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Liu H.,Chinese Academy of Sciences | Liu H.,Qingdao Institute of Animal Science and Veterinary Medicine | Zhou D.,Chinese Academy of Sciences
Animal Feed Science and Technology | Year: 2014

This study was conducted to evaluate the effects of the inclusion of sodium chloride (NaCl) in the diet on growth performance and nitrogen (N) balance in sheep and subsequent ammonia (NH3) and nitrous oxide (N2O) emissions from pasture treated with urine of sheep. A total of 30 Ujimqin male sheep (a local breed, average body weight 15±1.3kg) were equally divided into three groups. The control group (C) was fed a standard NaCl diet (3g NaCl/kg DM), and the treatments were a medium salt group (MS, 4.5g NaCl/kg DM) and high salt group (HS, 6g NaCl/kg DM). Urine samples collected from the control and HS groups were used for land application in a field experiment, which consisted of 5 treatments: soil group (S, soil without treatment), distilled water group 1 (W1, 326mL), distilled water group 2 (W2, 458mL), urine of sheep in control group (C, 326mL, 102kgN/ha), urine of sheep in HS group (HS, 458mL, 92kgN/ha). The supplementation of NaCl had no adverse effects on growth performance and N balance in sheep. The high NaCl diet increased total urine volume (P<0.05) and average urine volume (P<0.05) and reduced urinary pH (P<0.05) and the N concentration in the urine (P<0.05) compared with standard NaCl diet. Compared with the C treatment, the HS treatment resulted in a delay of the peak N2O emission and decreased the average emission rates of NH3 by 48% (P<0.05) and N2O by 26% (P<0.05) from soil. In conclusion, high NaCl diet of grazing sheep (6g/kg DM) seemed to be a feasible means of reducing emissions of NH3 and N2O from urine treated pasture by increasing urine volume and decreasing total N concentration in urine. © 2014 Elsevier B.V.

Wu W.,Nanjing Agricultural University | Wu W.,Huazhong Agricultural University | Ren Z.,Huazhong Agricultural University | Zhang L.,Huazhong Agricultural University | And 3 more authors.
Molecular and Cellular Biochemistry | Year: 2013

Sine oculis homeobox 1 (Six1) homeodomain transcription factor is implicated in the genesis of muscle fiber type diversity, but its regulatory mechanisms on the formation of muscle fiber type are still poorly understood. To elucidate the biological roles of Six1 gene in muscle fiber formation, we established C2C12 cell line overexpressing Six1 and determined the effects of forced Six1 expression on muscle-specific genes expression, cell proliferation, and cell cycles. Our results indicated that Six1 overexpression could significantly promote the expression of fast-type muscle genes Atp2a1, Srl, and Mylpf. Furthermore, Six1 overexpressing C2C12 cells displayed a relative lower proliferative potential, and cell cycle analysis showed that Six1 exerted its role in cell cycle primarily through the regulation of G1/S and G2/M phases. In conclusion, Six1 plays an essential role in modulation of the fast-twitch muscle fiber phenotype through up-regulating fast-type muscle genes expression, and it could suppress the proliferation of muscle cells. © 2013 Springer Science+Business Media New York.

Liu S.,Chinese Academy of Agricultural Sciences | Jia H.,Chinese Academy of Agricultural Sciences | Hou S.,Chinese Academy of Agricultural Sciences | Zhang G.,Chinese Academy of Agricultural Sciences | And 9 more authors.
Molecular Immunology | Year: 2014

