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A study was carried out to investigate the effects of dietary methionine source and level on plasma free amino acids patterns and the expression of genes involved in hepatic methionine metabolism in broiler breeders. A total of 2184 broiler breeders were assigned to 13 dietary treatments, with eight replicates per treatment. The 13 treatments included one control group and 12 additional treatments employing two sources and six levels (0.05, 0.10, 0.15, 0.20, 0.25 and 1.00%). Higher plasma methionine concentration was measured for DL-methionine (DLM) treated hens. Plasma alanine concentration was linearly increased as DLM or 2-hydroxy-4-(methylthio) butanoic acid (HMTBA) supplementation level increased. There was a linear increase in concentrations of tyrosine, valine, glycine and serine as dietary DLM supplementation level increased. Hens treated with DLM had higher relative expression of ADA than those fed HMTBA. The expression of MS, ADA, SAHH and MAT2A changed quadratically as HMTBA supplementation level increased, while the expression of GNMT and SAHH changed quadratically as DLM supplementation level increased. In conclusion, the effects of HMTBA on plasma free amino acid patterns and the expression of hepatic genes involved with methionine are different from DLM., (© 2017 The Authors. Animal Science Journal published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Animal Science.)

Background and Objectives: Damage to normal bone tissue following therapeutic irradiation (IR) represents a significant concern, as IR-induced bone microenvironment disruption can cause bone loss and create a more favorable environment for tumor metastases. The aim of the present study was to explore the cellular regulatory mechanism of IR-induced bone microenvironment disruption to effectively prevent radiotherapy-associated adverse effects in the future. Materials and Methods: In this study, a mouse model of local IR was established via local irradiation of the left hind limb of BALB/c mice with 12 Gy X-rays, and an in vitro osteocyte (OCY) model was established by exposing osteocyte-like MLO-Y4 cells to 2, 4, and 8 Gy irradiation to analyze multicellular biological injuries and cellular senescence. Small interfering RNA (siRNA) transfection at the cellular level and a selective antagonist intervention C-176 at the animal level were used to explore the potential role of the stimulator of interferon genes (STING) on IR-induced bone microenvironment disruption. Results: The results showed that 12 Gy local IR induces multicellular dysfunction, manifested as ascension of OCYs exfoliation, activation of osteoclastogenesis, degeneration of osteogenesis and fate conversion of adipogenesis, as well as cellular senescence and altered senescence-associated secretory phenotype (SASP) secretion. Furthermore, the expression of STING was significantly elevated, both in the primary OCYs harvested from locally irradiated mice and in vitro irradiated MLO-Y4 cells, accompanied by the markedly upregulated levels of phosphorylated TANK-binding kinase 1 (P-TBK1), RANKL and sclerostin (SOST). STING-siRNA transfection in vitro restored IR-induced upregulated protein expression of P-TBK1 and RANKL, as well as the mRNA expression levels of inflammatory cytokines, such as IL-1α, IL-6 and NF-κB, accompanied by the alleviation of excessive osteoclastogenesis. Finally, administration of the STING inhibitor C-176 mitigated IR-induced activation of osteoclastogenesis and restraint of osteogenesis, ameliorating the IR-induced biological damage of OCYs, consistent with the inhibition of P-TBK1, RANKL and SOST. Conclusions: The STING-P-TBK1 signaling pathway plays a crucial role in the regulation of the secretion of inflammatory cytokines and osteoclastogenesis potential in IR-induced bone microenvironment disruption. The selective STING antagonist can be used to intervene to block the STING pathway and, thereby, repair IR-induced multicellular biological damage and mitigate the imbalance between osteoclastogenesis and osteoblastgenesis. [ABSTRACT FROM AUTHOR]

In legumes, a LysM-RLK perception of rhizobial lipo-chitooligosaccharides (LCOs) known as Nod factors (NFs), triggers a signaling pathway related to the onset of symbiosis development. On the other hand, activation of LysM-RLKs upon recognition of chitin-derived short-chitooligosaccharides initiates defense responses. In this work, we identified the members of the LysM-RLK family in cultivated (Arachis hypogaea L.) and wild (A. duranensis and A. ipaensis) peanut genomes, and reconstructed the evolutionary history of the family. Phylogenetic analyses allowed the building of a framework to reinterpret the functional data reported on peanut LysM-RLKs. In addition, the potential involvement of two identified proteins in NF perception and immunity was assessed by gene expression analyses. Results indicated that peanut LysM-RLK is a highly diverse family. Digital expression analyses indicated that some A. hypogaea LysM-RLK receptors were upregulated during the early and late stages of symbiosis. In addition, expression profiles of selected LysM-RLKs proteins suggest participation in the receptor network mediating NF and/or chitosan perception. The analyses of LysM-RLK in the non-model legume peanut can contribute to gaining insight into the molecular basis of legume–microbe interactions and to the understanding of the evolutionary history of this gene family within the Fabaceae. [ABSTRACT FROM AUTHOR]

Yucca schidigera extract (YE) can decrease ammonia concentration in livestock housing, which could be associated with the inhibition of urease. The aim of this study was to investigate the other possible reasons of dietary YE supplementation reducing nitrogen emission in weaned piglets. A total of 14 crossbred weaned barrows were allotted into two groups fed the diets supplementing 0 and 120 mg/kg YE for 14 days. The YE administration decreased F/G ratio and hindgut NH3‐N production in weaned piglets (p < 0.05). Dietary YE supplementation decreased serum urea nitrogen levels, and increased nutrient digestibility, which could be related to the improvement of morphology, digestive and absorptive enzyme activities, and nutrient transporter mRNA expression in jejunal mucosa of weaned piglets (p < 0.05). The mRNA expression of tight junction proteins, mucins and apoptosis‐related genes was also improved by YE treatment in jejunal mucosa of weaned piglets (p < 0.05). In addition, dietary YE supplementation regulated the microbiota structure and volatile fatty acid content in distal intestine of weaned piglets (p < 0.05). These results suggest that YE administration can decrease hindgut NH3–N production in weaned piglets, which is associated with the increased nutrient utilization and gut‐barrier function. [ABSTRACT FROM AUTHOR]

