In this experiment, the primary intestinal epithelial cells (IECs) inflammation model was constructed to investigate the intestinal injury mechanism under high level addition of plant-derived proteins in hybrid grouper (Epinephelus fuscoguttatus female x E. lanceolatus male), and citric acid (CA) addition was used to investigate the protective effect in the primary IECs inflammation model. The vitro experiment was fed 30 fishes to three replicates of each group for 8 weeks. In the inflammation model of IECs, the addition of CA (1 mM) remarkably enhanced cell survival, and significantly reduced the secretion of inflammatory cytokines TNF-alpha and IL-1 beta (P < 0.05). CA activated phosphorylation of S6K and 4EBP-1 in the mTOR pathway and promoted the protein expression of tight junction proteins (ZO-1, E-cadherin, occludin and claudin-1) (P < 0.05). The supplementation of CA in the diet also could improve the expression of tight junction in intestine of hybrid grouper (P < 0.05). CA attenuated the protein expression of inflammatory cytokines (IL-1 beta, IL-6 and TNF-alpha) in inflammation models by regulating ERK and NF-kappa B pathways (P < 0.05). These findings suggest that citric acid enhances cell survival, strengthens the intestinal tight junction barrier and enhances intestinal immune function in the LPS-stimulated inflammatory model of IECs. Citric acid could serve as an effective means to counteract growth inhibition and reduced digestibility caused by high proportions of plant protein in fish feed.

Background: Astilbin is a bioactive flavonoid with documented anti-inflammatory properties; however, its sustained interactions with the gut microbiota remain poorly understood. Fecal samples from three healthy donors were pooled and fermented with astilbin in an in vitro human colonic model over 7 days. This exploratory study aimed to characterize the temporal ecological shifts associated with prolonged astilbin exposure. Methods: Time-resolved 16S rRNA gene sequencing, PICRUSt2 functional prediction, BugBase phenotypic inference, and pseudo-targeted metabolomics were integrated to track microbial-metabolic dynamics. Results: Astilbin exposure was associated with a highly coordinated, three-stage microbial succession. Day 3 (D3) emerged as a putative inflection point, where the enrichment of pioneer degraders (Flavonifractor, Bacteroides) was temporally correlated with the appearance of polyphenol cleavage intermediates. This transition featured an early decrease in markers of proteolytic fermentation alongside a transient in vitro lipid-stress response. By D7, the community shifted toward a stable configuration enriched in butyrogenic taxa (Roseburia, Subdoligranulum, Megamonas), with progressive depletion of potentially opportunistic pathogens (Escherichia-Shigella). Multi-omics integration suggested that these structural successions were strongly associated with marked metabolic shifts. Inflammatory lipid markers (e.g., leukotriene B4) showed a characteristic “D3-burst/D7-clearance” pattern, whereas potentially barrier-protective metabolites, particularly 3-indolepropionic acid (3-IPA, log2FC = 2.00) and urolithin B (log2FC = 1.18), accumulated substantially. Conclusions: This exploratory study provides valuable high-resolution insights into astilbin’s potential as a dynamic ecological modulator. It outlines a temporal framework illustrating how the gut microbiota may shift from proteolytic fermentation toward 3-IPA-associated homeostasis. Although limited by a pooled fecal model and the absence of a vehicle control, these hypothesis-generating findings offer a solid foundation for future in vivo studies and mechanistic validations across diverse human cohorts.

