Microbiome-metabolome analysis reveals alterations in the composition and metabolism of caecal microbiota and metabolites with dietary Enteromorpha polysaccharide and Yeast glycoprotein in chickens.

The intestinal microbiome is responsible for the fermentation of complex carbohydrates and orchestrates the immune system through gut microbiota-derived metabolites. In our previous study, we reported that supplementation of Enteromorpha polysaccharide (EP) and yeast glycoprotein (YG) in combination synergistically improved antioxidant activities, serum lipid profile, and fatty acid metabolism in chicken. However, the mechanism of action of these polysaccharides remains elusive. The present study used an integrated 16S-rRNA sequencing technology and untargeted metabolomics technique to reveal the mechanism of action of EP+YG supplementation in broiler chickens fed basal diet or diets supplemented with EP+YG (200mg/kg EP + 200mg/kg YG). The results showed that EP+YG supplementation altered the overall structure of caecal microbiota as evidenced by ß diversities analysis. Besides, EP+YG supplementation changed the microbiota composition by altering the community profile at the phylum and genus levels. Furthermore, Spearman correlation analysis indicated a significant correlation between altered microbiota genera vs serum cytokine levels and microbiota genera vs volatile fatty acids production. Predicted functional analysis showed that EP+YG supplementation significantly enriched amino acid metabolism, nucleotide metabolism, glycan biosynthesis and metabolism, energy metabolism, and carbohydrate metabolism. Metabolomics analysis confirmed that EP+YG supplementation modulates a myriad of caecal metabolites by increasing some metabolites, including pyruvic acid, pyridoxine, spermidine, spermine, and dopamine, and decreasing metabolites related to lipid metabolisms such as malonic acid, oleic acid, and docosahexaenoic acid. The quantitative enrichment analysis results further showed that glycolysis/gluconeogenesis, citric acid cycle, tyrosine metabolism, glycine, serine, and threonine metabolism, and cysteine and methionine metabolism were the most important enriched pathways identified with enrichment ratio >11, whereas, fatty acid biosynthesis and biosynthesis of unsaturated fatty acids pathways were suppressed. Together, the 16S-rRNA and untargeted metabolomics results uncovered that EP+YG supplementation modulates intestinal microbiota and their metabolites, thereby influencing the important metabolism pathways, suggesting a potential feed additive.

Enteromorpha polysaccharide and yeast glycoprotein mixture improves growth, antioxidant activity, serum lipid profile and regulates lipid metabolism in broiler chickens.

This study aimed to analyze the growth performance, antioxidant activity, serum lipid profile, meat quality, and lipid metabolism of broiler chickens fed mixtures containing Enteromorpha polysaccharide (EP) and yeast glycoprotein (YG). A total of 400 one-day-old broiler chickens were randomly divided into 4 treatment groups of 10 replicates with 10 birds each replicate. The dietary treatments consisted of the control group (fed basal diet), and diets supplemented with Enteromorpha polysaccharide (EP; 400 mg/kg), yeast glycoprotein (YG;400 mg/kg), and EP+YG (200 mg/kg EP + 200 mg/kg YG). Compared with the control group, EP+YG supplementation enhanced growth performance and significantly reduced (P < 0.05) serum total triglyceride (TG), cholesterol (CHOL), and low-density lipoprotein LDL levels, and increased high-density lipoprotein (HDL) levels. Besides, birds fed EP+YG supplemented diet exhibited higher (P < 0.05) serum catalase (CAT), total antioxidant capacity, superoxide dismutase (SOD), and lower malonaldehyde (MDA) activities, and upregulated expressions of related genes, such as nuclear factor-erythroid factor 2-related factor 2 (NRF2), SOD1, and glutathione peroxidase 4 (GPX4) in the liver and intestinal tissues than the control group. Interestingly, higher (P < 0.05) serum SOD and lower MDA contents were observed in the EP+YG group than in either EP or YG group, suggesting a synergetic effect. Breast meat from EP+YG supplemented group had significantly higher redness value (a*), and lower pH24, total saturated fatty acid profiles, C14:0, C16:0, C18:0 fatty acid, atherogenic index, and thrombogenicity index than meat from the control group (P < 0.05). Furthermore, the mRNA expressions of fatty acid synthesis genes were downregulated (P < 0.05), whereas lipid ß-oxidation-related genes were upregulated (P < 0.05) in the liver of the EP+YG supplemented group than in the control group. Overall, our data suggest that dietary EP+YG inclusion may have a synergistic effect, and therefore improve growth performance, regulate serum biochemical indexes, enhance antioxidant activity, and modulate lipid metabolism in broilers, indicating that it is a potential feed additive for chickens.