Objective. Accumulated evidence highlights the biological implications of exosomes in gastric cancer. Herein, we conducted the exosomal miRNA expression profiling and identified potential diagnostic markers for gastric cancer. Methods. Plasma exosomes were isolated and identified from three gastric cancer patients and three healthy participants. Microarrays of exosomal miRNAs were then analyzed. Differentially expressed exosomal miRNAs were screened with fold − change ≥ 2.0 and p ≤ 0.05. Among them, miR-4741, miR-32, miR-3149, and miR-6727 expressions were verified in tissues and plasma of patients and healthy subjects. ROC curves were conducted for evaluating the diagnostic performance. The roles of miR-32, miR-3149, miR-6727, and miR-4741 on gastric cancer progression were observed by cellular experiments. Results. Isolated exosomes were well characterized by Western blot and transmission electron microscopy as well as nanoparticle-tracking analyses. According to the microarrays, 142 exosomal miRNAs were upregulated, and 34 were downregulated in gastric cancer than healthy subjects. miR-4741 upregulation and miR-32, miR-3149, and miR-6727 downregulations were found in tissues and plasma of gastric cancer patients. The AUCs of miR-4741, miR-32, miR-3149, and miR-6727 were separately 0.8554, 0.9456, 0.7683, and 0.8923. Upregulated miR-32, miR-3149, and miR-6727 as well as downregulated miR-4741 lowered proliferative, migratory, and invasive capacities as well as elevated apoptotic levels of gastric cancer cells. Conclusion. Our study successfully isolated and verified exosomes from plasma of gastric cancer as well as proposed four exosomal miRNAs that could act as promising diagnostic markers and suppress gastric cancer progression. [ABSTRACT FROM AUTHOR]

Objective. Spontaneous intracerebral hemorrhage (ICH) is a blood clot arising in the brain parenchyma in the absence of trauma or surgery and accounts for 10% to 15% of all strokes, leading to higher rates of mortality and morbidity than either ischemic stroke or subarachnoid hemorrhage. We sought to investigate the potential association of DOCK1 with neurological deficits and outcomes in patients with spontaneous ICH. Methods. Identification of methylation-regulated differentially expressed genes (MeDEGs) between ICH patients and matched controls was performed by analyzing the raw data from the GSE179759 and GSE125512 datasets deposited in the Gene Expression Omnibus. A total of 114 patients who were admitted to our hospital for spontaneous ICH were retrospectively analyzed, with 108 healthy volunteers who had received physical examinations at the same period as controls. The mRNA expression of DOCK1 was determined by quantitative real-time polymerase chain reaction (qRT-PCR). The hematoma volume was calculated according to the Coniglobus formula. The severity of neurological deficits was evaluated using National Institutes of Health Stroke Scale (NIHSS) scores and function outcomes were evaluated by modified Rankin Scale (mRS) scores. Results. A total of 15 MeDEGs between ICH patients and matched controls were identified. The mRNA expression of DOCK1 was remarkably higher in the serum samples of patients with spontaneous ICH than in the healthy controls. According to hematoma volume after ICH attack, small (<10 mL), medium (10 to 30 mL), and large (>30 mL) groups were arranged. The proportions of male patients and patients aged ≥60 years were significantly higher in the large group than in the small and medium groups (P < 0.05). The mRNA expression of DOCK1 was significantly higher in the large group than in the small and medium groups (P < 0.05). According to NIHSS scores, mild (NIHSS scores ≤15), moderate (NIHSS scores from 16 to 30), and severe (NIHSS scores from 31 to 45) groups were classified. It was observed that the severe group had higher proportions of male patients and patients aged ≥60 years than the mild and moderate groups (P < 0.05). The severe group exhibited a higher mRNA expression of DOCK1 than the mild and moderate groups (P < 0.05). According to mRS scores, higher proportions of male patients and patients aged ≥60 years were observed in the unfavorable group than the favorable group (P < 0.05). The patients in the unfavorable group showed an elevated DOCK1 mRNA expression compared to those in the favorable group (P < 0.05). Conclusion. The study provided evidence that male gender, older age, and higher DOCK1 mRNA expression were related to higher admission hematoma volume, neurologic deterioration, and poor function outcomes in patients with spontaneous ICH. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors, and its prognosis is still poor. Mesencephalic astrocyte-derived neurotrophic factor (MANF) plays a key role in endoplasmic reticulum stress. ER stress plays a key role in HCC carcinogenesis. To confirm the clinical and prognostic value of MANF in HCC, we investigated the expression level of MANF in HCC as recorded in databases, and the results were verified by experiment. Survival analysis was probed by the Kaplan–Meier method. Cox regression models were used to ascertain the prognostic value of MANF in HCC tissue microarray. The diagnostic value of MANF in HCC was evaluated by receiver operating characteristic curve analysis. Potential correlation between MANF and selected genes was also analyzed. Results showed that MANF was overexpressed in HCC. Patients with high MANF expression levels had a worse prognosis and higher risk of tumor recurrence. Furthermore, the expression level of MANF had good diagnostic power. Correlation analysis revealed potential regulatory networks of MANF in HCC, laying a foundation for further study of the role of MANF in tumorigenesis. In conclusion, MANF was overexpressed in HCC and related to the occurrence and development of HCC. It is a potential diagnostic and prognostic indicator of HCC. [ABSTRACT FROM AUTHOR]

