• Journal of Ginseng Research
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• v.45 no.2
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• pp.273-286
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• 2021

Background: Prostate carcinoma is the second most common cancer among men worldwide. Developing new therapeutic approaches and diagnostic biomarkers for prostate cancer (PC) is a significant need. The Chinese herbal medicine Panax quinquefolius saponins (PQS) have been reported to show anti-tumor effects. We hypothesized that PQS exhibits anti-cancer activity in human PC cells and we aimed to search for novel biomarkers allowing early diagnosis of PC. Methods: We used the human PC cell line DU145 and the prostate epithelial cell line PNT2 to perform cell viability assays, flow cytometric analysis of the cell cycle, and FACS-based apoptosis assays. Microarray-based gene expression analysis was used to display specific gene expression patterns and to search for novel biomarkers. Western blot and quantitative real-time PCR were performed to demonstrate the expression levels of multiple cancer-related genes. Results: Our data showed that PQS inhibited the viability of DU145 cells and induced cell cycle arrest at the G1 phase. A significant decrease in DU145 cell invasion and migration were observed after 24 h treatment by PQS. PQS up-regulated the expression levels of p21, p53, TMEM79, ACOXL, ETV5, and SPINT1 while it down-regulated the expression levels of bcl2, STAT3, FANCD2, DRD2, and TMPRSS2. Conclusion: PQS promoted cells apoptosis and inhibited the proliferation of DU145 cells, which suggests that PQS may be effective for treating PC. TMEM79 and ACOXL were expressed significantly higher in PNT2 than in DU145 cells and could be novel biomarker candidates for PC diagnosis.

• Journal of Korean Society of Environmental Engineers
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• v.28 no.3
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• pp.329-336
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• 2006

Iron manufacturing process involves production of various by-product including slag, sludge, sintering and EAF(Electric Arc furnace dust). Some of the by-products such as EAF and sintering dust are disposed of as waste due to their high heavy metal contents. It has been notice for many years that the EAF dust also contain about 65% of Fe(0) and Fe(II) and then the possible utilization of the iron. One possibility is to apply the EAF as a lining material in conjunction with clay or HDPE liners, in waste landfill. The probable reaction between the leachate containing toxic elements such as TCE, PCE dioxine and $Cr^{6+}$ is reduction of the toxic materials in corresponding to the oxidation of the reduced iron and therefore diminishing the toxicity of the leachate. It is, however, prerequisite to evaluate the leaching characteristics of the EAF dust before application. Amelioration of the leachate would be archived only when the level of toxic elements in the treated leachate is less than that of in the untreated leachate. Several leaching techniques were selected to cover different conditions and variable environments including time, pH and contact method. The testing methods include availability test, pH-stat test and continuous column test. Cr and Zn are potentially leachable elements among the trace metals. The pH of the EAF dust is highly alkaline, recording around 12 and Zn is unlikely to be leached under the condition. On the contrary Cr is more leachable under alkaline environment. However, the released Cr should be reduced to $Cr^{3+}$ and then removed as $Cr(OH)_3$. Removal of the Cr is observed in the column test and further study on the specific reaction of Cr and EAF dust is underway.

• KSBB Journal
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• v.14 no.3
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• pp.284-290
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• 1999

Glucose fed-batch culture was studied to improve cellulose productivity by Acetobacter xylinum BRC5. When initial glucose concentrations in batch cultures were less than 20 g/L, yield coefficients of cellulose (Yp/s) remained a constant value of 0.21 g cellulose/g glucose. But a low yield coefficient, Yp/s=0.13 was obtained from an initial glucose concentration of 40 g/L. Since initial high glucose concentrations in batch culture resulted in low yields of cellulose, constant fed-batch cultures were carried out. The optimal feed rate for fed-batch culture was 2.22 g glucose/L.h. In constant fed-batch culture without DO control, 10 g/L of cellulose was obtained from 40 g/L of glucose with this feed rate, which was approximately two fold higher than that of the batch culture with the same initial glucose concentration. In DO stat plus fed-batch culture, the highest cellulose productivity could be obtained when dissolved oxygen level was controlled at 10% of air saturation, and cellulose productivity increased about 1.5 times compared with that of the culture without DO control.

• The Journal of Korean Medicine
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• v.31 no.2
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• pp.91-108
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• 2010