Dietary Enteromorpha Polysaccharide Enhances Intestinal Immune Response, Integrity, and Caecal Microbial Activity of Broiler Chickens.

Marine algae polysaccharides have been shown to regulate various biological activities, such as immune modulation, antioxidant, antidiabetic, and hypolipidemic. However, litter is known about the interaction of these polysaccharides with the gut microbiota. This study aimed to evaluate the effects of marine algae Enteromorpha (Ulva) prolifera polysaccharide (EP) supplementation on growth performance, immune response, and caecal microbiota of broiler chickens. A total of 200 1-day-old Ross-308 broiler chickens were randomly divided into two treatment groups with ten replications of ten chickens in each replication. The dietary treatments consisted of the control group (fed basal diet), and EP group (received diet supplemented with 400 mg EP/kg diet). Results showed that chickens fed EP exhibited significantly higher (P < 0.05) body weight and average daily gain than the chicken-fed basal diet. In addition, significantly longer villus height, shorter crypt depth, and higher villus height to crypt depth ratio were observed in the jejunal and ileal tissues of chickens fed EP. EP supplementation upregulated the mRNA expression of NF-κB, TLR4, MyD88, IL-2, IFN-α, and IL-1ß in the ileal and jejunal tissues (P < 0.05). Besides, we observed significantly higher (P < 0.05) short-chain volatile fatty acids (SCFAs) levels in the caecal contents of the EP group than in the control group. Furthermore, 16S-rRNA analysis revealed that EP supplementation altered gut microbiota and caused an abundance shift at the phylum and genus level in broiler chicken. Interestingly, we observed an association between microbiota and SCFAs production. Overall, this study demonstrated that supplementation of diet with EP promotes growth performance, improves intestinal immune response and integrity, and modulates the caecal microbiota of broiler chickens. This study highlighted the application of marine algae polysaccharides as an antibiotic alternative for chickens. Furthermore, it provides insight to develop marine algae polysaccharide-based functional food and therapeutic agent.

Uncover the Nuclear Proteomic Landscape with Enriched Nuclei Followed by Label-Free Quantitative Mass Spectrometry.

Treated with light pulse or under certain diurnal conditions, photoreceptors can translocate into nucleus followed by conformation change. Many critical components of light signaling pathways also majorly function in nucleus. Hence, it is beneficial to establish a combined method to uncover and compare the nuclear proteomic landscape among the mutants of light signaling components. Here we describe an optimized method to isolate nucleus with seedlings growing under light/dark cycles for further characterizing the nuclear proteome with label-free quantitation by liquid chromatography mass spectrometry (LC-MS).

The Effects of Endoplasmic-Reticulum-Resident Selenoproteins in a Nonalcoholic Fatty Liver Disease Pig Model Induced by a High-Fat Diet.

The present study aimed to investigate the intervention of selenium in the oxidative stress and apoptosis of pig livers, which were induced by a high-fat diet, and the effects of four endoplasmic reticulum (ER)-resident selenoproteins in the process. A 2×4 design trial was conducted that included two dietary fat levels (BD = basal diet and HFD = high-fat diet) and four dietary Se supplementation levels (0, 0.3, 1.0, and 3.0 mg/kg of the diet, in the form of sodium selenite (Na2SeO3)). Our results indicated that the HFD significantly increased the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum, as well as the degree of steatosis, the content of malondialdehyde (MDA), the apoptotic rate, and the level of mRNA caspase-3 in the liver compared to their BD counterparts (p < 0.05). Moreover, these parameters in the HFD groups were more significantly reduced (p < 0.05) for a Se concentration of 1.0 mg/kg than for the other concentrations. Further, for both the BD and HFD, the groups supplemented with 1.0 mg/kg Se showed the highest mRNA level of selenoprotein S. In conclusion, the consumption of an HFD can induce oxidative damage and apoptosis in the liver. This shows that the supplementation of Se at 1.0 mg/kg may be the optimum concentration against damage induced by HFD, and Sels may play a key role in this process.

