Dietary supplementation with a silicate clay mineral (palygorskite) alleviates inflammatory responses and intestinal barrier damage in broiler chickens challenged with Escherichia coli.

This experiment aimed to explore the protective effects of dietary palygorskite (Pal) supplementation on inflammatory responses and intestinal barrier function of broiler chickens challenged with Escherichia coli (E. coli). A 2 × 2 factorial arrangement was designed to assess the effects of Pal administration (0 or 5 g/kg of feed) and E. coli challenge (E. coli or bacterial culture medium) on broilers in a 21-d feeding trial. Birds were randomly assigned into one of the 4 groups, and each group had 8 replicates with ten birds each. The challenged chickens were orally gavaged with E. coli suspended in Luria-Bertani broth on 14 d of age, while unchallenged birds were administrated with an equivalent amount of culture medium. The sampling was performed at 21 d of age. Compared with the normal birds, an oral E. coli challenge reduced final body weight, and decreased feed intake, weight gain, and feed efficiency during the challenge period (P < 0.05). E. coli challenge promoted colonization of E. coli in cecal content and their translocation to internal organs (heart, liver, and spleen) (P < 0.05). E. coli infection also increased levels of pro-inflammatory cytokines in jejunum and ileum possibly through activating the toll-like receptor-4-mediated signaling pathway (P < 0.05). Moreover, E. coli administration increased intestinal mucosal permeability (higher serum D-lactate level and diamine oxidase activity, and lower intestinal mucosal disaccharidase activities), altered intestinal morphology, and downregulated the gene expression of intestinal tight junction proteins (P < 0.05). In contrast, Pal supplementation enhanced growth performance, inhibited colonization of E. coli, reduced intestinal inflammation, decreased intestinal permeability, restored intestinal morphology, and normalized the expression of genes responsible for inflammatory processes and maintenance of intestinal mucosal barrier (P < 0.05), and most of these beneficial effects resulting from Pal administration were independent of bacterial challenge. The results indicated dietary Pal incorporation was effective in improving growth performance and alleviating inflammation and intestinal mucosal barrier damage in broilers challenged with E. coli.

Protective effects of chlorogenic acid on the intestinal barrier of broiler chickens: an immunological stress model study.

This study was conducted to investigate the protective effects of chlorogenic acid (CGA) on inflammatory responses and intestinal health of lipopolysaccharide (LPS)-challenged broilers. One hundred and forty-four 1-day-old male broiler chicks were divided into 3 groups with 6 replicates of 8 birds each. The groups were as follows: 1) Control group: birds fed a basal diet; 2) LPS group: LPS-challenged birds fed a basal diet; 3) CGA group: LPS-challenged birds fed a CGA-supplemented diet. The LPS was intraperitoneally administered at a dose of 1 mg/kg of body weight. CGA increased the weight gain and feed intake of LPS-challenged birds by 37.05% and 24.29%, respectively (P < 0.05). CGA also alleviated LPS-induced inflammation, as evidenced by lower levels of pro-inflammatory cytokines in the serum and jejunum (tumor necrosis factor-α, interferon-γ, interleukin-1ß, and interleukin-6), and the decreased myeloperoxidase activity in the jejunum (P < 0.05). These effects were accompanied by a decrease in the mRNA abundance of toll-like receptor 4 and myeloid differentiation factor 88 and an inhibition of nuclear factor kappa-B translocation in the jejunum (P < 0.05). CGA reduced circulating diamine oxidase activity and levels of D-lactate and endotoxin, and positively regulated the expression of jejunal claudin-3 and zonula occludens-1 in LPS-challenged broilers (P < 0.05). Compared to the LPS group, CGA reduced the apoptotic rate of epithelial cells and cytochrome c concentration in the jejunum, and normalized the expression of genes responsible for proliferation and apoptosis in jejunal epithelial cells, including cysteine aspartate-specific protease-9, B cell lymphoma-2, and proliferating cell nuclear antigen (P < 0.05). Furthermore, CGA normalized the altered phosphorylation of protein kinase B and glycogen synthase kinase-3ß, as well as the translocation of nuclear ß-catenin in the jejunum of LPS-challenged broilers (P < 0.05). These results suggested that CGA supplementation improved growth performance, alleviated inflammation, and helped maintain intestinal integrity and barrier function in LPS-challenged broilers, possibly through the regulation of the toll-like receptor 4/nuclear factor kappa-B and protein kinase B/Wnt/ß-catenin pathways.

Enhanced CD19 activity in B cells contributes to immunodeficiency in mice deficient in the ICF syndrome gene Zbtb24.

Immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome is a rare autosomal recessive disorder characterized by DNA hypomethylation and antibody deficiency. It is caused by mutations in DNMT3B, ZBTB24, CDCA7, or HELLS. While progress has been made in elucidating the roles of these genes in regulating DNA methylation, little is known about the pathogenesis of the life-threatening hypogammaglobulinemia phenotype. Here, we show that mice deficient in Zbtb24 in the hematopoietic lineage recapitulate the major clinical features of patients with ICF syndrome. Specifically, Vav-Cre-mediated ablation of Zbtb24 does not affect lymphocyte development but results in reduced plasma cells and low levels of IgM, IgG1, and IgA. Zbtb24-deficient mice are hyper and hypo-responsive to T-dependent and T-independent type 2 antigens, respectively, and marginal zone B-cell activation is impaired. Mechanistically, Zbtb24-deficient B cells show severe loss of DNA methylation in the promoter region of Il5ra (interleukin-5 receptor subunit alpha), and Il5ra derepression leads to elevated CD19 phosphorylation. Heterozygous disruption of Cd19 can revert the hypogammaglobulinemia phenotype of Zbtb24-deficient mice. Our results suggest the potential role of enhanced CD19 activity in immunodeficiency in ICF syndrome.

Astragaloside IV Protects Against IL-1ß-Induced Chondrocyte Damage via Activating Autophagy.

BACKGROUND: Osteoarthritis (OA) is a chronic inflammatory condition that affects the articular cartilage. Astragaloside IV (AS-IV) constitutes the primary active component of the Chinese herbal medicine Huangqi (Radix Astragali Mongolici). AS-IV demonstrates anti-inflammatory and anti-apoptotic attributes, exhibiting therapeutic potential across various inflammatory and apoptosis-related disorders. Nevertheless, its pharmaceutical effects in OA are yet to be fully defined. OBJECTIVES: This study aimed to investigate the protective impact of AS-IV on rat chondrocytes treated with IL-1ß and ascertain whether autophagy plays a role in this effect. METHODS: Chondrocytes were isolated and cultivated from the knee joints of neonatal SD mice. The study included the blank control group, the model group, and the AS-IV concentration gradient group (50, 100, 200 µmol/L) to intervene with chondrocytes. The MTT assay was employed to assess cell viability at varying culture periods, enabling the determination of suitable concentration and duration. Subsequently, chondrocytes were treated with the optimal AS-IV concentration and divided into three groups: the model group replicated IL-1ß-induced inflammatory chondrocyte injury, the AS-IV group received a co-culture of AS-IV and IL-1ß, and a blank control group was established. Changes in cell morphology and structure were observed using ghost pen cyclic peptide staining. ELISA was used to measure TNF-α and GAG levels in cell supernatants. RT-qPCR assessed p62 and LC3 mRNA expression, while Western Blot evaluated p62 and LC3Ⅱ/Ⅰ protein expression. RESULTS: AS-IV promoted chondrocyte proliferation and concurrently inhibited cell apoptosis. An optimal AS-IV dose of 200 µmol/L and a suitable reaction time of 48 h were identified. Ghost pen cyclic peptide staining indicated that the model group's cytoskeleton exhibited fusiform changes with reduced immunofluorescence intensity, as opposed to the blank control group. The AS-IV group displayed more polygonal cytoskeletal morphology with increased immunofluorescence intensity. AS-IV reduced TNF-α levels and elevated GAG levels in the culture supernatant. Additionally, AS-IV lowered p62 mRNA and protein expression while increasing LC3 mRNA expression in cultured chondrocytes. CONCLUSION: Our findings suggest that AS-IV mitigates inflammatory chondrocyte injury, safeguarding chondrocytes through a potential autophagy suppression mechanism. These results imply that AS-IV could offer preventive advantages for OA.

Efficacy of two different dosages of prednisone for treatment of subacute thyroiditis: a single-centre, prospective, randomized, open-label, non-inferiority trial.

