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Song Z.-Q.,China Agricultural University | Yang L.-F.,China Agricultural University | Wang Y.-S.,China Agricultural University | Zhu T.,China Agricultural University | And 5 more authors.
CNS Neuroscience and Therapeutics | Year: 2014

Backgrounds and aims: Prion diseases are a group of infectious neurodegenerative diseases characterized by neuronal death and degeneration. Human leukocyte antigen-B-associated transcript 3 (BAT3) is an important apoptosis regulator. We therefore investigated the interactions between BAT3 and prion protein and the potential role of BAT3 in PrP106-126-induced apoptosis. Methods: BAT3 and prion protein were overexpressed in Hela, Neuro2A, or primary neuronal cells by transfection with BAT3-HA or PRNP-EGFP expression plasmids and their relationship studied by immunofluorescence and Western blotting. The effect of BAT3 on PrP106-126-induced cytotoxicity and apoptosis was detected by the CCK-8 assay and terminal-deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay. The expression of cytochrome c and Bcl-2 was examined by Western blotting. Results: BAT3 interacted with prion protein and enhanced PrP expression. After PrP106-126 peptide treated, BAT3 was transported from the nucleus to cytoplasm, increased cell viability, and protected neurons from PrP106-126-induced apoptosis through stabilizing the level of Bcl-2 protein and inhibiting the release of cytochrome c to cytoplasm. Conclusions: Our present data showed a novel molecular mechanism of PrP106-126-induced apoptotic process regulation through the overexpression of BAT3, which may be important for the basic regulatory mechanism of neuron survival in prion diseases and associated neurodegenerative diseases in vivo. © 2014 John Wiley & Sons Ltd.


Liu Y.,Inner Mongolia Agricultural University | Zhang Y.F.,Inner Mongolia Agricultural University | Sun X.L.,Inner Mongolia Agricultural University | Liu S.Y.,Inner Mongolia Agricultural University | Liu S.Y.,Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease
Genetics and Molecular Research | Year: 2016

The envelope protein (Env) of the Jaagsiekte sheep retrovirus (JSRV) is known to be a unique oncoprotein responsible for inducing ovine pulmonary adenocarcinoma (OPA). The objective of this study was to prepare a specific monoclonal antibody (mAb) against the JSRV Env protein using bioinformatic analysis. According to the structure and epitope prediction results of JSRV Env, the JSRV-Env572-615 antigen was prepared via peptide synthesis (amino acid sequence 572-615, denoted as JSRV-Env572-615). BALB/c mice were immunized to prepare the anti-JSRV-Env572-615 mAb. Spleen cells were fused with SP2/0 myeloma cells after being screened by indirect ELISA and cloned by limiting dilution. The specificity of mAb was evaluated by western blot analysis and immunohistochemistry assays. Western blot results showed that the JSRV Env protein was able to bind to mAb with high specificity. Immunohistochemistry assays demonstrated that the mAb was able to recognize JSRV Env in adenomatous hyperplasia of the lung. Furthermore, JSRV was detected in peripheral blood leukocytes during the pre-clinical period of OPA in 2 of the 25 sheep using this newly synthesized mAb. Therefore, this mAb may be a useful tool for the detection of JSRV in sheep. © 2016 The Authors.


Qi J.-W.,Inner Mongolia Agricultural University | Wu X.-L.,Inner Mongolia Agricultural University | Liu S.-Y.,Inner Mongolia Agricultural University | Liu S.-Y.,Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease | Cao G.-F.,Inner Mongolia Agricultural University
Virologica Sinica | Year: 2012