Resistant starch (RS) was recently approved to exert a powerful influence on gut health, but the effect of RS on the caecal barrier function in meat ducks has not been well defined. Thus, the effect of raw potato starch (RPS), a widely adopted RS material, on microbial composition and barrier function of caecum for meat ducks was determined. A total of 360 Cherry Valley male ducks of 1-d-old were randomly divided and fed diets with 0 (control), 12, or 24 % RPS for 35 d. Diets supplemented with RPS significantly elevated villus height and villus height:crypt depth ratio in the caecum. The 16S rRNA sequence analysis indicated that the diet with 12 % RPS had a higher relative abundance of Firmicutes and the butyrate-producing bacteria Faecalibacterium , Subdoligranulum , and Erysipelatoclostridium were enriched in all diets. Lactobacillus and Bifidobacterium were significantly increased in the 24 % RPS diet v. the control diet. When compared with the control diet, the diet with 12 % RPS was also found to notably increase acetate, propionate and butyrate contents and up-regulated barrier-related genes including claudin-1, zonula occludens-1, mucin-2 and proglucagon in the caecum. Furthermore, the addition of 12 % RPS significantly reduced plasma TNF- α , IL-1 β and endotoxin concentrations. These data revealed that diets supplemented with 12 % RPS partially improved caecal barrier function in meat ducks by enhancing intestinal morphology and barrier markers expression, modulating the microbiota composition and attenuating inflammatory markers. [ABSTRACT FROM AUTHOR]

DL-2-hydroxy-4(methylthio) butanoic acid (DL-HMTBA) exhibits a higher antioxidant capability in vitro as compared to DL-Met, but the mechanism is still not known. A total of 400 8-day-old broiler chicks were allotted to a 2 [low (12 to 14°C) vs. control temperature (thermoneutral, 24 to 26°C)] × 2 (0.17% or 0.51% of DL-HMTBA) factorial arrangement to investigate effects of DL-HMTBA on antioxidant capacity and its related gene expression in lung and liver of broilers exposed to low temperature. The hepatic glutathione (GSH), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activities were decreased, whereas protein carbonyl and malodndialdehyde contents in lung were increased in low temperature (P < 0.05). Dietary supplementation of 0.51% DL-HMTBA increased (P < 0.05) GSH and GSH-Px activity in liver and SOD activity in lung in spite of the temperature. The greater gene expression of GSH reductase and lower expression of GSH synthetase (P < 0.01) were observed in lung of broilers that were maintained at low temperature. Higher DL-HMTBA supplementation induced greater (P < 0.05) mRNA expression of glutathione-S transferase in lung, GSH synthetase in liver and lung, as well as lower expression of GSH reductase in lung, and this effect were more obvious for the chicks exposed to low temperature (interaction, P < 0.05). Chicks that were maintained under low temperature had the lower expression of cystathionine β-synthase (CβS), whereas those fed 0.51% DL-HMTBA increased the CβS expression in liver under low temperature (interaction, P ≤ 0.05). The γ-glutamylcysteine synthetase and Met adenosyltransferase 1 (MAT1) gene expression were downregulated (P < 0.05) by low temperature and the higher (P = 0.02) expression of MAT1 was observed in lung of chicks fed 0.51% DL-HMTBA. It indicated that under low temperature, DL-HMTBA supplementation at 0.51% upregulated gene expression of GSH synthesis and Met transsulfuration pathway in liver and lung to increase the antioxidant capacity, and then mitigate the negative effects of cold stress for broilers. [ABSTRACT FROM AUTHOR]

The experiment was conducted to investigate the bioavailability of manganese (Mn) from humate–Mn complex relative to Mn sulphate for the starter broilers fed a conventional corn–soya bean meal diet. A total of 560 1‐day‐old Arbor Acres male broiler chicks were randomly allotted to one of eight replicate cages (10 chicks per cage) for each of seven treatments in a completely randomized design involving a 2 × 3 factorial arrangement of treatments with two Mn sources (humate‐Mn and Mn sulphate) and three levels of added Mn (60, 120 or 180 mg Mn/kg) plus a Mn‐unsupplemented control diet containing 27.23 mg Mn/kg by analysis. At 14 days of age, the blood, liver, heart and tibia were collected for Mn analyses, and the activity and mRNA abundance of heart manganese superoxide dismutase (MnSOD). The results showed that humate‐Mn supplementation decreased feed intake from day 1 to day 14, whereas it did not influence (p > 0.20) body weight at day 14 as compared to Mn sulphate. The Mn source did not influence Mn concentration in the liver, heart and tibia, and the activity and mRNA abundance of heart MnSOD, while humate‐Mn decreased plasma Mn as compared to Mn sulphate. The Mn concentration in the plasma and heart, and the activity and mRNA abundance of heart MnSOD increased linearly as dietary Mn concentration increased. Based on slope ratios from multiple linear regressions of Mn concentrations in the plasma and heart, and the activity and mRNA abundance of heart MnSOD on daily intake amount of dietary analysed Mn, the bioavailability of humate–Mn complex relative to Mn sulphate (100%) was 82.8, 90.4, 82.8 and 81.9 respectively. These results indicated that the Mn from humate‐Mn complex was just as bioavailable as the Mn from Mn sulphate for the starter broilers (day 1–14). [ABSTRACT FROM AUTHOR]

Both genetic selection and increasing nutrient density for improving growth performance had inadvertently increased leg problems of meat ducks, which adversely affects animal welfare. We hypothesised that slowing weight gain with improving tibia quality probably enhanced tibial mechanical properties and alleviated leg deformities. Therefore, the present study aimed to evaluate the effect of graded Ca supplementation in a low-nutrient density (LND) diet on tibia composition and bone turnover in meat ducks. A total of 720 15-d-old male meat ducks were randomly assigned and fed a standard nutrient density positive control (PC) diet containing 0·9 % Ca, and four LND diets with 0·5, 0·7, 0·9 and 1·1 % Ca, respectively. Ducks fed the 0·5 % Ca LND diet and the PC diet had higher incidence of tibial dyschondroplasia (TD). When compared with the 0·5 % Ca LND diet, LND diets with ≥0·7 % Ca significantly improved tibia composition, microarchitecture and mechanical properties, and consequently decreased the incidence of TD. Furthermore, LND diets with ≥0·7 % Ca increased osteocyte-specific gene mRNA expression, blocked the expression of osteoblast differentiation marker genes including osteocalcin , collagenase-1 and alkaline phosphatase (ALP), and also decreased the expression of osteoclast differentiation genes, such as vacuolar-type H+-ATPase, cathepsin K and receptor activator of NF-κB. Meanwhile bone markers such as serum ALP, osteocalcin (both osteoblast markers) and tartrate-resistant acid phosphatase (an osteoclast marker) were significantly decreased in at least 0·7 % Ca treated groups. These findings indicated that LND diets with ≥0·7 % Ca decreased bone turnover, which subsequently increased tibia quality for 35-d-old meat ducks. [ABSTRACT FROM AUTHOR]