INTRODUCTION: The study aimed to explore the efficacy and safety of low-dose (LD) and regular-dose (RD) prednisone (PDN) for the treatment of subacute thyroiditis (SAT). MATERIAL AND METHODS: Patients were randomly allocated using the block randomization method to the 2 groups. The primary outcome was the time required for PDN treatment. Secondary outcomes included percentages of relapse, mean score for the Morisky Medication Adherence Scale-8© (MMAS-8), time required for symptoms to resolve, cumulative PDN dose (mg), and mean erythrocyte sedimentation rate (ESR) at 2 weeks and at baseline. RESULTS: The study cohort included 77 patients, randomized 74 participants, and 68 completed the study. There was no significant difference in the treatment duration between the LD and RD groups (55.31 ± 14.05 vs. 61.25 ± 19.95 days, p = 0.053). The mean difference in the time required for PDN treatment between the LD and RD groups was -1.86 [95% confidence interval (CI) = -10.64 to 6.92] days, which was within the non-inferiority margin of 7 days. There was a significant difference in the mean score for MMAS-8 between the LD and RD groups (5.84 ± 0.88 vs. 5.33 ± 1.12, p = 0.031). Also, there was a significant difference in the cumulative PDN dose between the LD and RD groups (504.22 ± 236.86 vs. 1002.28 ± 309.86, p = 0.046). The ESR at 2 weeks was statistically significant compared to baseline values in both groups, with pre-treatment and post-treatment ESRs of 49.91 ± 24.95 and 17.91 ± 12.60/mm/h, (p < 0.0001) in the LD group and 65.08 ± 21.77 and 17.23 ± 13.61/mm/h (p < 0.0001) in the RD group. CONCLUSION: Low-dose PDN therapy may be sufficient to achieve complete recovery and better outcomes for SAT. This study is registered with the Chinese Clinical Trial Registry (02/10/2021 ChiCTR2100051762).

Effects of dietary supplementation with different levels of palygorskite-based composite on growth performance, antioxidant capacity, and meat quality of broiler chickens.

This study was conducted to investigate the effects of different levels of palygorskite-based composite (PBC) on growth performance, antioxidant status, and meat quality of broilers. A total of 320 one-day-old mixed-sex Ross 308 broiler chicks were allocated to 1 of 5 groups with 8 replicates of 8 birds each, and given a basal diet supplemented with 0, 250, 500, 1,000, and 2,000 mg/kg PBC for a 42-day trial, respectively. PBC quadratically increased feed efficiency during the late and overall experimental periods (P < 0.05). Compared with the control group, 1,000 mg/kg PBC increased feed efficiency during the overall period (P < 0.05). PBC linearly increased serum total superoxide dismutase (T-SOD) activity at 21 d and glutathione peroxidase (GSH-Px) activity at both 21 d and 42 d (P < 0.05). Compared with the control group, PBC supplementation, regardless of its level, increased 21-day serum SOD activity (P < 0.05). The 21-day serum GSH-Px activity was increased by PBC when its level exceeded 250 mg/kg (P < 0.05). PBC linearly increased 42-day total antioxidant capacity (T-AOC) activity, but linearly decreased 42-day malondialdehyde level in liver (P < 0.05). An addition of PBC, irrespective of its level, increased 42-day hepatic T-AOC activity (P < 0.05). PBC quadratically increased 45-min yellowness value and linearly increased 24-h pH value, but quadratically decreased 24-h lightness value and linearly and quadratically reduced 24-h drip loss in breast muscle (P < 0.05). Compared with the control group, the 24-h drip loss of breast muscle was decreased by PBC, regardless of its dosage (P < 0.05). An addition of PBC linearly increased 42-day T-AOC and T-SOD activities of breast muscle (P < 0.05). Compared with the control group, muscle T-SOD activity was increased by PBC, regardless of its administration level (P < 0.05). These results suggested that PBC could improve growth performance, antioxidant capacity, and meat quality of broilers, and its recommended dosage is 1,000 mg/kg.

Characterization of a mouse model of ICF syndrome reveals enhanced CD19 activation in inducing hypogammaglobulinemia.

Immunodeficiency, centromeric instability and facial anomalies (ICF) syndrome is a rare autosomal recessive disorder characterized by DNA hypomethylation and antibody deficiency. It is caused by mutations in DNMT3B, ZBTB24, CDCA7 or HELLS . While progress has been made in elucidating the roles of these genes in regulating DNA methylation, little is known about the pathogenesis of the life-threatening hypogammaglobulinemia phenotype. Here we show that mice deficient for Zbtb24 in the hematopoietic lineage recapitulate major clinical features of patients with ICF syndrome. Specifically, Vav-Cre-mediated ablation of Zbtb24 does not affect lymphocyte development but results in reduced plasma cells and low levels of IgM, IgG1 and IgA. Zbtb24 -deficient mice are hyper- and hypo-responsive to T-dependent and Tindependent type 2 antigens, respectively, and marginal zone B cell activation is impaired. B cells from Zbtb24 -deficient mice display elevated CD19 phosphorylation. Heterozygous disruption of Cd19 can revert the hypogammaglobulinemia phenotype in these mice. Mechanistically, Il5ra (interleukin-5 receptor subunit alpha) is derepressed in Zbtb24 -deficient B cells, and elevated IL-5 signaling enhances CD19 phosphorylation. Our results reveal a novel link between IL-5 signaling and CD19 activation and suggest that abnormal CD19 activity contributes to immunodeficiency in ICF syndrome. SIGNIFICANCE STATEMENT: ICF syndrome is a rare immunodeficiency disorder first reported in the 1970s. The lack of appropriate animal models has hindered the investigation of the pathogenesis of antibody deficiency, the major cause of death in ICF syndrome. Here we show that, in mice, disruption of Zbtb24 , one of the ICF-related genes, in the hematopoietic lineage results in low levels of immunoglobulins. Characterization of these mice reveals abnormal B cell activation due to elevated CD19 phosphorylation. Mechanistically, Il5ra (interleukin-5 receptor subunit alpha) is derepressed in Zbtb24 -deficient B cells, and increased IL-5 signaling enhances CD19 phosphorylation.

