The current study investigated the efficacy of a probiotic mixture on ameliorating heat stress-induced impairment of intestinal microflora, morphology, and barrier integrity in broilers. The probiotic mixture contained Bacillus licheniformis, Bacillus subtilis, and Lactobacillus plantarum. Three hundred sixty 21-d-old Ross 308 male broilers were allocated in 4 experimental treatments, each of which was replicated 6 times with 15 broilers per replicate. A 2 × 2 factorial design was used in the study, and the main factors were composed of diet (basal diet or addition of 1.5 g/kg of probiotic mixture) and temperature (thermoneutral zone or heat stress). From d 22 to 42, birds were either raised in a thermoneutral zone (22°C) or subjected to cyclic heat stress by exposing them to 33°C for 10 h (from 0800 to 1800) and 22°C from 1800 to 0800. Compared with birds kept in the thermoneutral zone, birds subjected to heat stress had reduced ADG and ADFI; lower viable counts of Lactobacillus and Bifidobacterium and increased viable counts of coliforms and Clostridium in small intestinal contents; shorter jejunal villus height, deeper crypt depth, and lower ratio of villus height to crypt depth; decreased jejunal transepithelial electrical resistance and a higher level of jejunal paracellular permeability of fluorescein isothiocyanate dextran 4 kDa; and downregulated protein levels of occludin and zonula occludens-1 (P < 0.05). Supplemental probiotics increased (P < 0.05) small intestinal Lactobacillus and Bifidobacterium, jejunal villus height, protein level of occludin, and decreased (P < 0.05) feed to gain ratio and small intestinal coliforms. These results indicate that dietary addition of probiotic mixture was effective in partially ameliorating intestinal barrier function. But no temperature × diet interaction was observed in the present study, revealing that the supplemented probiotics had the same effect at both temperatures.
Long non-coding RNAs (lncRNAs) are important regulators of diverse biological processes. Here we report on functional identification and characterization of a novel long intergenic non-coding RNA with MyoD-regulated and skeletal muscle-restricted expression that promotes the activation of the myogenic program, and is therefore termed Linc-RAM (Linc-RNA Activator of Myogenesis). Linc-RAM is transcribed from an intergenic region of myogenic cells and its expression is upregulated during myogenesis. Notably, in vivo functional studies show that Linc-RAM knockout mice display impaired muscle regeneration due to the differentiation defect of satellite cells. Mechanistically, Linc-RAM regulates expression of myogenic genes by directly binding MyoD, which in turn promotes the assembly of the MyoD–Baf60c–Brg1 complex on the regulatory elements of target genes. Collectively, our findings reveal the functional role and molecular mechanism of a lineage-specific Linc-RAM as a regulatory lncRNA required for tissues-specific chromatin remodelling and gene expression.
Skeletal muscle stem cells, called satellite cells, are a quiescent heterogeneous population. Their heterogeneity is influenced by Pax7, a well-defined transcriptional regulator of satellite cell functions that defines two subpopulations: Pax7 Hi and Pax7 Lo . However, the mechanisms by which these subpopulations are established and maintained during myogenesis are not completely understood. Here we show that miR-431, which is predominantly expressed in the skeletal muscle, mediates satellite cell heterogeneity by fine-tuning Pax7 levels during muscle development and regeneration. In miR-431 transgenic mice, the Pax7 Lo subpopulation is enriched, enhances myogenic differentiation and accelerates muscle regeneration. Notably, miR-431 attenuates the muscular dystrophic phenotype in mdx mice and may be a potential therapeutic target in muscular diseases. miR-431 transgenic mice are a unique genetic model for investigating the cellular features and biological functions of Pax7 Lo satellite cells during muscle development and regeneration.