Purpose: Down-regulated expression of the putative tumor suppressor gene spleen tyrosine kinase ( SYK) is strongly associated with the development of various cancers, including colorectal cancer (CRC). SYK gene promoter polymorphisms have been shown to be involved in the pathogenesis of multiple malignant tumors. In this study, we investigated associations of SYK gene promoter polymorphisms with the susceptibility to colorectal cancer development in a Southern Han Chinese population. Methods: SNPs in the promoter region of the human SYK gene were identified using in silico analysis tools, linkage disequilibrium analysis, and a search for likely transcription factor binding sites via TFSEARCH in the NCBI SNP database (gene ID: 6850). Based on this information, −803A > T and −534T > C were selected as candidates for further analysis. TaqMan-MGB probe analyses were performed in 567 CRC patients and 569 age- and gender-matched healthy controls for SYK gene promoter genotyping. Associations between CRC risk and SNPs were estimated using an unconditional logistic regression model, and environmental risk factors were included in a multivariate logistic regression model for correction. Results: The frequencies of the TA and TT genotypes and the T allele of the -803A > T SNP were found to be significantly higher in the CRC patients compared to the healthy individuals of the control group ( P = 0.020, 0.023, and 0.013, respectively). Synergistic effects between -803A > T genotypes (i.e., TA + TT) and age (≤60 years; P = 0.039), male gender ( P = 0.011), smoking ( P = 0.005), drinking alcohol ( P = 0.002), and high BMI (≥24.0 kg/m; P = 0.009) were found to increase the risk to develop CRC by stratified analyses. Conclusions: The SYK −803 A > T genotypes TA and TT are independent risk factors for CRC development in Han Chinese in Southern China, and an association with TA + TT genotypes appears predominant among younger patients, male patients, patients with a high BMI, and patients who smoke or drink alcohol. [ABSTRACT FROM AUTHOR]

Simple sequence repeat ( SSR) markers and conserved-intron scanning primer ( CISP) markers developed from cereal expressed sequence tag sequences were successfully used to construct a linkage map for Lolium multiflorum Lam. using a L. multiflorum × Lolium temulentum L. × L. temulentum BC1 population consisting of 88 individuals. The genetic linkage map included 108 SSR and 84 CISP markers that covered 488.8 centimorgan (cM) in seven linkage groups, ranging in length from 42.2 to 98.9 cM. The average density of markers was 2.5 cM. Syntenic relationships between Lolium and rice ( Oryza sativa L.) based on the positions of the CISP markers largely agree with the previously reported relationships. The map constructed in the present study will be useful in developing a denser linkage map as well as for analysis of quantitative trait loci for important agronomic traits in Lolium species. [ABSTRACT FROM AUTHOR]

Background: Programmed death ligand-1 (PD-L1) has been identified as a factor associated with poor prognosis in a range of cancers, and was reported to be mainly induced by PTEN loss in gliomas. However, the clinical effect of PD-L1 and its regulation by PTEN has not yet been determined in colorectal cancer (CRC). In the present study, we verified the regulation of PTEN on PD-L1 and further determined the effect of PTEN on the correlation between PD-L1 expression and clinical parameters in CRC. Methods/Results: RNA interference approach was used to down-regulate PTEN expression in SW480, SW620 and HCT116 cells. It was showed that PD-L1 protein, but not mRNA, was significantly increased in cells transfected with siRNA PTEN compared with the negative control. Moreover, the capacity of PTEN to regulate PD-L1 expression was not obviously affected by IFN-γ, the main inducer of PD-L1. Tissue microarray immunohistochemistry was used to detect PD-L1 and PTEN in 404 CRC patient samples. Overexpression of PD-L1 was significantly correlated with distant metastasis (P<0.001), TNM stage (P<0.01), metastatic progression (P<0.01) and PTEN expression (P<0.001). Univariate analysis revealed that patients with high PD-L1 expression had a poor overall survival (P<0.001). However, multivariate analysis did not support PD-L1 as an independent prognostic factor (P = 0.548). Univariate (P<0.001) and multivariate survival (P<0.001) analysis of 310 located CRC patients revealed that high level of PD-L1 expression was associated with increased risks of metastatic progression. Furthermore, the clinical effect of PD-L1 on CRC was not statistically significant in a subset of 39 patients with no PTEN expression (distant metastasis: P = 0.102; TNM stage: P = 0.634, overall survival: P = 0.482). Conclusions: PD-L1 can be used to identify CRC patients with high risk of metastasis and poor prognosis. This clinical manifestation may be partly associated with PTEN expression. [ABSTRACT FROM AUTHOR]