Endogenous beta retroviruses (enJSRV) are highly homologous with Jaagsiekte sheep retrovirus (exJSRV)this exogenous retrovirus is the aetiological agent of ovine pulmonary adenocarcinoma (OPA). The aim of this study was to clarify the function of enJSRV and the immunological mechanisms of its corresponding antibody, that is undetectable in JSRV-infected ovine serum. The expression of enJSRV envelope protein and Hyal-2 mRNA in immune organs and lungs of ovine fetuses and lambs were analyzed by Real-Time reverse transcription PCR and In Situ Hybridization using specific probes. In Situ Hybridization results indicated that the enJSRV envelope protein and Hyal-2 mRNA were expressed in thymus, spleen, mesenteric lymph nodes and lungs at different times, while no positive signals were detected in the negative controls. On the other hand, results from Real-Time reverse transcription PCR analysis showed that in 130d fetuses and 3d newborn lambs the enJSRV mRNA levels were much higher in organs associated with the immune system than that in lungs, especially in the thymus and spleen, but levels of Hyal-2 mRNA expression was not significantly different in all collected tissue. These results provided evidence from an immunology point of view to understand why the circulating antibodies against exJSRV are undetectable in JSRV-infected ovine, and will help to unravel the pathogenesis of JSRV-infected ovine. © Wuhan Institute of Virology, CAS and Springer-Verlag Berlin Heidelberg 2012.


Liu D.-C.,Inner Mongolia Agricultural University | Liu D.-C.,Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease | Zhou X.-L.,Inner Mongolia Agricultural University | Zhou X.-L.,Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease | And 4 more authors.
Journal of Integrative Agriculture | Year: 2014

Isolated ruminal epithelia from caudal blind sacs of dairy goats were incubated with butyrate and insulin-like growth factor-1 (IGF-1) at different concentrations. Proportions of ruminal epithelium in different phases of the cell division cycle were determined by flow cytometric analysis. The proportion of epithelial cells in S phase and G2-M phase (PS&G2-M) increased significantly (P<0.01) whereas the proportion of epithelial cells in G0-G1 phase (PG0-G1) decreased after incubation with IGF-1. PS&G2-M decreased whereas PG0-G1 increased markedly (P<0.01) after incubation with sodium butyrate. PS&G2-M incubated with IGF-1 and butyrate sodium together increased more than that incubated with IGF-1 alone; PG0-G1, however, decreased significantly (P<0.01). Our results indicate that IGF-1 enhances whereas sodium butyrate inhibits the proliferation of rumen epithelial cells. Furthermore, butyrate and IGF-1, together, have a synergic effect on the proliferation of rumen epithelium. © 2014 Chinese Academy of Agricultural Sciences.


Li H.-J.,Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease | Li H.-J.,Inner Mongolia Agricultural University | Wang C.-Y.,Inner Mongolia Agricultural University | Mi Y.,Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease | And 6 more authors.
Theriogenology | Year: 2013

The factor associated suicide (Fas) and its ligand (FasL) signaling is an important regulatory pathway of apoptosis in mammalian follicles. However, whether apoptosis in bovine oocytes is regulated by the Fas-FasL signaling pathway remains unknown. In this study, localization of Fas and FasL in immature oocytes and FasL in cumulus cells were examined using immunofluorescence staining. In addition, exogenous FasL was added to an invitro culture system to investigate apoptotic changes in bovine oocytes, using annexin-V and terminal uridine nick-end labeling staining, and real-time quantitative polymerase chain reaction. In this study, Fas was expressed in immature oocytes, whereas FasL was expressed in cumulus cells, but not in immature oocytes; annexin-V- and terminal uridine nick-end labeling-positive rates of oocytes treated with 2, 10, or 50 ng/mL FasL were higher than those of control oocytes (P<0.05); and oocytes from the three treatment groups had higher expression levels of Fas and B cell lymphoma/leukemia-2 associated X than those in the control group (P < 0.05). Taken together, we concluded that the Fas-FasL signaling pathway was involved in regulation of bovine oocyte apoptosis, perhaps related to B cell lymphoma/leukemia-2 associated X upregulation. © 2013 Elsevier Inc.