Concentrated cottonseed protein (CPC) serves as an ideal protein replacement for fish meal in diets, though excessive use can adversely affect aquatic life. Organic trace element replacements can sustain the growth and metabolic functions of Litopenaeus vannamei, aiding in the conservation of high-quality protein resources such as fish meal. The purpose of this experiment is to explore the impact of substituting fish meal with CPC as a protein source, along with substituting amino acid trace element chelates, on the growth, immune antioxidant capacity, and intestinal microbiota of Litopenaeus vannamei. Four experimental diets were formulated: the control group FM (no CPC substitution, supplemented with inorganic trace elements), the inorganic trace element group C0 (CPC substituted for 40 % fish meal, supplemented with inorganic trace elements), and the experimental group C1 (CPC substituted for 40 % fish meal, amino acid trace element chelates substituted for 40 % inorganic trace elements). and C2 (CPC replacing 40 % fish meal, amino acid chelated trace minerals replacing 60 % inorganic trace minerals). Each group had three replicates. At the end of a 10-week feeding trial, the research results are as follows: Group C2 had significantly higher crude protein and crude lipid contents compared to Group C0 (P < 0.05). In comparison to Group C0, the FM group demonstrated much higher activities of CuZn-SOD, GPX, and CAT (P < 0.01), group C0 exhibited notably lower activities compared to Groups C1 and C2 (P < 0.05). Relative abundances of major intestinal microbiota species in Groups C1 and C2 underwent significant changes. Compared to Groups C0 and C2, the expression levels of lipid metabolism genes were much higher in Group C1 (P < 0.05). In fish meal replacement diets with CPC, using 40 % amino acid chelates can mitigate adverse effects of plant protein and enhance shrimp lipid metabolism, immune antioxidants, and intestinal flora.

This study investigated the effects of taurine supplementation in low-fishmeal diets on growth, antioxidant capacity and hepato-intestinal health of spotted sea bass (Lateolabrax maculatus). A high-fishmeal diet (HF, 30% fish meal) and four low-fishmeal diets (60% fish meal replaced by CAP) supplemented with 0%, 0.2%, 0.4% and 0.6% taurine were used. Each treatment had four replicates with 20 fish per tank (initial weight: 65.35±0.07 g), and the trial lasted 8 weeks. Results showed that weight gain rate (WGR) of the LF group was significantly lower than those of HF, T2 and T3 groups, while WGR in HF and T3 was significantly higher than in LF and T1 (P<0.05). Feed conversion ratio (FCR) of LF was significantly higher, and specific growth rate (SGR) and protein efficiency ratio (PER) were significantly lower than in other groups (P<0.05). Protein deposition rate (PDR) in HF was significantly higher than in LF and T2 (P<0.05). Taurine improved growth, feed efficiency and antioxidant status, reduced lipid peroxidation, and enhanced expression of immune-related genes (il-10, tlr2, myd88) in liver and intestine. Hepatic triglyceride content decreased with increasing taurine. Expression of srebp1 and fas was downregulated, and lpl was upregulated in T3 (P<0.05). Histology showed taurine alleviated hepatocyte vacuolization and inflammatory damage. In summary, 0.4%–0.6% taurine effectively mitigates low-fishmeal diet adverse effects and improves growth, antioxidant capacity, immunity and hepato-intestinal health in L. maculatus.

Chlorogenic acid (CGA) is a natural polyphenolic compound with antioxidant activity. We hypothesised that dietary CGA supplementation could improve the growth performance and selected muscle-related traits of Procambarus clarkii, possibly in association with changes in antioxidant-related indices and the expression of selected genes. To test this hypothesis, a 6-week feeding trial was conducted using five diets containing 0, 200, 400, 600, and 800 mg/kg CGA. Dietary supplementation with 400-600 mg/kg CGA improved growth performance and feed conversion ratio, and was associated with higher collagen-related indices, better texture in some parameters, lower freezing loss, and higher amino acid contents in some treatment groups. CGA supplementation was also associated with higher antioxidant enzyme activities, lower reactive oxygen species and malondialdehyde levels, and changes in the expression of genes related to antioxidant defence and muscle development. Overall, these results suggest that dietary CGA supplementation, particularly at 400-600 mg/kg, may improve growth performance and selected muscle-related traits of P. clarkii under the present experimental conditions. These findings support further evaluation of CGA as a dietary supplement in P. clarkii.

