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Analysis of Bone Marrow Micrometastasis Using RT-PCR in Patients with Small Cell Lung Carcinoma (소세포 폐암에서 역전사 중합효소 연쇄반응(RT-PCR)을 이용한 골수 미세전이(naicronaetastasis)의 분석)

  • Kim, Tae-You;Park, Jong-Kook;Ryoo, Baek-Ryeol;Im, Yung-Hyuck;Kang, Yoon-Koo
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.6
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    • pp.797-806
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    • 1999
  • Background: About 20% of small cell lung cancer(SCLC) patients have bone marrow(EM) metastasis at the time of diagnosis and the remaining patients are also considered with micrometastasis. In an attempt to detect EM micrometastasis, we used cytokeratin(CK)-20 as a molecular marker, which is specific for epithelial cells. Method: A sensitive RT-PCR assay was used to compare CK-20 expression both in SCLC cell line H209 and normal leukocyte and to evaluate EM aspirates of 28 SCLC patients. Result: H209 cell line showed CK-20 expression but normal leukocyte did not, suggesting CK-20 expression is lung tissue-specific. Of 28 patients(11 limited disease, 17 extensive disease), only 2(1/11, 1/17) samples tested revealed positive signal for CK-20. Two patients with CK-20 expression had EM metastasis or multiple bone involvement during follow-up. Conclusion: Although circulating tumor cells were detected in EM of small portion of patients with bone metastasis, CK-20 doesn't seem to be a reliable marker for the detection of micrometastasis in SCLC. This study emphasizes that identification of more specific marker for micrometastsis is mandatory prior to clinical application.

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Semi-quantitative Procalcitonin Assay in Critically ill Patients with Respiratory infections (중환자 호흡 감염에서 반정량적 Procalcitonin 분석)

  • Kim, Ji-Youn;Kim, Cheol-Hong;Park, Sunghoon;Lee, Chang-Youl;Hwang, Yong Il;Choi, Jeong-Hee;Shin, Taerim;Park, Yong-Bum;Jang, Seung-Hun;Lee, Jae Young;Park, Sang Myeon;Kim, Dong-Gyu;Lee, Myung-Goo;Hyun, In-Gyu;Jung, Ki-Suck
    • Tuberculosis and Respiratory Diseases
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    • v.67 no.3
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    • pp.205-211
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    • 2009
  • Background: Serum procalcitonin level has been considered prognostic during sepsis and septic shock. We investigated the significance of procalcitonin in critically ill patients with respiratory infections. Methods: The patients who had radiographically diagnosed diffuse lung infiltrations were enrolled on a prospective basis. Bronchoalveolar lavage (BAL) fluid for the purpose of quantitative cultures (${\geq}10^4$ cfu/mL) was obtained from all patients. Serum procalcitonin levels determined by PCT-Q kit were measured on BAL day and classified as follows; <0.5 ng/mL, 0.5~2.0 ng/mL, 2.0~10.0 ng/mL and >10.0 ng/mL. We analyzed the patient's characteristics according to outcome; favorable or unfavorable, defined as death. Results: Patients from the following categories were included: medical 17 (47.2%), surgical 9 (25%), and burned 10 (27.8%). APACHE II scores on admission to intensive care unit were 11.5${\pm}$6.89 and 11 (30.6%) had unfavorable outcomes. A procalcitonin level ${\geq}$0.5 ng/mL was in 17 (47.2%) of all. On univariate analysis, the frequencies of burn injury, mechanical ventilation, multiple organ failure, and a procalcitonin level ${\geq}$0.5 ng/mL were more often increased in patients with unfavorable outcomes than in those with favorable outcomes (p<.05). Also, a higher procalcitonin range and ventilator-associated pneumonia (VAP) were more closely associated with an unfavorable outcome (p<.05). However in multivariate analysis, a strong predictor of unfavorable outcome was burn injury (p<.05). A procalcitonin level ${\geq}$0.5 ng/mL was more sensitive in predicting VAP than unfavorable outcome. Conclusion: A higher procalcitonin level seems to be associated with VAP, but further study is required to know that procalcitonin would be a prognostic marker in critically ill patients with respiratory infections.