Effects of dietary supplementation with chlorogenic acid on growth performance, antioxidant capacity, and hepatic inflammation in broiler chickens subjected to diquat-induced oxidative stress.

This study was conducted to investigate the protective effects of chlorogenic acid (CGA) on broilers subjected to (DQ)-induced oxidative stress. In experiment 1, one hundred and ninety-two male one-day-old Ross 308 broiler chicks were distributed into 4 groups and fed a basal diet supplemented with 0, 250, 500, or 1,000 mg/kg CGA for 21 d. In experiment 2, an equivalent number of male one-day-old chicks were allocated to 4 treatments for a 21-d trial: 1) Control group, normal birds fed a basal diet; 2) DQ group, DQ-challenged birds fed a basal diet; and 3) and 4) CGA-treated groups: DQ-challenged birds fed a basal diet supplemented with 500 or 1,000 mg/kg CGA. The intraperitoneal DQ challenge was performed at 20 d. In experiment 1, CGA administration linearly increased 21-d body weight, and weight gain and feed intake during 1 to 21 d (P < 0.05). CGA linearly and/or quadratically increased total antioxidant capacity, catalase, superoxide dismutase, and glutathione peroxidase activities, elevated glutathione level, and reduced malondialdehyde accumulation in serum, liver, and/or jejunum (P < 0.05). In experiment 2, compared with the control group, DQ challenge reduced body weight ratio (P < 0.05), which was reversed by CGA administration (P < 0.05). DQ challenge increased serum total protein level, aspartate aminotransferase activity, and total bilirubin concentration (P < 0.05), which were normalized when supplementing 500 mg/kg and/or 1,000 mg/kg CGA (P < 0.05). DQ administration elevated hepatic interleukin-1ß, tumor necrosis factor-α, and interleukin-6 levels (P < 0.05), and the values of interleukin-1ß were normalized to control values when supplementing CGA (P < 0.05). DQ injection decreased serum superoxide dismutase activity, hepatic catalase activity, and serum and hepatic glutathione level, but increased malondialdehyde concentration in serum and liver (P < 0.05), and the values of these parameters (except hepatic catalase activity) were reversed by 500 and/or 1,000 mg/kg CGA. The results suggested that CGA could improve growth performance, alleviate oxidative stress, and ameliorate hepatic inflammation in DQ-challenged broilers.

Dietary supplementation with synbiotics improves growth performance, antioxidant status, immune function, and intestinal barrier function in broilers subjected to cyclic heat stress.

This study investigated the effects of synbiotics supplementation on growth performance, antioxidant status, immune function, and intestinal barrier function in broilers subjected to cyclic heat stress. One hundred and forty-four 22-day-old male broilers were randomly assigned to one of three treatment groups of six replicates each for a 21-day study, with eight birds per replicate. Broilers in the control group were reared at a thermoneutral temperature and received a basal diet. Broilers in the other two heat-stressed groups were fed a basal diet supplemented without (heat-stressed group) and with 1.5 g/kg synbiotic (synbiotic group). One and a half gram of the synbiotic consisted with 3 × 109 colony forming units (CFU) Clostridium butyricum, 1.5 × 109 CFU Bacillus licheniformis, 4.5 × 1010 CFU Bacillus subtilis, 600 mg yeast cell wall, and 150 mg xylooligosaccharide. Compared with the control group, heat stress increased rectal temperatures at 28, 35, and 42 days of age, respectively (P < 0.05). Birds subjected to heat stress had reduced weight gain, feed intake, and feed efficiency during 22 to 42 days (P < 0.05). In contrast, supplementation with the synbiotic decreased rectal temperature at 42 days of age and elevated weight gain of heat stress-challenged broilers (P < 0.05). Heat-stressed broilers exhibited a lower superoxide dismutase (SOD) activity in jejunal mucosa and a higher malondialdehyde accumulation in serum, liver and jejunal mucosa (P < 0.05), and the regressive SOD activity was normalized to control level when supplementing synbiotic (P < 0.05). Heat stress increased interleukin-1ß (IL-1ß) and interferon-γ (IFN-γ) levels in serum and IL-1ß content in jejunal mucosa of broilers (P < 0.05). Synbiotic reduced IL-1ß level in serum of broilers subjected to heat stress (P < 0.05). Compared with the control group, elevated serum diamine oxidase activity and reduced jejunal villus height were observed in broilers of the heat-stressed group (P < 0.05), and the values of these two parameters in the synbiotic group were intermediate (P > 0.05). Heat stress upregulated mRNA abundance of IL-1ß and IFN-γ and downregulated gene expression levels of occluding and zonula occluden-1 (ZO-1) in jejunal mucosa of broilers (P < 0.05). The alterations in the mRNA expression levels of jejunal IL-1ß and ZO-1 were reversed by the synbiotic (P > 0.05). In conclusion, dietary synbiotics could improve growth performance, antioxidant capacity, immune function, and intestinal barrier function in heat-stressed broilers.

