Cadmium (Cd), a pervasive environmental toxicant, induces hepatotoxicity via mitochondrial damage and dysregulated inflammation. Astragaloside IV (AS-IV), the primary bioactive constituent of Astragalus membranaceus with antioxidative/anti-inflammatory properties, has an undefined regulatory role in the cGAS-STING axis during Cd-induced hepatic injury. In this study, the cytoprotective mechanisms of AS-IV against Cd-induced hepatotoxicity were investigated. The results showed that Cd exposure significantly impaired hepatocellular viability, induced mitochondrial dysfunction, promoted mitochondrial DNA (mtDNA) release into the cytosol, and thereby activated the cGAS-STING signaling pathway, while AS-IV intervention effectively mitigated Cd-induced mitochondrial perturbations, suppressed mtDNA efflux, and inhibited cGAS-STING pathway activation by attenuating mtDNA-dependent STING activation via suppressing cytosolic mtDNA release.Collectively, AS-IV exerts robust hepatoprotection against Cd toxicity via preservation of mitochondrial integrity, inhibition of cytosolic mtDNA translocation, and suppression of cGAS-STING-driven innate immune hyperactivation. These findings nominate AS-IV as a viable therapeutic countermeasure against heavy metal-induced organ damage.

This study investigated the effects of supplementing diets for Shan Partridge Ducks (n = 720) with 1

Avian pathogenic Escherichia coli (APEC) causes significant economic losses to the global poultry industry, and its virulence mechanisms remain complex and not fully elucidated. Toxin-antitoxin (TA) systems have recently been implicated in bacterial virulence regulation beyond their traditional roles. This study is the first to reveal the critical role of the ecnAB TA system in APEC pathogenicity. Deletion of ecnAB significantly attenuated bacterial virulence in hosts. Transcriptomic and RT-qPCR analyses demonstrated that the absence of ecnAB led to marked downregulation of the neu gene cluster (including neuB, neuC, neuA, neuS, etc.) responsible for capsular sialic acid synthesis and transport, as well as the kpsM gene. Phenotypic assays confirmed that the ΔecnAB mutant exhibited a sharp reduction in capsular polysaccharide and sialic acid production, accompanied by loss of serum resistance and enhanced clearance by macrophages. Further mechanistic studies revealed that the EcnA antitoxin protein directly binds to the promoter regions of the neu operon and kpsM, positively regulating their transcription. In summary, this study identifies the ecnAB TA system as a novel regulatory hub that directly activates capsular sialic acid biosynthesis, promoting intact capsule formation and conferring critical immune evasion capabilities to APEC. This finding not only deepens the understanding of the functional diversity of TA systems but also provides potential targets for developing novel antimicrobial strategies against APEC infections.

In the original publication [...].