Effects of Solidago virga-aurea var. gigantea Miq. Root Extracts on the Activity and Differentiation of MC3T3- E1 Osteoblastic Cell (미역취(Solidago virga-aurea var. gigantea Miq.) 뿌리 추출물이 MC3T3-E1 조골세포의 활성과 분화에 미치는 영향)

  • Park, Jung-Hyun;Lee, Ji-Won;Kim, Hyun-Jeong;Lee, In-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.929-936
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    • 2005
  • The purpose of this study was to examine the ability of alkaline phosphatase (ALP) synthesis of MC3T3-E1 cells when above edible sources, Solidago virga-aurea var. gigantea Miq. root (SVR) extracts, were supplimented. MC3T3-E1 cells were cultured with $\alpha-MEM$(vehicle control), dexamethasone and genestein (positive control), and SVR extracts for 27 days. The effects of SVR MeOH extracts and its fractions on cell proliferation were measured by MTT assay. At 10, 100${\mu}g/mL$ of SVR methanol extract treated, that were elevated of cell proliferation to 140 and $120\%$ via vehicle control, respectively. And then ALP synthesis was measured by spectrophotometer for enzyme activity and by naphthol AS-BI staining for morphometry at 3, 9, 18, and 27th day. As the results, every extracts and fractions were promoted ALP activity by time course at 1, 10, 100${\mu}g/mL$, except n-hexane and chloroform fractions. Remarkably, the MeOH extracts were increased ALP activity more than 4.4 times compared with vehicle control, 2.2 times via positive control at 27th day (p<0.05). The SVR MeOH extracts treated cells, especially at a concentration of 10${\mu}g/mL$, showed remarkably higher than vehicle-treated control cells of mineralization which were checked by Alizarin red staining. These results indicate that SVR methanol extract have an induction ability of proliferation and differentiation on osteoblast.

Effects of Quinone Reductase Induction and Cytotoxicity of the Angelica radix Extracts (당귀 추출성분의 세포독성 효과와 Quinone Reductase 유도활성 효과)

  • 배송자;한은주;노승배
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.1
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    • pp.147-152
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    • 2000
  • Various lines of evidence suggest that dietary components protect the initiation of carcinogenesis. In this study, the ethanol extracts (AGE) and the methanol and hexane partition layers (AGEM, AGEH) of the Angelica radix were screened for their cytotoxic effects using the MTT assay on HepG2, HeLa, MCF7 and SW626 cells and for their ability to induce quinone reductase (QR) in HepG2 cells. AGEM and AGEH of the Angelica radix showed the strongest cytotoxic effects on HepG2 and HeLa cells. Cell growth was inhibited by 99.8% and 99.8% on HepG2 cells and 99.3% and 99.4% on HeLa cells, at dose of $100\;\mu\textrm{g}/ml$ of AGEM and AGEH extracts respectively. AGE and AGEH significantly induced QR activities in the HepG2 cells. The QR activities of HepG2 cells grown in the presence of AGE, AGEH, and AGEM at the concentration of $50\;\mu\textrm{g}/mL$ were 313.5, 273.3 and 133.3 nmol/min/mg protein, respectively. Therefore, based on these studies, Angelica radix may be developed into a potentially useful cancer chemopreventive agent.

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Antioxidant Properties of Red Yeast Rice (Monascus purpureus) Extracts (홍국쌀(Monascus purpureus) 추출물의 항산화 작용)

  • Kwon, Chong-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.4
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    • pp.437-442
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    • 2012
  • Red yeast rice (RER) has been used in China for centuries for its medicinal properties and is an increasingly popular alternative lipid-lowering treatment. This study was carried out to estimate the antioxidant properties of RER extracts. The ethyl acetate extract exhibited the DPPH radical scavenging activity of 85% at 0.2 mg/mL and $IC_{50}$ 0.13 mg/mL. A significant proportion of hydroxyl radicals in a cuvette were scavenged: 44.2% at 2.5 ${\mu}g$/mL, 74.1% at 5.0 ${\mu}g$/mL, and >100% at 10 ${\mu}g$/mL. The $HepG_2$ cells pre-treated with RER ethyl acetate extract reduced the hydroxyl radicals significantly compared to the control cells. Oxidative DNA damage was measured using a Comet assay. The RER ethyl acetate extract did not induce any DNA damage per se, and appeared to enhance the resistance to DNA damage caused by an oxidant challenge with $H_2O_2$, whereas lovastatin increased the level of DNA damage in the cells in both the unstressed (no oxidant) and those stressed with $H_2O_2$. The relative gene expression of the antioxidant enzymes in $HepG_2$ cells were also affected by the RER ethyl acetate extract. The $HepG_2$ cells were pre-incubated with the RER ethyl acetate extract, and then stressed with $H_2O_2$ or left unstressed (no oxidant). In the unstressed cells, superoxide dismutase (Cu/Zn SOD) and glutathione peroxidase (GPx) were increased significantly 3.25-fold and 2.67-fold, respectively, whereas in the stressed cells, the catalase (CAT) level was increased by 4.64-fold and 7.0-fold at 5 ${\mu}g$/mL and 10 ${\mu}g$/mL, respectively, compared to those of the control. From these results, RER appears to be effective in suppressing oxidative stress.

