ABSTRACT Blue eggshells have become economically valuable due to their higher popularity among consumers when compared with eggs of other colors, particularly in East Asia. However, eggshell colors vary widely and gradually become lighter with the increase of age. Therefore, to determine the association between DNA methylation and gene expression on the molecular mechanism of eggshell color variation in Xuefeng Black-bone chickens, we collected the shell glands from chickens that produced dark blue eggshells and light blue eggshells, respectively, and performed whole genome bisulfite sequencing (WGBS) and RNA sequencing (RNA-Seq) on them. The results showed that in the context of CG, DNA methylation levels were the highest compared to CHG and CHH contexts, and were negatively correlated with gene expression levels in three different gene regions in all samples, while there was no significant correlation in the contexts of CHG and CHH. Furthermore, when disregarding the location of coding genes, a total of 55 genes will show differences not only in expression levels but also in DNA methylation levels, and among the majority of differential expression genes (n=50), the DNA methylation levels in the gene body region were the highest. GO analysis and KEGG pathway analysis were performed in these genes and the results showed that the genes NR1H4 and HEPHL1, ABCA4 and ABCA12, and the pathways, cell adhesion molecular, ABC transporters, ECM-receptor interaction, and neuroactive ligand-receptor interaction, may affect the color variation of the blue eggshell of Xuefeng Black-bone chickens.

Shine Muscat, with its excellent quality and unique Muscat aroma, is emerging as one of the most profitable table grape cultivars in Asia. However, in-depth analysis of the cultivar's aroma profile before berries are harvest-ready is lacking. The aroma components and their concentrations undergo substantial changes during berry ripening, which provides an opportunity for producing some juice/wine products with special flavour. In this study, the evolution of free and bound aroma compounds in Shine Muscat grapes was analysed from EL-35 to EL-38. The main aroma attributes of harvest-ready berries were: 'green', 'sweet', 'floral' and 'fruity'. The headspace SPME-GC-MS analysis revealed that alcohols were the most abundant free and bound compounds in Shine Muscat. Furthermore, the free forms of hexanal, ß-damascenone, and (E, Z)-2,6-nonadienal determined the aroma characteristics of the grapes directly, while the bound forms of ß-damascenone, (E, Z)-2,6-nonadienal, and 2-hexanol determined the aroma of Shine Muscat that cannot to be smelled directly. Taking all indicators together, we believe that picking Shine Muscat grapes at EL-37 (when a minimum TSS of 17% has been reached) maintains its fine aroma characteristics. These results lay the foundation for the further development, utilisation and study of the aroma characteristics of Shine Muscat grapes.

ABSTRACT This study was conducted to investigate effects of vitamin A (VA) and vitamin K3 (VK3) on immune function and intestinal antioxidant capacity of aged laying hens. In a 3 × 3 factorial arrangement, the diets of 1080 Roman Pink laying hens (87 weeks old) was formulated with deficient, adequate and excess VA and VK3, including 0, 7000 and 14000 IU/kg VA and 0, 2.0 and 4.0 mg/kg VK3 for 8 weeks. Interactive effects between VA and VK3 were observed that VA and VK3 decreased the splenetic mRNA expression of inducible nitric oxide synthase (iNOS) and tumour necrosis factor α (TNF-α), but increased the plasma immunoglobulin G (IgG) content and jejunal mRNA expression of nuclear factor-like 2 (Nrf2). Hens fed adequate or excess VA had higher spleen index, mRNA expression of interleukin-10 (IL-10) in spleen, sIgA content, catalase (CAT), glutathione peroxidase and total dismutase (T-SOD) activity, and mRNA expression of polymeric immunoglobulin receptor (pIgR) in jejunum and lower mRNA expression of IL-1β in jejunum and iNOS, TNF-α in spleen. Furthermore, adequate or excess VK3 significantly increased plasma IgG content, the CAT, T-SOD and total antioxidant capacity activities, up-regulated the mRNA expression of pIgR, Nrf2, SOD1 and CAT in jejunum and down-regulated the mRNA expression of iNOS and TNF-α in spleen. In conclusion, dietary addition of adequate VA (7000 IU/kg) and VK3 (2.0 mg/kg) improved the immune function and intestine antioxidant capacity of aged laying hens and excess levels did not exhibit superior effects.