Effect of bovine lactoferrin supplementation on intestinal inflammatory factor expression in premature rats model of necrotizing enterocolitis / 中华实用儿科临床杂志

Objective:To investigate the protective effects of bovine lactoferrin (bLF) supplementation on intestinal mucosal tissue and its influence on of inflammatory factors in the premature rats model of necrotizing enterocolitis(NEC), and to provide the theoretical basis for prevention of NEC by bLF supplementation.Methods:Premature SD rats were randomly divided into 4 groups, 25 cases in each group.Control group: oral feeding; model group : oral feeding with lipopolysaccharides(LPS) gavage + hypoxic stimulation; high dose bLF intervention group: daily bLF (7 g/L) + oral feeding with LPS gavage + hypoxic stimulation; low dose bLF intervention group: daily bLF (2 g/L) + oral feeding with LPS gavage + hypoxic stimulation.Histopathological analysis was performed by HE staining.The expression levels of interleukin-1β(IL-1β)and interleukin-6(IL-6)in intestinal mucosa were detected by enzyme linked immunosorbent assay (ELISA).Results:(1) Morphological observation: the intestinal wall of model group was thin, and there were different degrees of pneumoconiosis and effusion in intestinal cavity.Under the microscopy, it could be observed that the intestinal tissue necrosis was serious, the intestinal villi fell off, glands arranged disorderly, epithelial edema was significant, the lamina propria and submucosa had severely edema and were separated, and there were a large number of inflammatory cells infiltrated.The above-mentioned manifestations were alleviated in the high-dose and low-dose bLF intervention groups, and no significant abnormalities were found in the control group.(2) The expression of IL-1β and IL-6 in intestinal tissue: the tissue concentration of IL-1β and IL-6 in the model group rats [(380.89±20.25) ng/L, (485.12±31.44) ng/L]were significantly higher than those in the control group[(270.69±45.58) ng/L, (212.62±89.46) ng/L]( q =9.785, 14.030, all P<0.01). The expression of IL-1β and IL-6 in mucosal tissue of ileum was significantly inhibited in hypoxic and LPS-stimulated rats fed with bLF(IL-1β: q=9.105, 8.761, all P<0.01; IL-6: q=8.175, 8.996, all P<0.01). There was no significant difference in the expression of IL-1β and IL-6 between high dose bLF(7 g/L) and low dose bLF (2 g/L) inter vention groups (IL-1β: q=-0.084, P>0.05; IL-6: q=-1.140, P>0.05). Conclusion:Enteral bLF supplementation can alleviate the damage of intestinal tissue in NEC model of premature SD rats, inhibit the expression of IL-1β and IL-6 inflammatory factors in intestinal tissue, and have a protective effect on intestinal tissue.

Effects of Dietary Selenium Deficiency or Excess on Selenoprotein Gene Expression in the Spleen Tissue of Pigs.

To evaluate the effects of dietary Se deficiency and excess on the mRNA levels of selenoproteins in pig spleen tissues, 20 healthy uncastrated boars (Duroc × Landrace × Yorkshire, 10 ± 0.72 kg) were randomly divided into four groups (5 pigs per group). The pigs were fed a Se deficient corn-soybean basal feed (Se content <0.03 mg/kg) or basal feed with added sodium selenite at 0.3, 1.0, or 3.0 mg Se/kg diet, respectively. The experiment lasted 16 weeks. The spleen tissue was collected to examine the mRNA expression levels of 24 selenoprotein genes at the end of the study. Compared with pigs in other groups, those fed with the 1.0 mg Se/kg diet had higher mRNA levels of glutathione peroxidase 1 (Gpx1), glutathione peroxidase 2 (Gpx2), deiodinase type II (Dio2), thioredoxin reductase 3 (Txnrd3), selenoprotein H (Selh), selenoprotein N, 1 (Sepn1), selenoprotein P1 (Sepp1), and selenoprotein V (Selv) in the spleen (p < 0.05). Dietary Se deficiency resulted in lower mRNA levels of Gpx1, Gpx2, glutathione peroxidase 3 (Gpx3), Dio2, thioredoxin reductase 2 (Txnrd2), Txnrd3, Selh, selenoprotein I (Seli), selenoprotein K (Selk), selenoprotein M (Selm), Sepn1, Sepp1, and Selv in the spleen than the other three groups. Dietary Se levels did not affect the mRNA levels of glutathione peroxidase 4 (Gpx4), deiodinase type I (Dio1), deiodinase type III (Dio3), selenophosphate synthetase 2 (Sephs2), thioredoxin reductase 1 (Txnrd1), selenoprotein O (Selo), selenoprotein S (Sels), selenoprotein W (Selw), selenoprotein X (Selx), and selenoprotein 15 (Sel15) in the spleen (p > 0.05). Dietary Se levels can affect the transcription levels of 14 selenoprotein genes in the spleen of pigs.