IntroductionEnterotoxic Escherichia coli (ETEC) is the main pathogen that causes diarrhea, especially in young children. This disease can lead to substantial morbidity and mortality and is a major global health concern. Managing ETEC infections is challenging owing to the increasing prevalence of antibiotic resistance. Berberine, categorized as a substance with similarities in “medicine and food,” has been used in China for hundreds of years to treat gastrointestinal disorders and bacteria-induced diarrhea. This study investigated the preventive effect of dietary berberine on the intestinal mucosal barrier induced by ETEC and the microbial community within the intestines of weaned piglets.MethodsTwenty-four piglets were randomly divided into four groups. Piglets were administered either a standard diet or a standard diet supplemented with berberine at concentrations of 0.05 and 0.1%. and orally administered ETEC or saline.ResultsDietary supplementation with berberine reduced diamine oxidase, d-lactate, and endotoxin levels in piglets infected with ETEC (P < 0.05). Berberine increased jejunal villus height, villus/crypt ratio, mucosal thickness (P < 0.05), and goblet cell numbers in the villi and crypts (P < 0.05). Furthermore, berberine increased the optical density of mucin 2 and the mucin 2, P-glycoprotein, and CYP3A4 mRNA expression levels (P < 0.05). Berberine increased the expressions of zonula occludins-1 (ZO-1), zonula occludins-2 (ZO-2), Claudin-1, Occludin, and E-cadherin in the ileum (P < 0.05). Moreover, berberine increased the expression of BCL2, reduced intestinal epithelial cell apoptosis (P < 0.05) and decreased the expression of BAX and BAK in the duodenum and jejunum, as well as that of CASP3 and CASP9 in the duodenum and ileum (P < 0.05). Berberine decreased the expression of IL-1β, IL-6, IL-8, TNF-α, and IFN-γ (P < 0.05) and elevated total volatile fatty acids, acetic acid, propionic acid, valeric acid, and isovaleric acid concentrations (P < 0.05). Notably, berberine enhanced the abundance of beneficial bacteria including Enterococcus, Holdemanella, Weissella, Pediococcus, Muribaculum, Colidextribacter, Agathobacter, Roseburia, Clostridium, Fusicatenibacter, and Bifidobacterium. Simultaneously, the relative abundance of harmful and pathogenic bacteria, such as Prevotella, Paraprevotella, Corynebacterium, Catenisphaera, Streptococcus, Enterobacter, and Collinsella, decreased (P < 0.05).DiscussionBerberine alleviated ETEC-induced intestinal mucosal barrier damage in weaned piglets models. This is associated with enhancement of the physical, chemical, and immune barrier functions of piglets by enhancing intestinal microbiota homeostasis.

Background: Chinese herbal medicine are rich in nutrients and active ingredients, with advantages such as immune function, antioxidant capacity, stress reduction, tumor inhibition and improved meat quality. The effects of Chinese herbal medicines on the muscle microstructure and meat quality of broiler chickens were investigated by feeding with compound Chinese herbal medicines composed of stir-fried Atractylodes macrocephala, licorice, astragalus, hawthorn fruit, poria cocos, isatis root, dried tangerine peel and common yam rhizome. Methods: The 800, 1-day-old, chickens were allocated to 4 treatments with 5 replicates, each replicate containing 40 birds. Birds were fed diets supplemented with 0.5%, 1.0% and 1.5% compound Chinese herbal medicines for a period of 7 weeks ad libitum. At 21 and 42 days old, the birds were slaughtered and the breast and thigh muscles were sampled from 4 birds per replicate to determine meat quality and microstructure of muscle. Result: The results revealed that 1.0% compound Chinese herbal medicines significantly increased water holding capacity of the thigh muscle and decreased drip loss, cooking loss and water loss rate of 21-day-old broiler chickens. Moreover, 1.0% compound Chinese herbal medicine significantly decreased the muscle fibers diameters of breast and thigh muscle. Microscopic observation showed that 0.5% and 1.0% compound Chinese herbal medicines decreased the fibers diameters of muscle fibers. The results indicated that the addition of 0.5% and 1.0% compound Chinese herbal medicines in the diets could promote muscle growth and improve the meat quality of broiler chickens.

