• Asian Pacific Journal of Cancer Prevention
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• 제16권10호
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• pp.4329-4333
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• 2015

Background: To investigate the change of frequency and immuno-inhibitory function of myeloid-derived suppressor cells (MDSCs) after treatment of cisplatin (DDP) in A375 human melanoma model. Materials and Methods: BALB/c nude mice were inoculated with A375 cells to establish the human melanoma model and randomly divided into control group given normal saline (NS) and experimental group treated with DDP (5 mg/kg). The percentages of MDSCs in the tumor tissue and peripheral blood after DDP treatment were detected by flow cytometry. The proliferation and interferon-${\gamma}$ (IFN-${\gamma}$) secretion of T cells co-cultured with MDSCs were analyzed through carboxyfluorescein succinimidyl ester (CFSE) labeling assay and enzyme-linked immunospot (ELISPOT) assay, respectively. Results: In A375 human melanoma model, DDP treatment could significantly decrease the percentage of MDSCs in the tumor tissue, but exerted no effect on the level of MDSCs in peripheral blood. Moreover, DDP treatment could attenuate the immuno-inhibitory function of MDSCs. T cells co-cultured with DDP-treated MDSCs could dramatically elevate the proliferation and production of INF-${\gamma}$. Conclusions: DDP can decrease the frequency and attenuate immuno-inhibitory function of MDSCs in A375 melanoma model, suggesting a potential strategy to augment the efficacy of combined immunotherapy.

• 한국가축번식학회지
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• 제16권2호
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• pp.87-102
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• 1992

본 연구는 우 casein을 신속, 정확하게 분석할 수 있는 특수 면역효소분석법을 개발하였다. Biotin이 연결된 casein과 peroxidase-conjugated avidin을 사용하였으며 면역화시킨 닭의 난황으로부터 추출한 항체를 이용하여 분석하였다. Sulfo-N-hydroxy succinimido biotin을 사용하여 casein에 biotin을 연결시키고 microplate에 고정한 뒤 peroxidase-conjugated avidin을 결합시켰다. 생산된 항체는 $\alpha$-와 $\beta$-casein에 특이적이었으며, 유청단백질, IgG, 우혈청 알부민과 교차반응은 면역효소분석법과 Western blot에서 나타나지 않았다. 본 분석법의 민감도는 2ng에서 20$\mu\textrm{g}$이었으며 Standard와 시료의 분석시 뚜렷한 평행곡선이 형성되었다. Intra-assay와 Inter-assay의 변이계수는 각각 5.5와 5.7%이었다. 그리고 비유 초기의 casein량을 조사한 결과 분만전 3일경부터 급격히 상승하는 것을 알 수 있었다.

• BMB Reports
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• 제30권5호
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• pp.308-314
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• 1997

We investigated biotinylation of nitrocellulose membrane for immuno-filtration capture assay. In order to enhance the efficiency of biotinylation, nitrocellulose membranes were pretreated with several chemicals for the purpose of suitable protein absorption through surface modification. As a signal generating enzyme, urease was used and the concentration of avidin was optimized for the efficient binding kinetics between urease-biotin in liquid phase and biotinylated membrane in solid phase. For effective biotinylation, bovine serum albumin-biotin complexes could be immobilized at a concentration of $370\;{\mu}g$/stick ($4.4\;cm^2$). Among tested chemicals, polylysine (0.25%) showed a significant effect in biotinylation. Polylysine is thought to enhance surface area by extending unbound residues into solution. Time of treatment over 30 min and higher molecular weight of polylysines (58,100 dalton) showed positive effect on the enhancement of biotinylation. The result from this study may be useful for developing a new biosensor and other biofunctional membranes for examining molecular recognition.

• KSBB Journal
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• 제24권3호
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• pp.253-258
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• 2009

본 연구에서는 ELISA와 면역-크로마토그래피 스트립기술을 결합하여 E. coli O157 : H7을 탐지할 수 있는 면역스트립 센서를 제작하기 위한 제작조건의 최적화 연구를 수행하였다. 포획항체 농도, 탐지항체 농도, 완충용액 첨가제 조성의 면역스트립 제작 또는 운전인자들의 최적화 조건을 결정하였다. 포획항체의 농도는 1 mg/mL를 최적 조건으로 선정하였고, 탐지항체의 최적 농도도 1 mg/mL로 결정하였다. 비특이적 결합을 방지하기 위한 시료 희석용 완충용액의 첨가제 조성으로는 0.5% Tween 20와 3% BSA 혼합 사용을 선정하였다.