This study revealed the dual effects of dietary selenium (Se) supplementation (added as sodium selenite SS, 0.71-3.3 mg/kg) in the spotted sea bass (Lateolabrax maculatus). Quadratic regression analysis identified an optimal supplemental level of SS at 1.39-1.53 mg/kg. This dose significantly enhanced growth performance, feed efficiency, and systemic antioxidant capacity, which was associated with upregulation of the nrf2 (nuclear factor erythroid 2-related factor 2)/ gpx2 (glutathione peroxidase 2) pathway and alterations in lipid metabolism involving ppar alpha (peroxisome proliferator-activated receptor alpha). Critically, high-dose SS (3.3 mg/kg) induced significant toxic effects, characterized by suppression of key antioxidant enzyme activities - GPx (glutathione peroxidase), SOD (superoxide dismutase), and CAT (catalase); along with downregulation of sod, nrf2, and ppar alpha. Concurrent with the toxicity, MDA (malondialdehyde) levels increased, confirming the occurrence of oxidative damage. The toxic cascade ultimately led to ROS (reactive oxygen species)-driven impairment of intestinal structural integrity (manifested as decreased muscular thickness) and growth suppression. These collective findings reveal a dose-response continuum: adverse signs (e.g., reduced intestinal muscular thickness and decreased antioxidant enzyme activities) emerged at 1.91 mg/kg SS, and progressively more pronounced toxic effects, including oxidative damage and intestinal structural impairment, became evident at 3.3 mg/kg SS, without a single discrete toxicity threshold.

BackgroundIschemic stroke is a leading cause of death and disability worldwide, yet the systemic metabolic mechanisms underlying poor functional recovery remain incompletely understood. Dysregulated lipid metabolism is a key pathogenic factor, and advances in metabolomics and lipidomics provide new opportunities for discovering pathophysiological signatures.MethodsIn this exploratory study, we integrated serum metabolomic and lipidomic profiling in 44 patients with acute ischemic stroke, stratified by favorable (n = 23) or unfavorable (n = 21) 90-day functional outcomes. Differential metabolites and lipids were identified, pathway-related molecular alterations were characterized, and candidate markers were evaluated to establish a proof-of-concept metabolic signature.ResultsPatients with unfavorable outcomes exhibited global metabolic reprogramming characterized by structural lipid disassembly (widespread reductions in membrane phospholipids, including phosphatidylcholines and ether phospholipids), mitochondrial energy failure (acylcarnitine abnormalities consistent with impaired β-oxidation), and antioxidant depletion (marked by decreased cis-caffeic acid). An exploratory three-marker panel—the C8:2-OH(3)/C18:2-OH(3) ratio, the free DHA/PC(44:12) ratio, and cis-caffeic acid—showed promising discriminatory potential in this derivation cohort (AUC = 0.950). Adding this metabolic panel to the initial NIHSS score improved the AUC to 0.975, suggesting incremental value beyond clinical assessment alone.ConclusionIntegrated serum metabolomics and lipidomics identified a metabolic signature reflecting lipotoxicity, mitochondrial dysfunction, and oxidative stress. These preliminary findings highlight systemic metabolic reprogramming as a potential driver of unfavorable outcome and provide a proof-of-concept for combining molecular phenotyping with conventional clinical scales. Without external validation and multiple-testing correction, these results should be viewed as hypothesis-generating.

This study evaluated the effects of dietary CpG oligodeoxynucleotide (CpG ODNs) on growth performance, immunity, digestive function, intestinal microbiota, and transcriptomic responses in Litopenaeus vannamei. Shrimp with an initial body weight of 0.30 ± 0.02 g were fed diets supplemented with 0, 0.1, 0.4, 1.6, 6.4, or 25.6 mg kg-1 CpG ODNs for 8 weeks. Dietary CpG ODNs did not significantly improve most growth parameters; however, shrimp fed the diet supplemented with 1.6 mg kg-1 CpG ODNs showed a lower feed conversion ratio. After white spot syndrome virus challenge, shrimp fed the diet supplemented with 25.6 mg kg-1 CpG ODNs showed higher survival than those in the control group, whereas survival after Vibrio parahaemolyticus challenge did not differ significantly among treatments. Dietary supplementation with CpG ODNs at levels above 0.4 mg kg-1 increased the activities of several serum immune-related and antioxidant enzymes, reduced malondialdehyde content, and enhanced intestinal trypsin and amylase activities. In addition, this treatment increased microbial richness indices, while reducing the relative abundances of Vibrio and Photobacterium. Transcriptomic analysis showed that several pathways, including the AMPK signaling pathway and JAK-STAT signaling pathway, were significantly enriched in the CpG ODNs (above 0.4 mg kg-1) groups compared with the control group. In conclusion, dietary supplementation with CpG ODNs did not significantly improve the growth performance of L. vannamei. However, dietary supplementation with CpG ODNs at levels above 0.4 mg kg-1 may help enhance immune and digestive functions and modulate the intestinal microbiota composition of L. vannamei.

