• Journal of Pharmaceutical Investigation
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• 제31권3호
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• pp.185-190
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• 2001

Human papillomavirus (HPV) infection is known to cause cervical cancers. Human papillomavirus-like particles (VLP) have been studied as preventive vaccines of cervical cancers. To develop VLP as a therapeutic gene carrier, we studied the method to encapsulate cytokine genes in virus-like particles. HPV type 16 capsid L1 genes were amplified by polymerase chain reaction and cloned into T vector. L1 gene was then inserted into baculovirus transfer vector. The clone of baculovirus encoding L1 gene was isolated and used to express L1 protein in Sf 21 insect cells. VLP were purified by CsCl density gradient and ultracentrifugation. VLP were disassembled to capsomer units by treatment of a reducing agent. Given that interleukin-2 (IL-2) genes have been used in anticancer gene therapy and as a molecular adjuvant, IL-2 cytokine plasmids were chosen as a model gene. IL-2 plasmids were incubated with the disassembled capsomer suspension. To reassemble the particles, the mixture of capsomers and cytokine plasmids was dialyzed. The disassembly and reassembly of VLP were confirmed by transmission electron microscopy. The entrapment of cytokine plasmids in reassembled VLP was tested by the stability of plasmids against DNase I. After treatment of reassembled virus-like particles with DNase I, discrete IL-2 DNA band was observed. Our results indicate that IL-2 cytokine plasmid (3.5 kb size) can be encapsulated in the virus-like particles, suggesting the potential of VLP as a gene delivery system. Moreover, VLP containing the adjuvant cytokine plasmids might function as more effective subunit vaccines.

• The Korean Journal of Physiology and Pharmacology
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• 제9권2호
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• pp.131-135
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• 2005

Potassium channels in human skin fibroblast have been studied as a possible site of Alzheimer disease pathogenesis. Fibroblasts in Alzheimer disease show alterations in signal transduction pathway such as changes in $Ca^{2+}$ homeostasis and/or $Ca^{2+}-activated$ kinases, phosphatidylinositol cascade, protein kinase C activity, cAMP levels and absence of specific $K^+$ channel. However, little is known so far about electrophysiological and pharmacological characteristics of large-conductance $Ca^{2+}$-activated $K^+$ ($BK_{Ca}$) channel in human fibroblast (CRL-1474). In the present study, we found Iberiotoxin- and TEA-sensitive outward rectifying oscillatory current with whole-cell recordings. Single channel analysis showed large conductance $K^{+}$ channels (106 pS of chord conductance at +40 mV in physiological $K^+$ gradient). The 106 pS channels were activated by membrane potential and $[Ca^{2+}]_i$, consistent with the known properties of $BK_{Ca}$ channels. $BK_{Ca}$ channels in CRL-1474 were positively regulated by adenylate cyclase activator ($10{\mu}M$ forskolin), 8-Br-cyclic AMP ($300{\mu}M$) or 8-Br-cyclic GMP ($300{\mu}M$). These results suggest that human skin fibroblasts (CR-1474) have typical $BK_{Ca}$ channel and this channel could be modulated by c-AMP and c-GMP. The electrophysiological characteristics of fibroblasts might be used as the diagnostic clues for Alzheimer disease.

• Journal of Microbiology and Biotechnology
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• 제30권7호
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• pp.1005-1012
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• 2020

Acid mine drainage (AMD) has been a serious environmental issue that threatens soil and aquatic ecosystems. In this study, an acid-tolerant sulfate-reducing bacterium, strain S4, was isolated from the mud of an AMD storage pond in Vietnam via enrichment in anoxic mineral medium at pH 5. Comparative analyses of sequences of the 16S rRNA gene and dsrB gene involved in sulfate reduction revealed that the isolate belonged to the genus Desulfovibrio, and is most closely related to Desulfovibrio oxamicus (with 99% homology in 16S rDNA sequence and 98% homology in dsrB gene sequence). Denaturing gradient gel electrophoresis (DGGE) analyses of dsrB gene showed that strain S4 represented one of the two most abundant groups developed in the enrichment culture. Notably, strain S4 was capable of reducing sulfate in low pH environments (from 2 and above), and resistance to extremely high concentration of heavy metals (Fe 3,000 mg/l, Zn 100 mg/l, Cu 100 mg/l). In a batch incubation experiment in synthetic AMD with pH 3.5, strain S4 showed strong effects in facilitating growth of a neutrophilic, metal sensitive Desulfovibrio sp. strain SR4H, which was not capable of growing alone in such an environment. Thus, it is postulated that under extreme conditions such as an AMD environment, acid- and metal-tolerant sulfate-reducing bacteria (SRB)-like strain S4 would facilitate the growth of other widely distributed SRB by starting to reduce sulfate at low pH, thus increasing pH and lowering the metal concentration in the environment. Owing to such unique physiological characteristics, strain S4 shows great potential for application in sustainable remediation of AMD.

