• 생약학회지
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• 제35권4호통권139호
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• pp.330-337
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• 2004

Based on the traditional application of Carthamus tinctorious L. (CF) as a component of Korean medicinal decoctions, in the present study, we investigated in vitro an immunomodulatory activity of water extract of CF(WECF). Water extract of CF significantly increased the in vitro proliferative responses of spleen cells (SPC). However, addition of WECF during anti-CD3 activation resulted in a significant decrease in SPC proliferation. Flow cytometric analysis showed that WECF addition chanced T and B cell frequencies in anti-CD3-activated spleen cell populations. Using purified cells, it was revealed that WECF is mitogenic to B cells but rather inhibitory to T cell Proliferation. Upon anti-CD3 stimulation, high concentration (1 mg/ml) of WECF significantly inhibited T cell proliferation until day 2 of stimulation. At day 3, anti-CD3-activated cells exposed to WECF recovered their proliferation to the level comparable to control. Although B cell proliferation was also inhibited in proliferation at day 1, it recovered sooner and then was rather augmented by WECF at day 3. These data indicate that WECF down-regulates lymphocyte proliferation at early phase of activation but T cells are more vulnerable than B cells to WECF, However, CD4+ and CD8+ T cells did not differ in WECF-mediated immunotoxicity. Data of propidium iodide (PI) staining showed that WECF accelerates activated T cell, but not B cell, apoptosis and WECF concurrently inhibited cytokine production of activated T cells. Taken together, WECF exhibits B cell mitogenic activity and differential toxicity more pronounced to T cells, suggesting a possible in vivo application of WECF for specific control of T cells without alteration of B cell activity.

• International Journal of Industrial Entomology and Biomaterials
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• 제7권2호
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• pp.197-201
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• 2003

The recombinant plasmids harboring a heterologous gene coding mouse endostatin were transfected and expressed stably in Trichoplusia ni Tn 5B1-4 cells and Bombyx mori BmN cells, respectively. Recombinant endostatin expressed in the stably transformed Tn 5B1-4 and BmN cells was secreted into the medium. BmN cells are relatively lower in maximum cell growth and recombinant endostatin production than Tn 5B 1-4 cells. Recombinant endostatin was also purified to homogeneity using a simple one-step ${Ni^2+}$ affinity fractionation method. Purified recombinant endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition $({ED_50})$ for recombinant endostatin was approximately 0.35 ${\mu}g$/ml.

• 생명과학회지
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• 제25권12호
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• pp.1425-1431
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• 2015

면역 시스템의 림프구는 B 림프구와 T 림프구 두 종류로 나눌 수 있다. B 림프구는 플라즈마 세포로 분화하여 항체를 생성하는 체액성 면역을 담당하며, T 림프구는 다른 세포나 세균을 죽이는 세포성 면역을 담당한다. 고전적으로 B 림프구와 T 림프구의 작용은 한 방향으로 이뤄졌다. T 림프구는 B 림프구의 분화를 촉진하고 면역글로불린종류의 전환을 조절한다. T 림프구가 부족한 경우 B 림프구의 부족을 초래함이 보고되어 있다. 그러나 최근에 역으로 B 림프구가 T 림프구의 분화와 활성을 조절할 수 있다는 보고가 있다. 예를 들어, B 림프구는 CD8+ T 림프구의 tolerance를 직접 조절할 수 있고, TGF-β의 분비를 통해 T 림프구의 anergy를 유도할 수 있다. 본 연구는 LPS에 의해 자극된 B 림프구가 수지상세포에서 IL-12의 분비를 억제하여 Th1 림프구의 분화를 억제할 수 있음을 보여준다. 이 억제는 B 림프구와 수지상세포의 직접적인 interaction에 의해 일어나는 것이 아니며 B 림프구가 수지상세포의 성숙을 조절하여 일어나는 것도 아니다. B 림프구에서 분비되는 soluble factor가 LPS에 의해 증가되는 수지상세포의 IL-12p35 transcription을 억제한다. 이 결과들은 B 림프구가 매개하는 새로운 면역억제 기전이 존재함을 보여준다. 이것은 고전적인 방향성을 가진 T 림프구에 의한 B 림프구 작용조절로 면역반응이 결정되는 것이 아니라 T 림프구와 B 림프구가 서로 작용을 하여 면역평형을 결정하는 기전이 존재함을 보여준다.

