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        Electroacupuncture-Modulated MiR106b-5p Expression Enhances Autophagy by Targeting Beclin-1 to Promote Motor Function Recovery After Spinal Cord Injury in Rats

        Shuhui Guo,Jianmin Chen,Ye Yang,Xiaolu Li,Yun Tang,Yuchang Gui,Jianquan Chen,Jianwen Xu 대한척추신경외과학회 2023 Neurospine Vol.20 No.3

        Objective: Electroacupuncture (EA) has a definite effect on the treatment of spinal cord injuries (SCIs), but its underlying molecular mechanism remains unclear. Meanwhile, MiR-106b-5p is an autophagy- and apoptosis-related microribonucleic acid, but whether it regulates the progression of autophagy and apoptosis in SCIs is yet undetermined. As such, this study aimed to elucidate the involvement of miR-106b-5p in the EA treatment of an SCI. Methods: The miR-106b-5p level was detected by quantitative real-time polymerase chain reaction. In vitro, SH-SY5Y cells were transfected with miR-106b-5p mimics or inhibitors to regulate the miR-106b-5p expression, while in vivo, SCI rats were treated with EA for 7 days at the bilateral Zusanli (ST36) and Jiaji (EX-B2) acupoints. The motor function was evaluated using the Basso-Beattie-Bresnahan (BBB) criteria. Further, autophagic vacuoles, pathological damage, and neuronal cell morphology were observed by transmission electron microscopy, as well as by hematoxylin and eosin and Nissl staining, respectively. Results: The miR-106b-5p level, which can interact directly with Beclin-1 by influencing its expression, as well as the expressions of P62, Caspase-3, and Bax, was upregulated after an SCI, but it decreased after EA. Moreover, the ratio of LC3-II to LC3-I was upregulated after EA. EA can enhance autophagy, reduce neuronal apoptosis, and minimize motor dysfunction and histopathological deficits after an SCI. More importantly, however, all the above effects induced by EA can be reversed after an injection of miR-106-5p agomir to produce an overexpression of miR-106b-5p. Conclusion: EA treatment could downregulate miR-106b-5p to alleviate SCI-mediated injuries by promoting autophagy and inhibiting apoptosis.

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        TsMIP6 enhances the tolerance of transgenic rice to salt stress and interacts with target proteins

        Linlin Sun,Guohong Yu,Xiaori Han,Shichao Xin,Xiaojing Qiang,Linlin Jiang,Shuhui Zhang,Xian-guo Cheng 한국식물학회 2015 Journal of Plant Biology Vol.58 No.5

        Aquaporins (AQPs), a large family of channel proteins in plants, play an important role in regulating the balance of osmotic potential in cells. We isolated an AQP gene, TsMIP6, from the halophyte Thellungiella salsuginea and functionally characterized it in transgenic rice (Oryza sativa). This gene belongs to a subfamily of tonoplast intrinsic proteins and is localized at the plasma membrane. Real-time PCR showed that expression of TsMIP6 in shoots or roots of T. salsuginea was markedly induced by salinity, whereas its ectopic expression in ‘Kitaake’ lines of rice significantly increased plant tolerance to salt stress. Physiological data suggested that TsMIP6 is involved in regulating ion homeostasis and water channel activity in salt-stressed transgenic rice. Heterologous expression analysis indicated that TsMIP6 specifically interacts with a member of the glycoside hydrolase family 64 protein #617 in yeast cells. This suggests that the relationship between TsMIP6 and #617 has a crucial role in mediating osmotic balance in plant cells. Moreover, TsMIP6 might help to modulate the transport of some neutral molecules and may function through a pathway regulating solute equilibrium to maintain osmotic potential.

      • Preparation and Characterization of Paclitaxel-loaded PLGA Nanoparticles Coated with Cationic SM5-1 Single-chain Antibody

        Kou, Geng,Gao, Jie,Wang, Hao,Chen, Huaiwen,Li, Bohua,Zhang, Dapeng,Wang, Shuhui,Hou, Sheng,Qian, Weizhu,Dai, Jianxin,Zhong, Yanqiang,Guo, Yajun Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.5

        The purpose of this study was to develop paclitaxel-loaded poly(lactide-co-glycolide) (PLGA) nanoparticles coated with cationic SM5-1 single-chain antibody (scFv) containing a polylysine (SMFv-polylys). SM5-1 scFv (SMFv) is derived from SM5-1 monoclonal antibody, which binds to a 230 kDa membrane protein specifically expressed on melanoma, hepatocellular carcinoma and breast cancer cells. SMFv-polylys was expressed in Escherichia coli and purified by cation-exchange chromatography. Purified SMFv-polylys was fixed to paclitaxel-loaded PLGA nanoparticles to form paclitaxel-loaded PLGA nanoparticles coated with SMFv-polylys (Ptx-NP-S). Ptx-NP-S was shown to retain the specific antigen-binding affinity of SMFv-polylys to SM5-1 binding protein-positive Ch-hep-3 cells. Finally, the cytotoxicity of Ptx-NP-S was evaluated by a non-radioactive cell proliferation assay. It was demonstrated that Ptx-NP-S had significantly enhanced in vitro cytotoxicity against Ch-hep-3 cells as compared with non-targeted paclitaxel-loaded PLGA nanoparticles. In conclusion, our results suggest that cationic SMFv-polylys has been successfully generated and may be used as targeted ligand for preparing cancer-targeted nanoparticles.

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