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Inhibitory Mechanisms of Cell Cycle Regulation Induced by Indole-3-carbinol in Hepatocellular Carci-noma HepG2 Cells. (간암 세포주에서의 Indole-3-Carbinol에 의해 유도되는 세포주기 억제 기전)

  • 김동우;이광수;김민경;조율희;이철훈
    • Microbiology and Biotechnology Letters
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    • v.29 no.3
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    • pp.181-185
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    • 2001
  • The naturally occurring chemical indole-3-carbinol (13C), found in vegetables of the Brassica genus, is a promising anticancer agent that was shown previ- ously to induce a Gl cell cycle arrest of human breast cancer cell lines, independent of estrogen receptor signaling. The anticancer activity of 13C and the possible mechanisms of its action were explored in a human hepatocellular carcinoma cell line, HepG2. Treatment of HepG2 cells with 13C suppressed the growth of the cells. The growth sup- pression caused by 13C ($IC_{50}$/: 444$\mu$M) was found to be partially due to its ability to stop the cell cycle in HepG2 cells. Western blot analysis for the Gl phase artiest demonstrated that the expression-levels of cyclin-dependent kinase (Cdk4, Cdk6) and cyclic D were reduced strongly after treatment of Hep72 cells with 13C (4007M) for 24- 72 hrs. Furthermore, I3C selectively abolished the expression of Cdk6 in a dose- and time-dependent manner, and accordingly, inhibited the phosphorylation of retinoblastoma. Interestingly, after the HepG2 cells reached their max- imal growth arrest, the level of the p21, a well-known Cdk inhibitor, increased significantly. Therefore, it could be considered that the Gl arrest of HepG2 cells treated with 13C was due to the indirect inhibition of Cdk4/6 activities by p21 Western blot analysis for G2/M phase arrest of demonstrated the levels of Cdc2 and cyclin Bl werer reduced dramatically after the treatment of HepG2 cells with 13C ($40\mu$M) for 24-72 hrs. flow cytometry of propidium iodide-stained HepG2 cells revealed that 13C induces a Gl (53%,72hr incubation) and G2 (25%,24hr incubation) cell cycle arrest. Thus, our observations have uncovered a previously undefined antiproliferative pathway for r3C that implicates Cdk4/6 and Cdc2 as a target for cell cycle control in human HepG2 cells. However, the 13C-medi- ated cell cycle arrest and repression of Cdk4/6 production did not affect the apoptotic induction of HepG2 cell.

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Inhibitory Effect of Hot-Water Extract of Paeonia japonica on Oxidative Stress and Identification of Its Active Components (백작약 열수추출물의 산화적 스트레스 억제효과 및 유효성분 동정)

  • Jeong, Ill-Yun;Lee, Joo-Sang;Oh, Heon;Jung, U-Hee;Park, Hae-Ran;Jo, Sung-Kee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.5
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    • pp.739-744
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    • 2003
  • This study was carried out to investigate the antioxidative activity and to identify the active components of hot-water extract of Paeoniajaponica (PJ), which was a main ingredient of a herb mixture preparation recently established as a potent candidate of radioprotector in our laboratory. The water extract was fractionated with CHCl$_3$, EtOAc and n-BuOH. The extract and its fractions showed very low activity in hydroxyl radical scavenging test. In lipid peroxidation test, the extract, EtOAc and water fractions showed moderate inhibition with the ratio above 50%. In DPPH radical scavenging test, the extract, EtOAc and water fraction showed high activity with the ratio above 80%, especially. EtOAc fraction scavenged the radicals as much as synthetic antioxidant (BHA), even at low concentration. It is suggested that mai or partition for antioxidative activity of Paeonia japonica was EtOAc fraction. Subsequently, two active compounds (PJE021-1 and JE024-1) from EtOAc fraction were isolated by using MCI gel and silica gel column chromatography The two compounds inhibited remarkedly the $H_2O$$_2$-induced DNA damage in human peripheral blood lymphocytes, measured by single-cell gel electrophoresis (SCGE). PJE021-1 protected the cells to almost negative control level, dose-dependently. PJE024-1 exhibited a potent inhibition with the ratio of 71% at even low concentration (0.5 $\mu\textrm{g}$/$m\ell$). Finally, their chemical structures were identified as gallic acid (PJE021-1) and (+)-catechin (PJE024-1), respectively, on the basis of the speculation of spectral and physical data.