Dietary taurine supplementation decreases fat synthesis by suppressing the liver X receptor α pathway and alleviates lipid accumulation in the liver of chronic heat-stressed broilers.

BACKGROUND: Chronic heat stress can enhance fat synthesis and result in lipid accumulation in the liver of broilers. To investigate the effects and molecular mechanisms of dietary taurine supplementation on fat synthesis and lipid accumulation in the liver of chronic heat-stressed broilers, 144 28 day-old chickens (Arbor Acres) were randomly distributed to normal control (NC, 22 °C, basal diet), heat stress (HS, consistent 32 °C, basal diet), or heat stress plus taurine (HS + T, consistent 32 °C, basal diet +5.00 g kg-1 taurine) groups for a 14-day feeding trial. RESULTS: Compared with those of the HS group, dietary taurine supplementation significantly decreased the level of very-low-density lipoprotein and the activity of aspartate aminotransferase in plasma and the relative weight of liver in the HS + T group. In addition, dietary taurine supplementation also significantly decreased the levels of triglyceride, acyl-CoA carboxylase (ACC) and fatty acid synthase (FAS), and suppressed the mRNA expression levels of liver X receptor α (LXRα), sterol response element-binding protein 1c, ACC and FAS in the liver of chronic heat-stressed broilers. Meanwhile, dietary taurine supplementation effectively alleviated lipid accumulation in the liver of broilers exposed to chronic heat stress. CONCLUSION: Chronic heat stress significantly increased fat synthesis and resulted in excess lipid deposition in the liver of broilers. Dietary taurine supplementation can effectively decrease fat synthesis by suppressing the LXRα pathway and alleviate lipid accumulation in the liver of chronic heat-stressed broilers. © 2019 Society of Chemical Industry.

Chronic heat stress alters hypothalamus integrity, the serum indexes and attenuates expressions of hypothalamic appetite genes in broilers.

The hypothalamus is crucial to ensure the functionality of the entire organisms, such as body temperature, feed intake and energy regulation. Exposing broilers to high ambient temperature usually induces lower feed intake and energy imbalance. We investigated the molecular mechanisms by which heat stress impairs the appetite via dysfunction in hypothalamus of the broilers. Broilers were allocated to three groups: the normal control (NC) group, and fed ad libitum; heat-stress (HS) group, and fed ad libitum; pair-fed (PF) group, which received the feed intake equal to HS group. Experiment lasted from the age of 28 to 42 d. The results showed that HS increased the head surface temperature of broiler and changed hypothalamic ultrastructure. HS treatment also increased the serum corticosterone in the broilers after 7 days of heat stress, elevated the FT4 and FT3 after 14 days of heat stress. Heat stress of 14 days showed a tendency to increase the leptin. However, the serum corticosterone in the HS group had no significant difference after 14 days of heat stress. In addition, HS treatment decreased the expression of orexigenic gene neuropeptide Y (NPY) after 14 days of heat stress, while HS treatment had no effect on the reactive oxygen species (ROS), as well as the gene expression of AMPKα1 and LKB1 in the hypothalamus. In conclusion, HS increased the surface temperature of head in broiler, and then altered the integrity of hypothalamus. Meanwhile, HS increased the serum corticosterone which may ascribe to the activation of HPA axis in the broilers. In addition, chronic heat stress decreased the expression of orexigenic gene NPY, which may cause the broiler to reduce feed intake.

