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      • KCI등재

        Development of Toxoplasma gondii Chinese I genotype Wh6 Strain in Cat Intestinal Epithelial Cells

        Gui-Hua Zhao,Lixin Zhang,Lisha Dai,Haozhi Xu,Chao Xu,Ting Xiao,Jin Li,Hui Sun,Beibei Zhou,Kun Yin 대한기생충학ㆍ열대의학회 2022 The Korean Journal of Parasitology Vol.60 No.4

        Felids are the unique definitive host of Toxoplasma gondii. The intestine of felid is the only site for initiating Toxoplasma gondii sexual reproduction. T. gondii excretes millions of infectious oocysts from the intestine, which are the primary source of infection. There are many difficulties in developing vaccines and drugs to control oocyst excretion due to the lack of an appropriate experimental model. Here, we established an in vitro feline intestinal epithelial cell (IEC) infection system and an efficient animal model of T. gondii Chinese 1 genotype, Wh6 strain (TgCtwh6). The Kunming mice brain tissues containing TgCtwh6 cysts were harvested 42-day post-infection. The bradyzoites were co-cultured with cat IECs in vitro at a ratio of 1:10. Five 3-month-old domestic cats were orally inoculated with 600 cysts each. The oocysts were detected by daily observation of cat feces by microscopy and polymerase chain reaction. We found that the parasite adhered and invaded cat IECs in vitro, transformed into tachyzoites, and then divided to form rose-like structures. These parasites eventually destroyed host cells, escaped, and finished the asexual reproduction process. Schizonts associated with sexual reproduction have not been observed during development in vitro cultured cells. However, schizonts were detected in all infected cat intestinal epithelial cells, and oocysts were presented in all cat feces. Our study provides a feasible cell model and an efficient infection system for the following studies of T. gondii sexual reproduction, and also lays a foundation to develop drugs and vaccines for blocking excretion and transmission of oocysts.

      • KCI등재

        Epithelial CST1 Promotes Airway Eosinophilic Inflammation in Asthma via the AKT Signaling Pathway

        Du Lijuan,Xu Changyi,Tang Kun,Shi Jia,Tang Lu,Lisha Xiao,Lei Chengcheng,Liu Huicong,Liang Yuxia,Guo Yubiao 대한천식알레르기학회 2023 Allergy, Asthma & Immunology Research Vol.15 No.3

        Purpose: Epithelial cystatin SN (CST1), a type 2 cysteine protease inhibitor, was significantly upregulated in asthma. In this study, we aimed to investigate the potential role and mechanism of CST1 in eosinophilic inflammation in asthma. Methods: Bioinformatics analysis on Gene Expression Omnibus datasets were used to explore the expression of CST1 in asthma. Sputum samples were collected from 76 asthmatics and 22 control subjects. CST1 mRNA and protein expression in the induced sputum were measured by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and western blotting. The possible function of CST1 was explored in ovalbumin (OVA)-induced eosinophilic asthma. Transcriptome sequencing (RNA-seq) was used to predict the possible regulated mechanism of CST1 in bronchial epithelial cells. Overexpression or knockdown of CST1 was further used to verify potential mechanisms in bronchial epithelial cells. Results: CST1 expression was significantly increased in the epithelial cells and induced sputum of asthma. Increased CST1 was significantly associated with eosinophilic indicators and T helper cytokines. CST1 aggravated airway eosinophilic inflammation in the OVA-induced asthma model. In addition, overexpression of CST1 significantly enhanced the phosphorylation of AKT and the expression of serpin peptidase inhibitor, clade B, member 2 (SERPINB2), while knockdown using anti-CST1 siRNA reversed the trend. Furthermore, AKT had a positive effect on SERPINB2 expression. Conclusions: Increased sputum CST1 may play a key role in the pathogenesis of asthma through involvement in eosinophilic and type 2 inflammation through activation of the AKT signaling pathway, further promoting SERPINB2 expression. Therefore, targeting CST1 might be of therapeutic value in treating asthma with severe and eosinophilic phenotypes.

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        Effect of Nano-CaCO3 on the Sealing Efficiency of Grouts in Flowing Water Grouting

        Yao-Hui Liu,Ping Yang,Ze-Bin Ouyang,Xiao Wei,Lisha Zhang 대한토목학회 2020 KSCE JOURNAL OF CIVIL ENGINEERING Vol.24 No.10

        Flowing water grouting is a big challenge in the tunneling and underground engineering. To enhance the early strength and grouting effectiveness of slurry for flowing water grouting, nano-CaCO3 and fly ash were mixed with cement based grout. A series of physical simulation tests were conducted to simulate the flowing water grouting process in rough rock fracture, and investigate the effect of nano-CaCO3 content on the fluid pressure and sealing efficiency of grouts. Results of viscosity tests show that the viscosity of grouts decreased with an increase of nano-CaCO3 content. Scanning electron microscope (SEM) tests indicate that nano-CaCO3can promote the formation of fibrous hydrates and enhance the flowing water resistance of grouts. Increasing nano-CaCO3 content resulted in the first increase while later decrease of maximal fluid pressure (MFP) and sealing efficiency (SE) of grouts. Reducing water cement ratio of grouts and incorporating fly ash can effectively improve the SE of flowing water grouting.

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