ABSTRACT The objectives of this study were to investigate the effects of tributyrin (TB) and coated sodium butyrate (CSB) on intestinal morphology, disaccharidase activity and intramuscular fat of broilers challenged with lipopolysaccharide (LPS). A total of 160 1-day-old healthy Cobb broilers were randomly allocated into four groups: (1) control; (2) LPS, in which broilers were fed a basal diet and intraperitoneally injected with 500 μg/kg LPS on days 38, 40 and 42; (3) TB, in which LPS-challenged broilers were fed basal diet supplemented with 500 mg/kg TB; and (4) CSB, in which LPS-challenged broilers were fed basal diet supplemented with 877 mg/kg CSB. Addition of TB and CSB inhibited (p<0.05) the decrease in villus height in the duodenum and ileum of LPS-challenged broilers, respectively. Both TB and CSB increased (p<0.05) activity of maltase in the small intestine, and TB increased (p<0.05) activity of isomaltase in the ileum. Additionally, dietary addition of TB and CSB decreased (p<0.05) the content of intramuscular fat. In conclusion, dietary supplementation of TB was more effective than CSB in improving intestinal morphology and disaccharidase activity of LPS-challenged broilers, and they both reduced intramuscular fat in the breast and legs.

OBJECTIVE: To evaluate the overall diagnostic performance of the p16 methylation for diagnosing malignant pleural effusion (MPE). METHODS: All published literature in English and Chinese were reviewed. Sensitivity, specificity, likelihood ratio and diagnostic odds ratio (DOR) were pooled by using random-effects model or fixed-effects model. Summary receiver operating characteristic (SROC) curve was used to evaluate the overall diagnostic value. RESULTS: Six studies were included with a total of 378 cases. The sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR) and DOR of p16 methylation in the diagnosis of MPE were 0.41 [95% confidence interval (CI) 0.35, 0.48], 0.97 [95% CI 0.93, 0.99], 9.57 [95% CI 4.53, 20.20], 0.61 [95% CI 0.45, 0.82] and 19.82 [95% CI 8.35, 47.04], respectively. The area under the curve (AUC) was 0.864. CONCLUSION: Pleural p16 methylation test plays a useful role in the diagnosis of MPE.

OBJETIVO: Evaluar el rendimiento diagnóstico general de la metilación p16 para el diagnóstico del derrame pleural maligno (DPM). MÉTODOS: Se revisó toda la literatura publicada en inglés y chino. La sensibilidad, especificidad, razón de verosimilitud, y el odds-ratio diagnóstico (DOR) fueron agrupados mediante el modelo de efectos aleatorios o el modelo de efectos. La curva de las características operativas de resumen del receptor (SROC) fue usada para evaluar el valor diagnóstico general. RESULTADOS: Se incluyeron seis estudios con un total de 378 casos. La sensibilidad, especificidad, razón de verosimilitud positiva (PLR), razón de verosimilitud negativa (NLR) y el DOR de la metilación p16 en el diagnóstico de DPM, fueron 0.41 [95% intervalo de confianza (IC) 0.35 0.48], 0.97 [95% IC 0.93, 0.99], 9.57 [95% IC 4.53, 20.20], [95% IC 0.45, 0.82] 0.61 y 19.82 [95% IC 8.35, 47.04], respectivamente. El área bajo la curva (AUC) fue 0.864. CONCLUSIÓN: La prueba de metilación p16 pleural desempeña un papel útil en el diagnóstico del DPM.

The tissue inhibitor of metalloproteinases (TIMP)-1 is a multifunctional protein which is not only an inhibitor of matrix metalloproteinases (MMPs) but also to have a possible "cytokine-like" action. Here, we first compared mRNA expression of TIMP-1 and MMP-9 in BEL-7402 (a hepatocellular carcinoma cell line), L-02 (a normal liver cell line) and QSG-7701 (a cell line derived from peripheral tissue of liver carcinoma) using real-time quantitative RT-PCR. By evaluating the variation of the MMP-9/TIMP-1 ratio as an index of reciprocal changes of the expression of the two genes, we observed that the MMP-9/TIMP-1 ratio was about 13- and 5-fold higher in BEL-7402 than in L-02 and QSG-7701, respectively. Significantly, overexpression of TIMP-1 decreased the MMP-9/TIMP-1 ratio in BEL-7402 and then inhibited the cell growth to 60% and reduced the migration to about 30%. Meanwhile, our data showed that interleukin-6 (IL-6) (100 ng/mL) could also inhibited the cell growth of BEL-7402. Further studies indicated that TIMP-1 mediated the inhibitory effect of IL-6 on BEL-7402 cell proliferation in a STAT3-dependent manner, which could further accelerate the expression of the cyclin-dependent kinase inhibitor p21. A dominant negative STAT3 mutant totally abolished IL-6-induced TIMP-1 expression and its biological functions. The present results demonstrate that TIMP-1 may be one of the mediators that regulate the inhibitory effect of IL-6 on BEL-7402 proliferation in which STAT3 signal transduction and p21 up-regulation also play important roles.