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RISS 인기검색어
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- 저자 : Wanhong Li (검색결과 4 건)
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Gallic acid caused cultured mice TM4 Sertoli cells apoptosis and necrosis
Wanhong Li,Xiangpeng Yue,Fadi Li 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.5
Objective: The study was designed to determine the cytotoxic effect of gallic acid (GA), obtained by the hydrolysis of tannins, on mice TM4 Sertoli cells apoptosis. Methods: In the present study, non-tumorigenic mice TM4 Sertoli cells were treated with different concentrations of GA for 24 h. After treatment, cell viability was evaluated using WST-1, mitochondrial dysfunction, cells apoptosis and necrosis was detected using JC-1, Hoechst 33342 and propidium iodide staining. The expression levels of Cyclin B1, proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (BAX), and Caspase-3 were also detected by quantitative real-time polymerase chain reaction and Western-blotting. Results: The results showed that 20 to 400 μM GA inhibited viability of TM4 Sertoli cells in a dose-dependent manner. Treatment with 400 μM GA significantly inhibited PCNA and Cyclin B1 expression, however up-regulated BAX and Caspase-3 expression, caused mitochondrial membrane depolarization, activated Caspase-3, and induced DNA damage, thus, markedly increased the numbers of dead cells. Conclusion: Our findings showed that GA could disrupt mitochondrial function and caused TM4 cells to undergo apoptosis and necrosis.
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Cooperation between Human DAF and CD59 in Protecting Cells from Human Complement-mediated Lysis
Xu, Li,Wu, Wenlan,Zhao, Zhouzhou,Shao, Huanjie,Liu, Wanhong,Liu, Hui,Li, Wenxin Korean Society for Biochemistry and Molecular Biol 2006 Journal of biochemistry and molecular biology Vol.39 No.6
The complement (C) regulatory proteins decay accelerating factor (DAF, CD55) and CD59 could protect host cells using different mechanisms from C-mediated damage at two distinct levels within the C pathway. Co-expression of DAF and CD59 would be an effective strategy to help overcome host C-induced xenograft hyperacute rejection. In this study, we made a construct of recombinant expression vector containing DAF and CD59 cDNA and the stable cell lines were obtained by G418 selection. Extraneous genes integration and co-expression were identified by PCR, RT-PCR and Western blot analysis. Human c-mediated cytolysis assays showed that NIH/3T3 cells transfected stably with pcDNA3-CD59, pcDNA3-DAF, and pcDNA3-CD59DAF-DP were protected from C-mediated damage and that synchronously expressed human CD59 and DAF provided the most excellent protection for host cells as compared with either human CD59 or DAF expressed alone. Therefore, the construct represents an effective and efficacy strategy to overcome C-mediated damage in cells and, ultimately, in animals.
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Expression and Detection of Retinol-Binding Protein-4 Gene of Pig in E. coli
LiNa Sun,WanHong Li,ShuXiong Chen,Chao Chen,XiaoFeng Hou,Yun Zhao,Lu Chen,ChunJin Li,Xu Zhou 한국동물생명공학회(구 한국동물번식학회) 2014 한국동물번식학회 한중일 심포지엄 Vol.2014 No.1
Retinol-Binding Protein-4 (RBP-4) is a low molecular weight lipocalin, which mainly functions as a carrier for vitamin A. Though liver is the main machinery for synthesis of this protein, it is also detectable in other extrahepatic tissues, for example, ovary, uterus, and placenta. Recent evidences have shown that RBP-4 plays important roles in animal reproduction, for example, promoting the development of uterus and embryo. To the best of our knowledge, our laboratory firstly reported that high level of RBP-4 existed in follicular fluid from follicular cysts in sows. Moreover, we have also found that RBP-4 could be secreted by granulosa cells, and RBP-4 receptor was detected in granulosa cells. However, there is no any evidence on the role of RBP-4 in regulating the follicular development. Therefore, cloning and expression of RBP-4 and preparation of polyclonal antibody could help us to explore the role of RBP-4 in follicular development. The aim of this work was to construct prokaryotic expression system of swine RBP-4 gene. The total RNA was extracted from swine’s normal ovarian tissue. The sequence including the whole length of RBP-4 was amplified by RT-PCR and inserted into pEASY-E1.Then transformed into E. coli BL21(DE3)pLysS after gene sequencing. Three hours later, adding IPTG with the final concentration of 1mmol/L and inducing five hours. After centrifugation, the supernatant was discarded. By adding Glucose to Luria-Bertani broth, the expressions of protein were increased. SDS-PAGE showed that the RBP-4 gene expressed in the form of inclusion body with a molecular weight of 21KD. Western-Blot results showed that the target protein could be specifically recognized by mouse anti-human monoclonal antibody. Prokaryotic expression vector of RBP-4 gene was successfully established, and the gene was successfully expressed n E. coli, which is ready for purification and RBP-4 polyclone antibody. Meanwhile, these results were beneficial to investigate the function of RBP-4 in follicular development.
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