Antioxidant Activities and Induction of Apoptosis by Methanol Extracts from Avocado (아보카도 추출물의 Apoptosis 유도와 항산화 활성)

  • Lee, Sung-Gyu;Yu, Mi-Hee;Lee, Sam-Pin;Lee, In-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.3
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    • pp.269-275
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    • 2008
  • The avocado is a widely grown and consumed fruit that is high in nutrients and low in calories, sodium, and fats. In this study, antioxidant activities and induction of apoptosis by methanol extracts from sarcocarp, seed and peel of avocado were investigated in vitro. Contents of total polyphenols in methanol extracts from sarcocarp, seed and peel were 13.89, 137.12 and $223.45{\mu}g/mg$ respectively. Radical-scavenging activities of the methanol extracts were examined by using ${\alpha},{\alpha}$-diphenyl-${\beta}$-picrylhydrazyl (DPPH) radicals and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) assay. The methanol extracts from the peel of avocado showed higher scavenging activities against DPPH, ABTS than those from sarcocarp and seed. Apoptosis in MDA-MB-231 cells mediated by the methanol extracts of avocado was associated with the increase of activation of caspase-3 and caspase-3 target protein, PARP. Therefore, with more researches on identification and action mechanism of active compounds, the methanol extracts from peel and seed of avocado is expected to be a natural source for the developments of functional food and medical agents to prevent human breast cancer.

Manufacturing and Establishment of the 2nd National Standard for Varicella Vaccine (수두생바이러스백신 국가표준품 (2차) 제조 및 확립에 관한 연구)

  • Kim, Yeon-Hee;Kim, Do-Keun;Sohn, Yeo-Won;Han, Eui-Ri;Kim, Seok-Hwan;Lim, Jong-Mi;Won, Yun-Jung;Yoon, Heui-Seong;Jo, Moon-Hee;Kim, Kwan-Soo;Kim, Jae-Ok
    • KSBB Journal
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    • v.25 no.6
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    • pp.572-576
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    • 2010
  • Biological products, such as live varicella vaccine, are composed of biological substances derived from biological organisms. It is very difficult to identify these biologics' characteristics by analysis of simple physical and chemical methods alone. So the reference material is essential in order to evaluate the quality of bilogics. The 1'st national standard for varicella live vaccine was manufactured, established in 2002 and 2003, and have been used for the manufacturer's quality control and national lot release since then. As the lack of its availability and the decrease of its stability, this study was initiated by National Institute of Food and Drug Safety Evaluation (NiFDS) in 2008 to manufacture and establish the 2nd national standard for varicella live vaccine. The candidate material was manufactured from one of domestic manufacterers and the joint research of the NiFDS and manufacturers of varicella live vaccine was conducted to estimate of the reliable virus content. In the collaborative study, 3 laboratories including NiFDS performed the virus content test more than 7 times and all assay results were statistically analyzed. The mean coefficient of variation (CV) was 1.24%, and the geometric mean titre (GMT) variation range of each laboratory was low. On the basis of the results of this study, the candidate material of 2nd national standard for varicella live vaccine was assigned a potency of 4.26 log10 pfu/0.5 mL, when reconstituted in 0.7 mL.

Plasma Transforming Growth Factor-$\beta$1 Levels of Cancer Patients (암 환자의 혈장 Transforming Growth Factor-$\beta$1 농도)

  • 전지현;이시은;이수진;박찬후;장정순;하우송;박순태;박병규
    • Biomedical Science Letters
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    • v.5 no.2
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    • pp.181-190
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    • 1999
  • To evaluate the usefulness of transforming growth factor-$\beta$1 (TGF-$\beta$1) as a new tumor marker, we determined the plasma TGF-$\beta$1 levels using sandwich ELISA assay in cancer patients. Patients with three most common adult cancers in Korea (stomach, liver and breast cancer) and children's cancers (leukemia and two kinds of solid tumor) were enrolled for the study. Furthermore, 39 individuals were subjected to age and sex-stratified plasma TGF-$\beta$1 analysis. No statistical difference was demonstrated with respect to age or sex. The mean plasma TGF-$\beta$1 level (16.0 ng/ ml) of stomach cancer patients was significantly higher than that (8.3 ng/ml) of controls. However, there was no difference among the mean plasma TGF-$\beta$1 levels of liver, breast cancer patients and controls. Seven of 16 patients (43.7%) with stomach cancer, one of 8 (12.5%) with liver cancer, and one of 7 (14.3%) with breast cancer showed higher TGF-$\beta$1 levels compared to controls. Plasma TGF-$\beta$1 concentrations of five leukemic children remained in the normal range regardless of the remission state. In contrast, initial high TGF-$\beta$1 levels from two children with solid tumors returned to normal range on surgical resection of tumors. From the above results, we could conclude that plasma TGF-$\beta$1 levels of apparently healthy individuals seem to be rather constant irrespective of difference in age or sex, and the plasma TGF-$\beta$1 has the limited value as a screening test for the diagnosis of aforementioned adult cancers because of its low sensitivity. Finally, additional studies need to be pursed for the large number of stomach cancer and pediatric solid tumor patients in order to reach a secure conclusion on the usefulness of plasma TGF-$\beta$1 as a tumor marker in these patients.