Effects of dietary taurine supplementation on growth performance, jejunal morphology, appetite-related hormones, and genes expression in broilers subjected to chronic heat stress.

This study was aimed to elucidate effects of taurine supplementation on growth performance, jejunal histology, and appetite-related genes expressions of broilers under heat stress. A total of 144 broilers on 28 d were allocated to three groups with 6 cages each group, 8 broilers per cage. The experiment period is from 28 to 42 d of age. In normal control (NC) group, chickens were held at 22°C ambient temperature (thermoneutral) and fed a basal diet. In the heat stress (HS) group, chickens were raised to constant HS at 32°C and received a basal diet. In the HS+ taurine group, chickens were fed a basal diet with 5 g/kg taurine supplementation. The results showed that HS group had lower average daily feed intake, average daily gain, higher feed/gain ratio compared with the NC group (P < 0.05), while taurine addition did not ameliorate the lowered growth performance. Cloacal temperatures and respiration rates in the HS and heat taurine group were higher (P < 0.05) than in the NC group. Heat stress treatment elevated (P < 0.05) the concentrations of ghrelin and cholecystokinin (CCK) in serum and intestine, together with peptide YY and somatostatin (SS) in the intestine after 7 or 14 d of heat exposure. In addition, HS damaged the jejunal morphology by shortening villus height and deepening crypt depth (P < 0.05), upregulated (P < 0.05) the mRNA expression of taste receptor type 1 member 1 (T1R1), taste receptor type 1 member 3 (T1R3), CCK and ghrelin in the intestine. Taurine supplementation significantly mitigated the impairment of jejunal morphology, decreased the concentrations of serum ghrelin, increased the concentrations of somatostatin and peptide YY in the duodenum, elevated the mRNA expression levels of CCK in the jejunum compared with the HS group. In conclusion, taurine exerted no positive effects on the growth performance, while mitigated the impairment of jejunal morphology, increased some anorexic hormones secretion and mRNA expression of appetite-related genes in the intestine of broilers subjected to HS.

Dietary taurine supplementation improves breast meat quality in chronic heat-stressed broilers via activating the Nrf2 pathway and protecting mitochondria from oxidative attack.

BACKGROUND: Chronic heat stress can induce oxidative impairment and decrease breast meat quality in broilers. Taurine is a ß-amino acid with antioxidant properties. To investigate the alleviative effects and molecular mechanisms of taurine supplementation on breast meat quality in broilers exposed to chronic heat stress, 144 28-day-old chickens (Arbor Acres) were randomly distributed to thermoneutral (TN, 22 °C, basal diet), heat stress (HS, consistent 32 °C, basal diet), or heat stress plus taurine (HS + T, consistent 32 °C, basal diet + 5.00 g kg-1 taurine) groups for a 14-day trial. RESULTS: Chronic heat stress did not affect the contents of moisture, crude protein and crude fat in breast muscle, but impaired breast meat quality in broilers. Taurine supplementation significantly alleviated the increase in lightness and drip loss and the decrease in pH45 min and shear force of breast meat in chronic heat-stressed broilers. Compared with the HS group, taurine supplementation significantly decreased the levels of reactive oxygen species and malonaldehyde and increased the messenger RNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), NAD(P)H quinone dehydrogenase 1 and heme oxygenase 1 in the HS + T group. Meanwhile, taurine supplementation effectively alleviated mitochondrial damage caused by chronic heat exposure. CONCLUSION: Dietary taurine supplementation can effectively improve the quality of breast meat in chronic heat-stressed broilers via activating the Nrf2 pathway and protecting mitochondria from oxidative attack. © 2018 Society of Chemical Industry.

