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Iso-nitrogenous and iso-lipidic diets containing 0%, 3%, 6%, 9%, and 12% hydrolyzed porcine mucosa (namely, HPM0, HPM3, HPM6, HPM9, and HPM12) were prepared to evaluate their effects on the growth performance, muscle nutrition composition, texture property, and gene expression related to muscle growth of hybrid groupers ( Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂). Groupers were fed to apparent satiation at 08:00 and 16:00 every day for a total of 56 days. It was found that the weight gain percentage in the HPM0, HPM3, and HPM6 groups did not differ ( P > 0.05). The cooking loss and drip loss of the dorsal muscle in the HPM3 group were lower than those in the HPM6 and HPM9 groups ( P < 0.05). The hardness and chewiness of the dorsal muscle in the HPM3 group were higher than those in the HPM0, HPM9, and HPM12 groups ( P < 0.05). The gumminess in the HPM3 group was higher than that in the HPM9 and HPM12 groups ( P < 0.05). The total essential amino acid content of the dorsal muscle in the HPM12 group was higher than that in the HPM0 group ( P < 0.05). The contents of total n-3 polyunsaturated fatty acid and total n-3 highly unsaturated fatty acid, as well as the ratio of n-3/n-6 polyunsaturated fatty acid in the dorsal muscle was higher in the HPM0 group than in all other groups ( P < 0.05). The relative expressions of gene myogenic factor 5, myocyte enhancer factor 2c, myocyte enhancer factor 2a, myosin heavy chain, transforming growth factor-beta 1 ( TGF - β1 ), and follistatin ( FST ) were the highest in the dorsal muscle of the HPM3 group. The results indicated that the growth performance of hybrid grouper fed a diet with 6% HPM and 27% fish meal was as good as that of the HPM0 group. When fish ingested a diet containing 3% HPM, the expression of genes TGF-β1 and FST involved in muscle growth were upregulated, and then the muscle quality related to hardness and chewiness were improved. An appropriate amount of HPM could be better used in grouper feed.

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In this study, five iso-nitrogenous and iso-lipidic experimental low-FM diets were formulated with enzyme-digested hydrolysed porcine mucosa (EH) at concentrations of 0 g/kg (EH0, controls), 30 g/kg (EH3), 60 g/kg (EH6), 90 g/kg (EH9) and 120 g/kg (EH12). Juvenile hybrid groupers (n = 450; initial body weight 7.50 ± 0.02 g) were randomly divided into 15 tanks and fed the experimental diets twice daily for 56 days. The weight gain rate, specific growth rate, protein efficiency rate, feed conversion rate and feed intake rate of group EH3 were not significantly different from those of controls. The villus length and muscle thickness of the distal intestine (DI) in group EH12 and the villus width of the proximal intestine (PI) in group EH6 were significantly higher than those in controls. The Shannon index was significantly higher in group EH6 than in other groups. Group EH6 exhibited 17 pathway subclasses with significant differences to the other groups, comprising organismal systems (4), human diseases (6), cellular processes (2), metabolism (4) and environmental information processing (1). The expressions of CAT1, CAT2, LAAT1 and SNAT3 in the PI, and PAT1, B 0 AT3 and B0, +AT in the mid intestine (MI), and EAAT1 and PepT1 in the PI and MI in group EH6 were significantly higher than those in controls. The expressions of CAT1, CAT2, LAAT1, PAT1, SNAT3, EAAT1, B 0, + AT and PepT1 in the DI of group EH3 were significantly greater than those in controls. The maximum amount of EH that could be added to low-FM feed to improve the digestive enzyme activity and amino acid and small peptide transport ability of the DI without obviously affecting the growth performance of hybrid groupers was 30 g/kg. EH contents ≥ 60 g/kg had negative impacts on growth performance and feed efficiency.

Neutrophils are primary effector cells of the innate immune system. Emerging evidence has consistently shown that activated neutrophils produce and release neutrophil extracellular traps (NETs) that play roles in immunity and non-infectious diseases. NETs are composed of DNA and proteins and serve as a structural platform for pathogen sequestration and degradation. In contrast to their protective role during pathogenic infection, NETs are pathologically involved in cardiovascular disease (CVD). In this review, we introduce the formation, release, and clearance of NETs and the regulatory mechanisms of NETs formation, followed by an overview of the clinical evidence for the involvement of NETs in CVD. Because atherosclerosis is a fundamental part of the pathogenesis of CVD, we chose to focus on the mechanisms by which NETs promote endothelial cell damage and collaborate with macrophages and platelets to accelerate plaque progression and thrombosis. Finally, we present options for clinical intervention to inhibit NETs production and release in the treatment of CVD. In conclusion, this review integrates the latest findings and provides new insights into NETs, which represent a novel biomarker and therapeutic target in clinical practice.