Atherosclerotic cardiovascular disease can progress despite intensive lipid lowering, highlighting the need to identify local mechanisms that remodel established plaques. Intermittent fasting may engage metabolic, circadian, and immune pathways relevant to this process. We used genome-wide association study structural equation modeling to construct circulatory plaque burden (CPB) and integrated multi-omic data to prioritize candidate pathways. We then tested intermittent fasting in a baseline-controlled Ldlr⁻/⁻ mouse induction–intervention model and evaluated the BMAL1–MERTK efferocytosis axis in macrophages. Systems-genetic analyses linked systemic atherosclerotic burden to nutrient-response, circadian, and efferocytosis-related pathways. In mice, intermittent fasting reduced plaque size relative to both the ad libitum-fed and week-14 Baseline groups and improved structural features of plaque stability without additional endpoint lipid lowering. Aortic and single-cell analyses prioritized macrophage BMAL1–MERTK signaling. BMAL1 activated the Mertk promoter and occupied the tested promoter region. Mertk re-expression rescued the efferocytosis defect caused by Bmal1 knockdown, whereas MERTK inhibition reversed the benefit of BMAL1 overexpression. In this mouse model, intermittent fasting promoted remodeling of established atherosclerotic plaques, with macrophage BMAL1–MERTK signaling and efferocytosis identified as contributing mechanisms. These findings provide mechanistic insight and highlight this lifestyle-linked axis as a candidate for future translational investigation alongside established lipid-lowering therapy. Cardiovascular disease is often driven by atherosclerosis, in which fat, dead cells, and inflammatory debris accumulate within artery walls. Cholesterol-lowering medicines are effective but do not always stop established plaques from progressing. In mice, intermittent fasting reduced plaque size and improved features of stability despite endpoint blood lipid levels similar to those in freely fed controls. Macrophages act as "garbage collectors" by removing dead cells from plaques. Our experiments showed that BMAL1 increased MERTK expression and supported this clearance process; restoring Mertk rescued impaired clearance when Bmal1 was reduced. By integrating human genetics with animal and cell experiments, our study suggests that intermittent fasting may promote plaque remodeling partly through macrophage efferocytosis. These preclinical findings require further testing in humans.

This study investigated how defatted silkworm pupae meal (DSPM) modulates flesh quality in pearl gentian grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂). A total of 360 size-uniform fish were randomly assigned to four dietary treatments for 8 weeks: D0, the fishmeal (FM) based control diet; and D1, D2, and D3, in which DSPM substituted 25%, 50%, and 100% of dietary FM, respectively, with three replicates per treatment and 30 fish per replicate. Compared with D0, D1 and D2 improved final body weight, weight gain rate, specific growth rate, and feed conversion ratio, whereas D3 reduced survival and feed utilization. For flesh quality, D2 showed the most favorable phenotype, characterized by higher crude protein, collagen, flavor-associated amino acids, pH, texture attributes, and muscle fiber density, together with lower ether extract, cooking loss, freezing loss, and shear force. DSPM substitution also increased unsaturated and polyunsaturated fatty acid proportions and improved lipid health indices. Moderate substitution, especially D2, enhanced oxidative stability by increasing SOD and CAT activities and reducing MDA accumulation. Non-targeted metabolomics combined with qPCR analysis of selected flesh quality-related genes suggested that these improvements were associated with osmotic regulation, collagen remodeling, purine metabolism, membrane lipid homeostasis, nutrient sensing, myogenesis, and antioxidant defense. Complete FM substitution impaired muscle structure, water retention, and redox balance. Thus, 50% FM substitution with DSPM effectively optimized grouper flesh quality in this study.