Intestinal Microbiota in Early Life and Its Implications on Childhood Health / 基因组蛋白质组与生物信息学报·英文版

Trillions of microbes reside in the human body and participate in multiple physiological and pathophysiological processes that affect host health throughout the life cycle. The microbiome is hallmarked by distinctive compositional and functional features across different life periods. Accumulating evidence has shown that microbes residing in the human body may play fundamental roles in infant development and the maturation of the immune system. Gut microbes are thought to be essential for the facilitation of infantile and childhood development and immunity by assisting in breaking down food substances to liberate nutrients, protecting against pathogens, stimulating or modulating the immune system, and exerting control over the hypothalamic-pituitary-adrenal axis. This review aims to summarize the current understanding of the colonization and development of the gut microbiota in early life, highlighting the recent findings regarding the role of intestinal microbes in pediatric diseases. Furthermore, we also discuss the microbiota-mediated therapeutics that can reconfigure bacterial communities to treat dysbiosis.

Arabidopsis O-GlcNAc transferase SEC activates histone methyltransferase ATX1 to regulate flowering.

Post-translational modification of proteins by O-linked ß-N-acetylglucosamine (O-GlcNAc) is catalyzed by O-GlcNAc transferases (OGTs). O-GlcNAc modification of proteins regulates multiple important biological processes in metazoans. However, whether protein O-GlcNAcylation is involved in epigenetic processes during plant development is largely unknown. Here, we show that loss of function of SECRET AGENT (SEC), an OGT in Arabidopsis, leads to an early flowering phenotype. This results from reduced histone H3 lysine 4 trimethylation (H3K4me3) of FLOWERING LOCUS C (FLC) locus, which encodes a key negative regulator of flowering. SEC activates ARABIDOPSIS HOMOLOG OF TRITHORAX1 (ATX1), a histone lysine methyltransferase (HKMT), through O-GlcNAc modification to augment ATX1-mediated H3K4me3 histone modification at FLC locus. SEC transfers an O-GlcNAc group on Ser947 of ATX1, which resides in the SET domain, thereby activating ATX1. Taken together, these results uncover a novel post-translational O-GlcNAc modification-mediated mechanism for regulation of HKMT activity and establish the function of O-GlcNAc signaling in epigenetic processes in plants.

Proteomic discovery of H2O2 response in roots and functional characterization of PutGLP gene from alkaligrass.

MAIN CONCLUSION: Hydrogen peroxide-responsive pathways in roots of alkaligrass analyzed by proteomic studies and PutGLP enhance the plant tolerance to saline-, alkali- and cadmium-induced oxidative stresses. Oxidative stress adaptation is critical for plants in response to various stress environments. The halophyte alkaligrass (Puccinellia tenuiflora) is an outstanding pasture with strong tolerance to salt and alkali stresses. In this study, iTRAQ- and 2DE-based proteomics approaches, as well as qRT-PCR and molecular genetics, were employed to investigate H2O2-responsive mechanisms in alkaligrass roots. The evaluation of membrane integrity and reactive oxygen species (ROS)-scavenging systems, as well as abundance patterns of H2O2-responsive proteins/genes indicated that Ca2+-mediated kinase signaling pathways, ROS homeostasis, osmotic modulation, and transcriptional regulation were pivotal for oxidative adaptation in alkaligrass roots. Overexpressing a P. tenuiflora germin-like protein (PutGLP) gene in Arabidopsis seedlings revealed that the apoplastic PutGLP with activities of oxalate oxidase and superoxide dismutase was predominantly expressed in roots and played important roles in ROS scavenging in response to salinity-, alkali-, and CdCl2-induced oxidative stresses. The results provide insights into the fine-tuned redox-responsive networks in halophyte roots.

Chronic heat stress affects muscle hypertrophy, muscle protein synthesis and uptake of amino acid in broilers via insulin like growth factor-mammalian target of rapamycin signal pathway.