• 약학회지
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• 제35권3호
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• pp.154-164
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• 1991

These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.102-102
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• 2018

Protaetia Brevitarsis Seulensis(white grub) has been traditionally used as medicinal stuff to treat blood stasis, occlusion of menstruation, tetanus and liver cancer in Asian countries (Korea, Japan, China, Taiwan, India and Myanmar). Especially, Donguibogam, which is traditional korean medicinal book, described the white grub as traditional medicine to treat hepatic diseases and vascular disorders. The white grub has been considered as highly nutritional food. The major constituents of white grub are rich in protein, healthy fats, iron, calcium. Recent studies announced that white grub has hepatoprotective effect and anti-microbacterial effect. However, the immuno-enhancing effect of white grub extracts in RAW 264.7 macrophage cells has not been studied yet. In this study, the various concentrations of white grub extract were examined to find immuno-enhancing effects on RAW 264.7 cells. Cytotoxicity was determined by MTT assay and immuno-enhancing effect of white grub extract was investigated by measuring nitric oxide (NO) production compared with only lipopolysaccharide (LPS) treatment. White grub extracts (0.001 - 10 mg/ml) did not show cytotoxicity. Additionally, white grub extracts (0.001 - 1mg/ml) had Immuno-enhancing effect on RAW 264.7 cells compared with only LPS treated group. These results might be provided proof to develop beneficial immuno-enhancing material for human health.

• 동의생리병리학회지
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• 제17권6호
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• pp.1468-1474
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• 2003

Taklidanggui-tang was a drug that treated carbuncle and cellulitis. So, the purpose of this Study was to investigate effect of Taklidanggui-tang on the anti-cancer and proliferation of immunocytes, nitric oxide(NO) production of peritoneal macrophages. We used Taklidanggui-tang extract(TDT) with freeze-dried, 8wks-old male mice, and L1210 cell lines for this Study. The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). The results of this Study were obtained as follow ; TDT was showed cytotoxicity on the L1210 cell lines, increased significantly proliferation of thymocytes and splenocytes in vitro. TDT inhibited significantly proliferation of L1210 cells, increased significantly proliferation of immunocytes in L 1210 cells transplanted mice. And TDT was extended significantly mean survival days in 5-180 cells transplanted mice, but TDT did not increased NO production from peritoneal microphages in L1210 cells transplanted mice. This results suggest that TDT has anti-cancer and immuno-action.

• BMB Reports
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• 제31권1호
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• pp.101-105
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• 1998

Immunoassay of botulinum toxin (BTX) B type was investigated using two typed of biosensors: light addressable potentiometric sensor (LAPS) and surface plasmon resonance (SPR) sensor. Urease-tagged and immuno-filtration capture method have been used for LAPS. Tag-free and direct binding real-time detection method have been used for SPR sensor. The detection limit of sandwich assay format with LAPS was 10 ng/ml, which was the lowest among methods tested. SPR has the advantage of being more convenient because tag-free direct binding assay can be used and reaction time was reduced, regardless of low sensitivity. This result shows that sandwich assay format with LAPS can be used as an alternative method of BTX mouse bioassay which is known as the most sensitive method for the detection of BTX.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1046-1052
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• 2007

Transferrin은 저장된 철분자를 운반하고 살아있는 유기체에서 철의 항상성을 유지한다. 호랑나비의 종령 유충 혈림프내 철 운반 단백질인 transferrin을 KBr 밀도구배 초원심분리와 Superose 6 HR을 이용한 fast protein liquid chromatography법으로 분리 정제하였으며, transferrin의 조직에 따른 분포는 면역화학적 방법에 따라 확인하였다. 정제된 transferrin의 아미노산 조성은 아스파르트산(Asp), 발린(Val), 루이신(Leu), 글루타민(GIu)이 많이 존재하였으며, subunit의 분자량은 78, 80 kDa으로 확인되었다. Immuno-diffusion을 통해 각 조직에서 분포를 확인한 결과 혈림프와 지방체에서 transferrin이 전시기에 걸쳐 뚜렷한 동질성을 나타냈다. 또한 rocket immuno-electrophresis법에 의해 transferrin의 양적 분포를 살펴보면 혈림프에서 용화 3일과 용화 5일에 증가하였으며, 지방체에서는 전용기와 용화직후에 양적인 증가를 나타냈다.

• 한국식생활문화학회지
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• 제32권5호
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• pp.453-464
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• 2017

Vegetable soup has been reported to have anti-inflammatory, anti-oxidative and anti-cancer effects. In this study, five kinds of vegetable soup were developed using a new manufacturing process and compositional changes in raw material, and anti-cancer and immuno-stimulatory activities were evaluated. Cytotoxicity tests based on MTT assay revealed that all vegetable soups had strong inhibitory effects against CT26 mouse colon cancer cells, with soups including Solomon's seal being most effective based on comparison of $IC_{50}$ values. Apoptosis in response to vegetable soup was occurred by 3-5 fold on cancer cells compared to normal cells. Mouse splenocytes increased by 266-541% in response to addition of vegetable soup in an in vitro proliferation experiment. In co-culture with splenocytes and CT26 cancer cells, splenocytes increased by more than 280% in every vegetable soup treatment, while cancer cells decreased by about 60% and cytokines such as $IFN- {\gamma}$ and IL-12 were secreted from splenocytes in high levels only in response to vegetable soup including Solomon's seal. In conclusion, all vegetable soups developed in this study had anti-cancer effects, and vegetable soup including Solomon's seal showed the strongest anti-cancer and immuno-stimulatory effects. These results suggest that functionality of vegetable soup could be increased by changes in manufacturing processes and raw materials composition.