Improving feed efficiency in Tianchang Sanhuang chickens is essential for reducing production costs and environmental burden. The objective of this study was to integrate transcriptomic and metabolomic analyses to identify key regulatory genes, metabolites, and pathways associated with residual feed intake (RFI) and feed efficiency. In this study, 650 Tianchang Sanhuang laying hens with similar body weights at 36 weeks of age were evaluated for daily feed intake (DFI), RFI, and feed conversion ratio (FCR). The chickens were classified by RFI (mean ± 0.5 SD) into high-RFI (HRFI, n = 165) and low-RFI (LRFI, n = 158) groups. Phenotypes, serum biochemistry, antioxidant indices, and intestinal traits were compared in subsets (n = 8 per group). Duodenal transcriptomes (RNA-seq) and serum metabolomes (LC-MS/MS) were profiled in independent subsets (n = 4 per group). Compared with HRFI, LRFI hens showed lower RFI, FCR, and DFI (P < 0.01), with no differences in expected feed intake (EFI), metabolic body weight (MBW), daily egg mass (DEM), or average daily gain (ADG) (P > 0.05). The LRFI group showed increased breast muscle redness (a)* (P < 0.05), higher leg muscle drip loss (P < 0.01), and significantly lower levels of triglycerides (TG), cholesterol (CHO), low-density lipoprotein cholesterol (LDL-C), and malondialdehyde (MDA) (P < 0.05). The intestinal morphology and molecular analyses revealed enhanced nutrient absorption and intestinal barrier function in the LRFI group. Transcriptomic analysis identified 237 differentially expressed genes (P < 0.05, |log2FC| ≥ 1) enriched in pathways related to digestion, energy metabolism, and appetite regulation. Metabolomic analysis detected 101 differentially expressed metabolites (VIP ≥ 1, |log2FC| ≥ 1), indicating that RFI is closely associated with protein and lipid metabolism. Integrated analysis identified candidate biomarkers for low RFI individuals selection, including genes such as ACSM5, AHSG, CTRB1, PLA2G1B, AMY2A, CPA1, CCKAR and metabolites including taurine, uridine, L-phenylalanine, D-glucose 6-phosphate and 5‑hydroxy-L-tryptophan. Overall, LRFI hens maintain production while achieving lower intake, potentially via reduced inflammation/oxidative stress and enhanced digestion, barrier integrity, appetite, and energy metabolism, offering targets for marker-assisted improvement of feed efficiency in local breeds.

Enterotoxigenic Escherichia coli (ETEC), present in contaminated food, water, and environments, can induce hepatic injury via the gut-liver axis, posing a serious threat to ecological systems and public health. Linarin, a flavonoid extracted from Chrysanthemum indicum, exhibits anti-inflammatory and antioxidant properties, but its protective effects against ETEC-induced hepatic injury remain unclear. In this study, 24 weaned piglets were randomly assigned to four groups: BD+NB (basal diet + nutrient broth), LN+NB (basal diet + 150 mg/kg linarin + nutrient broth), BD+ETEC (basal diet + ETEC challenge), and LN+ETEC (basal diet + 150 mg/kg linarin + ETEC challenge). Dietary linarin significantly increased ADFI and the genes related to oxidative damage and bile acid metabolism, while decreasing F:G ratio, liver index, serum liver function-related parameters, and the genes related to inflammatory response and apoptosis. It also significantly altered the relative abundances of gut microbiota, which were closely associated with key hepatic metabolic pathways, including nicotinate and nicotinamide metabolism and fatty acid biosynthesis. Our study suggests that linarin alleviated ETEC-induced hepatic inflammation and apoptosis, enhanced antioxidant capacity, and regulated bile acid metabolism. The potential mechanism involves linarin modulating gut microbiota-mediated key hepatic metabolic pathways to exert protective effects. In contrast to previous flavonoid-ETEC studies that primarily focused on the gut, this study, based on the gut-liver axis, investigates the potential mechanisms by which linarin is associated with the alleviation of ETEC-induced hepatic injury through integrated analysis of gut microbiome metagenomics and liver metabolomics.

This study aimed to investigate the preventive effects of dietary linarin supplementation on Enterotoxigenic coli (ETEC) induced small intestinal barrier dysfunction in weaned piglets via gut microbiota modulation. Twenty-four weaned piglets were randomly assigned to four experimental groups. The four treatments were as follows: BD + NB (basal diet and orally administered nutrient broth), LN + NB (basal diet supplemented with 150 mg/kg linarin, and orally administered nutrient broth), BD + ETEC (basal diet and orally administered ETEC), LN + ETEC (basal diet supplemented with 150 mg/kg linarin, and orally administered ETEC). The results showed that linarin lowers the serum levels of diamine oxidase (DAO), endotoxin, and D-lactate (P < 0.05). Linarin significantly increased villus height and villus/crypt ratio (P < 0.01), while decreasing crypt depth in the duodenum. jejunum, and ileum (P < 0.05). Additionally, Linarin increased the number of goblet cells within villus-crypt units in the duodenum. jejunum, and ileum (P < 0.05). Linarin significantly enhanced intestinal barrier function, upregulated detoxification pathways, reduced epithelial apoptosis, and improved nutrient transporter expression in the small intestine." Linarin decreased the relative abundances of Actinobacillus, Romboutsia, Enterococcus, and Terrisporobacter, consequently modulating key metabolic pathways, including arginine and proline metabolism, steroid biosynthesis, and cysteine and methionine metabolism. Dietary linarin supplementation mitigates ETEC-induced intestinal mucosal barrier dysfunction and enhances nutrient assimilation via targeted modulation of microbial communities and associated metabolic networks, thereby providing a new strategy for the prevention and treatment of ETEC diarrhea.