This study investigated the effects of dietary n-3/n-6 highly unsaturated fatty acid (HUFA) ratios on growth performance, hepatic health, antioxidant capacity, and immune function in hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂). Six isonitrogenous and isolipidic diets were formulated with graded n-3/n-6 HUFA ratios of 25.29 (Y1), 19.54 (Y2), 13.27 (Y3), 7.74 (Y4), 3.96 (Y5), and 1.90 (Y6), and fed for 8 weeks. Optimal specific growth rate and protein efficiency ratio were observed at ratios 7.74–13.27, indicating superior nutrient utilization within this range. Hepatic lipid accumulation was lowest at a ratio of 19.54, as evidenced by minimal triglyceride content and lipid droplet area, whereas both excessively high and low ratios stimulated fatty acid synthase activity and promoted steatosis. The strongest antioxidant defense was observed at 7.74, characterized by upregulated glutathione peroxidase (gpx) and catalase (cat) gene expression, elevated glutathione peroxidase activity, induction of heat shock proteins (hsp70/hsp90), and the lowest malondialdehyde levels. Hepatocellular integrity was best preserved at 13.27, reflected by minimal glutamate pyruvate transaminase activity. Furthermore, this ratio enhanced humoral immunity—marked by the highest serum IgM—and shifted the inflammatory balance toward anti-inflammatory dominance through downregulation of interleukin 6 (il-6) and interleukin 8 (il-8) and upregulation of interleukin 10 (il-10) and transforming growth-factor β (tgf-β). Collectively, dietary n-3/n-6 HUFA ratios modulate growth and liver health in hybrid grouper by coordinately regulating protein utilization, lipid metabolism, redox homeostasis, and immune responses, with a functional optimum between 7.74 and 13.27 for integrated physiological performance.

BACKGROUND AND AIMS:Excessive hepatic lipid production drives disrupted lipid homeostasis. Although deubiquitinases have emerged as key modulators of metabolic enzymes, their involvement in de novo lipogenesis (DNL) remains elusive. RESULTS:Hepatic USP20 is identified by the multi-screening system based on biological assays of the constructed 105 DUB-plasmids library, combined with transcript analysis of multi-species with MASLD. USP20 activates the DNL pathway through multi-omics integrated analysis and isotope-metabolic flux tracing. The most potential USP20-interacting protein, FASN is screened by IP-MS. USP20 interacts with FASN and deubiquitinates K48-linked FASN to block proteasome-mediated degradation. Consequently, downstream lipogenic genes are activated in DNL pathway and excessive PA occurs in hepatocytes. Through rank by virtual screening, based on the pocket alignment of USP20 and molecular dynamics simulation, the small molecule compound-C2 screened markedly inhibits USP20, thereby suppressing DNL to alleviate MASLD. CONCLUSIONS:Our finding establishes USP20 as a central driver of DNL via deubiquitinating FASN. A promising therapeutic strategy targeting-USP20 is provided for MASLD.

Heat stress is a major environmental challenge in aquaculture that disrupts multiple physiological processes in fish, including gastrointestinal function. The intestine is increasingly recognized as a heat-sensitive organ. Chlorogenic acid (CGA) has intestinal microbiota-modulating properties, but its protective role against heat stress-induced intestinal barrier dysfunction remains unclear. In this study, hybrid grouper (Epinephelus fuscoguttatus♀ ×Epinephelus lanceolatus♂) were fed diets with or without CGA (400 mg/kg) for 56 days, after which they were exposed to either 28°C (control) or 35°C (heat stress), resulting in four treatment groups as CON, CGA, CONHT and CGAHT, respectively. Compared with the control group, dietary CGA supplementation alone significantly increased intestinal villus height, upregulated the expression of tight junction genes and proteins, elevated anti-inflammatory cytokine levels, and improved the intestinal microbiota composition. Heat stress markedly reduced intestinal villus height from 641 to 343 μm, whereas dietary CGA supplementation increased villus height to 488 μm. Heat stress also significantly downregulated the expression of intestinal tight junction genes and proteins, and disrupted tight junction structure; these changes were alleviated by CGA supplementation. In addition, heat stress increased serum D-lactic acid, endothelin-1, and lipopolysaccharide levels, while decreasing diamine oxidase activity, whereas CGA supplementation reversed these effects. Heat stress also reduced intestinal trypsin (from 2098.16 to 1746.55 U/mg pro) and amylase activities, while CGA supplementation restored both. Moreover, heat stress increased pro-inflammatory gene and protein expression but decreased anti-inflammatory gene expression. CGA supplementation suppressed pro-inflammatory responses and enhanced anti-inflammatory gene levels. Heat stress also reduced the Simpson index and beneficial bacteria while increasing harmful taxa; these alterations were ameliorated by CGA supplementation. Overall, dietary CGA mitigated heat stress-induced intestinal barrier injury in hybrid grouper by improving digestive function, reducing inflammation, preserving structural integrity, and modulating intestinal microbiota, indicating its potential as a dietary strategy to alleviate heat stress-related intestinal damage.