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Photoprotection and Anti-inflammatory Effects of Chinese Medical Plants (약용식물추출물의 광보호 효과와 항염증 효과 연구)

  • Jin-Hwa, Kim;Sung-Min, Park;Gwan-Sub, Sim;Bum-Chun , Lee;Hyeong-Bae, Pyo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.227-233
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    • 2004
  • Chronic exposure to solar radiation, particularly ultraviolet (UV) light, causes a variety of adverse reactions on human skin, such as sunburn, photoaging and photocarcinogenesis. Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental facts play critical roles in cellular damage. And, repeated-UV irradiation activated the expression of the matrix metalloproteinase (MMP) and induced skin irritation. Therefore, the development of effective and safe photoprotectants that can reduce and improve the skin damage has been required. The purpose of this study was to investigate the photo-protective effect of several chinese medical plants (Juniperus chinensis) on the UV -induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. UVA induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. Expression of prostaglandin E$_2$ (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay(EIA) using PGE$_2$ monoclonal antibody. In the human skin we tested anti-irritation effect on the SLS-induced damage skin after appling the extract containing emulsion. We found that Juniperus chinensis extract had potent radical scavenging effect by 98% at 100$\mu\textrm{g}$/mL. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25$\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. In the test of proinflammatory cytokines of human keratinocytes Juniperus chinensis extract decreased expression of interleukin 6 about 30%. The amount of PGE$_2$ by HaCaT keratinocytes was significantly increased at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB (p < 0.05). At the concentrations of 3.2-25$\mu\textrm{g}$/mL of this extract, the production of PGE$_2$ by HaCaT keratinocytes (24 h after 10mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p < 0.05). In SLS-induced skin irritation model in vivo, we found to reduce skin erythema and improve barrier recovery after appling Juniperus chinensis extract containing emulsion when compared to irritated non-treated and placebo-treated skin. Our results suggest that Juniperus chinensis extract can be effectively used for the prevention of UV and SLS-induced adverse skin reactions and applied as anti-aging and anti-irritation cosmetics.

Antioxidant Activity and Component Analysis of Fermented Lavandula angustifolia Extracts (라벤더 발효추출물의 항산화 활성과 성분 분석)

  • Park, Soo-Nam;Ahn, You-Jin;Won, Bo-Ryoung;Kang, Myung-Kyu;Kim, Jai-Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.2
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    • pp.125-134
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    • 2009
  • In this study, the antioxidative effects, inhibitory effects on tyrosinase, and component of non-fermented and fermented Lavandula angustifolia extracts were investigated. The ethyl acetate fraction of fermented extract (5.95 ${\mu}g/mL$) showed the most prominent the free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of L. angustifolia extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction of fermented extract (1.45 ${\mu}g/mL$) showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of L. angustifolia on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The L. angustifolia extracts suppressed photohemolysis in a concentration dependent manner (1 ${\sim}$ 50 ${\mu}g/mL$). The inhibitory effect of L. angustifolia extracts on tyrosinase was investigated to assess their whitening efficacy. Inhibitory effects ($IC_{50}$) on tyrosinase were determined with ethyl acetate fraction of L. angustifolia extract (144.80 ${\mu}g/mL$) and ethyl acetate fraction of fermented extract (122.40 ${\mu}g/mL$). Fractions of ethyl acetate and fermented extracts showed both 3 band in TLC and 3 peaks, 2 peaks in HPLC (340 nm), respectively. In each chromatography, fractions of ethyl acetate both from non-fermented and fermented L. angusfifolia have rosmarinic acid in common. These results indicate that the component and content of non-fermented and fermented extracts of L. angustifolia are different. Both of the extract of L. angustifolia can be used as an antioxidant.