Dietary chitooligosaccharide supplementation improves mineral deposition, meat quality and intramuscular oxidant status in broilers.

BACKGROUND: The present study aimed at evaluating the in vitro adsorption capability of chitooligosaccharide (COS) with some metal elements (Fe, Zn, Cd, Pb) at different pH values along with potential effects of dietary COS supplementation on growth performance, mineral content, meat quality and oxidant status in broilers. Day-old male chicks were randomly distributed into two groups and offered a basal diet supplemented with or without 30 mg kg-1 COS for 42 days. RESULTS: In vitro trials demonstrated that Fe levels were higher (P < 0.001) in the COS-treated group compared with the non-treated group at pH of 2.5. However, these levels became lowered when pH values were raised to 5 (P < 0.01) or 6 (P < 0.001). Similarly, COS adsorbed more (P < 0.05) Zn at pH values of 2.5 and 6, and Cd contents at pH of 2.5 for 70 min when compared with the control. For in vivo trial, the feed-to-gain ratio, serum Cu (P < 0.01), hepatic Mn, Cr (P < 0.05) and intramuscular Cd (P < 0.01) were lower in response to COS treatment. Supplementation of COS improved (P < 0.05) meat quality of broilers in terms of lower drip loss, cooking loss and malondialdehyde content with a concomitant increase (P < 0.01) in the pH of breast meat at 24 h post mortem. CONCLUSION: COS adsorbed heavy metal ions not only in vitro but also in broilers, and dietary supplementation with 30 mg kg-1 COS improved growth performance, breast meat quality and oxidant status in broilers. © 2022 Society of Chemical Industry.

An evaluation of the protective effects of chlorogenic acid on broiler chickens in a dextran sodium sulfate model: a preliminary investigation.

This study was conducted to investigate the protective effects of chlorogenic acid (CGA) on broilers subjected to dextran sodium sulfate (DSS)-induced intestinal damage. One hundred and forty-four 1-day-old male Arbor Acres broiler chicks were allocated into one of 3 groups with 6 replicates of eight birds each for a 21-d trial. The treatments included: 1) Control group: normal birds fed a basal diet; 2) DSS group: DSS-treated birds fed a basal diet; and 3) CGA group: DSS-treated birds fed a CGA-supplemented control diet. An oral DSS administration via drinking water was performed from 15 to 21 d of age. Compared with the control group, DSS administration reduced 21-d body weight and weight gain from 15 to 21 d, but increased absolute weight of jejunum and absolute and relative weight of ileum (P < 0.05). DSS administration elevated circulating D-lactate concentration and diamine oxidase activity (P < 0.05), which were partially reversed when supplementing CGA (P < 0.05). The oral administration with DSS decreased villus height and villus height/crypt depth ratio, but increased crypt depth in jejunum and ileum (P < 0.05). Compared with the control group, DSS administration increased serum glutathione level and jejunal catalase activity and malonaldehyde accumulation, but decreased jejunal glutathione level (P < 0.05). In contrast, feeding a CGA-supplemented diet normalized serum glutathione and jejunal malonaldehyde levels, and increased jejunal glutathione concentration in DSS-administrated birds (P < 0.05). Additionally, CGA supplementation reduced ileal malonaldehyde accumulation in DSS-treated birds (P < 0.05). DSS challenge increased levels of serum interferon-γ and interleukin-6, jejunal interleukin-1ß, tumor necrosis factor-α, and interleukin-6, and ileal interleukin-1ß and interleukin-6 when compared with the control group (P < 0.05). The elevated serum interferon-γ and ileal interleukin-6 levels were normalized to control values when supplementing CGA (P < 0.05). The results suggested that CGA administration could partially prevent DSS-induced increased intestinal permeability, oxidative damage, and inflammation in broilers, although it did not improve their growth performance and intestinal morphology.

