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Li X.,Huazhong Agricultural University | Hu X.,Wuhan Institute of Animal Husbandry and Veterinary Science | Jin X.,Hubei Provincial Institute of Veterinary Drug Control | Zhou X.,Huazhong Agricultural University | And 3 more authors.
Xenobiotica | Year: 2015

1. The expression and the activity of cytochromes P450 (CYPs) can be elevated by the activation of nuclear receptors. The pregnane X receptor (PXR, or nuclear receptor NR1I2) is a ligand-activated transcription factor that mediates responses to diverse xenobiotics and endogenous chemicals. Here we investigated the regulatory role of PXR in IFN-γ-mediated CYP3A29 expression in pig liver microsomes, primary porcine hepatocytes, and a cultured hepatocyte cell line. 2. IFN-γ significantly up-regulated CYP3A29 and PXR expressions at mRNA and protein levels in a dose-dependent manner. IFN-γ treatment significantly increased the metabolism of nifedipine. PXR and IFN-γ treatments significantly enhanced the activity of CYP3A29 promoter and the upstream region from -1473 to -1021 of CYP3A29 might be PXR-binding site. Moreover, the IFN-γ-induced CYP3A29 expression was blocked by PXR knockdown, whereas CYP3A29 mRNA and protein expression levels were dramatically elevated by PXR overexpression. 3. The regulatory effect of IFN-γ on CYP3A29 expression is mediated via PXR. © 2014 Informa UK Ltd. All rights reserved: reproduction in whole or part not permitted. Source

Huang H.-J.,Wuhan Institute of Animal Husbandry and Veterinary Science | Peng X.,Wuhan Institute of Animal Husbandry and Veterinary Science | Deng B.,Wuhan Institute of Animal Husbandry and Veterinary Science | Huang C.,Huazhong Agricultural University | And 12 more authors.
Cytotechnology | Year: 2014

Despite the powerful impact gene expression markers like the green fluorescent protein (GFP) or enhanced GFP (EGFP) exert on linking the expression of recombinant protein for selection of high producers in recent years, there is still a strong incentive to develop more economical and efficient methods for isolating mammalian cell clones secreting high levels of recombinant proteins. Here we present a new method based on the co-expression of EGFP that allows clonal selection in standard 96-well cell culture plates. The genes encoding the EGFP protein and the related protein are linked by an internal ribosome entry site and thus are transcribed into the same mRNA in an independent translation process. Since both proteins arise from a common mRNA, the EGFP expression level correlates with the expression level of the therapeutic protein in each clone. By expressing recombinant porcine β-defensin 1 in Marc 145 cells, we demonstrate the robustness and performance of this technique. The method can be served as an alternative to identify high-producer clones with various cell sorting methods. © 2014 Springer Science+Business Media Dordrecht Source

Gong P.,Huazhong Agricultural University | Gong P.,Wuhan Institute of Animal Husbandry and Veterinary Science | Yang Y.,Huazhong Agricultural University | Yang Y.,Wuhan Institute of Animal Husbandry and Veterinary Science | And 4 more authors.
Acta Biologica Hungarica | Year: 2012

New technologies in gene transfer combined with experimental embryology make the chicken embryo an excellent model system for gene function studies. The techniques of in ovo electroporation, in vitro culture for ex ovo electroporation and retrovirus-mediated gene transfer have already been fully developed in chicken. Yet to our knowledge, there are no definite descriptions on the features and application scopes of these techniques. The survival rates of different in vitro culture methods were compared and the EGFP expression areas of different gene transfer techniques were explored. It was that the optimal timings of removing embryo for EC culture and Petri dish system was at E1.5 and E2.5, respectively; and optimal timing of injecting retrovirus is at E0. Results indicated that the EC culture, in ovo electroporation, the Petri dish system and retrovirus-mediated method are, respectively, suitable for the very early, early, late and whole embryonic stages in chicken. Comparison of different gene transfer methods and establishment of optimal timings are expected to provide a better choice of the efficient method for a particular experiment. © 2012 Akadémiai Kiadó, Budapest. Source

Yang Y.,Huazhong Agricultural University | Yang Y.,Wuhan Institute of Animal Husbandry and Veterinary Science | Gong P.,Wuhan Institute of Animal Husbandry and Veterinary Science | Feng Y.P.,Huazhong Agricultural University | And 5 more authors.
Acta Biologica Hungarica | Year: 2013

Doublesex and mab-3-related transcription factor 1 (Dmrt1) is a Z-linked gene that putatively determines the phenotype of gonads in birds. The sex differential expression of Dmrt1 was examined using wholemount in situ hybridization (WISH) in the urogenital systems during embryogenesis. The results revealed that Dmrt1 showed dimorphic expression in chicken gonads, which increased from day 6.5 to day 10.5. The expression of Dmrt1 in male (ZZ) gonads was not twice as much as in female (ZW) gonads, suggesting the existence of other regulatory mechanisms in addition to Z chromosome dosage effect. © 2013 Akadémiai Kiadó, Budapest. Source

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