Background & Objective: The aim of this study was to investigate the effect of P. radix on the inflammatory related gene expression in IL-$1{\beta}$-stimulated primary human gingival fibroblast using Whole Transcript Sense Target (WT-ST). Method: Human gingival fibroblast was incubated with P. radix [100 or $200\;{\mu}g/ml$], and IL-$1{\beta}$ [$1ng/m{\ell}$] added an hour later. After 24h, total RNA was extracted using RNeasy Mini Kit and the whole gene expression patterns were performed using WT-ST Labeling $Assay^{(R)}$. Result: In the DEG results, 782 genes were up-regulated in the IL-$1{\beta}$-treated group as compared to control and among those, 43 genes were associated with inflammation. 981 genes were down-regulated after treatment with IL-$1{\beta}$ and of those 7 genes were associated with inflammation. 1439 genes were up-regulated after treatment with P. radix plus IL-$1{\beta}$-treated when compared to IL-$1{\beta}$-treated alone group and 1225 genes were down-regulated in the same condition. Among the down-regulated genes, 5 were associated with inflammation- and inhibitor genes such as GDF15 and LIF. In the analysis of the P. radix plus IL-$1{\beta}$-treated group, the most significant pathways were the cytokine-cytokine receptor interaction, toll-like receptor signaling, JAK-STAT signaling and tyrosine metabolism. The gene expression patterns in the P. radix $200{\mu}g/m{\ell}$ plus IL-$1{\beta}$-treated group appear to be more involved in the metabolism-related pathways than in the $100{\mu}g/m{\ell}$ plus IL-$1{\beta}$-treated group. Conclusion & Discussion: By microarray analysis of gene expression data, we are able to identify gene expression patterns associated with not only anti-inflammation effect but also transcription function of P. radix.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.273-278
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• 2002

Two-step fed-batch fermentations were carried out to overproduce Bacillus licheniformis maltogenic amylase (BLMA) in recombinant Escherichia coli. The first step was to increase the cell mass by controlling the feeding of a glucose solution, while the second step was designed to improve the amylase expression efficiency by supplementing organic nitrogen sources. The linear gradient feeding method was successfully adopted to maintain the glucose concentration below 0.2 g/l during the fed-batch mode, as effectively minimizing acetic acid formation. When the dissolved oxygen (DO) level became limiting, an accumulation of acetic acid and drastic decrease in specific BLMA productivity were observed. Glucose and organic nitrogen sources consisting of yeast extract and casein hydrolysate were simultaneously supplied in the pH-stat mode to further increase the specific BLMA expression efficiency. An organic nitrogen source consisting of 200 g/1 yeast extract and 100 g/1 casein hydrolysate was found to be the best among the various combinations tested. The feeding of an organic nitrogen source in the second-step fed-batch period was highly beneficial in enhancing the BLMA production. The optimized two-step fed-batch culture resulted in 78 g/l maximum dry cell mass and 443 U/ml maximum BLMA activity, corresponding to 1.5-fold increase in the dry cell mass and 3.7-fold enhancement in BLMA production, compared with the simple fed-batch fermentation.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.82-82
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• 2002

Catalase(CAT)는 주요한 활성산소(ROS)의 일종인 과산화수소($H_2O$$_2$)가 Hydroxyl 유리기 생성에 관여하지 못하도록 $H_2O$$_2$를 $H_2O$ 및 $O_2$로 대사시켜 주는 효소계 항산화제의 한 종류이다. 따라서 본 실험에서는 CAT가 5% $O_2$및 20% $O_2$조건하에서 1-세포기 돼지체외수정란의 배발달에 미치는 영향을 조사하였다. 돼지난포란을 10% PFF, 0.1mg/ml cysteine, 10IU/m1 PMSG, 10IU/m1 hCG 및 10ng/m1 EGF가 첨가된 NCSU23 배양액에서 22시간 동안 배양을 실시하고, 성선자극호르몬이 배제된 배양액에서 추가로 22시간을 배양하여 체외성숙을 유도하였다. 체외성숙이 유기된 난자는 난구세포를 제거하고, 2.5mM caffeine과 0.1% BSA가 첨가된 mTBM 배양액에 정자를 1.25 $\times$ $10^{5}$cells/ml의 농도로 5-6시 동안 공동배양을 실시하여 체외수정을 유도하였다. 체외수정후 1-세포기의 수정란을 0.4mg/ml BSA가 첨가된 NCSU23 배양액에 CAT를 각각 0, 100, 500 및1,000uni1 첨가하여 30 embryos/ 50u1 소적으로하여 38.8$^{\circ}C$, 5% $CO_2$및 5%$CO_2$, 5% $O_2$90% $N_2$조건하의 배양기에서 각각 7일간 배양을 실시하였다. 조사된 결과는 SAS/STAT 6.03 Package를 이용하여 통계분석을 실시하였다. 체외배양 48시간에 난할률을 조사하였을 때, 대조구와 처리구간 차이가 인정되지 않았으나, 배양 7일째 배반포형성률에 있어서는 각각 22.7$\pm$2.7, 22.1$\pm$3.9, 18.7$\pm$4.9, 및 15.1$\pm$2.5%로서 처리구가 대조구보다 유의적으로 낮은 결과를 나타냈다. 그리고 산소농도에 따른 배반포형성률은 5% $O_2$ 조건(22.1$\pm$2.4%)이 20% $O_2$조건(17.2$\pm$2%)보다 유의적(P<0.05)으로 높은 것으로 나타났다. 총세포수에 있어서는 각각 44.4$\pm$4.0, 43.3$\pm$36, 25.4$\pm$2.4 및 13.4$\pm$1.5로서 처리구가 대조구보다 세포수가 적은 것으로 나타났으며, 산소농도에 따른 차이는 인정되지 않았다. 따라서, 체외생산된 돼지초기수정란을 배양할 때, CAT의 첨가는 효과가 없는 것으로 나타났다.다.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.81-81
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• 2002