Effects of dietary palygorskite supplementation on the growth performance, oxidative status, immune function, intestinal barrier and cecal microbial community of broilers.

The present study aimed to investigate the effects of palygorskite (PAL) as an alternative to antibiotic on the growth performance, oxidative status, immune function, intestinal barrier and cecal microbial community of broilers. A total of 360 1-day-old male Ross-308 broilers were randomly allotted to three treatments with eight replicates. Broilers in the three groups were designated as follows: basal diet (CON group), basal diet+50 mg/kg chlorotetracycline (ANT group), and basal diet+ 10 g/kg PAL (PAL group). Supplementing PAL reduced feed to gain ratio in broilers during 22 to 42 days of age (P < 0.05), with its value being similar to that of the ANT group (P > 0.05). Broilers fed a PAL-supplemented diet exerted decreased contents of interferon-γ (IFN-γ) and interleukin-1ß in serum, and the same reduction was found in jejunal IFN-γ level, when compared to the CON group (P < 0.05). Moreover, compared with the CON group, broilers after PAL treatment had a lower malondialdehyde content in jejunal mucosa (P < 0.05). Supplementing PAL elevated jejunal villus height (VH) and ratio of VH to crypt depth compared with the ANT group (P < 0.05). Cecal microbiota communities among the three groups were significant different, as demonstrated by distinct clusters from partial least squares discriminant analysis, although dietary treatments had no significant effects on the bacterial richness and diversity indices (P > 0.05). At genus level, the addition of PAL increased the relative abundance of norank_f__Barnesiellaceae and decreased that of unclassified_f__Oscillospiraceae in cecal digesta compared with those in the CON group (P < 0.05); the proportion of genus norank_f__Barnesiellaceae was increased by PAL treatment when compared with the ANT group (P < 0.05). Moreover, spearman's correlations showed that the modulation of cecal microflora composition by PAL supplementation was closely correlated with the promotion of growth performance (feed to gain ratio) and intestinal health-related (contents of malondialdehyde and IFN-γ, and VH value in jejunum) variables of broilers (P < 0.05). Taken together, dietary PAL could improve the growth performance, antioxidant capacity, and immune status, as well as intestinal barrier function in broilers, which might be partially associated with the alteration of cecal microbiota. Moreover, dietary PAL may be a promising alternative to antibiotic growth promoter for broilers.

An evaluation of graded levels of beta-sitosterol supplementation on growth performance, antioxidant status, and intestinal permeability-related parameters and morphology in broiler chickens at an early age.

This study was designed to examine the effects of different levels of beta-sitosterol (BS) supplementation on growth performance, serum biochemical indices, redox status, and intestinal permeability-related parameters and morphology of young broilers. Two hundred and forty male Arbor Acres broiler chicks were allocated into 5 groups of 6 replicates with 8 birds each, and fed a basal diet supplemented with 0, 25, 50, 75, and 100 mg/kg BS for 21-d, respectively. The BS quadratically decreased feed conversion ratio during 1 to 14 d and 1 to 21 d, with its effect being more prominent at 25 or 50 mg/kg (P < 0.05). The BS linearly and quadratically reduced 14-d plasma diamine oxidase activity and D-lactate level, and this effect was more pronounced when its supplemental level was 25 or 50 mg/kg (P < 0.05). The BS linearly increased duodenal villus height (VH) and quadratically increased jejunal VH and ratio of VH and crypt depth (CD) at 14 d, and these effects in 25 mg/kg group were more remarkable (P < 0.05). Similarly, BS linearly or quadratically increased VH and ratio of VH and CD, but decreased CD in the jejunum and ileum at 21 d, with these effects being more pronounced at 50 mg/kg (P < 0.05). The BS supplementation especially at 50 or 75 mg/kg linearly or quadratically reduced 14-d serum and 21-d hepatic malondialdehyde concentration, and increased serum glutathione peroxidase and catalase activities at 14 and 21 d (P < 0.05). Moreover, the BS administration linearly and/or quadratically increased glutathione peroxidase, catalase, and superoxide dismutase activities and glutathione level, and reduced malondialdehyde accumulation in the intestinal mucosa at 14 and/or 21 d, and these consequences were more significant in 50 to 100 mg/kg BS-supplemented groups (P < 0.05). The results demonstrated that BS administration could improve growth performance, intestinal barrier function, and antioxidant status of broilers at an early age, with these effects being more pronounced at a level of 50 mg/kg.