Two hundred four broilers (1-d-old) were randomly allocated to 4 treatments, each of which had 3 pens of 17 chicks per pen and were used to investigate the effects of beta-mannanase (Hemicell) on growth performance and immunity. The chicks received the same basal diet based on corn-soybean meal and Hemicell was added to the basal diet at 0, 0.025, 0.05, and 0.075%, respectively. Weight of each replicate was determined at wk 0, 3, and 6 of age. There were no significant differences in average feed intake in the 0- to 3-wk and 0- to 6-wk periods, and no differences in serum IgA, or IgG concentrations. However, the addition of Hemicell significantly increased (P < 0.05) weight gain in the 4- to 6-wk and 0- to 6-wk periods. Feed conversion for the 0.025 and 0.05% groups was significantly greater (P < 0.05) than for the control group in the 4- to 6-wk and 0- to 6-wk periods. Hemicell significantly increased (P < 0.05) the serum IgM concentration in 3- and 6-wk-old broilers. Proliferation of T lymphocytes in 6-wk-old broilers for the 0.05% group was also improved (P < 0.05) significantly. The results indicate that Hemicell may improve growth performance and immunity of broilers.
The effects of supplemental dietary threonine (Thr) on laying performance, expression of intestinal mucin 2 (MUC2) and secretory IgA (sIgA), and intestinal microbiota of laying hens fed a low CP diet were investigated. A total of 240 Lohmann Brown laying hens, 28 wk of age, was allocated to 3 dietary treatments, each of which included 5 replicates of 16 hens. Hens were fed a control diet (16% CP), a low CP diet (14% CP), or a low CP diet supplemented with 0.3% L-Thr for 12 weeks. Chemical analyses of the diets for Thr are 0.49, 0.45, and 0.69%, respectively. Lowering dietary CP impaired egg production and egg mass of laying hens. Dietary Thr supplementation to the low CP diet increased (P < 0.05) egg production and egg mass. In addition, ileal sIgA contents and MUC2 and sIgA mRNA expression were increased (P < 0.05) by dietary Thr addition. Dietary CP reduction reduced (P < 0.05) intestinal bacterial diversity, whereas dietary Thr supplementation to the low CP diet recovered the bacteria diversity and significantly increased the abundance of potential beneficial bacteria. In conclusion, dietary Thr supplementation to a low CP diet could affect intestinal health and hence productivity via regulating intestinal mucin and sIgA expression, and microbial population of laying hens.
The topical application of exosomes secreted by mesenchymal stem cells (MSC-Exos) on the skin is a very new and interesting topic in the medical field. In this study, we aimed to investigate whether marine sponge Haliclona sp. spicules (SHSs) could effectively enhance the skin delivery of human umbilical cord-derived MSC-Exos (hucMSC-Exos), and further evaluate the topical application of hucMSC-Exos combined with SHSs in rejuvenating photoaged mouse skin. Materials and Methods: SHSs were isolated from the explants of sponge Haliclona sp. with our proprietary method, and hucMSC-Exos were prepared from the conditioned medium of hucMSCs using ultracentrifugation. The effects of SHSs on the skin penetration of fluorescently labeled hucMSC-Exos were determined using confocal microscopy in vitro (porcine skin) and in vivo (mouse skin). The therapeutic effects of hucMSC-Exos coupled with SHSs against UV-induced photoaging in mice were assessed by using microwrinkles analysis, pathohistological examination and real-time RT-PCR. We also tested the skin irritation caused by the combination of hucMSC-Exos and SHSs in guinea pigs. Results: In vitro results showed that hucMSC-Exos could not readily penetrate through porcine skin by themselves. However, SHSs increased the skin absorption of exosomes by a factor of 5.87 through creating microchannels. Similar penetration enhancement of hucMSC-Exos was observed after SHSs treatment in mice. The combined use of hucMSC-Exos and SHSs showed significant anti-photoaging effects in mice, including reducing microwrinkles, alleviating histopathological changes, and promoting the expression of extracellular matrix constituents, whereas hucMSC-Exos alone produced considerably weaker effects. Skin irritation test showed that the combination of hucMSC-Exos and SHSs caused slight irritation, and the skin recovered shortly. Conclusion: SHSs provide a safe and effective way to enhance the skin delivery of MSC-Exos. Moreover, the combination of MSC-Exos and SHSs may be of much use in the treatment of photoaging.
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