• 한국미생물·생명공학회지
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• 제31권2호
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• pp.111-116
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• 2003

Xylan 분해 균주인 Bacillus stearothermophilus No. 236 분리균의 $\beta$-xylosidase 생산 유전자(xylA)의 염기 서열 및 transcription start site를 결정한 이전 연구 결과에 의하면 xylA 유전자는 매우 특이하게 UUG codon에서 translation이 시작되며 initiation codon 15dp 윗쪽에는 promoter로 추정되는 염기 서열을 가지고 있는 것으로 분석되었다. 이와 같은 xylA 유전자 promoter region의 구조는 E. coli에 클로닝된 xalA 유전자를 이용한 실험 결과로도 확인되었다. xalA promoter의 -10 element는 CATAAT로서 6개의 염기 중 5개가 그리고 -35 element의 경우는 TTGTTA로서 6개의 염기 중 4개가 consensus sequence와 일치되었으나 두 hexamer 사이의 거리가 최적 거리에서 크게 벗어난 12 bp인 것으로 분석되었다. 본 연구에서는 $\beta$-xylosidase의 대량 생산을 위한 연구의 일환으로 xalA promoter sequence의 체계적 구조 변화에 의한 promoter strength에 미치는 효과를 E. coli와 B. subtilis두 숙주 세포에서 조사 분석해 본 결과, 첫째로 두 promoter elements사이의 거리를 최적거리인 17 bp로 바꾸었을 때 xalA의 발현율은 E. coli에서는 1.6배, B. subtilis에서는 2.5배 정도 증가함을 보여주었다. 그리고 -35 element는 consensus sequence와 같이 5'쪽에서 네번째 위치에 있는 T$\longrightarrow$A로 변이 시켰을 때 E. coli경우 2.3배, 특히 B. subtilis에서는 35배나 되는 가장 높은 promoter 활성의 증가를 보였다. 그러나 -10 sequence의 경우 consensus sequence와 같이 5' 쪽에서 첫번째 위치에 있는 C$\longrightarrow$T로 transition시켰을 때 예상외로 오히려 발현율이 5~15배까지 낮아지는 특이한 결과를 얻었다. 따라서 본 연구 결과 xalA promoter의 경우 -10 sequence인 CATAAT의 C와 -35 element의 두 염기가 promoter활성에 있어 가장 중요한 염기임을 알 수 있었다.

• Journal of Ginseng Research
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• 제35권1호
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• pp.86-91
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• 2011

Ginsenoside (G)-F1 is an enzymatic metabolite generated from G-Rg1. Although this metabolite has been reported to suppress platelet aggregation and to reduce gap junction-mediated intercellular communication, the modulatory activity of G-F1 on the functional role of skin-derived cells has not yet been elucidated. In this study, we evaluated the regulatory role of G-F1 on the cellular responses of B16 melanoma cells. G-F1 strongly suppressed the proliferation of B16 cells up to 60% at 200 ${\mu}g/mL$, while only diminishing the viability of HEK293 cells up to 30%. Furthermore, G-F1 remarkably induced morphological change and clustering of B16 melanoma cells. The melanin production of B16 cells was also significantly blocked by G-F1 up to 70%. Interestingly, intracellular signaling events involved in cell proliferation, migration, and morphological change were up-regulated at 1 h incubation but down-regulated at 12 h. Therefore, our results suggest that G-F1 can be applied as a novel anti-skin cancer drug with anti-proliferative and anti-migration features.

