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      • KCI등재

        Effect of NaCl on the Stability of Oncolytic Vaccinia Virus

        Seong-Geun Kim(김성근),Gui Shao Ran(계소연),Hyuk-Chan Kwon(권혁찬),Tae-Ho Hwang(황태호) 한국생명과학회 2016 생명과학회지 Vol.26 No.1

        Pexa-Vec (JX-594)은 암특이적 암용해 면역치료제인 백시니아 바이러스이다. 본 연구의 목적은 Pexa-Vec의 안정성을 극대화하기 위한 방법을 개발하는 것이다. 단기안정성 실험에서 바이러스의 활성은 4℃와 실온에서 감소하였으나, 초음파처리와 회전처리로 완전히 회복되었다. Pexa-Vec의 장기안정성 시험은 (A) 30 mM Tris/pH 7.6, (B) 30 mM Tris/pH 8.6, (C) 30 mM Tris/pH 7.6, 150 mM NaCl, 15% sucrose, (D) 30 mM Tris/pH 7.6, 15% sucrose, (E) 30 mM Tris/pH 8.6, 15% sucrose 조건 하에서 수행하였다. 제형 A는 4℃에서 4-8주 후, 실온에서 1주일 후에 2로그 이하로 바이러스활성이 감소되었다. 반면 제형 B의 경우 4℃와 실온에서 바이러스 활성이3일 후 감소되는 것으로 관찰되어 중성 산도가 바이러스 안정성을 유지하는데 필수적이다. 제형A에 15%의 슈크로즈 수크로오스를 추가했을 때(제형D), -20℃, 4℃와 실온에서 바이러스성 안정성이 크게 증가 하였고, 제형 E (pH 7.6)에서 다시 한번 확인되었다. 제형 D (pH 7.6)에 150 mM 염화나트륨을 추가한 제형 C에서 바이러스 안전성을 증가시키는 슈크로즈 수크로오스 효과를 더욱 향상시켜, 4℃와 실온에서 바이러스 활성이 각각 1.5년과 1-2주 동안 유지되는 결과를 보였다. 결론적으로, 우리는 제형C가 항암 백시니아 바이러스를 적절히 저장하기 위한 충분한 조건을 제공할 수 있다고 제안한다. Pexa-Vec (JX-594) is a specific cancer-targeted oncolytic and immunotherapeutic vaccinia virus. The purpose of this study was to develop methods to maximize the stability of Pexa-Vec. In short-term instability testing, viral activity was rapidly decreased both at 4°C and at room temperature (RT), but it was completely restored after sonication followed by vortex. Long-term stability testing of Pexa-Vec in the following liquid formulations was performed: (A) 30 mM Tris/pH 7.6, (B) 30 mM Tris/pH 8.6,(C) 30 mM Tris/pH 7.6, 150 mM NaCl, 15% sucrose, (D) 30 mM Tris/pH 7.6, 15% sucrose, and (E) 30 mM Tris/pH 8.6, 15% sucrose. Viral activity decreased less than 2 log10 at 4°C, and RT was observed in 3 days in B, while viral activity was not decreased even after 4–8 weeks at 4°C and at 1 week in RT in A, suggesting that neutral pH may be essential to maintain virus stability. The addition of 15% sucrose into A (D) significantly increased viral stability at -20°C, 4°C, or RT, and it was also observed at pH 8.6 (E). The addition of 150 mM NaCl into D (C) significantly increased viral stability in addition to the sucrose effect at 4°C or RT. Accordingly, the viral activity in formulation C was maintained for 1.5 years at 4°C, and for 1-2 weeks in RT. In conclusion, we propose that formulation C can provide the most adequate condition for the proper storage of vaccinia oncolytic virus.

      • SCOPUSKCI등재

        Original Articles : Effect of aldosterone on the amplification of oncolytic vaccinia virus in human cancer Lines

        ( Hyun Ju Lee ),( Ja Sung Rho ),( Shao Ran Gui ),( Mi Kyung Kim ),( Yu Kyoung Lee ),( Yeon Sook Lee ),( Jeong Eun Kim ),( Eu Na Cho ),( Mong Cho ),( Tae Ho Hwang ) 대한간학회 2011 Clinical and Molecular Hepatology(대한간학회지) Vol.17 No.3

        Background/Aims: JX-594 is an oncolytic virus derived from the Wyeth vaccinia strain that causes replication-dependent cytolysis and antitumor immunity. Starting with a cross-examination of clinical-trial samples from advanced hepatocellular carcinoma patients having high levels of aldosterone and virus amplification in JX-594 treatment, we investigated the association between virus amplification and aldosterone in human cancer cell lines. Methods: Cell proliferation was determined by a cell-counting-kit-based colorimetric assay, and vaccinia virus quantitation was performed by quantitative polymerase chain reaction (qPCR) and a viral plaque assay. Also, the intracellular pH was measured using a pH-sensitive dye. Results: Simultaneous treatment with JX-594 and aldosterone significantly increased viral replication in A2780, PC-3, and HepG2 cell lines, but not in U2OS cell lines. Furthermore, the aldosterone treatment time altered the JX-594 replication according to the cell line. The JX-594 replication peaked after 48 and 24 hours of treatment in PC-3 and HepG2 cells, respectively. qPCR showed that JX-594 entry across the plasma membrane was increased, however, the changes are not significant by the treatment. This was inhibited by treatment with spironolactone (an aldosterone-receptor inhibitor). JX-594 entry was significantly decreased by treatment with EIPA [5-(N-ethyl-N-isopropyl)amiloride; a Na+/H+-exchange inhibitor], but aldosterone significantly restored JX-594 entry even in the presence of EIPA. Intracellular alkalization was observed after aldosterone treatment but was acidified by EIPA treatment. Conclusions: Aldosterone stimulates JX-594 amplification via increased virus entry by affecting the H+ gradient. (Korean J Hepatol 2011;17:213-219)

      • KCI등재

        Ionic liquid extraction of silkworm pupa protein and its biological characteristics

        Zeng Qing-Lei,Zhang Ning,Zhang Yue-Yue,Xin Xiang-Dong,Attaribo Thomas,Shao Ying,Tang Liu-Mei,Zhang Ran,이광식,진병래,Gui Zhongzheng 한국응용곤충학회 2021 Journal of Asia-Pacific Entomology Vol.24 No.1

        Silkworm (Bombyx mori) pupa protein (SPP) is a high-quality source of animal protein with substantial nutri tional benefits and health value. To develop an efficient extraction method for SPP that is environmentally friendly, we selected choline hydroxide ionic liquid (CH-IL) as the extraction solvent and performed orthogonal experiments to optimize the extraction conditions. We demonstrated that 3% CH-IL, a solid-to-liquid ratio (g/mL) of 1:30, an extraction temperature of 40 ◦ C, and an extraction time of 1 h facilitated the most efficient extraction. Compared to the conventional alkali solubilization–acid precipitation method, the CH-IL extraction increased protein content by 12.14%. Protein structure analysis showed that the β-sheet content increased by 10.98% and that of disulfide bonds reduced by 16.4%. The processing properties of the CH-IL extracted protein showed that the solubility, emulsification, and foaming capacity were enhanced by 82.87%, 15.44%, and 18.97%, respec tively. The physical properties of SPP remarkably improved relative to the increased stretching of the poly peptide chains. The findings of this study provide technical knowledge that will enhance the processing performance of pupal proteins.

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