Enterotoxigenic Escherichia coli (ETEC) causes diarrhea and triggers intestinal inflammation posing a significant threat to children which is a global health concern. Berberine (BBR) has diverse pharmacological properties can alleviate diarrhea and intestinal damage. This metabolomics study analyzed the effects of berberine on the hepatic function, lipid metabolism, and inflammatory responses in weaned piglet model infected with ETEC. A total of 18 piglets were randomly divided into three groups: basal diet (BD) + Saline, BD + ETEC, and BBR + ETEC. Compared with the BD + Saline group, ETEC infection significantly increased the contents of aspartate aminotransferase (AST), AST/alanine aminotransferase (ALT), total protein, globulin, triglycerides, tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), IL-6, IL-8, IL-12, interferon γ (IFN-γ), IL-4, IL-10, and transforming growth factor β (TGF-β) and significantly decreased the contents of total cholesterol, low-density lipoprotein (LDL), and high-density lipoprotein (HDL) in the serum of weaned piglets. There were 14 metabolites upregulated and 21 metabolites downregulated after ETEC infection, and total cholesterol, LDL, IL-1β, IL-8, and TGF-β were correlated with fructose and mannose metabolism. Compared with the BD + ETEC group, dietary berberine significantly increased the contents of total cholesterol, LDL, and HDL and significantly decreased the contents of AST, AST/ALT, total protein, globulin, triglycerides, TNF-α, IL-1β, IL-6, IL-8, IL-12, IFN-γ, and IL-4 in the serum of weaned piglets. There were 111 metabolites upregulated and 48 metabolites downregulated after berberine addition, and total protein, IL-1β, IL-6, IL-8, and IL-12 were correlated with cholinergic synapse. Both ETEC infection and berberine addition can induce changes in taste transduction and affect the physiological functions of weaned piglets through metabolic pathways.

Ochratoxin A (OTA) is a prevalent contaminant in feed and poses a serious threat to the poultry industry and public health. The liver is the primary target of OTA, and oxidative stress alongside consequent inflammation, is considered the main contributor to OTA-induced liver damage. Astragaloside IV (AS-IV), a key constituent of the traditional Chinese medicinal herb Astragalus membranaceus, exhibits diverse pharmacological properties, including anti-inflammatory, antioxidant, immunoregulatory, and organ-protective effects. However, whether AS-IV can ameliorate OTA-induced liver damage remains uncertain. In the current investigation, we investigated the effect of AS-IV on OTA-induced liver damage in chicks and elucidated the underlying mechanisms. The results revealed that AS-IV inhibited OTA-induced increases in alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (AKP) activities. Additionally, AS-IV reversed the OTA-induced decrease in glutathione peroxidase (GSH-Px) and total superoxide dismutase (T-SOD) activities, as well as the increase in malondialdehyde (MDA) content (P < 0.05). The results demonstrated that AS-IV is capable of mitigating mitochondrial damage and reducing the elevation of reactive oxygen species (ROS) in the chicken liver cell line LMH induced by OTA. Moreover, AS-IV was discovered to reverse the OTA-induced decrease in nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase 1 (HO-1), NADPH quinone oxidoreductase 1 (NQO1) expression, and the increase in Kelch-like ECH-associated protein 1(Keap1) expression. Meanwhile, AS-IV also counteracted OTA-induced NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome expression. Taken together, our findings suggested that AS-IV ameliorated OTA-induced hepatotoxicity in chicks by regulating the Nrf2 and NLRP3 signaling pathways.

