• Korean Journal of Microbiology
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• v.38 no.3
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• pp.153-161
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• 2002

Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.42 no.2
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• pp.111-118
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• 2016

Acne vulgaris is a chronic inflammatory condition characterized by comedo, papule, cyst, nodule and postinflammatory hyperpigmentation. Meanwhile, it is also induced by adverse event of drugs. Among them, acneiform folliculitis is a side effect of epidermal growth factor receptor (EGFR) inhibitor, which is an anticancer agent, and its incidence may occur in upward of 75 ~ 100% of cases. The main method of acne vulgaris treatment is oral antibiotics, retinoids, topical medication and so on. However, it is limitation that teratogenicity caused by retinoids and antibiotic resistance increased by using antibiotics. In this study, we aimed to evaluate the clinical efficacy and safety of topical recombinant human (rh) EGF in treating facial acne vulgaris. Twenty three Koreans (age: 10 ~ 29 years) with mild to moderate acne vulgar participated in the study and applied topical rhEGF cream (trouble control EGF) with 3 products (trouble control clarifying cleansing foam, trouble control all-clear filling toner, redroll calming moisture) on their face twice daily for four weeks. Several assessment methods were applied: Acne lesion counts score by investigator's global assessment, efficacy and satisfaction score by subjects. Skin sebum output level, hydration level and redness level were also measured at each visit. At the final visit, skin sebum level, transepidermal water loss, skin redness statistically decreased and acne lesions (comedone, papule) were statistically reduced. No severe side effects were observed during the study. In conclusion, topical rhEGF seems to be an effective and safe adjuvant treatment option for mild acne vulgaris.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.4
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• pp.524-531
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• 2015

Antioxidant activities and in vitro anticancer effects of bamboo salt doenjang on HT-29 human colon cancer cells were studied. 3Y3B-D (three-year fermentation using three-time baked bamboo salt doenjang), 3Y9B-D (three-year fermentation using nine-time baked bamboo salt doenjang), 6Y3B-D (six-year fermentation using three-time baked bamboo salt doenjang), and 6Y9B-D (six-year fermentation using nine-time baked bamboo salt doenjang) were compared to C-D (commercial doenjang) and 3B-S (cooked soy beans prepared using three-time baked bamboo salt). There were no differences between experimental groups in pH, amino-type nitrogen, or ammonia-type nitrogen levels. 6Y9B-D showed the highest antioxidative effect, followed by 6Y3B-D, 3Y9B-D, and 3Y3B-D, in order. 6Y9B-D showed the highest total polyphenol concentration. 6Y9B-D showed the highest anticancer effect, as determined by MTT assay, as well as levels of the pro-inflammatory cytokines including TNF-${\alpha}$, IL-6, iNOS, and COX-2, followed by 6Y3B-D, 3Y9B-D, and 3Y3B-D, in order. From the results above, 6Y9B-D showed the highest antioxidative and anticancer effects, followed by 6Y3B-D, 3Y9B-D, 3Y3B-D, C-D, and 3B-S.

• Clinical and Experimental Pediatrics
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• v.51 no.1
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• pp.73-77
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• 2008

Purpose : p16 gene, mapped to the 9p21 chromosomal region, has emerged as a candidate tumor suppressor gene in human neoplasm. It is an inhibitor of cyclin-dependent kinase and inhibits Rb phosphorylation. In a variety of tumors including childhood acute lymphoblastic leukemia (ALL), deletion and/or mutation of the p16 gene has been found. Despite their high frequency, the prognostic importance of p16 alterations is still controversial in ALL and has been reported to be either unfavorable or similar to that of other patients. We studied the correlation between loss of p16 protein confirmed by immunohistochemical staining and clinical outcomes of patients diagnosed as ALL. Methods : We performed an immunohistochemical staining for p16 protein in 74 cases of bone marrow biopsy slide initially diagnosed as ALL between January 1998 and December 2006. We reviewed the clinical manifestations, laboratory findings, treatment outcomes retrospectively. Results : Of 74 slides, 12 were negative for p16 protein. Seven were males and 5 were females with a median age at diagnosis was 5.8 (1.3-18.8) years. Initial WBC were 17,225 $(500-403,300)/{\mu}L$. By immunologic surface marker analysis, 7 patients were early pre-B CALLA (+) and 5 patients were T-cell ALL. Two patients of intermediate risk group had relapsed and died. Three patients had family history of breast cancer. Four patients died and overall survival rates were $53.5{\pm}18.7%$. Conclusion : Loss of p16 protein is supposed to be an independent risk factor of childhood ALL associated with poor outcomes. In clinical setting, the clinician must take into account p16 status, not only at the genomic but also at the protein level. Further clinical experience on thoroughly investigated cases will help a better understanding between p16 status and clinical outcomes.