The TB10.4 antigen of Mycobacterium bovis/. Mycobacterium tuberculosis induces a strong Th1 CD4+ T-cell response. Thus, it is currently under intensive study as a possible vaccine candidate. However, how TB10.4 activates innate immune cells is unclear. How TB10.4 interacts with toll-like receptors (TLRs) and signaling pathways responsible for active inflammation have also not been fully elucidated. Here, as stimulated RAW264.7 cells with recombinant TB10.4 (rTB10.4), derived from M. bovis, increased TNF-α, IL-6 and IL-12 p40 secretin in a dose-dependent manner. Blocking assays showed that TLR2-, but not TLR4-neutralizing antibody reduced expression of TNF-α, IL-6 and IL-12 p40 in RAW264.7 cells. rTB10.4 stimulation activated p38 kinase (p38) and extracellular-regulated kinase (ERK) was TLR2-dependent, whereas inhibition of p38 and ERK activity significantly reduced TNF-α, IL-6 and IL-12 p40 production. Furthermore, rTB10.4 stimulation of RAW264.7 cells resulted in TLR2-mediated activation of NF-κB and induced translocation of NF-κB p65 from the cytoplasm to the nucleus via IκBα degradation. rTB10.4-induced TNF-α, IL-6 and IL-12 p40 release was attenuated by the specific IκB phosphorylation inhibitor, BAY 11-7082. These findings indicate that the M. bovis-derived rTB10.4 induced production of TNF-α, IL-6 and IL-12 p40 involves p38, ERK and NF-κB via the TLR2 pathway. © 2014 Elsevier Ltd.

Ruan J.,Chinese Academy of Agricultural Sciences | Ruan J.,Jilin University | Li H.,Chinese Academy of Agricultural Sciences | Li H.,Qingdao Institute of Animal Science and Veterinary Medicine | And 10 more authors.
Scientific Reports | Year: 2015

Transgenic pigs play an important role in producing higher quality food in agriculture and improving human health when used as animal models for various human diseases in biomedicine. Production of transgenic pigs, however, is a lengthy and inefficient process that hinders research using pig models. Recent applications of the CRISPR/Cas9 system for generating site-specific gene knockout/knockin models, including a knockout pig model, have significantly accelerated the animal model field. However, a knockin pig model containing a site-specific transgene insertion that can be passed on to its offspring remains lacking. Here, we describe for the first time the generation of a site-specific knockin pig model using a combination of CRISPR/Cas9 and somatic cell nuclear transfer. We also report a new genomic "safe harbor" locus, named pH11, which enables stable and robust transgene expression. Our results indicate that our CRISPR/Cas9 knockin system allows highly efficient gene insertion at the pH11 locus of up to 54% using drug selection and 6% without drug selection. We successfully inserted a gene fragment larger than 9 kb at the pH11 locus using the CRISPR/Cas9 system. Our data also confirm that the gene inserted into the pH11 locus is highly expressed in cells, embryos and animals. © 2015 Scientific Reports.

Liu N.,Qingdao Agricultural University | Li H.,Qingdao Agricultural University | Liu K.,Qingdao Institute of Animal Science and Veterinary Medicine | Yu J.,Qingdao Agricultural University | And 8 more authors.
Molecular Biology Reports | Year: 2014

Sheep are valuable resources for the wool industry. Wool growth of Aohan fine wool sheep has cycled during different seasons in 1 year. Therefore, identifying genes that control wool growth cycling might lead to ways for improving the quality and yield of fine wool. In this study, we employed Agilent sheep gene expression microarray and proteomic technology to compare the gene expression patterns of the body side skins at August and December time points in Aohan fine wool sheep (a Chinese indigenous breed). Microarray study revealed that 2,223 transcripts were differentially expressed, including 1,162 up-regulated and 1,061 down-regulated transcripts, comparing body side skin at the August time point to the December one (A/D) in Aohan fine wool sheep. Then seven differentially expressed genes were selected to validated the reliability of the gene chip data. The majority of the genes possibly related to follicle development and wool growth could be assigned into the categories including regulation of receptor binding, extracellular region, protein binding and extracellular space. Proteomic study revealed that 84 protein spots showed significant differences in expression levels. Of the 84, 63 protein spots were upregulated and 21 were downregulated in A/D. Finally, 55 protein points were determined through MALDI-TOF/MS analyses. Furthermore, the regulation mechanism of hair follicle might resemble that of fetation. © 2014 Springer Science+Business Media.

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