products of three AAV serotypes (AAV8, AAV2/8, and AAV2). This bioluminescent method meets regulatory potency assay requirements and serves as an effective QC tool for process development, batch release, and stability evaluation of rAAV-hCYP4V2 products. Full article

Hyperlipidemia (HL) is a major metabolic disorder and a critical risk factor for cardiovascular diseases, closely associated with oxidative stress, inflammation, and disrupted cholesterol homeostasis. Freeze-dried Poecilobdella manillensis powder (FDPMP), a traditional medicinal product, has shown therapeutic potential against hyperlipidemia; however, its underlying mechanisms remain largely unclear. In this study, HL was induced in ApoE^−/− mice by feeding a high-fat diet (HFD) for eight weeks, during which FDPMP or simvastatin (positive control) was orally administered daily. Concurrently, an in vitro foam cell model was established by exposing RAW264.7 macrophages to oxidized low-density lipoprotein (ox-LDL, 80 μg/mL) for 24 h, with FDPMP pretreatment applied 30 min prior to ox-LDL stimulation. Following intervention, serum lipid profiles, hepatic oxidative stress markers, histopathological changes, and cholesterol metabolism-related gene and protein expression were systematically evaluated. FDPMP administration significantly improved serum lipid profiles by reducing triglycerides, total cholesterol, and low-density lipoprotein cholesterol, while increasing high-density lipoprotein cholesterol levels. Additionally, FDPMP alleviated histopathological damage in the liver, kidney, and heart, enhanced antioxidant enzyme activities, and attenuated oxidative stress. Untargeted metabolomic analysis revealed that FDPMP markedly modulated key metabolic pathways, including choline metabolism, glycerophospholipid metabolism, and arachidonic acid metabolism. Mechanistically, FDPMP restored cholesterol homeostasis through dual regulation of cholesterol metabolism, characterized by upregulation of cholesterol 7α-hydroxylase (CYP7A1) to promote bile acid-mediated cholesterol excretion, alongside downregulation of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) and synthase (HMGCS1) to inhibit cholesterol biosynthesis. In vitro, FDPMP effectively suppressed ox-LDL-induced foam cell formation, reduced intracellular lipid accumulation, and mitigated oxidative stress in macrophages. Collectively, these findings demonstrate that FDPMP ameliorates hyperlipidemia through coordinated regulation of cholesterol synthesis and excretion, coupled with systemic metabolic reprogramming and antioxidative effects. This study provides mechanistic insights supporting FDPMP as a promising natural therapeutic candidate for hyperlipidemia and related metabolic disorders. Full article

Aspergillus fumigatus is the predominant pathogenic fungus responsible for aspergillosis. In recent years, the global detection rate of azole-resistant A. fumigatus has continuously increased, and the extensive application of agricultural azole fungicides has been recognized as a crucial driving factor for the emergence and spread of resistance mutations in environmental A. fumigatus. Previous investigations conducted by our research team in karst vegetable fields of Guizhou Province revealed that the triazole resistance rate of local A. fumigatus was only 0.49%, which was markedly lower than those reported in most previous studies in China and outside of China. To supplement the prevalence data of azole resistance across different habitats in this region, a total of 191 environmental A. fumigatus strains were isolated from nine tea plantations across Guizhou. In this study, two clinically prevalent azole drugs, itraconazole and voriconazole, were used for antifungal susceptibility testing, and the triazole target gene cyp51A of all isolates was sequenced and analyzed. Antifungal susceptibility results demonstrated that the MIC ranges of the tea plantation A. fumigatus population were 0.015–0.5 μg/mL for itraconazole and 0.031–0.25 μg/mL for voriconazole, with no evidence of triazole resistance. Genetic analysis identified ten different gene mutations among 29 isolates, all of which were classified as non-resistance-associated mutations. Among these mutations, four were synonymous mutations, including 267G→A, 540G→A, 1074A→G, and 1362T→C, while six were non-synonymous mutations, including 137T→A, 514A→G, 743A→C, 744T→A, 765C→G, and 1279G→A. These non-synonymous mutations resulted in five amino acid substitutions in 25 strains, namely F46Y, M172V, N248T/K, D255E, and E427K. The N248T/K mutation exhibited the highest mutational frequency of 0.1309 (25/191) and was distributed across all nine sampling sites. Correlation analyses indicated that no significant correlations were observed between all detected variant loci and MICs of isolates to itraconazole and voriconazole. Phylogenetic analysis revealed that the six sequence types of cyp51A in Guizhou tea plantations were broadly intermixed with those from other parts of China and outside of China. We discussed the implications of these results in the management of antifungal resistance. Full article

