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        The effects of a short sequence enhancer (50-GTGAAATAAATGCAAATAAAGT) and its derived sequences on green fluorescent protein expression

        Zhihong Ma,Zhanjun Lv,Xiu-Fang Wang,Xiangyang Jing,Jianjun Cheng 한국유전학회 2014 Genes & Genomics Vol.36 No.4

        Cis-regulatory elements are regions of DNAthat regulate the expression of genes located on that samemolecule of DNA. Though these elements are important forgene expression regulation, the functions of cis elementsremain largely unknown. To explore the mechanisms ofgene activation by short sequence enhancers, we examinedthe enhancer activity of short sequence DNA and itsderived sequences using a GFP expression system. Wefound that AA sequence (50-GTGAAATAAATGCAAATAAAGT) induced strong GFP gene expression, while7pieA (50-GTGAAAAAAATGCAAAAAAAGT) did not. We mutated the five T bases of the AA sequence to A, C orG. Our findings indicated that sequences retaining the 7thand/or 17th Ts possessed strong enhancer activity. RT-PCRand RNA synthesis inhibition analysis using actinomycin Drevealed that the enhanced GFP gene expression inducedby the AA sequence occurred at the transcriptional level. To determine whether the AA sequence formed a secondarystructure via atypical complementation, PAGE methodwas used, and the results showed that the AA sequenceformed a secondary structure. Our results support previousevidence that AATAAA is an important composition of ciselements (enhancer/promoter), and suggest that the formationof an unstable stem-loop structure via atypicalcomplementation may be a new mechanism of enhanceractivity.

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        The mechanism of mesna in protection from cisplatin-induced ovarian damage in female rats

        Xiaohuan Li,Shu Yang,Xiangyang Lv,Haimei Sun,Jing Weng,Yuanjing Liang,Deshan Zhou 대한부인종양학회 2013 Journal of Gynecologic Oncology Vol.24 No.2

        Objective: Cisplatin is a widely used chemotherapeutic agent in the treatment of cancers in clinic; but it often induces adverse effects on ovarian functions such as reduced fertility and premature menopause. Mesna could attenuate the cisplatin-induced ovarian damages; however, the underlying mechanism is still unknown. This study aimed to figure out the underlying mechanism of the protection of mesna for ovaries against cisplatin therapy in cancers. Methods: We performed female adult Sprague-Dawley rats into normal saline control (NS), low-dose cisplatin (CL), high-dose cisplatin (CH), CL plus mesna (CL+M), and CH plus mesna (CH+M) groups and detected anti-Müllerian hormone (AMH)-positive follicle, oxidative stress status and anti-oxidative capability in ovaries. Results: AMH-positive follicles were significantly decreased after cisplatin administration, which was significantly reversed when mesna was co-administered with cisplatin. The end product of lipid peroxidation, malondialdehyde (MDA), was significantly increased, but the anti-oxidative enzymatic activity of superoxide dismutase (SOD) and glutathione (GSH) were significantly decreased in cisplatin groups when compared with NS group. In contrast, after co-administration of cisplatin with mesna, MDA was significantly decreased whereas the activity of SOD and the concentration of GSH were increased. Moreover, mesna did not decrease the anti-tumor property of cisplatin in HePG2 cell lines. Conclusion: Cisplatin damages the granulosa cells by oxidative stress to deplete the ovarian reserve and mesna could protect ovarian reserve through anti-oxidation. These results might highlight the mechanism of the protection of mesna for ovarian reserve and open an avenue for the application of mesna as a protective additive in cisplatin chemotherapy in clinical practise.

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