Introduction:Enterotoxigenic Escherichia coli (ETEC) is a globally recognized gastrointestinal pathogen and a major cause of diarrhea in neonatal and post-weaning animals, leading to significant economic losses in pig production. Premature weaning disrupts colonic morphology and barrier integrity, resulting in diarrhea, dehydration, growth retardation, and increased mortality. Linarin, a natural flavonoid derived from wild chrysanthemum, exhibits antioxidant, sedative, and anti-osteoporotic properties, demonstrating potential as a therapeutic agent and functional food ingredient. Methods:24 healthy 21-day-old weaned piglets (Duroc × Landrace × Large Yorkshire) were randomly assigned to four groups fed a basal diet (BD) or linarin-supplemented diet (LN) with oral infusion of 10 mL nutrient broth (NB) or 10⁹ colony-forming units/mL ETEC. Following a 3-day acclimation period, piglets were fed the corresponding diet for 21 days; infusion with ETEC or NB was performed for 3 days on days 8 and 18. Colonic morphology, diarrhea incidence, gene expression, short-chain fatty acids (SCFAs), microbiota composition, and metabolomic profiles were assessed. Results:Linarin supplementation significantly ameliorated colonic crypt hyperplasia, increased goblet cell numbers, and decreased diarrhea incidence following ETEC infusion. It downregulated pro-apoptotic and pro-inflammatory gene expression while upregulating barrier-associated genes. Linarin also significantly increased the concentrations of short-chain fatty acids (acetic, propionic, valeric, and isovaleric acids) in the colon. Integrated analysis of 16S rRNA gene sequencing and non-targeted metabolomics revealed that linarin modulated the intestinal microbiota by altering the relative abundance of key bacterial taxa (Pedosphaera, Fusicatenibacteria, Tyzerella, Sporobacteria, Limosilactobacillus, Senegalimassilia, Catenibacillus, and Bryobacteria), and associated metabolic pathways, including purine and pyrimidine metabolism; steroid, porphyrin, and vitamin biosynthesis; various amino acid and nucleotide metabolic processes; unsaturated fatty acid biosynthesis; and the citric acid cycle. Discussion:These findings indicate that linarin restores colonic barrier function and intestinal microbiota homeostasis, enhancing resistance to ETEC infection along with the development and well-being of piglets after weaning. This study offers a new mechanistic understanding of how linarin confers protection against ETEC, which can promote its widespread application as a natural feed additive to replace antibiotics.

As a core strategy for antibiotic replacement, probiotics have two advantages insofar as they enhance both animal productivity and pathogen suppression. In this study, we screened the intestines of antibiotic-naïve chickens for broad-spectrum antimicrobial lactic acid bacteria (LAB) with natural adaptability, based on the host–microbiota coevolution theory, and systematically evaluated their potential for development as poultry probiotics. We isolated a LAB strain, Lactiplantibacillus plantarum Y300, from traditional native free-range chickens, which showed strong inhibitory activity against avian pathogenic Salmonella, Escherichia coli, and Staphylococcus aureus. In vitro experiments indicated that the Lpb. plantarum strain Y300 had no hemolytic activity; excellent acid-producing ability;an outstanding tolerance to bile salts, low-pH environments, and simulated gastrointestinal fluids; a positive hydrophobic interaction with xylene, and good auto-aggregation characteristics. It also displayed a relatively high antioxidant capacity. Whole-genome sequencing revealed that the genome of Lpb. plantarum Y300 was approximately 3.05 mb, with a GC content of 44.74%. The main carbohydrate-active enzyme and bacteriocin genes were predicted in the Y300 genome, and no virulence genes or drug-resistance genes were detected. In summary, this study suggests that Lpb. plantarum Y300 has potential utility as a probiotic, and lays the theoretical foundation for the further development of microecological preparations of avian-sourced LAB.