Chronic kidney disease is associated with neurovascular complications including cognitive impairment, potentially involving blood–brain barrier (BBB) impairment. The protein-bound uremic toxin indoxyl sulfate (IS) promotes endothelial injury, but its effects on human brain microvascular endothelial cells remain incompletely understood. Human cerebral microvascular endothelial cells (hCMEC/D3) were exposed to IS 200 and 900 μM. BBB integrity was assessed by the FITC-dextran (4 kDa) transwell permeability assay and claudin-5 immunofluorescence. Transcriptional responses were quantified by qPCR for aryl hydrocarbon (AhR) target genes, oxidative stress-associated and inflammatory markers, senescence, and junction-associated genes. Senescence-associated phenotypic changes were evaluated by SA-β-galactosidase staining and cytokine array profiling. IS increased endothelial permeability at 24 and 48 h (~1.5-fold relative to control) without evidence of cytotoxicity and reduced claudin-5 staining intensity. IS strongly upregulated AhR target genes, including CYP1A1, CYP1B1, and CYP1A2. NFE2L2 and IDO1 increased, while NFKB1 remained unchanged. SA-β-gal positivity increased, accompanied by elevated GM-CSF and G-CSF secretion, while CDKN1A decreased at IS 900 µM and CDKN2A remained unchanged. CDH5 was downregulated, TJP1 increased at 900 µM, and CLDN5 remained unchanged. These findings indicate that IS exposure is associated with impaired BBB integrity, AhR-related transcriptional responses, oxidative stress-associated transcriptional changes, junctional remodeling, and senescence-like endothelial features. However, causal attribution to individual pathways requires inhibition or knockdown studies. Full article

Background: The bean flower thrips, Megalurothrips usitatus, is a destructive agricultural pest with increasing resistance to pyrethroid insecticides. Most mechanistic studies emphasize mutations in the pore-forming voltage-gated sodium channels α-subunit, whereas the contribution of insect-specific sodium-channel auxiliary subunits remains unclear. Methods: We used dsRNA feeding, pyrethroid bioassays, RNA sequencing, and RT-qPCR validation to evaluate the role of temperature-induced paralytic E (TipE) in M. usitatus. Results: The optimized RNAi condition was 400 μg/mL of dsTipE for 48 h, which reduced TipE expression by 32.55%. TipE knockdown increased adult mortality after exposure to λ-cyhalothrin and permethrin, indicating increased pyrethroid-associated susceptibility under laboratory conditions. Transcriptome sequencing identified 205 differentially expressed genes, with enrichment in metabolism-related pathways. RT-qPCR confirmed reduced expression of CYP307a1, an ecdysteroid-biosynthesis-related P450 gene, and SCD, a lipid-metabolism gene, while Na_v α-subunit transcript abundance did not change significantly. Conclusions: These results identify TipE as an auxiliary-subunit factor associated with pyrethroid susceptibility in M. usitatus and provide a transcriptome-based working model linking sodium-channel auxiliary regulation with endocrine- and lipid-metabolism-related responses. This study expands the candidate space for pyrethroid resistance research beyond Na_v α-subunit mutations, although the causal roles of CYP307a1 and SCD require further validation. Full article

Background and Objectives: Cornerstone therapies for metastatic prostate cancer include androgen deprivation and androgen receptor pathway inhibition, but cognitive impairment is a recognized, life-altering potential adverse effect of this treatment. Abiraterone acetate (AA), an androgen receptor pathway and CYP17A1 inhibitor, suppresses androgen synthesis and may contribute to cognitive changes. This cell culture-based study uses the BE(2)M17 human male neuroblastoma model to investigate AA-induced alterations in gene and protein expression that may underlie cognitive decline, laying the foundation for a mechanistic investigation aimed at identifying molecular targets to mitigate cognitive impairment in men with prostate cancer receiving androgen-directed therapies. Materials and Methods: BE(2)M17 cells were pretreated for 12 h with dihydrotestosterone (DHT; 5 nM) or vehicle control, then exposed to AA (0, 5, 10 µM, 24 h). RNA and protein were analyzed by qRT-PCR and Western blot for markers of amyloid processing, neuronal health, and mitochondrial function. Results: AA significantly altered multiple neurobiological markers. BACE1 mRNA increased in DHT + 10 µM AA compared to control and DHT alone (p = 0.0416 and p = 0.0118). However, BACE1 protein decreased in 10 µM AA + DHT versus DHT alone (p = 0.0132). Immunoblot revealed reduced amyloid precursor protein (APP) in 10 µM AA versus control (p = 0.0057) and 10 µM versus 5 µM (p = 0.0263). APP was reduced in 10 µM AA + DHT versus control (p = 0.0015) and versus DHT alone (p = 0.0467). LRP1 and BDNF were significantly reduced with 10 µM AA versus 5 µM AA (p = 0.0092 and p = 0.0081), while synaptophysin decreased in 10 µM AA + DHT versus DHT alone (p = 0.0049). BDNF also declined in 10 µM AA + DHT compared to 5 µM AA + DHT (p = 0.0301). PGC1α mRNA increased in AA + DHT versus DHT alone (p = 0.0332). MitoTracker analysis showed reduced fluorescence with 5 µM AA alone but increased fluorescence with 5 µM AA + DHT relative to control and DHT (p = 0.0019; p < 0.0001), while 10 µM AA + DHT reduced fluorescence compared to 5 µM AA + DHT (p = 0.0003). Conclusions: AA, alone or combined with DHT, disrupts key pathways involved in neuronal health, amyloid processing, and mitochondrial function. These findings suggest a potential mechanistic link between AA treatment and cognitive impairment. Full article

