ABSTRACT Chicken coccidiosis is a common and severe parasitic disease caused by infection from Eimeria spp., which affects the integrity of the intestinal mucosa. TGF-β has been shown to play an important role in the healing of intestinal mucosas, immunity, and the maintenance of bowel mucosa integrity. Very little is known about the presence of the components of TGF-β/Smads signaling pathway of chicken at different times following coccidian infection. In the present study, we measured the levels of TGF-β2, 3, 4, receptor TβRI, II, down-stream Smad 2, 3, 7 in cecum and spleen of chicken at different times after inoculation with Eimeria tenella using quantitative real-time PCR. The results showed that the TGF-β/Smads signaling pathway was not activated in cecum in the early stage of infection. However, on the 8th day, the expression of TGF-β2, 4, down-stream protein Smad 2, 7 were significant up-regulated in the spleen, which indicated that the TGF-β/Smads signaling was changed in the E. tenella infection and was differentially expressed in various tissues in the early stages of infection.

ABSTRACT This experiment aimed to investigate whether Escherichia coli (E. coli) infection could affect the TGF-β/smads signaling pathway in the jejunal tissue of chickens. One-day-old Cobb 500 broilers were randomly divided into 2 groups and treated with intraperitoneal E. coli or broth injection. Clinical signs of the birds were assessed every day. Spleen and bursa of Fabricius of the birds, post-infection (pi), were collected to evaluate immune organ index. Jejunal tissues were collected to ascertain the expression of TGF-βs, TβRs, and Smads. The results showed that the infected birds had significantly higher index of the spleen (24hrs and 48hrs pi) compared with birds in the control group (p<0.05). The relative gene expression of TGF-β4 increased (p<0.05), while the expression of Smad7 down-regulated in the E. coli group (p<0.01). There was no significant difference in TGF-β2, TGF-β3, TβR I, TβR II, Smad2, Smad3 expression (p>0.05). In conclusion, TGF-β/Smad signaling pathway was associated with the immune response of broilers in E. coli infection and TGF-β4 was the main subtype interacting with E. coli infection.

ABSTRACT Heparan sulfate proteoglycans (HSPGs) are present on the cell surface and in the extracellular matrix in all metazoans. HSPGs interact with growth factors and receptors through heparan sulfate (HS) chains. The sulfation pattern of heparan sulfate chains influences signaling events mediated by heparan sulfate proteoglycans located on the cell surface. SULF1 and SULF2 are two endo-sulfatases that can cleave specific 6-O-sulfate groups within the heparan chains. To determine their possible roles in tissues and satellite cells in vitro, their expression pattern was examined in tissues from 40-day-old chickens and in satellite cells from the breast muscles of 1-week-old and 2-week-old chickens using RT-PCR and immunocytochemistry analyses. The SULF1 and SULF2 transcripts were widely distributed in various tissues. Upon increasing culture times in chicken´s primary skeletal muscle satellite cells, SULF1 and SULF2 expression in 1-week-old chickens was significantly higher than in 2-week-old chickens, suggesting that sulfatases play a key role in satellite cell development. Therefore, our findings increase our knowledge of sulfatase expression diversity and provide a solid basis for further research concerning this molecular mechanism.

Due to the special climate conditions in the Qingtongxia region, grapes are high in sugar and low in titratable acidity from the stages of ripening. Therefore, the common methods used for determining the maturity of grapes, which depend on the ratio of sugar and titratable acidity in other regions, are inappropriate in Qingtongxia. This research was done in order to seek for a simple and convenient method of determining the optimal harvest time of grapes, further providing some theoretical basis for improving the quality of wine in Qingtongxia. Phenolic contents and some basic physico-chemical parameters of Merlot and Pinot Noir were evaluated during different ripening stages. The results showed that a different harvest time significantly affects the phenolic contents and physico-chemical parameters of Merlot and Pinot Noir. The total contents of anthocyanins in skins and total contents of phenolic in seeds was screen out as two important indexes to evaluate the maturity of polyphenols, in order to better improve the quality of grape and wine.

Due to the special climate conditions in the Qingtongxia region, grapes are high in sugar and low in titratable acidity from the stages of ripening. Therefore, the common methods used for determining the maturity of grapes, which depend on the ratio of sugar and titratable acidity in other regions, are inappropriate in Qingtongxia. This research was done in order to seek for a simple and convenient method of determining the optimal harvest time of grapes, further providing some theoretical basis for improving the quality of wine in Qingtongxia. Phenolic contents and some basic physico-chemical parameters of Merlot and Pinot Noir were evaluated during different ripening stages. The results showed that a different harvest time significantly affects the phenolic contents and physico-chemical parameters of Merlot and Pinot Noir. The total contents of anthocyanins in skins and total contents of phenolic in seeds was screen out as two important indexes to evaluate the maturity of polyphenols, in order to better improve the quality of grape and wine.