• 한국식품영양학회지
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• 제35권2호
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• pp.127-136
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• 2022

Comparing the quality characteristics of kimchi were measured and anticancer effects using AGS human gastric cancer cells were observed. Five kinds of kimchi samples were made of Kanghwa Baek kimchi (KB), Kangwha Turnip kimchi (KT), Turnip: Chinese cabbage = 1:1 Baek kimchi (T1B1), Turnip:Chinese cabbage = 4:1 Baek kimchi (T4B1), Turnip mul kimchi (T). As a result T kimchi showed the best fermentation characteristics among the five samples. T kimchi had a lower percentage of the total number of aerobic bacteria, while the number of lactobacillus was higher than that of other samples. The mRNA and protein expression levels of apoptosis-related factors found that T kimchi significantly increases the mRNA expression levels of caspases-3 and caspases-9 in AGS human gastric cancer cells as compared to the other kimchi samples. It showed high anticancer effects in the order of T, T1B1, and KB kimchi. As the anticancer effect of Turnip mul kimchi made only of turnip was higher, the higher the turnip content, the higher the anticancer effect. These results show that there were changes in fermentation characteristics such as pH, acidity, number of lactic acid bacteria, and anticancer effects according to the ratio of turnip and cabbage.

• 한국동물생명공학회지
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• 제35권4호
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• pp.347-356
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• 2020

Gangliosides are glycolipids in which oligosaccharide is combined with sialic acids. Our previous studies have suggested an interplay between ganglioside GD1a/GT1b and meiotic maturation capacity in porcine oocyte maturation. Furthermore, ganglioside GD1a and GT1b are known for its antioxidant activity, but it is still unclear whether possible antioxidant role of GD1a and GT1b is involved in porcine embryos development competence during in vitro culture (IVC). Here, the effects of ganglioside GD1a and GT1b on the embryonic developmental competence during in vitro culture of porcine were investigated. The effects of ganglioside GD1a and GT1b on the expression of ST3GAL2 were confirmed during embryos development (2-cell, 4-cell, 8-cell and blastocyst) using immunofluorescent staining (IF). As a result, the fluorescent expression of ST3GAl2 was higher in embryos at 4-8 cells stage than blastocysts. Blastocyst development rate significantly increased in only 0.1 μM GD1a and GT1b treated groups compared with control group. To investigate the cellular apoptosis, we analyzed TUNEL assay. In case of only 0.1 μM GD1a and GT1b treated groups, the total number of cells in blastocyst compared with control group, but there was no significant difference in the rate of apoptotic cells. We identified the intracellular ROS levels using DCF-DA staining. According to the result, ROS production significantly decreased in blastocysts derived from the 0.1 μM GD1a and GT1b treated groups. These results suggest that ganglioside GD1a and GT1b improve the developmental competence of porcine embryos via reduction of intracellular ROS during preimplantation stage.

• 동의생리병리학회지
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• 제35권1호
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• pp.15-21
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• 2021

Donggwaja (Benincasae Semen), the seed of Benincasa hispida (Thunb.) Cogn., has been used in Korean traditional medicine to control the body heat and water retention caused by various diseases. Both the symptoms targeted by the herbal medicine in clinic and studies with disease mouse models support the potential anti-inflammatory effect of Donggwaja. However, it is less understood how Donggwaja exerts its possible anti-inflammatory effect. Here, we present evidence that Donggwaja suppresses macrophage inflammatory reactions via expressing tumor necrosis factor a-induced protein 3 (TNFAIP3 or A20) and suppressing NF-kB activity. The ethanol extract of Donggwaja (EED) showed no toxicity when added to RAW 264.7 cells less than 100mg/ml. When treating the cells for 16 h, EED significantly suppressed the nuclear localization of NF-kB, suggesting that EED suppresses NF-kB activity. Concordantly, a semi-quantitative RT-PCR analysis showed that EED decreased the expression of prototypic pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-a, IL(interleukin)-6, and IL-1b. EED induced in RAW 264.7 cells the expression of A20, a ubiquitin modulator that suppresses inflammatory signaling cascades initiated from TLR4 and TNF and IL-1 receptors, while not affecting the induction of Nrf2, an anti-inflammatory factor that could suppress the effect of NF-kB. These results suggest that EED exerts its suppressive effect on inflammation, at least in part, by expressing anti-inflammatory factor A20 and suppressing pro-inflammatory factor NF-kB activity.