Background: Rotavirus (RV) is a major cause of diarrhea in young children and animals, especially piglets, leading to substantial economic losses in the global pig industry. Isoleucine (Ile), a branched-chain amino acid, plays an important role in regulating nutrient metabolism and has been shown to improve diarrhea. This study aimed to evaluate the effects of Ile supplementation on the mucosal immune barrier of the small intestine in RV-infected weaned piglets. Methods: Forty-eight 21-day-old weaned piglets were randomly divided into three dietary treatments (each treatment was subdivided into two groups, eight replicates per group), with 0%, 0.5%, or 1% Ile added for 15 days, and then, one group from each treatment was challenged with RV. Results: The results showed that 1% Ile added to the diet promoted the healthy development of the intestinal mucosa. Ile could restore the reduced villus height in the ileum and the goblet cell number in the duodenum and ileum to normal levels, improving the intestinal epithelial tight junctions in RV-infected piglets. Additionally, Ile increased the activity of lipase, amylase, and sucrase, as well as superoxide dismutase (SOD) and glutathione (GSH), along with the expression of SIgA, DEFβ1, and DEFβ2 in parts of the small intestine. Conclusions: The addition of Ile to the diet mitigated the effects of RV infection on intestinal morphology and mucosal barrier function, as well as the physiological functions of weaned piglets, and improved the antioxidant and immune functions of the piglets to some extent. These findings offer valuable insights, contributing to a deeper understanding of the role of Ile in supporting intestinal health.

Hard yolk (HY), commonly found in cooked, salted eggs, has recently become a great concern in the commercial production of salted eggs because of its negative impact on the taste of yolk. Herein, the chemical and structural characteristics of HY and the underlying mechanism of HY were investigated. Although there were similar nutritional compositions between HY and non-hard yolk (NHY), HY exhibited higher hardness, springiness, gumminess and chewiness than NHY, suggesting the lower acceptability of HY. In HY, a large amount of fibrillike substance was wrapped around the surface of yolk particles, whereas this was not observed in NHY. HY demonstrated higher beta-sheet content and surface hydrophobicity but lower alpha-helices than NHY, indicating that alpha-helices of yolk protein change into beta-sheet. In addition, HY exhibited higher disulphide bonds than NHY (13.5% vs. 11%). A higher level of protein aggregates was observed in HY compared with that in NHY, whereas a reducing agent, dithiothreitol (DTT), decreased the level. Similarly, the addition of tea polyphenols as a reducing agent in the pickling solution reduced HY formation and improved the taste texture of salted eggs. The results of this study show that the occurrence of HY compromised the taste acceptability of salted egg. Disulphide bond-mediated protein aggregation plays an important role in mediating HY formation through which the reducing agent suppresses HY formation.