Cadmium (Cd), a widespread aquatic pollutant, induces dose-dependent gill epithelial damage in hybrid grouper (Epinephelus fuscoguttatus × Epinephelus lanceolatus). However, the toxicological mechanisms underlying its effects on intestinal mucosal barrier function remain unclear. This study examined intestinal mucosal barrier injury in hybrid grouper exposed to three Cd concentrations: 0 mg/L (control, CON), 0.0363 mg/L (Cd1%), and 0.363 mg/L (Cd10%). Cd exposure significantly inhibited intestinal trypsin and lipase activities compared with the control group. In addition, Cd exposure reduced villus height and muscularis propria thickness, disrupted tight junction structures, and induced mitochondrial vacuolization. Cd exposure also significantly downregulated the expression of occludin1, ZO-1, and claudin7a. Compared to the control group, the Cd10% group showed fewer intestinal goblet cells, accompanied by mitochondrial calcium deposition. Analysis of the intestinal microbiota revealed that Cd exposure significantly decreased the diversity of the intestinal microbiota. Specifically, compared with the control group, Cd exposure significantly increased the relative abundance of Pseudomonadota, while concurrently decreasing the relative abundances of Bacteroidota and Bacillota. Additionally, Cd exposure reduced Prevotellaceae (including Prevotella ruminicola) but increased Vibrionaceae (particularly Vibrio and V. vulnificus CMCP6). Functional prediction analysis revealed that Cd exposure significantly suppressed the digestive system pathways in the intestinal microbiota. Intestinal gene expression analysis further showed that the Cd10% group exhibited downregulation of MHC, TGF, and NRF2 and upregulation of IL-1β relative to the control group. In summary, Cd exposure impairs digestive function, induces structural damage to intestinal tissues, disrupts intestinal microbiota homeostasis, compromises intestinal barrier integrity, and elevates inflammation risk in hybrid grouper. The findings establish a theoretical framework for both ecological risk assessment of Cd exposure and sustainable practices in aquaculture.

The present study investigated the effects of dietary n-3 highly unsaturated fatty acids (n-3 HUFA) on the growth, antioxidant capacity, and ovarian development of female Litopenaeus vannamei broodstock. Five experimental diets with varying levels of n-3 HUFA (2.00 %, 2.50 %, 3.50 %, 5.00 %, and 7.00 %) were formulated and fed to female shrimp over a 10-week period. The growth performance, biochemical indices in the hepatopancreas, ovary, and serum, as well as antioxidant enzyme activities, were measured. The results indicated that moderate n-3 HUFA supplementation (3.50-5.00 %) significantly enhanced weight gain (WG), specific growth rate (SGR), and gonadosomatic index (GSI), while higher or lower levels of n-3 HUFA did not show significant improvement in these parameters. Antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px), showed optimal activities in shrimp fed diets with 3.50-5.00 % n-3 HUFA. Furthermore, serum and ovarian biochemical indicators, such as total cholesterol (TC), triglycerides (TG), and glucose (Glu), were positively influenced by dietary n-3 HUFA. Hormonal analysis revealed that estradiol (E2) and vitellogenin (VTG) levels in the serum and ovaries were significantly higher in the 5.00 % n-3 HUFA group, supporting enhanced reproductive performance. We recommend an n-3 HUFA level of 5.28-5.48 % in feed, based on the SGR and GSI fitting curves.