$\beta$-Mercaptoethanol($\beta$-ME)은 일반적으로 황화합물(thiol compounds)의 일종으로, 배양액 중에서 이황화결합(disulfide bonds)을 분해하여 일정한 물질의 산화.환원반응에 관여하며, 특히 cysteine이 cystine으로 산화되는 것을 차단함으로서 cysteine의 이용능력을 증대시키고, GSH의 합성을 촉진 및 증대시키는 것으로 알려져 있고, 각종 활성산소로부터 세포를 보호하는 역할을 수행하는 것으로 보고되었다. 특히, 돼지수정란의 체외배양체계에 유의적인 영향을 미치는 것으로 보고되었다(Abebydeera 등, Theriogenol., 50:747-756, 1998). 따라서 본 실험에서는 돼지난포란의 체외성숙시 $\beta$-ME의 첨가배양이 체외수정과 배발달에 미치는 영향에 관하여 조사하였다. 돼지난포란을 10% PFF, 0.1mg/ml cysteine, 10IU/m1 PMSG, 10IU/m1 hCG 및 10ng/m1 EGF가 첨가된 NCSU23 배양액에 $\beta$-ME를 각각 0, 25, 50 및 100uM을 처리하여 22시간 동안 배양을 실시하고, 성선자극호르몬이 배제된 배양액에서 추가로 22시간을 배양하여 체외성숙을 유도하였다. 체외성숙이 유기된 난자는 난구세포를 제거하고, 2.5mM caffeine과 0.1% BSA가 첨가된 mTBM배양액에 정자를 1.25 $\times$ $10^{5}$cells/ml의 농도로 5-6시간 동안 공동배양을 실시하여 체외수정을 유도하였다. 체외수정후 일부의 수정란은 12시간에 난자 급속 염색방법으로 염색하여 다정자침입률 및 자.웅전핵형성률 등을 확인하였다. 그리고 나머지1-세포기의 수정란은 0.4mg/ml BSA가 함유된 NCSU23 배양액에 30 embryos/50ul 소적으로하여 38.8$^{\circ}C$, 5% $CO_2$의 탄산가스 배양기에서 각각 7일간 배양을 실시하였다. 조사된 결과는 SAS/STAT를 이용하여 통계분석을 실시하였다. 체외수정 12시간 후에 난자 급속 염색법으로 염색을 실시한 결과, 모든 처리구에서 핵성숙률(76.4~95.2%), 정자침투율(51.1~66.9%), 웅성전핵형성률(95.2~100%), 다정자침입률(18.2~25.6%) 및 평균침입정자수(1.2~l.4개)에서 유의적인 차이가 인정되지 않았다. 체외배양 48시간 난할률을 조사한 결과, 처리구별 차이(53.9~67.9%)는 인정되지 않았으나, 배양 7일째 배반포형성률은 각각 14.5, 25.4, 17.3 및 12.4%로서 25uM의 $\beta$-ME처리구가 유의적(P<0.05)으로 높은 배발달률을 나타내었고, 총세포수에 있어서는 대조구와 처리구간 유의적인 차이가 인정되지 않았다. 따라서 돼지 난포란을 성숙배양할 때, 25uM $\beta$-ME를 첨가배양하는 것이 양질의 돼지체외수정란을 생산하는 하나의 방법으로 조사되었다.다.

• The Korean Journal of Physiology and Pharmacology
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• v.17 no.3
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• pp.203-208
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• 2013

As the abnormal proliferation of vascular smooth muscle cells (VSMCs) plays a critical role in the development of atherosclerosis and vascular restenosis, a candidate drug with antiproliferative properties is needed. We investigated the antiproliferative action and underlying mechanism of a newly synthesized naphthoquinone derivative, 5,8-dimethoxy-2-nonylamino-naphthalene-1,4-dione (2-nonylamino-DMNQ), using VSMCs treated with platelet-derived growth factor (PDGF). 2-Nonylamino-DMNQ inhibited proliferation and cell number of VSMCs induced by PDGF, but not epidermal growth factor (EGF), in a concentration-dependent manner without any cytotoxicity. This derivative suppressed PDGF-induced $[^3H]$-thymidine incorporation, cell cycle progression from $G_0/G_1$ to S phase, and the phosphorylation of phosphor-retinoblastoma protein (pRb) as well as the expression of cyclin E/D, cyclin-dependent kinase (CDK) 2/4, and proliferating cell nuclear antigen (PCNA). Importantly, 2-nonylamino-DMNQ inhibited the phosphorylation of PDGF receptor${\beta}$(PDGF-$R{\beta}$) enhanced by PDGF at $Tyr^{579}$, $Tyr^{716}$, $Tyr^{751}$, and $Tyr^{1021}$ residues. Subsequently, 2-nonylamino-DMNQ inhibited PDGF-induced phosphorylation of STAT3, ERK1/2, Akt, and $PLC{\gamma}1$. Therefore, our results indicate that 2-nonylamino-DMNQ inhibits PDGF-induced VSMC proliferation by blocking PDGF-$R{\beta}$ autophosphorylation, and subsequently PDGF-$R{\beta}$-mediated downstream signaling pathways.