Abstract: Aquaporin 3 (AQP3) and c-Met are both overexpressed in human gastric carcinoma and highly associated with its metastasis and invasion. However, it still remains unknown whether c-Met and AQP3 correlate with each other. Herein, we demonstrated that c-Met expression in gastric cancer tissues significantly correlated with differentiation, lymph node metastasis and lymphovascular invasion, and c-Met exhibited marked association with AQP3 expression. Immunoblotting assays showed that hHGF phosphorylated c-Met in SGC7901 and AGS cells and upregulated AQP3 expression in a dose- or time-dependent way. RNAi against c-Met reduced total c-Met levels by about two thirds in both AGS and SGC7901 cells and attenuated hHGF-induced AQP3 expression significantly. In vitro migration and proliferation assays showed that siRNA against AQP3 noticeably restrained HGF-promoted migration and proliferation of these cells. Furthermore, Immunoblotting studies revealed that HGF induced phosphorylation of ERK, and pre-treatment with U0126, a MAPK/ERK inhibitor, partially inhibited hHGF-induced increase in AQP3 expression. Together, these data provide initial evidence that c-Met regulates the expression of AQP3 via the ERK signalling pathway in gastric carcinoma. These findings assist in understanding the mechanism of growth and invasion of gastric carcinoma, and provide a possible strategy for the inhibition of gastric tumor metastasis. [Copyright &y& Elsevier]

The high stocking density is a major stress factor that adversely affects the health and performance of poultry. Therefore, the object of this study was conducted to explore whether dietary 25-hydroxyvitamin D (25-OH-D 3) could improve gut health of laying hens reared under high stocking density. A 2 × 2 factorial design was used in this 16-week study, in which 800 45-week-old Lohmann laying hens were allocated into two levels of dietary 25-OH-D 3 levels (0 and 69 µg/kg) and two rates of stocking densities [506 (low density, LD) and 338 (high density, HD) cm2/hen]. Compared with the layers with LD, the layers with HD had lower crypt depth in duodenum (P (Density) < 0.05), lower short chain fatty acid (propionic and butyric acid) contents in cecum (P (Density) < 0.05), and lower mRNA expression of intestinal barrier associated protein (claudin-1, mucin-1 and mucin-2). Exposed layer to HD also led to lower intestinal antioxidative capacity [superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (T-AOC), and higher malondialdehyde (MDA) content] in small intestine (P (Density) < 0.05), lower bacterial abundance of Bacteroidetes (phylum), Spirochaetes (phylum) and Bacteroides (genus; P (Density) < 0.05), higher bacterial enrichment of Lactobacillaceae (genus) and Firmicutes/Bacteroidetes ratio (P (Density) < 0.05) in cecum. Dietary 25-OH-D 3 increased the villus height in duodenum and jejunum (P (25-OH-D3) < 0.05), decreased Chao 1 and ACE indexes in cecum (P (25-OH-D3) < 0.05), and it also up-regulated the mRNA expression of claudin-1, mucin-1 and mucin-2 (P (25-OH-D3) < 0.05). Layers treated with 25-OH-D 3 led to an enhanced antioxidative enzyme activity of CAT (P (25-OH-D3) < 0.05). Additionally, the effect of 25-OH-D 3 reversed the effect of HD on T-AOC and MDA content (P (Interaction) < 0.05). In HD layers, 25-OH-D 3 administration decreased the enrichment of Bacteroidetes (phylum), increased Firmicutes (phylum), and Firmicutes/Bacteroidetes ratio (P (Interaction) < 0.05). These results suggest that supplementing 25-OH-D 3 in diets may elevate gut health through the improvement of intestinal barrier function, antioxidant capacity and cecal microbiota composition in laying hens with high stocking density. [ABSTRACT FROM AUTHOR]

Background: Peanut is an important legume crop growing worldwide. With the published allotetraploid genomes, further functional studies of the genes in peanut are very critical for crop improvement. CRISPR/Cas9 system is emerging as a robust tool for gene functional study and crop improvement, which haven't been extensively utilized in peanut yet. Peanut plant forms root nodules to fix nitrogen through a symbiotic relationship with rhizobia. In model legumes, the response of plants to rhizobia is initiated by Nod factor receptors (NFRs). However, information about the function of NFRs in peanut is still limited. In this study, we applied the CRISPR/Cas9 tool in peanut hairy root transformation system to explore the function of NFR genes. Results: We firstly identified four AhNFR1 genes and two AhNFR5 genes in cultivated peanut (Tifrunner). The gene expression analysis showed that the two AhNFR1 and two AhNFR5 genes had high expression levels in nodulating (Nod+) line E5 compared with non-nodulating (Nod-) line E4 during the process of nodule formation, suggesting their roles in peanut nodulation. To further explore their functions in peanut nodulation, we applied CRISPR technology to create knock-out mutants of AhNFR1 and AhNFR5 genes using hairy root transformation system. The sequencing of these genes in transgenic hairy roots showed that the selected AhNFR1 and AhNFR5 genes were successfully edited by the CRISPR system, demonstrating its efficacy for targeted mutation in allotetraploid peanut. The mutants with editing in the two AhNFR5 genes showed Nod- phenotype, whereas mutants with editing in the two selected AhNFR1 genes could still form nodules after rhizobia inoculation. Conclusions: This study showed that CRISPR-Cas9 could be used in peanut hairy root transformation system for peanut functional genomic studies, specifically on the gene function in roots. By using CRISPR-Cas9 targeting peanut AhNFR genes in hairy root transformation system, we validated the function of AhNFR5 genes in nodule formation in peanut. [ABSTRACT FROM AUTHOR]