Heat stress markedly impairs the growth performance of broilers, such as the reduction of breast muscle mass and yield. The aim of this study was to examine the molecular mechanism of depressed muscle mass and yield caused by heat stress. A total of 144 (28-day-old) male broilers were allocated randomly into 3 treatment groups: (1) the normal control group (environment temperature was 22°C), (2) the heat stress group (environment temperature was 32°C), (3) the pair-fed group (environment temperature was 22°C and pair-fed to heat stress group). The experiment lasted for 14 d (from the age of 28 to 42 d). After 14 d of heat exposure, heat stress decreased (P < 0.05) broiler average daily gain, breast muscle mass, and muscle yield, and increased (P < 0.05) feed to gain ratio. After 14 d of heat exposure, heat stress increased (P < 0.05) the activities of aspartate aminotransferase and the concentrations of uric acid and most amino acids in serum, and reduced (P < 0.05) the concentration of insulin like growth factor 1 (IGF-1) in serum. Additionally, heat stress decreased (P < 0.05) the mRNA expressions of IGF-1, IGF-1 receptor, insulin receptor substrate 1, mammalian target of rapamycin (mTOR), the 70 kD ribosomal protein S6 kinase, myogenic differentiation, myogenin, solute carrier family 38 member 2, solute carrier family 7 member 5, and solute carrier family 3 member 2 of the breast muscle. In conclusion, chronic heat stress resulted in lower breast muscle mass and yield, and decreased muscle protein synthesis and amino acid transportation by downregulating IGFs-mTOR signal pathway. These findings have important practical significance in discovering effective means to alleviate muscle loss caused by chronic heat stress.

Chronic Heat Stress Damages Small Intestinal Epithelium Cells Associated with the Adenosine 5'-Monophosphate-Activated Protein Kinase Pathway in Broilers.

Heat-stressed broilers usually reduce their feed intake, leading to energy imbalance and disturbing the homeostasis in the small intestine. This study was aimed to explore heat-stress-mediated physiological features that may be ascribed to impairments in the intestinal tract of broilers. The results revealed that heat exposure increased the activities of trypsin and Na+/K+-ATPase, while it decreased the activities of amylase, lipase, and maltase as well as the proliferating cell nuclear antigen cells in the jejunum after 14 days of heat exposure. Meanwhile, heat stress upregulated the mRNA expressions of AMPKα1, LKB1, and HIF-1α and protein expressions of p-AMPKαThr172 and p-LKB1Thr189 in the small intestine after 7 or 14 days of heat exposure. In conclusion, chronic heat exposure impeded the development of digestive organs, disordered the activities of intestinal digestive enzymes, and impaired the intestinal epithelial cells by increasing the cell apoptosis and declining cell proliferation, which might be correlated with the adenosine 5'-monophosphate-activated protein kinase signaling pathway. Additionally, heat stress upregulated the gene expression of HIF-1α, which indicated that heat stress may disturb the homeostasis in the intestine.

Serum metabolomics study of nutrient metabolic variations in chronic heat-stressed broilers.

To investigate the effects of heat stress on broiler metabolism, we assigned 144 broilers to normal control (NC), heat stress (HS) or pair-fed (PF) groups and then monitored the effects using growth performance, carcass characteristics, biochemical assays and GC-MS-based metabolomics. The up-regulation of cloacal temperature confirmed that our experiment was successful in inducing chronic heat stress. The average daily gain and average daily feed intake of the HS group were significantly lower than those of the NC group, by 28·76 and 18·42 %, respectively (P1 and P<0·05). The greater feed:gain ratio of the HS group was significantly positively correlated with the leg, abdominal fat, subcutaneous fat and intramuscular fat proportions and levels of some free amino acids (proline, l-cysteine, methionine and threonine) but was negatively correlated with breast proportion and levels of some NEFA (stearic acid, arachidonic acid, palmitic acid and oleic acid). These findings indicated that the heat-stressed broilers were in negative energy balance and unable to effectively mobilise fat, thereby resulting in protein decomposition, which subsequently affected growth performance and carcass characteristics.

Effects of chronic heat exposure on growth performance, intestinal epithelial histology, appetite-related hormones and genes expression in broilers.