Background/Objectives: Cancer is a multifactorial disease in which drug resistance and limited selectivity remain major therapeutic challenges, highlighting the need for novel anticancer agents. As a privileged scaffold for multitarget anticancer drug discovery, quinazolin-4(3H)-one was selected for the design, synthesis, and evaluation of new derivatives as potential anticancer agents, together with investigation of their mechanisms of action and molecular targets. Methods: Fifteen new quinazolin-4(3H)-one derivatives were synthesized and screened using the NCI-60 human cancer cell line panel. The mechanism of action of the most active compound was investigated through cell cycle, apoptosis, and RT-qPCR analyses. A potential molecular target was identified from transcriptomic data in the Human Protein Atlas, focusing on highly expressed cancer-implicated genes in the most responsive cell lines, followed by molecular docking, molecular dynamics simulations, and in vitro kinase studies. Safety and pharmacokinetic properties were evaluated using an MTT cytotoxicity assay in normal WI-38 fibroblasts and in silico ADME analyses. Results: Compound 3e emerged as the most active and tumor-selective derivative, exhibiting GI₅₀ values ranging from 2.63 to 17.12 µM across 31 cancer cell lines. In A549 cells, selected as a representative responsive model, 3e (GI₅₀ = 10.8 µM, 72 h) induced G2/M cell-cycle arrest (59.58% vs. 26.96% in control), increased early apoptosis (43.94% vs. 0.11% in control), reduced viable cells (49.71% vs. 98.66%), elevated the Bax/Bcl-2 ratio (7.91), and upregulated the expression of caspase-9 and caspase-3 by 2.5- and 4.6-fold, respectively. Integrated target identification studies and an in vitro kinase assay (IC₅₀ = 21.34 nM) suggested TGF-βRI (ALK5) as a plausible molecular target. Compound 3e also showed low cytotoxicity toward WI-38 fibroblasts (IC₅₀ = 88.3 µM) and favorable predicted pharmacokinetic properties; nevertheless, high plasma protein binding and potential CYP2C9 inhibition are anticipated. Conclusions: Compound 3e is a promising tumor-selective anticancer lead with potential TGF-βRI inhibitory activity. Its antiproliferative effects in A549 cells appear to be mediated through G2/M cell-cycle arrest and activation of the intrinsic apoptotic pathway, supporting further development and pharmacokinetic optimization of this scaffold for anticancer therapy. Full article

Invasive infections caused by Aspergillus flavus are more common in tropical and subtropical countries. The emergence of azole resistance in A. flavus complicates the management of aspergillosis, as azoles are the first-line and empirical therapy. The aim of this study was to investigate the molecular mechanisms underlying azole resistance in A. flavus, focusing on the cyp51C gene. We screened 34 molecularly confirmed A. flavus isolates obtained from patients with invasive aspergillosis for cyp51C gene expression by real-time RT-qPCR and for mutations by PCR sequencing. Molecular modeling and docking studies were performed using SWISS-MODEL, SwissDock, and I-TASSER software. Susceptibility testing revealed that 14.71% and 8.82% of isolates were resistant to itraconazole and posaconazole, respectively, with 5.88% exhibiting cross-resistance. The mRNA expression of cyp51C was upregulated (>2.5-fold) in five of the six resistant strains (83.33%). Hyperexpression of cyp51C was significantly more frequent among resistant isolates than among susceptible isolates (Fisher’s exact test, p = 0.014). Sequencing identified ten point mutations, including six synonymous and four non-synonymous substitutions. The non-synonymous mutations M54T and S240A were detected in the protein sequences of both resistant and susceptible isolates. Notably, D254N and I285V were observed exclusively in resistant isolates and in susceptible isolates with itraconazole MICs near the epidemiological threshold. Homology modeling and 3D structure prediction of the mutated Cyp51C protein demonstrated interactions with itraconazole, posaconazole, and voriconazole. Importantly, I-TASSER analysis indicated that the I285V substitution is located near the itraconazole binding site. Simultaneous overexpression of the cyp51A, cyp51B and cyp51C genes was observed in 33.33% of resistant isolates. These findings suggest that multiple target genes and mechanisms may act concurrently to confer azole resistance in A. flavus. Overall, this study supports the hypothesis that azole resistance in A. flavus is multifactorial and highlights the potential value of combining mutation analysis, gene expression profiling, and structural modeling for improved molecular surveillance and antifungal resistance monitoring. Full article