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.315-322
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• 2006

FS11052, a novel microbial metabolite from Streptomyces spp. was identified as a small molecular substance and shown inhibition activities for the release of neurotransmitter from rat hippocampal neuron and PC12 cells. FS11052 is an inhibitor of tritiated norepinephrine ($[^3H]-NE$) release in high $K^+$ buffer solution containing ionomycin, indicating that FS11052 inhibits neurotransmitter release after the influx of $Ca^{2+}$ ions. When examined the effect of FS11052 on glucuronidase release from guinea pig neutrophils, FS11052 inhibited glucuronidase release: when treated with $5{\mu}g/ml$ of FS11052, which was not induced cellular cytotoxicity. The fact that the glucuronidase release in neutrophil and norepinephrine release in neuron was inhibited suggests the similarity in the locations and the mechanisms of FS11052 action targets. When treated with $5{\mu}g/ml$ of FS11052, $[^3H]-NE$ release and neurite extension for both rat hippocampal neurons and PC12 cells were prevented. These observations of FS11052 functioning as an inhibitor of neurotransmitter release suggest that FS11052 has an important role in synaptic transmission in neuron.

• Journal of Life Science
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• v.25 no.9
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• pp.961-969
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• 2015

Epigallocatechin gallate (EGCG), the main catechin in green tea, has been shown to have some beneficial effects against various human diseases, including diabetes, neurodegenerative disorders, cancer, cardiovascular disease and obesity. To investigate the mechanism of the suppressive effects of EGCG on inflammatory response in macrophages, alterations on the levels of nitric oxide (NO) regulatory factors and inflammatory cytokines were measured in lipopolysaccharide (LPS)-activated RAW264.7 cells. No significant toxicity was detected in RAW264.7 cells treated with 100–400 μM EGCG. Moreover, the optimal concentration of LPS was determined to be 1 μg/ml based on the results of cell viability assay, NO assay and IL-6 enzyme-linked immunsorbent assay (ELISA). Furthermore, NO levels decreased significantly by 68.2% in the 400 μM EGCG/LPS treated group, while the level of inducible nitric oxide synthase (iNOS) expression decreased by 12-17% in the 200 and 400 μM EGCG/LPS treated group. A significant decrease in transcription of pro-inflammatory cytokines (TNF- α and IL-1β) and anti-inflammatory cytokine (IL-10) was also detected in the EGCG/LPS treated group. However, IL-6 transcript and protein was maintained at a constant level when in the LPS treated group relative to the EGCG/LPS treated group. Overall, these results suggest that the differential regulation of inflammatory cytokines is an important factor influencing the suppressive effects of EGCG against LPS-activated inflammatory response in RAW264.7 cells.

• Journal of Korean Society of Environmental Engineers
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• v.30 no.7
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• pp.721-728
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• 2008

DBPs(Disinfection By-Products) are most formed through the reactions between chlorine and NOM(Natural Organic Matter) in water treatment. In this study, occurrence of DBPs including THMs(Trihalomethanes) is evaluated. Also, influencing factors by the seasons and raw water quality were investigated for correlation among them and the characteristics of THMs formation by prechlorination process. This study investigated the operation condition for THMs removal depending on raw water quality. Water treatment plant from intake station to gauging well flows for about 10 hours in Y Water Supply Office. It is limited to control of THMs formation because of excessive reaction time between chlorine and THMs precursors in the intake station. Therefore, as multi-points prechlorination from intake station to gauging well, THMs formation was decreased in the water treatment, and it was willing to prevent overdosage of chlorine. The concentration of THMs was 0.021 mg/L in the summer, 0.015 mg/L in the winter, respectively. Also, THMs formation showed that 0.013 mg/L in the water of gauging well after prechlorination, 0.014 mg/L in the flocculation/sedimentation/filtration, 0.016 mg/L in the water after postchlorination, respectively. THMFP(Trihalomethane Formation Potential) removed 42.7% and 50% through the flocculation/sedimentation and filtration, respectively, and it is similar TOC removal efficiency. In this study, multi-points prechlorination from intake station to gauging well decreases in contact time and concencrations of NOM and chlorine. Also, it decreases in THMs and amount of chlorine uesd. In the result of multi-points prechlorination in the summer, the concentration of THMs was 0.013mg/L in the treated water. In view of these facts, THMs formation can be decreased approximately 50%.