Background: The SWEET (Sugars Will Eventually be Exported Transporters) gene family is a recently identified group of sugar transporters that play an indispensable role in sugar efflux, phloem loading, plant-pathogen interaction, nectar secretion, and reproductive tissue development. However, little information on Saccharum SWEET is available for this crop with a complex genetic background. Results: In this study, 22 SWEET genes were identified from Saccharum spontaneum Bacterial Artificial Chromosome libraries sequences. Phylogenetic analyses of SWEETs from 11 representative plant species showed that gene expansions of the SWEET family were mainly caused by the recent gene duplication in dicot plants, while these gene expansions were attributed to the ancient whole genome duplication (WGD) in monocot plant species. Gene expression profiles were obtained from RNA-seq analysis. SWEET1a and SWEET2s had higher expression levels in the transitional zone and maturing zone than in the other analyzed zones. SWEET1b was mainly expressed in the leaf tissues and the mature zone of the leaf of both S. spontaneum and S. officinarum, and displayed a peak in the morning and was undetectable in both sclerenchyma and parenchyma cells from the mature stalks of S. officinarum. SsSWEET4a\4b had higher expression levels than SWEET4c and were mainly expressed in the stems of seedlings and mature plants. SWEET13s are recently duplicated genes, and the expression of SWEET13s dramatically increased from the maturing to mature zones. SWEET16b's expression was not detected in S. officinarum, but displayed a rhythmic diurnal expression pattern. Conclusions: Our study revealed the gene evolutionary history of SWEETs in Saccharum and SWEET1b was found to be a sucrose starvation-induced gene involved in the sugar transportation in the high photosynthetic zones. SWEET13c was identified as the key player in the efflux of sugar transportation in mature photosynthetic tissues. SWEET4a\4b were found to be mainly involved in sugar transportation in the stalk. SWEET1a\2a\4a\4b\13a\16b were suggested to be the genes contributing to the differences in sugar contents between S. spontaneum and S. officinarum. Our results are valuable for further functional analysis of SWEET genes and utilization of the SWEET genes for genetic improvement of Saccharum for biofuel production. [ABSTRACT FROM AUTHOR]

Abstract: Chronic IFN-α treatment as an antiviral or anti-cancer therapy can lead to severe psychiatric complications, including depression and anxiety in patients. In many animal models of IFN-α-induced behavioral dysfunction, the opposite results have frequently been reported. In an attempt to overcome the limitation of pharmacological studies, IFN-α-transgenic mice with CNS-targeted expression of the IFN-α transgene were used to study depression and anxiety-like behaviors by Porsolt swim and elevated plus-maze assays, respectively. Interestingly, chronic stimulation of IFN-α signaling in mouse brains did not cause depression or anxiety as measured by these tests in comparison with wild-type littermates. This observation suggests that factors other than IFN-α may be necessary for the development of psychiatric complications following IFN-α therapy in patients. [Copyright &y& Elsevier]

Knocking out of the global regulator LaeA in Aspergillus flavipes leads to substantial up-regulation of the expression of two NRPS-like biosynthetic gene clusters in the ΔflLaeA mutant. With a scaled-up fermentation of the ΔflLaeA strain, five compounds, including two previously undescribed piperazine derivatives flavipamides A and B (1 and 2), and three known compounds (3 – 5), were obtained and identified by LC-MS guided isolation and extensive spectroscopic analysis. [Display omitted] • The Knocking out The expression of two NRPS-like BGCs was up-regulated in the LaeA mutant strain. • Two piperazine derivatives, flavipamides A and B (1 and 2), were characterized. • The convergent biosynthetic pathway of 1 and 2 was tentatively proposed. Genome sequencing on an intertidal zone-derived Aspergillus flavipes strain revealed its great potential to produce secondary metabolites. To activate the cryptic compounds of A. flavipes , the global regulator flLaeA was knocked out, leading to substantial up-regulation of the expression of two NRPS-like biosynthetic gene clusters in the ΔflLaeA mutant. With a scaled-up fermentation of the ΔflLaeA strain, five compounds, including two previously undescribed piperazine derivatives flavipamides A and B (1 and 2), along with three known compounds (3 – 5), were obtained by LC-MS guided isolation. The new compounds were elucidated by spectroscopic analysis and electronic circular dichroism (ECD) calculations, and the biosynthetic pathway was proposed on the bias of bioinformatic analysis and 13C isotope labeling evidence. This is the first report to access cryptic fungi secondary metabolites by inactivating global regulator LaeA and may provide a new approach to discovering new secondary metabolites by such genetic manipulation. [ABSTRACT FROM AUTHOR]

Cryptocarya irritans is an obligate parasite causing the "white spot disease." This disease often leads to the death of a large number of cultured marine fish. This study aimed to compare the effects of cuprous oxide (Cu 2 O) and nano‑copper on the morphology and hatchability of tomonts to investigate the effects of copper derivatives on C. irritans tomonts. The potential mechanistic difference was also explored using the transcriptome. The results showed that Cu 2 O seriously damaged the internal cell structure of tomonts, and the hatching rate of the tomonts in the Cu 2 O-treated group was lower than that in the control group within 10 days. On the contrary, the effect of nano‑copper on tomonts was not significant. The cytoplasm of tomonts treated with nano‑copper dissolved, the internal structure was slightly loose, and the hatching rate of tomonts was even slightly higher than control group within 5 days. In the transcriptome analysis, 850 differentially expressed genes (DEGs), including 332 upregulated genes and 518 downregulated genes, were identified in the Cu 2 O-treated groups. However, only 56 DEGs, including 34 upregulated genes and 22 downregulated genes, were identified in the nano‑copper-treated groups. This indicated that the effect of Cu 2 O on tomonts was greater than nano‑copper. In the enrichment pathway analysis, DEGs in the Cu 2 O-treated group were mostly enriched in the ribosome and apoptosis pathways. Further analysis showed that genes (RPL18e , RPLP0 , RPL10 , RPL17 , RPL26 , RPL35 , and RPL37) related to cell stress and defense were significantly upregulated and those (P53 and PKA) related to cell proliferation and development were downregulated, indicating that the killing effect was mainly mediated by different ribosomal and structural proteins. However, the DEGs (KCNMA1, CACNA1C, CNGA1) in the nano‑copper-treated group were enriched in the insulin secretion and cAMP signaling pathways, which were activate part of tomonts' innate immune response, completely different from those in the Cu 2 O-treated group. The results provided more information on the mechanism of killing parasites by copper derivatives. • Copper derivatives had significant differences in the killing effects of the C. irritans tomonts. • Compared with nano‑copper, Cu2O significantly affected the cell morphology, hatchability, and gene expression of tomonts. • The killing effect of Cu2O was mainly mediated by different ribosomal and structural proteins. [ABSTRACT FROM AUTHOR]