Zhang Y.,Inner Mongolia Agricultural University | Shi J.,Inner Mongolia Agricultural University | Liu S.,Inner Mongolia Agricultural University | Liu S.,Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease
BioMed Research International | Year: 2016

The primary sheep trophoblast cells (STCs) have a finite lifespan in culture. This feature limits the scope for long-term in vitro studies with STCs. This study was an attempt to establish and characterize a telomerase-immortalized sheep trophoblast cell line. STCs were isolated and purified by using Percoll and specific immunoaffinity purification, respectively. The purified STCs were transfected with a plasmid carrying sequences of human telomerase reverse transcriptase (hTERT) to create immortalized sheep trophoblast cell line (hTERT-STCs). hTERT-STCs showed a stable expression of hTERT gene, serially passaged for a year, and showed active proliferation without signs of senescence. Cytokeratin 7 (CK-7), secreted human chorionic gonadotrophin subunit β (CG-β), placental lactogen (PL), and endogenous jaagsiekte sheep retrovirus (enJSRV) envelope genes were expressed in hTERT-STCs. Transwell cell invasion assay indicated that hTERT-STCs still possessed the same invasive characteristics as normal primary sheep trophoblast cells. hTERT-STCs could not grow in soft agar and did not develop into tumors in nude mice. In this study, we established a strain of immortalized sheep trophoblast cell line which could be gainfully employed in the future as an experimental model to study trophoblast cells with secretory function, invasive features, and probable biological function of enJSRV envelope genes. © 2016 Yufei Zhang et al.


Jirintai S.,Jichi Medical University | Jinshan,Inner Mongolia University | Jinshan,Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease | Tanggis,Jichi Medical University | And 7 more authors.
Virus Research | Year: 2012

Rabbit hepatitis E virus (HEV) strains have recently been isolated in several areas of China and in the US and France. However, the host range, distribution and zoonotic potential of these HEV strains remain unknown and their propagation in cultured cells has not yet been reported. A total of 211 4-month-old rabbits raised on a farm in Inner Mongolia were tested for the presence of anti-HEV antibodies and HEV RNA. Overall, 121 rabbits (57.3%) tested positive for anti-HEV antibodies, and 151 (71.6%) had detectable HEV RNA. The 174 HEV strains recovered from these viremic rabbits, including two distinct strains each from 23 rabbits, differed from each other by up to 13.6% in a 412-nucleotide (nt) sequence within ORF2, and were 89.3-95.9% identical to the reported rabbit HEV strains in other provinces of China. Three representative Inner Mongolian strains, one each from three phylogenetic clusters, whose entire genomic sequences were determined, shared 79.6-96.7% identities with reported rabbit HEV strains within the entire or 242- to 1349-nt partial genomic sequence. Rabbit HEV strains recovered from liver tissues of rabbits with a high HEV load propagated efficiently in human cell lines (A549 and PLC/PRF/5 cells), suggesting the potential zoonotic risk of rabbit HEV. © 2012 Elsevier B.V.


PubMed | Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease
Type: Journal Article | Journal: Theriogenology | Year: 2013

The factor associated suicide (Fas) and its ligand (FasL) signaling is an important regulatory pathway of apoptosis in mammalian follicles. However, whether apoptosis in bovine oocytes is regulated by the Fas-FasL signaling pathway remains unknown. In this study, localization of Fas and FasL in immature oocytes and FasL in cumulus cells were examined using immunofluorescence staining. In addition, exogenous FasL was added to an in vitro culture system to investigate apoptotic changes in bovine oocytes, using annexin-V and terminal uridine nick-end labeling staining, and real-time quantitative polymerase chain reaction. In this study, Fas was expressed in immature oocytes, whereas FasL was expressed in cumulus cells, but not in immature oocytes; annexin-V- and terminal uridine nick-end labeling-positive rates of oocytes treated with 2, 10, or 50 ng/mL FasL were higher than those of control oocytes (P < 0.05); and oocytes from the three treatment groups had higher expression levels of Fas and B cell lymphoma/leukemia-2 associated X than those in the control group (P < 0.05). Taken together, we concluded that the Fas-FasL signaling pathway was involved in regulation of bovine oocyte apoptosis, perhaps related to B cell lymphoma/leukemia-2 associated X upregulation.

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