ABSTRACT The melanocortin 1 receptor (MC1R) gene plays a key role in controlling the deposition of melanin. In mammals, the MC1Rgene is regarded as a major candidate gene in the control of melanin formation. In domestic animals, the MC1R gene mainly controls the expression of coat, skin, and plumage color in mammals and birds. In order to breed chickens with dark-green shank faster, we screened the molecular markers for shank color in a HS chicken population by exploring the relationship between polymorphism of the MC1R gene and three different shank colors (light green, dark green and yellow). Two primer pairs for code region of the MC1R gene were designed in the basic of chicken genomic sequence. DNA sequencing was performed to detect the polymorphisms and PCR was used to amplify DNA fragment. Sequences analysis indicated that 7 SNPs were predominant the three HS chicken populations with different shank color, including g.18,287,945C>T, g.18,288,088T>C, g.18,288,150G>A, g.18,288,303A>G, g.18,288,512G>A, g.18,288,513T>C, and g.18,288,520A>C. Association analysis revealed that the dark-green shank population showed moderate polymorphism, whereas the light-green shank population showed low polymorphism among overall 7 SNPs and that SNP6 (g.18,288,513T>C) may be significantly associated with three different shank colors in HS chickens. The haplotype CTGGACA had the largest haplotype frequencies, accounting for 56.22%, and the haplotype combination H1H1 is mainly distributed in the dark-green shank population, and may be used as molecular maker for marker-assisted selection of shank color in HS chickens.

ABSTRACT There is increasing evidence that bone morphogenetic protein 6 (BMP6) plays critical roles in regulating various stages of ovarian follicle development in mammals. However, the mechanisms of regulation of BMP6 in the chicken ovary remain unclear. In this study, mRNA and protein expression level of BMP6 in chicken ovarian follicles at different development stages were determined by qRT-PCR and western blot separately. Different concentrations of BMP6 protein and FSH were added to the culture medium, and the effects to proliferation of granulose cells were detected, further effect on expression pattern of progesterone synthesis associated genes were also analyzed by qRT-PCR and Western blotting and the secretion of progesterone was detected by ELISA. The results showed that mRNA and protein expression level of BMP6 increased significantly in the follicle with the development of follicle (p<0.05) and reached a peak at F1 follicle. Adding concentration of 50ng/ml and 100ng/ml of BMP6 protein promoted significantly the proliferation of granulosa cells (p<0.05), as well as up-regulated the expression of Steroid hormone synthesis acute regulatory protein (StAR) and 3β-hydroxysteroid dehydrogenase (3β-HSD) genes in mRNA and protein level. Meanwhile, the secretion of progesterone was significantly higher in the group that added BMP6 and FSH separately than blank control group (p<0.05) and reached a peak in the group that both added BMP6 and FSH. Collectively, these findings highlight that BMP6 is associated with proliferation of follicular cells and the synthesis of progesterone, which indicated that it took an important role in the follicular development of chicken.

ABSTRACT In this study, a method utilizing PCR-restriction fragment length polymorphism (PCR-RFLP) of a mitochondrial gene was developed for the identification of chicken (Gallus gallus), quail (Coturnix coturnix), and common pigeon (Columba livia) meat. PCR products of ~440 bp were obtained from the 12S rRNA gene of these three birds using a pair of universal primers. The three terrestrial birds can be distinguished using one restriction endonuclease, Alu I, which was selected based on species-specific variations in the mt 12S rRNA gene sequence using 9 newly-obtained and 44 published chicken, quail and pigeon sequences. This method was also successfully used to identify commercial quail and pigeon meat products, which were found to be adulterated with chicken meat. Additionally, our method had relatively high sensitivity for detecting a meat mixture. Ten percent of chicken meat in the mixed quail and pigeon sample was detectable. This assay can be useful for the accurate identification of meats from terrestrial birds, avoiding mislabeling or fraudulent species substitution in meat products.

Abstract Brønsted acidic ionic liquids composed of 1-butyl-3-methylimidazolium, triethylbutylammonium, and 1-butylpyridinium cation counterparts and HSO4‾ and H2PO4‾ anion counterparts were used as the catalysts for the selective chlorination of glycerol with hydrogen chloride to 3-chloro-1,2-propandiol. From the perspective of the glycerol conversion and product yields, the catalytic activities of the ionic liquids containing HSO4‾ were higher than the ionic liquids containing H2PO4‾. As compared with the carboxylic acid catalysts, Brønsted acidic ionic liquids favored the catalytic chlorination of glycerol to 3-chloro-1,2-propandiol. When the glycerol chlorination was catalyzed by the Brønsted acidic ionic liquids at 110 ºC for 12 h with the catalyst loading of 0.75 mol·kg‾1 glycerol, the 3-chloro-1,2-propandiol yield was more than 81% at the complete conversion of glycerol.

ABSTRACT BMP6, a member of the subfamilies of the morphogenetic proteins (BMPs), plays a crucial role in osteogenic and chondrocyte differentiation in vitro and stimulates chondrogenesis, making chondrocytes differen-tiate on their terminal stage. The objective of this study is to explore the relationship between polymorphism of BMP6 gene and slaughter traits in chicken respectively. We screened the exonic and intronic regions of BMP6 gene by DNA pool construction and amplified DNA fragment by PCR, and finally, we got nine SNPs. Association analysis revealed that BMP6 had no significant association among all slaughter traits in Yellow bantam chicken. However, BMP6 had a significant difference with femur weight, tibia weight, femur length (p<0.05), and was extremely significant with tibia length (p<0.01) in Avian chicken. Moreover, femur perimeter also had significant correlation with BMP6 in Avian chicken. These results provide useful information for further investigation on the function of chicken BMP6 gene.