Background: This study was conducted to identify genetic risk variants associated with nicotine addiction in the CHRNA5, HTR2A, DRD4, and CYP2A6 genes among electronic cigarette users who also smoke combustible cigarettes (dual users), and to assess potential genotoxic and cytotoxic damage in the oral mucosal cells of the study population. Methods: We included dual e-cig users (ECIG, n = 70), combustible cigarette smokers (CCU, n = 24), and non-smokers and non-e-cig users (NS, n = 110). Genetic variants in CHRNA5, HTR2A, DRD4, and CYP2A6 were genotyped. Micronucleus analysis was performed on oral mucosal cells to detect cellular abnormalities. Results: The ECIG group demonstrated greater nicotine addiction on the Fagerström Test for Nicotine Dependence (FTND, 5.5 vs. 1, p = 0.023). Salivary cotinine levels were significantly higher in the ECIG group compared to the CCU group (39 vs. 12 ng/mL, p < 0.001). The carriers of the A allele (rs16969968/CHRNA5) had higher FTND scores, carriers of the C allele (rs1800955/DRD4) used electronic cigarettes more frequently each day, and carriers of the T allele (rs4105144/CYP2A6) started using nicotine products at a younger age. The number of micronuclei and cellular abnormalities in the oral mucosa was higher in the ECIG and CCU groups compared to the NS group. Conclusions: Salivary cotinine levels and FTND are higher in dual e-cigarette users than in combustible cigarette users. Dual users exhibit risk alleles in the CHRNA5, DRD4, and CYP2A6 genes, which are associated with traits linked to increased nicotine addiction. Dual e-cigarette use poses comparable genotoxic risks to combustible smoking. Full article

Introduction: The temperature of rivers in the Iberian Peninsula has increased due to global warming. In addition, these rivers are polluted by contaminants of emerging concern, such as polycyclic aromatic hydrocarbons (PAHs). Higher temperatures and pollution concurrently impose threats to the Iberian Peninsula’s endemic species, including the brown trout (Salmo trutta), a cold-water species widely used in ecotoxicological studies. Because the liver is the main biotransformation organ, and is particularly sensitive to both chemical and temperature changes, in vitro liver models may represent valuable alternatives for assessing combined stressor effects, complying with the 3Rs principle. Objective: In line with the above, the present study aimed to evaluate the combined effects of a 4 °C temperature increase and the model PAH benzo[k]fluoranthene (B[k]F) on fish liver cells using a primary brown trout hepatocyte culture as a model. Methodology: Primary hepatocytes were seeded in 6-well plates at a density of 1.0 × 10⁶ cells/mL and exposed for 48 h to 1, 10, and 20 µM B[k]F at 18 °C (normothermia) and 22 °C (warming scenario). Cell viability was assessed using trypan blue, alamarBlue, and lactate dehydrogenase (LDH) assays. Cytochrome P450 (CYP)1A was evaluated in terms of its gene expression by RT-qPCR and its protein expression through immunocytochemistry (ICC). The immunostaining was quantified using a score system which considered five intensity staining levels. Results: Exposure to B[k]F and to the higher temperature increased LDH leakage without interaction effects. In contrast, the other viability assays did not show significant differences across conditions. Regarding CYP1A, both gene and protein expression increased with all B[k]F concentrations in relation to the controls, but were not influenced by temperature. Notably, the lowest B[k]F concentration (1 µM) elicited the highest CYP1A gene expression, suggesting a non-monotonic response. Conclusions: Overall, the model was responsive to both temperature (4 °C) increase and to B[k]F, validating its usefulness for assessing liver pollutant effects in the context of global warming. These findings support the application of fish primary hepatocyte models as relevant tools in ecotoxicology under environmentally realistic multi-stressor scenarios. Full article