• Journal of Chest Surgery
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• v.43 no.4
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• pp.345-355
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• 2010

Background: Extracellular and intracellular pH ($pH_o$ and $pH_i$), which can be changed in various pathological conditions such as hypoxia, affects vascular contractility. To elucidate the mechanism to alter vascular contractility by pH, the effects of pH on reactivity to vasocontracting agents, intracellular $Ca^{2+}$ influx, and $Ca^{2+}$ sensitivity in vascular smooth muscle were examined. Material and Method: Isometric contractions in rat superior mesenteric arteries (SMA) were observed. Intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) was recorded by microfluorometer using Fura-2/acetoxylmethyl ester in muscle cells. $pH_o$ was increased from 7.4 to 7.8 or decreased to 6.9 or 6.4. $pH_i$ was decreased by applying $NH_4^+$ or propionic acid or modulated by changing $pH_o$ after increasing membrane permeability using $\beta$-escin. Result: Decreases in $pH_o$ from 7.4 to 6.9 or 6.4 shifted concentration-response curve by norepinephrine (NE) or serotonin (SE) to the right and significantly increased half maximal effective concentration (EC50) to NE or SE. Increase in $pH_o$ from 7.4 to 7.8 shifted concentration-response curve by norepinephrine (NE) or serotonin (SE) to the left and significantly reduced EC50 to NE or SE. NE increased $[Ca^{2+}]_i$ in cultured smooth muscle cells from SMA and the increased $[Ca^{2+}]_i$ was reduced by decreases in $pH_o$. NE-induced contraction was inhibited by $NH_4^+$, whereas the resting tension was increased by $NH_4^+$ or propionic acid. When the cell membrane of SMA was permeabilized using ${\beta}$-escin, SMA was contracted by increasing extracellular $Ca^{2+}$ concentration from 0 to $10{\mu}M$ and the magnitude of contraction was decreased by a decrease in $pH_o$ and vice versa. Conclusion: From these results, it can be concluded that a decrease in $pH_o$ might inhibit vascular contraction by reducing the reactivity of vascular smooth muscle to vasoactive agents, $Ca^{2+}$ influx and the sensitivity of vascular smooth muscle to $Ca^{2+}$.

• Tuberculosis and Respiratory Diseases
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• v.52 no.6
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• pp.616-626
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• 2002

Background : The present study was performed to further improve our understanding of molecular mechanisms involved in the activation of NFkB, a major transcriptional factor involved in the inflammatory response in the lung, by particulate matter in lung epithelial cells with an aerodynamic diameter of less than $2.5{\mu}m$(PM2.5). Materials and Methods : Immediate production of reactive oxygen species (ROS) and nitrogen species (RNS), with the PM2.5 induced expression of inducible nitric oxide synthase (iNOS), $I{\kappa}B$ degradation and $NF{\kappa}B$-dependent transcriptional activity, in 549 cells, were monitored. Addition, we also examined the effect of the iNOS inhibitor, L-N6-(1-iminoethyl) lysine hydrochloride (L-NIL), on the PM2.5-induced $NF{\kappa}B$ activation in A549 cells. Results : The rapid degradation of $I{\kappa}B$ and the increase of transcriptional activity of the $NF{\kappa}B$-dependent promotor were observed in A549 cells exposed to PM2.5. The immediate production of ROS in response to PM2.5 in A549 cells was not clearly detected, although immediate responses were observed in RAW264.7 cells. A 549 cells, cultured in the presence of PM2.5, produced an increase in NO, which was noticeably significant after 15 min of exposure with the expression of iNOS mRNA. The addition of L-NIL, an iNOS inhibitor, significantly inhibited the PM2.5-induced $I{\kappa}B$ degradation and the increase of the $NF{\kappa}B$-dependent transcriptional activity. Conclusion : These results suggest that PM2.5 stimulates the immediate production of RNS, leading to the activation of $NF{\kappa}B$ in the pulmonary epithelium.