IntroductionEnterotoxigenic Escherichia coli (ETEC) is the main diarrhea-causing pathogen in children and young animals and has become a global health concern. Berberine is a type of “medicine and food homology” and has a long history of use in China, particularly in treating gastrointestinal disorders and bacterial diarrhea.MethodsIn this study, we explored the effects of berberine on growth performance, intestinal inflammation, oxidative damage, and intestinal microbiota in a weaned piglet model of ETEC infection. Twenty-four piglets were randomly divided into four groups—a control group (fed a basal diet [BD] and infused with saline), a BD+ETEC group (fed a basal diet and infused with ETEC), a LB+ETEC group (fed a basal diet with 0.05% berberine and infused with ETEC infection), and a HB+ETEC group (fed a basal diet with 0.1% berberine and infused with ETEC).ResultsBerberine significantly improved the final body weight (BW), average daily gain (ADG), and average daily feed intake (ADFI) (P<0.05) of piglets, and effectively decreased the incidence of diarrhea among the animals (P<0.05). Additionally, berberine significantly downregulated the expression levels of the genes encoding TNF-α, IL-1β, IL-6, IL-8, TLR4, MyD88, NF-κB, IKKα, and IKKβ in the small intestine of piglets (P<0.05). ETEC infection significantly upregulated the expression of genes coding for Nrf2, CAT, SOD1, GPX1, GST, NQO1, HO-1, GCLC, and GCLM in the small intestine of the animals (P<0.05). Berberine significantly upregulated 12 functional COG categories and 7 KEGG signaling pathways. A correlation analysis showed that berberine significantly increased the relative abundance of beneficial bacteria (Gemmiger, Pediococcus, Levilactobacillus, Clostridium, Lactiplantibacillus, Weissella, Enterococcus, Blautia, and Butyricicoccus) and decreased that of pathogenic bacteria (Prevotella, Streptococcus, Parabacteroides, Flavonifractor, Alloprevotella) known to be closely related to intestinal inflammation and oxidative stress in piglets. In conclusion, ETEC infection disrupted the intestinal microbiota in weaned piglets, upregulating the TLR4/MyD88/NF-κB and Nrf2 signaling pathways, and consequently leading to intestinal inflammation and oxidative stress-induced damage.DiscussionOur data indicated that berberine can optimize intestinal microbiota balance and modulate the TLR4/MyD88/NF-κB and Nrf2 signaling pathways, thus helping to alleviate intestinal inflammation and oxidative damage caused by ETEC infection in weaned piglets.

Boron is an essential trace element with roles in growth, development, and physiological functions; however, its mechanism of action is still unclear. In this study, the regulatory roles of the PI3K/Akt signaling pathway on boron-induced changes in barrier function, proliferation, and apoptosis in rat intestinal epithelial cells were evaluated. Occludin levels, the proportion of cells in the G2/M phase, cell proliferation rate, and mRNA and protein expression levels of PCNA were higher, while the proportions of cells in the G0/G1 and S phases, apoptosis rate, and caspase-3 mRNA and protein expression levels were lower in cells treated with 0.8 mmol/L boron than in control IEC-6 cells (P < 0.01 or P < 0.05). However, 40 mmol/L boron decreased ZO-1 and Occludin levels, the proportion of cells in the G2/M phase, cell proliferation rate, and mRNA and protein levels of PCNA and increased the apoptosis rate and caspase-3 mRNA expression (P < 0.01 or P < 0.05). After specifically blocking PI3K and Akt signals (using LY294002 and MK-2206 2HCL), 0.8 mmol/L boron had no effects on Occludin, PCNA level, apoptosis rates, and caspase-3 levels (P < 0.05); however, the proliferation rate and PCNA levels decreased significantly (P < 0.01 or P < 0.05). The addition of 40 mmol/L boron did not affect ZO-1 and Occludin levels and did not affect the apoptosis rate or PCNA and caspase-3 levels. These results suggested that the PI3K/Akt signaling pathway mediates the effects of low-dose boron on IEC-6 cells.

Hepatotoxicity is frequently observed following acute cadmium (Cd) exposure in chicken. Oxidative stress and subsequent inflammation are regarded as the main reasons for cadmium-induced liver injury. NOD-like receptor (NLR) family pyrin domain-containing 3 (NLRP3) inflammasome-induced pyroptosis is involved in various inflammatory diseases, including liver injury. Poultry are more susceptible to harmful effects of heavy metals. However, the mechanism of cadmium-induced liver injury in chicken is still elusive. In this study, the effect of cadmium on chicken liver cells and the underlying mechanisms were investigated. The results showed mitochondria was damaged and excessive reactive oxygen species (ROS) were generated in chicken liver cell line LMH after cadmium exposure. Furthermore, cadmium-induced NLRP3 inflammasome activation and the cell membrane rupture indicated LMH cells pyroptosis. The ROS scavengers, acetylcysteine (NAC) and Mito-TEMPO prevented pyroptosis in LMH cells, suggesting that ROS were responsible for the activation of the NLRP3 inflammasome induced by cadmium. Additionally, anti-oxidative transcription factor Nrf2 was inhibited after cadmium exposure, explaining the excessive ROS generation. In summary, our study showed that cadmium leads to ROS generation by inducing mitochondrial damage and inhibiting Nrf2 activity, which promotes NLRP3 inflammasome activation and eventually induces pyroptosis in LMH cells.