The yak lung functions as a vital adaptive organ in cold, low-oxygen environments. Hypoxia can induce pathological remodeling in yak lung tissue, so we need to understand the molecular mechanisms of this remodeling. For this study, we used cross-omics comparative approaches, including transcriptomics, label-free proteomics, and untargeted metabolomics, to examine both normal and diseased yak lung tissues. From histological observations, the disease phenotype was identified as pulmonary emphysema. Our results showed a significant up-regulation of differentially expressed genes such as MTTP, CXCL8, RETN, and NNAT, while genes like SLC45A1, IL10, SDSL, and COL12A1 were clearly down-regulated. In the differential protein analysis, proteins such as RASSF4, EDC4, CTSC, and FECH were notably up-regulated, whereas CYP27A1, FKBP9, RAD23A, and PLSCR2 were significantly down-regulated. Metabolomic profiling revealed that palmitoyl-L-carnitine, decanoyl-L-carnitine, and o-acetylcarnitine were significantly higher in emphysematous lung tissue, whereas racemethionine and L-methionine S-oxide were significantly much lower. Also, when we compared of bulk RNA-seq, label-free proteomics, and untargeted metabolomics data revealed enrichment in three common pathways: the asthma pathway, the linoleic acid metabolism pathway, and the gastric acid secretion pathway. Of note, histamine levels were higher in both the asthma and gastric acid secretion pathways. While the mRNA expression level of BoLA-DQB was increased in the asthma pathway, its protein expression level was decreased. This study offers some initial cross-omics evidence about what happens. These findings give us a scientific basis for developing effective prevention and control strategies, which in turn can help the protection of yak health and the sustainable development of plateau animal husbandry. Full article

Rhinogobio nasutus, an endangered fish species endemic to the upper and middle reaches of the Yellow River in China, lacks essential genomic information on gonadal development, hindering research into its reproductive biology. To address this, mRNA-seq and miRNA-seq datasets derived from adult testis (n = 3) and ovary (n = 3) were integrated to characterize sex-biased expression profiles and potential regulatory mechanisms. A total of 34,813 genes and 68,623 transcripts were detected, and 16,105 differentially expressed genes (DEGs) were identified between testis and ovary, including 9365 testis-biased and 6740 ovary-biased genes. Small-RNA profiling identified 51 differentially expressed miRNAs (DEMs: 31 testis-biased; 20 ovary-biased). The sex-biased mRNA profiles highlighted conserved candidate genes associated with germ-cell maintenance, somatic regulation, ovarian differentiation, and oocyte maturation, including vasa, piwi, dmrt1, amh, cyp19a1a, zar1, zar1l, and rbpms2. Integrated miRNA–mRNA analysis further predicted potential interactions involving key sex-related genes, suggesting that DEMs may contribute to post-transcriptional regulation during gonadal differentiation. Functional enrichment (GO and KEGG analyses) highlighted pathways associated with gonadal differentiation, germline maintenance, and signal transduction pathways. qRT-PCR validation of nine mRNAs and nine miRNAs showed expression patterns consistent with the sequencing results. Collectively, these results provide an integrated mRNA and miRNA resource for R. nasutus gonads and identify candidate genes and miRNAs for future studies on sex-biased gonadal development, reproductive regulation, and artificial propagation. Full article

Gatifloxacin (GTFX), a fourth-generation fluoroquinolone, causes metabolic disturbances in mammals, but its hepatotoxic mechanisms in aquatic vertebrates remain unclear. This study investigated whether GTFX induces liver injury in zebrafish larvae through oxidative stress or alternative pathways. Larvae at 3 days post-fertilization were exposed to 0.2–2.3 mg/mL GTFX for 48 h. Liver morphology, histopathology, intracellular reactive oxygen species (ROS), and expression of lipid metabolism (pparg) and xenobiotic biotransformation genes (cyp1a, cyp1b1) were assessed. GTFX exposure caused concentration-dependent reductions in liver area, increased hepatic opacity, delayed yolk sac absorption, and hepatocyte swelling with cytoplasmic vacuolization. Despite these structural changes, ROS levels did not differ significantly from those of controls. In contrast, transcriptional analysis revealed significant upregulation of pparg, cyp1a, and cyp1b1, indicating disrupted lipid homeostasis and enhanced detoxification responses. Acute high-dose GTFX exposure induced a metabolism-associated hepatotoxic response in zebrafish larvae, which occurred without a statistically significant change in bulk ROS levels. Together, these findings offer mechanistic insight into fluoroquinolone-associated liver injury. Full article