• Journal of Oriental Neuropsychiatry
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• v.15 no.2
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• pp.149-158
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• 2004

만성적이거나 반복적인 스트레스는 우울중이나 심신병 등의 발병 기전과 밀접한 관련이 있다. 전기적 자극(electric foot shock), 한랭 자극, 속박(immobilization), 투쟁(fighting), 에테르 노출 등과 같은 심한 스트레스 자극이 뇌 조직의 시상 및 시상하부, 편도체 등에서의 노르아드레날린(noradrenalin) 방출을 유의 하게 감소시키는 동시에 혈청 corticosterone을 증가시키는 것으로 알려져 있다. 한의학에서는 스트레스 인자에 대한 신체반응을 기(氣)의 변화로 인식하고 있고, 이러한 자극요인들은 신체에 대하여 병적 요인을 제공하여 제반 질환을 야기한다. 또한 질병 발생에 대한 근거로 정기(正氣)와 사기(邪氣)의 상호 관계 위주로 인식하는데, 내부의 기(氣)의 변조(變調)는 질병의 발생을 야기하는 기초가 된다. 실험에 사용된 연근(蓮根)은 연꽃과에 속한 수생초본인 연꽃의 뿌리부분으로 우(藕) 또는 우절(藕節)이라는 명칭으로 불리기도 한다. 성미(性味)는 감(甘) 삽(澁) 평(平) 무독(無毒)하고 폐(肺) 위(胃) 간경(肝經)으로 들어가며 수렴지혈(收斂止血) 화어(化瘀)의 효능이 있어 토혈(吐血) 객혈(喀血) 뇨혈(尿血) 편혈(便血) 등의 각종 출혈 증상에 사용되어져 왔다. 최근 연꽃의 부위별 추출물이 흰쥐의 지질 과산화 생성을 효과적으로 억제하는 것으로 보고되고 있어 연근(蓮根)에 대한 다양한 약리 효능의 검색이 필요할 것으로 생각되며 연꽃의 씨앗인 연자육(蓮子肉)이 양심안신(養心安神)의 효능이 있어 스트레스성 질환에 많이 응용되고 있는 것에 착안하여 본 연구를 착수하였다. 실험 동물은 ICR계 마우스를 이용하였으며, 사회 심리적 스트레스는 옆쪽 cage에서 다른 마우스의 신체에 가해지는 전기 충격을 하루 1시간 동안 지켜보게 하는 것으로 유발하였으며 이 상태에서 약물을 투여한 그룹을 실험군, 그렇지 않은 그룹을 대조군으로 하였다. 정상군은 아무런 자극 없이 하루 1시간 동안 일정 공간에 가두어 두는 것으로 하였다. 실험 결과 사회 심리적 스트레스를 받은 경우에 아무런 처치를 하지 않은 대조군에 비해 연근 추출물을 100mg/kg/day 용량으로 5 일간 투여한 실험군에서 혈청 중 corticosterone 함량이 유의하게 감소하였으므로 매우 효과적으로 스트레스를 해소하였음을 알 수 있다. 뇌세포 중에서 신경전달물질로 분비되는 노르아드레날린의 분비량은 전반적으로 증가되는 경향을 나타내었으나 유의성은 없었다. 신체적 또는 사회 심리적 스트레스가 간 조직 내 지질의 과산화를 유발하는 것으로 나타났으며 연근 추출물이 이 결과에는 영향을 주지 못하였다. 사회 심리적 스트레스로 인하여 간 조직 내 지질과산화 정도가 증가하였으므로 혈청 내 ALT 함량도 따라 증가할 것으로 추정되었으며 이에 대한 연근 추출물 경구 투여가 간 조직을 보호할 수 있는지를 확인하기 위해 분리한 혈청으로부터 ALT 함량을 측정한 결과 대조군에 비하여 유의한 감소를 나타내었다. 또한 연근 추출물이 혈청 내 지질 과산화물의 생성을 억제할 수 있다면 질병의 예방과 치료에 효과적일 것으로 추정할 수 있으므로 그 생성량을 측정하여 보았으나 대조군과의 차이가 나타나지 않았다. 이상의 결과들을 종합하여 보면 스트레스가 부하된 5일 동안 연근(蓮根) 추출물을 함께 투여한 결과 혈청 corticosterone 함량을 유의하게 감소시켰고 뇌 조직내 noradrenaline 함량을 증가시키는 경향을 나타내어 스트레스 해소에 도움이 될 수 있음을 시사하였다. 또한 혈청 내 ALT 함량을 유의하게 감소시켜 스트레스로 인해 발생하는 간 기능의 손상도 어느 정도 억제시키는 것을 확인할 수 있었는데 앞으로 연근(蓮根)의 이러한 작용에 대한 보다 자세한 연구들이 필요한 것으로 생각된다.