• Food Science and Biotechnology
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• 제16권1호
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• pp.163-166
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• 2007

The cytotoxic effects of partially purified substances from Bacillus polylfermenticus SCD toward a variety tumor cell lines were studied. Cytotoxic activity was determined with regard to the A549 (human lung carcinoma), AGS (human stomach adenocarcinoma), DLD-1 (human colon adenocarcinoma), HEC-1-B (human uterus adenocarcinoma), SW-156 (human kidney carcinoma), and NIH/3T3 (murine normal fibroblast) cell lines using the MTT assay. Cytotoxic substances were partially purified through Diaion HP-20 columns and extracted with methanol or other organic solvents (n-hexane, chloroform, ethylacetate, and butanol). B. polyfermenticus SCD supernatant showed up to 60% inhibition of cell viability fer all five human cancer cell lines tested. When treated with 10 mg/mL of n-hexane, chloroform, ethylacetate, and butanol extract, HEC-1-B cells showed a 25,62,35, and 63% rate of inhibition respectively, and AGS cells showed a 72, 61, 44, and 67% rate of inhibition, respectively. At a concentration of 10 mg/mL, 100% methanol Diaion HP-20 extracts showed inhibition rates of 97.0% toward A-549 cells, 98.1% toward AGS cells, 81.6% toward DLD-1 cells, 83.5% toward HEC-1-B cells, and 92.7% toward SW-156 cells. These results indicate that partially purified fractions from B. polyfermenticus SCD have the potential to inhibit not only colon cancer cells, but also lung, stomach uterus, and kidney cancer cells. Further studies are needed to characterize the cytotoxic substances released in B. polyfermenticus SCD cultures.

• 미생물학회지
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• 제37권3호
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• pp.209-213
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• 2001

Bacillus spp. 이용하여 B3공법을 사용하는 하수 종말 처리장의 포기조에서 Bacillus sup. 변화를 FSIH방법과 포자염색 방법으로 관찰하였다. DAPI로 측정한 세균수는 $3.2{\times}10^8 ~ 4.4{\times}10^8 $ cells/ml 범위로 큰 변화가 없었으나, Bacillus sp. 균수는 $0.3{\times}10^8 ~ 2.5{\times}10^8 $ cells/ml 범위로 지점별, 조사시기별로 큰 차이가 있었다. 포자수는 $0.3{\times}10^7 ~ 8.4{\times}10^7 $ cells/ml 범위로 3월에는 세 번째 포기조, 5월에는 두 번째 포기조에서 각각 8.5, $5.5{\times}10^7$cells/ml으로 높은 값을 보였다. 총세균수는 DAPH를 측정한 값과 포자수를 합하여 사용하였고, Bacillus속 세균수는 FISH로 계수된 수와 포자수를 합하여 사용하였다. 충세균수에 대한 Bacillus sp.균의 비율은 약 8~60% 범위로 첫 번째 및 세 번째 포기조에서 각각 50%, 35% 이상의 높은 비율을 보였다. B3 공법에서는 Bacillus가 우점하고 있음이 FISH 방법으로 확인되었으며, 반송 슬러지와 하수가 유입되는 첫 번째포기 조에서 그 비율이 높았고, 다음 포기조로 가면서 그 비율이 줄어들고 있었다. Bacillus의 포자수는 뒤의 포기조로 가면서 그 수와 비율이 높아지고 있었다.