Abstract Ethnopharmacological relevance Involucrum castaneae (IC)is used in Chinese folk medicine to treat various lung diseases, as well as for its reducing phlegm and anti-inflammatory properties. Aim of the study The purpose of this experiment is to verify the effect of IC on airway inflammation, responsiveness in ovalbumin (OVA)-induced asthmatic guinea pigs. The main chemical components of IC were also analyzed. Materials and methods The potential of the ethanol extract of Involucrum castaneae (EEIC) to protect against OVA-induced allergic airway response in guinea pigs was investigated. The latency of asthma in guinea pigs were recorded after the allergic asthma induced. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of immunoglobulin E (IgE), interleukin-5 (IL-5), nerve growth factor (NGF) and interferon-γ (IFN-γ) in asthma allergy. Reverse transcription-PCR (RT-PCR) was used to detect the expression of IL-5 mRNA in asthmatic guinea pig lungs. Paraffin sections of lung tissue were used to analyze pathological changes. The total flavonoid content was determined and the chemical components were analyzed by LC-MS/MS. Results It was found that EEIC was able to reduce the number of eosinophil (EOS) in bronchoalveolar lavage fluid (BALF) and peripheral blood (PB) in the guinea pig model of OVA -induced asthma. Meanwhile, it also significantly reduced the levels of inflammation-related factors IgE and IL-5, decreased the expression of IL-5 mRNA in lung tissue, and increased the level of IFN-γ. Pathological examination of paraffin section of lung tissue showed that EEIC can reduce the thickening of bronchial smooth muscle and reduce the infiltration damage of tissues by various inflammatory cells. The presence of flavonoids, terpenoids and phenolic compounds in EEIC might be responsible for these activities. Conclusion IC alleviated airway inflammation and smooth muscle thickening in guinea pigs with OVA-sensitized allergic asthma. The paper explains the traditional efficacy and material basis of IC and lays a foundation for further development. Graphical abstract fx1 [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Castanea mollissima shell (CMS) has been used for wound healing in China as traditional medicine. The shell is directly applied on the wounded skin as fine powder or as water maceration. Aim of the study To investigate the wound healing activity of CMS and the potential mechanism of anti-inflammatory activity. Materials and methods The effects of ethanol extract of CMS (ECMS) on nitricoxide (NO), tumor necrosis factor (TNF)-α and interleukin (IL)− 6 productions in lipopolysaccharide (LPS)-treated RAW 264.7 cells were explored by enzyme linked immunosorbent assay (ELISA) in vitro. To study wound healing properties of ECMS in vivo, excision and incision wound models were performed on rats. Inflammatory cytokines from wound biopsies such as NO, TNF-α and IL-6 production were tested by ELISA. mRNA levels of iNOS, cyclooxygenase (COX) −2 and TNF-α were detected by real-time Polymerase Chain Reaction (PCR), and protein levels of IL-1β and Heme Oxygenase (HO) −1 were analyzed by Western blotting. Results ECMS potently inhibited LPS-induced production of IL-6, NO and TNF-α in RAW 264.7 cells. The presence of quercetin, kaempferol, ursolic acid and gallic acid in ECMS might be responsible for its anti-inflammatory activity. 3% and 5% w/w ECMS significantly accelerated the wound healing process in both wound models, evidenced by the faster rate of wound contraction, epithelialization, increased hydroxyproline content, high tensile strength, decreased level of inflammatory markers compared to the control group. Histopathological studies also revealed the amelioration of wound healing by re-epithelialization, collagenation and vascularization of wounded skin sample in ECMS treated groups. Conclusion The experimental data revealed that CMS had ability to prevent exaggerated inflammation and accelerates wound epithelialization and might be beneficial for healing dermal wounds. [ABSTRACT FROM AUTHOR]

The underlying mechanism between the gut microbiota and reproductive function is not yet well-known. This study was conducted to investigate the effect of the administration of fecal microbiota transplantation (FMT) from highly laying rate donors on the cecal microbiota, intestinal health and ovarian function in broiler breeders. A total of 60 broiler breeders (53 wk of age) were selected by their laying rate [high (HP , 90.67 ± 0.69%; n = 10) and low (LP , 70.23 ± 0.87%; n = 20)]. The LP breeders were then be transplanted with fecal microbiota from HP hens (FMTHP ; n = 10) or the same dosage of PBS (FMTCON ; n = 10) for 28 d. The results revealed that FMT from HP donors increased egg-laying rate and serum hormone levels [17β-estradiol (E2), anti-Müller hormone], also decreased proinflammatory cytokine levels (interleukin-6, interleukin-8, tumor necrosis factor-α) of LP breeders (P < 0.05). The FMTHP group breeders had higher villus height, villus height/crypt depth ratio, and upregulated mRNA expression of jejunum barrier-related gene ( ZO-2 and mucin-2) and estrogen, follicle-stimulating hormone (FSH) and anti-Müller hormone (AMH) receptor genes ( ESR1, ESR2, FSHR, AMHR ) (P < 0.05) than FMTCON group. FMT from HP donors led to higher mRNA expression of Bcl2 and sirtuin1 ( SIRT1 ), while it downregulated the proapoptotic genes (Bax, caspase-3, caspase-8 , and caspase-9) mRNA expressions in ovary compared with the FMTCON breeders (P < 0.05), and this pattern was also observed in HP donors. Also, HP breeder had higher observed_species and alpha-diversity indexes (Chao1 and ACE) than FMTCON group, while FMTHP can increase observed_species and alpha-diversity indexes (Chao1 and ACE) than FMTCON group (P < 0.05). The bacteria enrichment of Firmicutes (phylum), Bacteroidetes (phylum), Lactobacillus (genus), Enterococcus (genus), and Bacteroides (genus) were increased by FMTHP treatment. The genera Butyricicoccus, Enterococcus , and Lactobacillus were positively correlated with egg-laying rate. Therefore, cecal microbiomes of breeders with high egg-laying performance have more diverse activities, which may be related to the metabolism and health of the host; and FMT from high-yield donors can increase the hormone secretion, intestinal health, and ovarian function to improve egg-laying performance and the SIRT1 -related apoptosis and cytokine signaling pathway were involved in this process. [ABSTRACT FROM AUTHOR]

