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      • KCI등재

        Oncomodulin/Truncated Protamine-Mediated Nogo-66 Receptor Small Interference RNA Delivery Promotes Axon Regeneration in Retinal Ganglion Cells

        Cui, Zhili,Kang, Jun,Hu, Dan,Zhou, Jian,Wang, Yusheng Korean Society for Molecular and Cellular Biology 2014 Molecules and cells Vol.37 No.8

        The optic nerve often suffers regenerative failure after injury, leading to serious visual impairment such as glaucoma. The main inhibitory factors, including Nogo-A, oligodendrocyte myelin glycoprotein, and myelin-associated glycoprotein, exert their inhibitory effects on axonal growth through the same receptor, the Nogo-66 receptor (NgR). Oncomodulin (OM), a calcium-binding protein with a molecular weight of an ~12 kDa, which is secreted from activated macrophages, has been demonstrated to have high and specific affinity for retinal ganglion cells (RGC) and promote greater axonal regeneration than other known polypeptide growth factors. Protamine has been reported to effectively deliver small interference RNA (siRNA) into cells. Accordingly, a fusion protein of OM and truncated protamine (tp) may be used as a vehicle for the delivery of NgR siRNA into RGC for gene therapy. To test this hypothesis, we constructed OM and tp fusion protein (OM/tp) expression vectors. Using the indirect immunofluorescence labeling method, OM/tp fusion proteins were found to have a high affinity for RGC. The gel shift assay showed that the OM/tp fusion proteins retained the capacity to bind to DNA. Using OM/tp fusion proteins as a delivery tool, the siRNA of NgR was effectively transfected into cells and significantly down-regulated NgR expression levels. More importantly, OM/tp-NgR siRNA dramatically promoted axonal growth of RGC compared with the application of OM/tp recombinant protein or NgR siRNA alone in vitro. In addition, OM/tp-NgR siRNA highly elevated intracellular cyclic adenosine monophosphate (cAMP) levels and inhibited activation of the Ras homolog gene family, member A (RhoA). Taken together, our data demonstrated that the recombinant OM/tp fusion proteins retained the functions of both OM and tp, and that OM/tp-NgR siRNA might potentially be used for the treatment of optic nerve injury.

      • KCI등재

        Oncomodulin/Truncated Protamine-Mediated Nogo-66 Receptor Small Interference RNA Delivery Promotes Axon Regeneration in Retinal Ganglion Cells

        Zhili Cui,Jun Kang,Dan Hu,Jian Zhou,Yusheng Wang 한국분자세포생물학회 2014 Molecules and cells Vol.37 No.8

        The optic nerve often suffers regenerative failure after injury, leading to serious visual impairment such as glaucoma. The main inhibitory factors, including Nogo-A, oligodendrocyte myelin glycoprotein, and myelin-associated glycoprotein, exert their inhibitory effects on axonal growth through the same receptor, the Nogo-66 receptor (NgR). Oncomodulin (OM), a calcium-binding protein with a molecular weight of an ~12 kDa, which is secreted from activated macrophages, has been demonstrated to have high and specific affinity for retinal ganglion cells (RGC) and promote greater axonal regeneration than other known polypeptide growth factors. Protamine has been reported to effectively deliver small interference RNA (siRNA) into cells. Accordingly, a fusion protein of OM and truncated protamine (tp) may be used as a vehicle for the delivery of NgR siRNA into RGC for gene therapy. To test this hypothesis, we constructed OM and tp fusion protein (OM/tp) expression vectors. Using the indirect immunofluorescence labeling method, OM/tp fusion proteins were found to have a high affinity for RGC. The gel shift assay showed that the OM/tp fusion proteins retained the capacity to bind to DNA. Using OM/tp fusion proteins as a delivery tool, the siRNA of NgR was effectively transfected into cells and significantly down-regulated NgR expression levels. More importantly, OM/tp-NgR siRNA dramatically promoted axonal growth of RGC compared with the application of OM/tp recombinant protein or NgR siRNA alone in vitro. In addition, OM/tp-NgR siRNA highly elevated intracellular cyclic adenosine monophosphate (cAMP) levels and inhibited activation of the Ras homolog gene family, member A (RhoA). Taken together, our data demonstrated that the recombinant OM/tp fusion proteins retained the functions of both OM and tp, and that OM/tp-NgR siRNA might potentially be used for the treatment of optic nerve injury.