Multi-modal molecular programs regulate melanoma cell state.

Melanoma cells display distinct intrinsic phenotypic states. Here, we seek to characterize the molecular regulation of these states using multi-omic analyses of whole exome, transcriptome, microRNA, long non-coding RNA and DNA methylation data together with reverse-phase protein array data on a panel of 68 highly annotated early passage melanoma cell lines. We demonstrate that clearly defined cancer cell intrinsic transcriptomic programs are maintained in melanoma cells ex vivo and remain highly conserved within melanoma tumors, are associated with distinct immune features within tumors, and differentially correlate with checkpoint inhibitor and adoptive T cell therapy efficacy. Through integrative analyses we demonstrate highly complex multi-omic regulation of melanoma cell intrinsic programs that provide key insights into the molecular maintenance of phenotypic states. These findings have implications for cancer biology and the identification of new therapeutic strategies. Further, these deeply characterized cell lines will serve as an invaluable resource for future research in the field.

Construction of lncRNA-miRNA-mRNA network based on ceRNA mechanism reveals the function of lncRNA in the pathogenesis of gout.

OBJECTIVE: To identify differentially expressed lncRNA, miRNA, and mRNA during the pathogenesis of gout, explore the ceRNA network regulatory mechanism of gout, and seek potential therapeutic targets. METHOD: First, gout-related chips were retrieved by GEO database. Then, the analysis of differentially expressed lncRNAs and mRNAs was conducted by R language and other software. Besides, miRNA and its regulated mRNA were predicted based on public databases, the intersection of differentially expressed mRNA and predicated mRNA was taken, and the lncRNA-miRNA-mRNA regulatory relationships were obtained to construct the ceRNA regulatory network. Subsequently, hub genes were screened by the STRING database and Cytoscape software. Then the DAVID database was used to illustrate the gene functions and related pathways of hub genes and to mine key ceRNA networks. RESULTS: Three hundred and eighty-eight lncRNAs and 758 mRNAs were identified with significant differential expression in gout patient, which regulates hub genes in the ceRNA network, such as JUN, FOS, PTGS2, NR4A2, and TNFAIP3. In the ceRNA network, lncRNA competes with mRNA for miRNA, thus affecting the IL-17 signaling pathway, TNF signaling pathway, Oxytocin signaling pathway, and NF-κB signaling pathway through regulating the cell's response to chemical stress. The research indicates that five miRNAs (miR-429, miR-137, miR-139-5p, miR-217, miR-23b-3p) and five lncRNAs (SNHG1, FAM182A, SPAG5-AS1, HNF1A-AS1, UCA1) play an important role in the formation and development of gout. CONCLUSION: The interaction in the ceRNA network can affect the formation and development of gout by regulating the body's inflammatory response as well as proliferation, differentiation, and apoptosis of chondrocytes and osteoclasts. The identification of potential therapeutic targets and signaling pathways through ceRNA network can provide a reference for further research on the pathogenesis of gout.

Dietary palygorskite-based antibacterial agent supplementation as an alternative to antibiotic improves growth performance, intestinal mucosal barrier function, and immunity in broiler chickens.

The aim of this study was to evaluate effects of palygorskite-based antibacterial agent (PAA) as an alternative to antibiotic on growth performance, intestinal barrier function, and immunity in broilers. Three hundred and eighty-four mixed-sex 1-day-old Ross 308 broiler chicks were allocated into 6 groups of 8 replicates with 8 birds each. Birds were given a basal diet, an antibiotic diet (50 mg/kg chlortetracycline), and the basal diet supplemented with 250, 500, 1,000, and 2,000 mg/kg PAA for 42 d, respectively. Compared with control group, supplementing 1,000 mg/kg PAA reduced overall feed conversion ratio (P < 0.05), with its value being similar to that of antibiotic group (P > 0.05). However, a higher level of PAA (2,000 mg/kg) increased feed conversion ratio during the late period (P < 0.05). The 1,000 and 2,000 mg/kg PAA decreased plasma endotoxin and D-lactate levels at 42 d (P < 0.05) to comparable values (P > 0.05). The 1,000 mg/kg PAA decreased jejunal crypt depth, while 500 and 1,000 mg/kg PAA increased the ratio between jejunal villus height and crypt depth at 42 d (P < 0.05), with their values being similar to antibiotic group (P > 0.05). The highest level of PAA increased 42-d jejunal mucosal secretory immunoglobulin A and immunoglobulin M concentrations (P < 0.05). The 1,000 and 2,000 mg/kg PAA reduced 21-d interleukin-1ß and tumor necrosis factor-α (TNF-α) levels in serum and ileal mucosa and 42-d interferon-γ level in serum and jejunal mucosa (P < 0.05), which did not differ from antibiotic group (P > 0.05). Moreover, PAA administration, regardless of its dosage, reduced 42-d serum TNF-α concentration, and 500 to 2,000 mg/kg PAA decreased 21-d and 42-d jejunal and 42-d ileal mucosal TNF-α levels (P < 0.05), with their values being comparable with antibiotic group (P > 0.05). The results suggested that PAA as an alternative to antibiotic could improve growth performance, intestinal barrier function, and immunity of broilers, and its optimal dosage was 1,000 mg/kg.