Highlights: [•] Gene expression and miRNA arrays of nonresponders’ and responders’ fresh-frozen tissues were compared. [•] We find 59 genes and 17 miRNAs differentially expressed by at least two folds. [•] Expression of AQP9, SATB2, and WIF1 are lower in the nonresponders and modulated by differentially expressed miRNAs. [•] RT-PCR validates lower expression of AQP9, but not the other two genes, in nonresponders. [ABSTRACT FROM AUTHOR]

Abstract: Objectives: The present study was aimed to explore the prognostic strength of Sox2 and Oct4A in hepatocellular carcinoma (HCC). Design and methods: We investigated the expression of Sox2 and Oct4A in five hepatoma cell lines, one immortalized normal liver cell line, HCC tissues with matched nontumorous liver tissues and normal liver tissues by reverse transcription-polymerase chain reaction. Results: Sox2 and Oct4A mRNA were overexpressed in hepatoma cell lines and tumor tissues. Sox2 or Oct4A positive expression was significantly associated with an aggressive phenotype. Both univariate and multivariate analyses revealed that Sox2 or Oct4A was an independent prognostic factor for HCC. When using subgroup analysis, the patients with a co-expression of Sox2/Oct4A had the poorest prognosis. Further analysis demonstrated that Sox2 alone or Sox2/Oct4A could stratify outcome in HCC patients with early stage. Conclusions: Sox2 and Oct4A can be novel predictors of poor prognosis for patients undergoing resection of HCC. [Copyright &y& Elsevier]

This study was conducted to explore whether dietary pectic oligosaccharide (POS) supplementation could improve gut health of broiler breeders with different egg-laying rates. A 2 × 2 factorial design was used in this study. Two hundred fifty-six Arbor Acres broiler breeders (48 wk of age), including 128 average egg-laying rate and 128 low egg-laying rate (LELR) birds, were randomly fed with the diets supplemented with or without 200 mg kg−1 of POS (n = 8). The trial lasted for 8 wk. Compared with average egg-laying rate broiler breeders, LELR broiler breeders had lower laying rate and qualified egg rate (P < 0.05), higher egg weight and feed conversion ratio (P < 0.05), higher malondialdehyde (MDA) levels in the jejunum (P < 0.05), higher IL-6 (P < 0.05) and tumor necrosis factor α ( TNF-α ) (P = 0.07) mRNA expressions in the jejunal mucosa, and lower microflora diversity in cecal digesta. Dietary POS supplementation increased egg weight of broiler breeders (P < 0.05), enhanced superoxide dismutase activity in the jejunum (P < 0.05), decreased MDA level in the jejunum (P < 0.05), upregulated zonula occluden 1 mRNA expression in the jejunal mucosa (P < 0.05), downregulated IL-6 and TNF-α mRNA expressions in the jejunal mucosa (P < 0.05), and regulated relative abundance of some microbiota (including the phylum and genus, P < 0.05). In addition, in LELR broiler breeders, POS administration enhanced villus height (P = 0.08) and ZO-2 mRNA expression (P = 0.09) in the jejunal mucosa, alleviated the increasing MDA level in the jejunum (P < 0.05) and IL-6 and TNF-α mRNA expressions in the jejunal mucosa (P < 0.05), and regulated relative abundance of some microbiota (including the phylum and genus, P < 0.05). These results suggest that supplementing POS in diets may elevate gut health via improvement of intestinal barrier function, antioxidant capacity, and microbiota composition in broiler breeders with different egg-laying rates. [ABSTRACT FROM AUTHOR]

• Dietary inclusion of 12 % resistant potato starch did not alleviate skin inflammatory response and feather growth depression caused by high stocking density (32 ducks /m2 (d 1−14); 16 ducks /m2 (d15−35)) in Pekin ducks. • Dietary resistant starch supplementation had a positive effect on the performance of ducks at normal stocking density during 1 to 14 d of age. • High stocking density had a negative effect on growth and feather performance, as well as meat deposition and even caused skin inflammation of meat duck. This study investigated whether dietary resistant potato starch (RPS) inclusion alleviated the stress response of high stocking density (HSD) based on growth performance, carcass traits, immune organ index, serum corticosterone concentration, and skin inflammatory cytokine gene expression in meat ducks. A total of 576 1-d-old male ducklings were assigned to 4 groups of 8 replicates and were under the following conditions: normal stocking density (NSD; 16 birds/m2(d 1−14); 8 birds/m2(d 15−35)) and HSD (32 birds/m2(d 1−14); 16 birds/m2(d 15−35)). The ducks were fed the basal diet or the basal diet supplemented with 12 % RPS for 35d reared under HSD and NSD to form a 2 × 2 factorial experimental design. The results showed that HSD significantly depressed (P < 0.05) the growth performance, breast meat yield, feather and spleen development, and increased (P < 0.05) the pH value in proventriculus, serum corticosterone as well as mRNA abundance of interleukin (IL)-1β and IL-6 in the skin of the pelvic back in meat ducks compared to NSD. However, dietary RPS inclusion only markedly increased (P < 0.05) BW (d 14) and BWG (d 1−14), breast meat yield, and percentage of spleen in body weight (d 35) of ducks reared under NSD. Regardless of stocking density, dietary RPS inclusion significantly increased (P < 0.05) bursa of Fabricius and spleen percentages in BW and decreased (P < 0.05) feather scoring of 35-day-old ducks in comparison with no RPS inclusion. These results suggested that HSD had a negative effect on growth and feather performance, as well as meat deposition and even caused skin inflammation of meat ducks; however, RPS supplementation did not alleviate these negative effects caused by HSD. [ABSTRACT FROM AUTHOR]