      • KCI등재

        Effect of sintering temperature on properties of lightweightporous ceramics prepared by foam impregnation method

        Zhili Cui,Shiming Xiao,Xianli Luo,Yunxuan Liu,Ming Liu,Yuyun Zeng,Xiaoli Zhong,Hong Zheng,Haifeng Guo 한양대학교 청정에너지연구소 2023 Journal of Ceramic Processing Research Vol.24 No.5

        In this paper, cheap mineral materials were used as the base materials of lightweight porous ceramics prepared through foamimpregnation method. The effect of the sintering temperature on the properties of the prepared porous ceramics was studied. The porous ceramic was mainly composed of amorphous silicon oxide, crystalline cordierite and mullite phases, and a smallamount of alumina phase. As the sintering temperature increased, the porosity of porous ceramics gradually decreased from94% to 92%, and the bulk density increased from 0.173 gcm-3 at 1100 ℃ to 0.194 gcm-3 at 1200 ℃. The best sinteringtemperature was 1180 ℃. The porosity of the porous ceramics sintered at 1180 ℃ was 92.14%, the volume weight was 0.189gcm-3, the shrinkage rate was 15.80%, the compressive strength was 0.79 MPa, and the thermal conductivity was 0.295 Wm-1k-1. The lightweight porous ceramic has high porosity, low density and good thermal insulation, as well as low cost, having greatpotential for application in fields such as thermal insulation, adsorption, and environmental protection.

      • KCI등재

        Cellular uptake mechanism and comparative <i>in vitro</i> cytotoxicity studies of monomeric LMWP-siRNA conjugate

        Ye, Junxiao,Pei, Xing,Cui, Hui,Yu, Zhili,Lee, Hyukjin,Wang, Jianxin,Wang, Xu,Sun, Lu,He, Huining,Yang, Victor C. THE KOREAN SOCIETY OF INDUSTRIAL AND ENGINEERING 2018 JOURNAL OF INDUSTRIAL AND ENGINEERING CHEMISTRY -S Vol.63 No.-

        <P><B>Abstract</B></P> <P>The covalent attachment of CPPs to siRNA molecules offers great potential for CPP-mediated siRNA delivery. We recently reported a concise and high-yield synthesis strategy of the cell-permeable, cytosol-dissociable LMWP-siRNA covalent conjugate. Herein, cell uptake mechanism and cellular toxicity studies of this conjugate were performed to evaluate the potential of LMWP-siRNA conjugate for clinical translation. Cellular uptake mechanism study indicated that the conjugate could be taken up by cells via multiple pathways, including direct penetration of the plasma membrane and clathrin- and caveolae-independent endocytosis. <I>In vitro</I> cytotoxicity study revealed that the conjugation promoted internalization in a low-toxic fashion.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

      • KCI등재

        Cellular uptake mechanism and comparative in vitro cytotoxicity studies of monomeric LMWP-siRNA conjugate

        Junxiao Ye,Xing Pei,Hui Cui,Zhili Yu,이혁진,Jian-Xin Wang,Xu Wang,Lu Sun,Huining He,Victor C. Yang 한국공업화학회 2018 Journal of Industrial and Engineering Chemistry Vol.63 No.-

        The covalent attachment of CPPs to siRNA molecules offers great potential for CPP-mediated siRNA delivery. We recently reported a concise and high-yield synthesis strategy of the cell-permeable, cytosol-dissociable LMWP-siRNA covalent conjugate. Herein, cell uptake mechanism and cellular toxicity studies of this conjugate were performed to evaluate the potential of LMWP-siRNA conjugate for clinical translation. Cellular uptake mechanism study indicated that the conjugate could be taken up by cells via multiple pathways, including direct penetration of the plasma membrane and clathrin- and caveolae-independent endocytosis. In vitro cytotoxicity study revealed that the conjugation promoted internalization in a low-toxic fashion.

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