• Environmental Engineering Research
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• 제19권4호
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• pp.331-338
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• 2014

Descriptive statistics methods were used to study the spatiotemporal variations and sources of ambient particulate matter ($PM_{10}$) in Luzhou, China. The analyzed datasets were collected from four national air quality monitoring stations: Jiushi (S1), Xiaoshi (S2), Zhongshan (S3), Lantian (S4) over the period of 2003-2012. This city was subjected serious $PM_{10}$ pollution, and the long-term annual average $PM_{10}$ concentrations varied from 76 to $136{\mu}g/m^3$. The maximum concentration was more than 3-fold of the annual average ($40{\mu}g/m^3$) issued by EPA-China for the ambient air quality. General temporal pattern was characterized by high concentrations in winter and low concentrations in summer, and general spatial gradient was in the reduction order of S2 > S4 > S3 > S1, which were both due to different particulate contributors and special meteorological conditions. The source apportionment indicated that vehicular emissions, road dusts, coal burning and chemical dusts were the major contributors of the identified $PM_{10}$ pollution, and the vehicular emissions and the road wear re-suspended particles dominated the heavy $PM_{10}$ pollution in recent years. Two other potential sources, agricultural and celebration activities could decrease the air quality in a short term. Finally, some corresponding suggestions and measures were provided to improve the air quality.

• Journal of Microbiology and Biotechnology
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• 제23권11호
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• pp.1617-1626
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• 2013

The Bacterial community structure and its complexity of the enrichment culture during the isolation and screening of imidacloprid-degrading strain were studied using denaturating gradient gel electrophoresis analysis. The dominant bacteria in the original tea rhizosphere soil were uncultured bacteria, Rhizobium sp., Sinorhizobium, Ochrobactrum sp., Alcaligenes, Bacillus sp., Bacterium, Klebsiella sp., and Ensifer adhaerens. The bacterial community structure was altered extensively and its complexity reduced during the enrichment process, and four culturable bacteria, Ochrobactrum sp., Rhizobium sp., Geobacillus stearothermophilus, and Alcaligenes faecalis, remained in the final enrichment. Only one indigenous strain, BCL-1, with imidacloprid-degrading potential, was isolated from the sixth enrichment culture. This isolate was a gram-negative rod-shaped bacterium and identified as the genus Ochrobactrum based on its morphological, physiological, and biochemical properties and its 16S rRNA gene sequence. The degradation test showed that approximately 67.67% of the imidacloprid (50 mg/l) was degraded within 48 h by strain BCL-1. The optimum conditions for degradation were a pH of 8 and $30^{\circ}C$. The simulation of imidacloprid bioremediation by strain BCL-1 in soil demonstrated that the best performance in situ (tea soil) resulted in the degradation of 92.44% of the imidacloprid (100 mg/g) within 20 days, which was better than those observed in the ex situ simulations that were 64.66% (cabbage soil), 41.15% (potato soil), and 54.15% (tomato soil).

• 대한환경공학회지
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• 제36권9호
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• pp.624-628
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• 2014

최근 해수와 담수의 염분농도차를 이용하여 발전하는 역전기투석(reverse electrodialysis, RED)은 잠재량이 크고 지속적 전력생산이 가능한 친환경적이며 미래지향적 신재생에너지로 인식되어 연구가 활발히 진행되고 있다. 이러한 RED 기술의 상용화를 위해서는 최적화된 운전조건을 찾는 것이 중요하며 특히, 스택 내부의 저항을 최소화하는 연구가 절실한 상황이다. 본 연구에서는 RED의 중요한 운전조건인 해수와 담수의 공급유량이 내부저항에 미치는 영향을 조사하고, 해수와 담수의 유량비에 따른 내부저항과 전력밀도의 관계를 고찰하여 최적의 운전조건을 실험적으로 파악하였다. 그 결과 전체유량 80 mL/min에서 최적의 해수/담수 유량비는 1.7이었으며, 이때 전력밀도는 $1.30W/m^2$을 얻을 수 있었다.