This study evaluated the effects of replacing inorganic trace elements with organic forms in Tenebrio molitor-based diets on the growth performance, antioxidant capacity, immune response, and intestinal microbiota of Pacific white shrimp (Litopenaeus vannamei). A total of 480 juvenile shrimp (initial body weight 0.26 +/- 0.01 g) were randomly assigned to four dietary treatments with three replicates per treatment for a 10-week feeding trial. A fishmeal-based diet supplemented with inorganic trace elements served as the control. In the negative control, 40 % of fishmeal was replaced with T. molitor. Based on this diet, inorganic iron, manganese, and zinc were replaced with 40 % or 60 % organic forms. Shrimp fed the diet containing 60 % organic trace elements showed significantly lower final body weight, weight gain rate, and specific growth rate, as well as a higher feed conversion ratio, compared with the 40 % replacement group. Although fishmeal replacement with T. molitor reduced antioxidant capacity, supplementation with organic trace elements significantly increased the activities of superoxide dismutase, catalase, and glutathione peroxidase. The activities of acid phosphatase and alkaline phosphatase and the expression of immune-related genes were significantly enhanced in shrimp fed the diet containing 40 % organic trace elements. In addition, organic trace element supplementation partially improved intestinal microbiota composition by reducing potential pathogenic bacteria. In conclusion, the 40 % replacement of inorganic iron, manganese, and zinc with organic forms was identified as the optimal proportion, as it enhanced antioxidant and immune responses without compromising growth performance in L. vannamei.

Leopard coral grouper (Plectropomus leopardus) is a high-value marine species increasingly produced under intensive farming conditions; however, its dietary lipid requirement remains poorly defined, hindering the development of cost-effective formulated feeds. This study aimed to determine the optimal dietary lipid level for juvenile P. leopardus by assessing growth performance, hepatic lipid metabolism, and antioxidant capacity in response to graded lipid inclusion. Juveniles (initial body weight [IBW]: 13.94 ± 0.07 g) were fed six isonitrogenous diets (53% crude protein) containing 6%, 8%, 10%, 12%, 14%, or 16% lipid for 9 weeks (three replicate tanks per diet; 27 fish per tank). Weight gain (WG) and specific growth rate (SGR) increased with dietary lipid up to 10% and then declined. Intestinal lipase activity increased with dietary lipid level. Dietary lipid at 8%-12% improved hepatic lipid metabolic balance, as indicated by reduced activities of lipogenic enzymes (malate dehydrogenase [MDH], glucose-6-phosphate dehydrogenase [G6PD], and fatty acid synthase [FAS]) and increased lipoprotein lipase (LPL) activity (p < 0.05). Dietary lipid at 8%-12% also enhanced antioxidant defenses, reflected by higher glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities in serum, liver, and hindgut, without an increase in malondialdehyde (MDA). In contrast, high-lipid diets (14%-16%) induced hepatic lipid metabolic imbalance followed by pronounced hepatic lipid deposition (histological staining and increased hepatic crude lipid; p < 0.05), elevated serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and significantly increased MDA levels in serum, liver, and hindgut (p < 0.05), while antioxidant enzyme activities tended to decrease, indicating heightened oxidative stress. Overall, growth performance was highest at ~10% dietary lipid under the present experimental conditions, and quadratic regression analysis suggested an approximate optimum near 9.3%. Moderate dietary lipid (8%-12%) promoted growth by improving hepatic lipid metabolism and antioxidant capacity, whereas excessive lipid inclusion (≥14%) induced excessive hepatic lipid accumulation and oxidative damage in P. leopardus.

The cardiac burden associated with doxorubicin (DOX) significantly limits its application in cancer treatment. Therefore, it is essential to identify effective strategies to protect the heart from cardiotoxic damage caused by chemotherapy. As sex is among the risk factors associated with DOX-induced cardiotoxicity, whether the cardiac beneficial effects observed from male mice can be applied to female mice remains unknown. We established a two-week DOX-induced cardiotoxicity model, in which the cumulative DOX dose administered to mice was comparable to that used in previous research. This model effectively induces cardiotoxicity and fibrosis while allowing for a sufficiently long monitoring period to evaluate the chemotherapeutic effects of DOX on tumors, without imposing an excessive physiological burden on the mice from prolonged tumor growth. Utilizing this tumor-bearing murine model, we employed TC-1 cancer cells, which express HPV16-E6 and HPV16-E7 proteins, to investigate the cardioprotective effects of circ-ZNF609 inhibition in DOX-treated tumor-bearing female mice. Our findings indicate that cardiac inhibition of circ-ZNF609 protects against DOX-induced cardiotoxicity without compromising the anti-tumor efficacy of DOX in females. These results suggest that targeting circ-ZNF609 in the heart may represent a promising and viable therapeutic strategy for preventing DOX-induced cardiotoxicity.