The experiment aimed to investigate the effects of plant polysaccharides combined with boric acid on digestive function, immune function and harmful gas and heavy metal contents in the faeces of fatteners. For this study, 90 healthy crossbred fatteners were selected and randomly divided into five groups: the control group was fed with a basal diet (Con); experimental group I was fed with basal diet + 40 mg/kg boric acid (BA); experimental group II was fed with basal diet + 40 mg/kg boric acid + 400 mg/kg Astragalus polysaccharides (BA+APS); experimental group III was fed with basal diet + 40 mg/kg boric acid + 200 mg/kg Ganoderma lucidum polysaccharides (BA+GLP); and experimental group IV was fed with basal diet + 40 mg/kg boric acid + 500 mg/kg Echinacea polysaccharides (BA+EPS). Compared with Con, the average daily gain (ADG), the trypsin activities in the duodenum and jejunum, the IL-2 levels in the spleen, the T-AOC activities and GSH-Px contents in the lymph node of fattening were increased in the BA group (p < 0.05), but malondialdehyde content in the lymph and spleen, and the contents of NH3, H2S, Hg, Cu, Fe and Zn in the feces and urine were decreased (p < 0.05). Compared with the BA, the ADG, gain-to-feed ratio (G/F), the trypsin and maltase activities in the duodenum and jejunum were increased in the BA+APS (p < 0.05), and the T-SOD activities in the spleen and T-AOC activities in the lymph node were also increased (p < 0.05), but the H2S level was decreased in the feces and urine (p < 0.05). Compared with the BA, the ADG, G/F and the trypsin and maltase activities in the duodenum were increased in the BA+GLP and BA+EPS (p < 0.05), the activities of maltase and lipase in the duodenum of fatteners in the BA+GLP and the activities of trypsin, maltase and lipase in the BA+EPS were increased (p < 0.05). Gathering everything together, our findings reveal that the combined addition of boric acid and plant polysaccharides in the diet of fatteners synergistically improved their growth performance and immune status. That may be achieved by regulating the activity of intestinal digestive enzymes, improving the antioxidant function and then promoting the digestion and absorption of nutrients. Furthermore, the above results reduce the emission of harmful gases and heavy metals in feces and urine.

The regulatory role of estrogen receptor beta (ERβ) and the JNK signaling pathway in the effect of high-dose boron on rat splenic lymphocytes has been examined by measuring proliferation, apoptosis, and immune function. The results showed that, compared with the control group, the addition of high-dose boron (40 mmol/L) reduced the proportion of CD3+, CD4+, and CD8+ T lymphocytes, the concentrations of IgG, IL-2, IFN-γ, and IL-4, the proliferation rate of splenic lymphocytes, and the expression levels of PCNA and Bcl-2 mRNA (P < 0.01 or P < 0.05), and increased the apoptosis rate of splenic lymphocytes, caspase-3, and BAX mRNA expression levels (P < 0.01 or P < 0.05). After specific blocking of ERβ, the addition of high-dose boron could not reduce the proportion of CD8+ T lymphocytes, the concentrations of IgG and cytokines IL-2, IFN-γ, and IL-4, the lymphocyte proliferation rate, or PCNA mRNA expression levels, nor could it increase BAX mRNA expression levels. After specific blocking of JNK, the addition of high-dose boron could not increase BAX mRNA expression levels. However, after specific blocking of ERβ and JNK, the addition of high-dose boron could neither reduce the proportion of CD4+ and CD8+ T lymphocytes, the concentrations of IgG and IL-2 (P = 0.05), nor increase caspase-3 and BAX mRNA expression levels (P < 0.01). The results suggest that the ERβ-mediated JNK signaling pathway participates in regulating the effects of high-dose boron on the expression of genes related to the proliferation and apoptosis of rat splenic lymphocytes.