Effects of Zinc Oxide Nanoparticles on Growth, Intestinal Barrier, Oxidative Status and Mineral Deposition in 21-Day-Old Broiler Chicks.

This experiment was to study the effects of zinc oxide nanoparticles (ZnO-NPs) on growth, intestinal barrier, oxidative status, and mineral deposition. In total, 256 one-day-old chicks were randomly allotted to 4 dietary groups and fed with basal diet plus 80 mg/kg ZnSO4 (ZnSO4 group) or plus 40, 80, and 160 mg/kg ZnO-NPs, respectively, for 21 days. Compared with the ZnSO4 group, dietary 40, 80, and 160 mg/kg ZnO-NPs did not alter growth (average daily gain, body weight, and gain to feed ratio), and serum activities of glutamic-pyruvic transaminase, alkaline phosphatase and glutamic oxalacetic transaminase (P > 0.05). However, dietary 80 and 160 mg/kg ZnO-NPs linearly decreased serum D-lactate content and diamine oxidase activity (P < 0.01). Moreover, 80 mg/kg ZnO-NPs enhanced zonula occludens-1 (ZO-1) mRNA expression in jejunal mucosa (P = 0.02). Dietary ZnO-NPs increased total antioxidant capacity activity (P = 0.01), and 80 mg/kg ZnO-NPs decreased malondialdehyde content in jejunal mucosa as compared to the ZnSO4 group (P = 0.02). In contrast, dietary ZnO-NPs did not alter mRNA expressions of superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, heme oxygennase-1 (HO-1) and NAD (P)H: quinone oxidoreductase 1 (NQO1) (P > 0.05). No significant difference was found in selected mineral concentrations (Mn, Cu, Fe and Zn) in the liver among ZnSO4 and 3 ZnO-NP groups (P > 0.05). However, 160 mg/kg ZnO-NPs increased fecal contents of Zn, Fe and Cu (P < 0.01), but did not affect fecal Mn level (P > 0.05). Therefore, results suggested that ZnO-NPs could be an additive to enhance the intestinal barrier and antioxidant capacity of broiler chicks, whereas the inclusion of 80 mg/kg would be more efficient.

The Xiaogu San Attenuates Pain and Cartilage Damage in Rats with Monosodium Iodoacetate Induced Osteoarthritis.

BACKGROUND: Osteoarthritis (OA) is a degenerative joint disease with an increasing incidence associated with increased life expectancy. The application of traditional Chinese medicine in the treatment of OA has become a research hotspot. OBJECTIVE: This study investigated the effects of XGS externally applied to osteoarthritic joints and analyze its effect on pain in monosodium iodoacetate (MIA)-induced OA rats. This study also evaluates potential mechanisms behind the anti-osteoarthritic effects of XGS. METHODS: A total of 24 Sprague Dawley rats were evenly and randomly divided into three separate groups, including the normal control (NC), OA and XGS groups. MIA (50 µL, 10 mg/mL) was injected into the left knee joints of the rats to induce OA. After 7 days, The rats of XGS group were given XGS (0.45 g) that was externally applied to the left knee joint, were fixed with gauze, and continuously administered XGS for 28 days. Morphological changes in tissues and organs were examined using H&E staining. Biochemical indicators were measured using an automatic biochemical analyzer. Inflammatory cytokines were detected using ELISA kits and immunohistochemistry. RNA-based high-throughput sequencing (RNA-seq) was performed to detect differential expression of mRNAs in normal and MIA-induced OA rats. RESULTS: Stride of the left leg was extended in rats, matrix increased on cartilage tissue surfaces, and inflammatory cytokines were reduced when treated with XGS. RNA-seq results revealed that the PI3K-Akt signaling pathway is activated in the OA model. The qRT-PCR showed that the expression levels of Tnn, Col6a6, Igf1 and Lamb1 were up-regulated by XGS. CONCLUSION: Altogether, this work demonstrated the potential therapeutic effects of XGS in rats with OA induced by MIA. The XGS may be considered an alternative therapy to manage OA.