BACKGROUND: Heat stress often occurs in the modern poultry industry, which impairs growth performance, particularly via reducing appetite. This study was aimed to investigate the mechanism of attenuating appetite by chronic heat exposure in broilers. A total of 144 broilers (28 days old) were allocated to normal control (NC, 22 °C), heat stress (32 °C), and pair-fed (22 °C) groups. RESULTS: Chronic heat exposure significantly increased cloacal temperatures and respiration rates, decreased the average daily feed intake and average daily gain, increased feed-to-gain ratio compared with the NC group, and elevated the concentrations of ghrelin and cholecystokinin (CCK) both in serum and intestine, as well as peptide YY and somatostatin in intestine on 35- or 42-day-old broilers. Moreover, heat exposure decreased villi height (VH) and the ratio of VH to crypt depth (CD), while it increased CD in the jejunum on 35- and 42 day-olds, increased (P < 0.05) the concentrations of valine and isoleucine in plasma on 42 days, and upregulated (P < 0.05) the expression of taste receptor type 1 members 1 and 3 (T1R1 and T1R3), CCK, and ghrelin in the intestine on 35- or 42-day-old broilers. CONCLUSION: Chronic heat exposure impairs the performance, intestinal morphology and appetite, which may be correlated with the increased secretion or gene expression of appetite-related hormones and genes, and the higher expression of nutrient-sensing receptors (T1R1 and T1R3) in broilers. © 2018 Society of Chemical Industry.

A simplified method for isolating murine primary pancreatic acinar cells / 中华胰腺病杂志

Objective To explore a simplified and economical method to isolate the murine primary pancreatic acinar cells.Methods The collagenase and trypsin inhibitor dissolving in DMEM solution were used to digest the murine pancreas,and 4％ BSA dissolving in DMEM solution was used to purify and isolate primary pancreatic acinar cells from pancreas.CCK-8 method was applied to check the ability of pancreatic acinar cells to secret amylase.Results After digestion,shaking in the water bath,resuspension,filtration and precipitation,murine primary pancreatic acinar cells could be obtained within 2 hours.Pancreatic acinar cells in good conditions appeared in clusters,and their basolateral domains were round and devoid of blebs,and the cytoplasm appeared clear.Their apical domain were surrounded by hundreds of zymogen granules which looked darker.The nucleus was located in the basal area of the vesicular region.The basal level of amylase release as a percent of total release from pancreatic acinar cells was around 2.5％ in CCK8-unstimulated group.This rate started to increase after CCK-8 stimulation and reached its peak [(12.83 ± 1.04) ％] at a concentration of 50 pmol/L of CCK-8,but the ratio of the amylase level secreted by the pancreatic acinar cells to the total amylase level displayed a decreasing trend with the increase of CCK-8 concentration.Conclusions This optimized method had the advantage of being fast and simple,low technical difficulty and good repetition.It was a new simplified and cheap method for isolating murine pancreatic acinar cells.

Effects of ApoC3 on the severity of acute pancreatitis in mice / 中华胰腺病杂志

Objective To explore the effects of ApoC3 gene on the severity of hypertriglyceridemiainduced acute pancreatitis (AP).Methods ApoC3 transgenetic mice and C57BL/6J mice AP model was induced by cerulein intraperitoneal injection,and ApoC3 transgenetic mice and C57BL/6J mice injected by normal saline solution in equal volume served as control group.Serum triglyceride and cholesterol were detected,and the pathological changes of the pancreas were observed.RT PCR method was used to examine the changes of the inflammatory factor including IL-1β,IL-6,α-SMA and TNF-α mRNA levels,which reflected the severity of the inflammation.Results Serum triglyceride and cholesterol were higher in ApoC3 transgenetic mice than in C57BL/6J mice [(3.434 ± 0.931) mmol/L vs (0.766 ± 0.120) mmol/L,(2.553 ±0.178) mmol/L vs (1.996 ± 0.080) mmol/L],and the differences were statistically different (P ＜ 0.05).The pathological changes of the pancreas were more severe in ApoC3 transgenetic AP mice than in C57BL/6J AP mice,and the IL-1β,IL-6 and α-SMA mRNA levels in the pancreatic tissue were obviously higher in ApoC3 transgenetic AP mice than in C57BL/6J mice (1.72 ± 0.07vs 0.78 ± 0.09,1.58 ± 0.09vs 0.87 ±0.04,0.83 ± 0.05vs 0.44 ± 0.04),and the differences were statistically significant (P ＜ 0.05),while there was no statistical difference on TNF-αmRNA level (0.70 ± 0.09vs 0.65 ± 0.08,P ＞ 0.05).Conclusions ApoC3 gene could aggravate the severity of the inflammation in hypertriglyceridemia-induced AP.