Selenium biofortification in soybean is one approach to addressing widespread selenium deficiency in human populations, yet the temporal dynamics of gene expression and antioxidant responses following selenium enrichment remain poorly characterized. Here, we examined the physiological and transcriptional responses of the HK88 soybean variety to selenium-enriched nutrient solution treatment, with seedlings sampled at 1, 24, and 48 h post-treatment. Total tissue selenium rose from 0.01 mg/kg at 1 h to 0.33 mg/kg at 48 h, with parallel increases in both inorganic and organic fractions, consistent with active biotransformation, though this interpretation remains to be confirmed experimentally. Antioxidant responses followed a distinct temporal pattern: superoxide dismutase (SOD) and peroxidase (POD) activities were initially lower in treated plants at 1 h relative to controls but were elevated at 24 and 48 h, while catalase (CAT) activity remained comparatively low across all time points. Malondialdehyde (MDA) levels were lower in selenium-treated plants at 1 h, suggesting early membrane stabilization, though this difference was no longer apparent by 48 h. RNA sequencing of 18 libraries identified 6,793 differentially expressed genes (DEGs) at 1 h, peaking at 13,196 (approximately 18% of the 72,513 annotated genes) at 24 h, then declining to 8,996 at 48 h. A Venn diagram analysis identified 565 DEGs shared across all three time points, comprising 196 consistently up-regulated and 369 consistently down-regulated genes. Up-regulated genes were enriched for nucleotide transmembrane transport functions, including ATP, ADP, and purine transport, with associated ABC transporter activity. Down-regulated genes were predominantly associated with primary carbon metabolism, including monosaccharide biosynthesis, gluconeogenesis, and the Calvin cycle. Gene Set Variation Analysis (GSVA) indicated positive enrichment scores for nucleotide transport pathways at 24 and 48 h in treated plants, contrasting with negative scores in controls. Mantel tests revealed significant associations between gene set activity profiles and measured physiological traits, particularly for gene sets related to molecular function and selenium accumulation. These findings suggest that selenium biofortification in HK88 is associated with a coordinated metabolic shift, in which primary carbon fixation is reduced while nucleotide transport capacity is enhanced, supporting antioxidant defense during selenium assimilation.

Lead (Pb) is a well-known toxic heavy metal that induces oxidative stress, tissue damage, and neurotoxicity in several aquatic species including Litopenaeus vannamei. However, the effective mitigation strategies remain limited. Astragalus polysaccharide (APS) is recognized for its antioxidant and detoxifying properties. In this study, the mitigating role of dietary APS against chronic Pb-induced toxicity we investigated in the L. vannamei from the aspects of oxidative stress, metabolism dysfunction, and intestinal microbiota. Shrimp with an initial weight of 0.40 +/- 0.01 g were reared for 56 days under four treatments: no Pb and no APS (CG), Pb exposure without APS (P0), Pb exposure with 0.2 g/kg APS (P0.2), and Pb exposure with 0.4 g/kg APS (P0.4). The results showed that Pb exposure caused hepatopancreatic damage and reduced survival, whereas dietary supplementation with 0.4 g/kg APS effectively alleviated hepatopancreatic injury and improved survival. Pb exposure significantly suppressed intestinal amylase and lipase activities, as well as hepatic acetylcholinesterase and glutathione Stransferase activities. Pb exposure also increased hepatic oxidative stress indicators, including catalase, superoxide dismutase, and glutathione peroxidase activities, and elevated reactive oxygen species and malondialdehyde levels. Notably, APS supplementation reversed these Pb-induced changes. Pb exposure significantly decreased the Shannon, Simpson, Chao, and ACE indices and reduced the relative abundance of beneficial bacteria (including Psychrobacter, Bacillus, and Demequina), while increasing harmful taxa (including Vibrionaceae and Flavobacteriaceae, as well as Photobacterium and Vibrio). These alterations were also reversed by APS supplementation. Metabolomic analysis indicated that Pb exposure increased intestinal gamma-aminobutyric acid and acetylcholine levels and decreased D-erythro-imidazolylglycerol phosphate and phenylpyruvate levels, whereas these metabolites were normalized by dietary APS at 0.4 g/kg. In conclusion, dietary supplementation with 0.4 g/kg APS effectively alleviated Pb-induced oxidative stress, neurotoxicity, metabolic dysfunction, and intestinal dysbiosis in L. vannamei.