• The Korean Journal of Physiology and Pharmacology
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• 제19권6호
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• pp.485-489
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• 2015

Diabetic nephropathy (DN) is the leading cause of end-stage failure of the kidney, but the efficacy of currently available strategies for the prevention of DN remains unsatisfactory. In this study, we investigated the effects of free anthraquinones (FARs) extract, which was extracted from the rhubarb and purified by macroporous resin DM130 with gradient mixtures of ethanol/water as the lelution solvents, in high glucose-cultured glomerular mesangial cells (MCs). The cell proliferation was determined by CCK-8 assay, the levels of TGF-${\beta}1$, CTGF, ColIV and FN proteins in the supernatant of MCs were measured by ELISA assays, and the mRNA levels of these four genes were detected by RT-PCR. The results showed that the increased proliferation of MCs, the mRNA levels and protein expression of TGF-${\beta}1$, CTGF, ColIV and FN induced by high glucose were inhibited after the treatment with the FARs extract. This indicated that FARs extract could inhibit cell proliferation and the expression of main extracellular matrix induced by high glucose in MCs. The FARs extract exhibited potential values for prophylaxis and therapy of DN.

• 한국고분자학회:학술대회논문집
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• 한국고분자학회 2006년도 IUPAC International Symposium on Advanced Polymers for Emerging Technologies
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• pp.333-333
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• 2006

We synthesized two novel polynorbornene derivatives, chiral poly(norbornene acid methyl ester) (C-PNME) and racemic poly(norbornene acid n-butyl ester) (R-PNME), which are potential low dielectric constant materials for applications in advanced microelectronic and display devices. Thin films of these polymers deposited on substrates were investigated by structural analyses using synchrotron grazing incidence X-ray scattering, specular reflectivity and ellipsometry. These analyses provided important information on the structure, electron density gradient across film thickness, chain orientation, refractive index and thermal expansion of the polymers in substrate-supported thin films. The structural characteristics and properties of the thin films were first dependent on the polymer chain' tacticity and further influenced by film thickness and thermal annealing.

• Asian Pacific Journal of Cancer Prevention
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• 제14권1호
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• pp.309-314
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• 2013

Aim: Brain tumors almost universally have fatal outcomes; new therapeutics are desperately needed and will only come from improved understandins of glioma biology. Methods: Exosomes are endosomally derived 30~100 nm membranous vesicles released from many cell types. Examples from GL26 cells were here purified using density gradient ultracentrifugation and monitored for effects on GL26 tumor growth in C57BL/6j mice (H-2b). Lactate dehydrogenase release assays were used to detect the cytotoxic activity of CD8+T and NK cells. Percentages of immune cells producing intracellular cytokines were analyzed by FACS. Results: In this study, exosomes from murine-derived GL26 cells significantly promoted in vivo tumor growth in GL26-bearing B6 mice. Then we further analyzed the effects of the GL26 cells-derived exosomes on immune cells including CD8+T, CD4+T and NK cells. Inhibition of CD8+T cell cytotoxic activity was demonstrated by CD8+T cell depletion assays in vivo and LDH release assays in vitro. The treatment of mice with exosomes also led to a reduction in the percentages of CD8+T cells in splenocytes as determined by FACS analysis. Key features of CD8+T cell activity were inhibited, including release of IFN-gamma and granzyme B. There were no effects of exosomes on CD4+T cells and NK cells. Conclusion: Based on our data, for the first time we demonstrated that exosomes from murine derived GL26 cells promote the tumor growth by inhibition of CD8+T cells in vivo and thus may be a potential therapeutic target.

• 한국지반환경공학회 논문집
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• 제3권1호
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• pp.27-36
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• 2002

본 연구는 동제련슬래그를 지반개량재로써 활용하기 위하여 배수에 대한 적용가능성을 평가한 것이다. 모래말뚝의 모래 대체재로써의 동제련슬래그 활용방안을 검토하였고, 동제련슬래그의 재료특성, 동제련슬래그말뚝이 설치된 복합지반에서의 압밀배수 특성, 배수 필터재로써의 특성 등을 파악하였다. 연구결과, 동제련슬래그는 일반적으로 쓰여지는 모래와 유사한 연직배수효과를 나타내었으므로, 모래말뚝의 모래 대체재로써의 적용은 충분히 가능할 것으로 판단된다. 더욱, 분사현상에 대한 안정성 및 입자 유실에 대한 내적안정성을 검토한 결과, 일반적인 사질토에 비해 한계동수경사가 크게 나타나고, 내적안정성이 우수하게 평가되었다. 따라서 동제련슬래그는 지반조건에 따라 배수 필터기능을 충분히 발휘할 수 있으며, 내적으로도 안정된 배수 필터재로써 사용이 가능한 것으로 판단된다.