Change of hemostatic markers according to the clinical state in Kawasaki disease (가와사끼병의 임상경과에 따른 지혈성 표지의 변화와 임상적 의의)

  • Kim, Yong Beom;Yoon, You Sook;Lee, Sang Yun;Kil, Hong Ryang
    • Clinical and Experimental Pediatrics
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    • v.50 no.12
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    • pp.1247-1251
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    • 2007
  • Purpose : Pathologically, Kawasaki disease (KD) is associated with widespread vascular endothelial damage in the acute phase. The vasculitis induced endothelial injury leads to coagulation abnormalities. Abnormalities of endothelial function, platelet activation, and fibrinolysis are present during acute phase and long after the onset of KD. The aim of study is to evaluate the change of hemostatic markers in the clinical stages of KD and to assess the hemostatic markers to be a useful indicator of the development of coronary artery lesion (CAL). Methods : Seventy four KD patients diagnosed in Chungnam National University Hospital from November 2004 to June 2007. Eleven febrile control and eleven healthy children were selected for healthy control. All blood samples were collected before and after Intravenous gammaglobulin (IVGG), $2^{nd}$ week, and $4^{th}-8^{th}$ week of illness of KD. Results : Initial D-dimer level of Kawasaki disease showed meaningful difference compared to control group (P<0.05). D-dimer and fibrinogen degradation products (FDP) before IVGG increased compared with normal control group and decreased after IVGG administration. It is normalized until 2 weeks later, and continue to decreasing. D-dimer and FDP were significantly different according to the CAL before IVGG. Conclusion : The hemostatic markers may change to the clinical stage of KD, which may suggest the degree of endothelial injury. Increased some hemostatic markers may be the predictors for development of CAL.

Neuroprotective effects of resveratrol via anti-apoptosis on hypoxic-ischemic brain injury in neonatal rats (신생 백서의 저 산소 허혈 뇌손상에서 항세포사멸사를 통한 resveratrol의 신경보호 효과)

  • Shin, Jin Young;Seo, Min Ae;Choi, Eun Jin;Kim, Jin Kyung;Seo, Eok Su;Lee, Jun Hwa;Chung, Hai Lee;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1102-1111
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    • 2008
  • Purpose : Resveratrol, extracted from red wine and grapes, has an anti-cancer effect, an antiinflammatory effect, and an antioxidative effect mainly in heart disease and also has neuroprotective effects in the adult animal model. No studies for neuroprotective effects during the neonatal periods have been reported. Therefore, we studied the neuroprotective effect of resveratrol on hypoxic-ischemic brain damage in neonatal rats via anti-apoptosis. Methods : Embryonic cortical neuronal cell culture of rat brain was performed using pregnant Sprague-Dawley (SD) rats at 18 days of gestation (E18) for the in vitro approach. We injured the cells with hypoxia and administered resveratrol (1, 10, and $30{\mu}g/mL$) to the cells at 30 minutes before hypoxic insults. In addition, unilateral carotid artery ligation with hypoxia was induced in 7-day-old neonatal rats for the in vivo approach. We injected resveratrol (30 mg/kg) intraperitoneally into animal models. Real-time PCR and Western blotting were performed to identify the neuroprotective effects of resveratrol through anti-apoptosis. Results : In the in vitro approach of hypoxia, the expression of Bax, caspase-3, and the ratio of Bax/Bcl-2, indicators of the level of apoptosis, were significantly increased in the hypoxia group compared to the normoxia group. In the case of the resveratrol-treated group, expression was significantly decreased compared to the hypoxia group. And the results in the in vivo approach were the same as in the in vitro approach. Conclusion : The present study demonstrates that resveratrol plays neuroprotective role in hypoxic-ischemic brain damage during neonatal periods through the mechanism of anti-apoptosis.

A Study on the Relationship between Adiponectin, BDNF and Leptin with Abdominal Fat Thickness in Male Workers (남성 근로자의 복부지방두께와 adiponectin, BDNF 및 leptin의 관련성)

  • Ko, Kyung-Sun;Choi, Yoon-Jung
    • Journal of radiological science and technology
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    • v.36 no.3
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    • pp.209-217
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    • 2013
  • Adiponectin (AdipoN), brain-derived nerotrophic factor (BDNF) and leptin (LeP) are mainly secreted from adipose tissue and are known to be involved in regulation of the development of obese. However, there are not many studies on the association between abdominal fat and neuropeptides such as AdipoN, BDNF and LeP. The aim of this study was undertaken to investigate the association between abdominal fat thickness, neuropeptides and cardiovascular disease (CVD) risk factors. The participants in the study were 138 male employees without CVD. This study was approved by the Institutional Review Board of Occupational Safety and Health Research Institute. Written informed consent for the participants in this study was obtained from all individuals. We obtained subcutaneous fat thickness (SFT) and visceral fat thickness (VFT) by using ultrasonography and neuropeptides levels were measured with ELISA kit according to the method suggested by kit manufacturer. The mean SFT and VFT were $1.58{\pm}0.51$ and $4.52{\pm}1.44$ cm. The mean concentrations of AdipoN, BDNF and LeP were $3.14{\pm}3.52$ ng/ml, $24.11{\pm}8.52$ pg/ml and $4.27{\pm}2.38$ ng/ml, respectively. VFT were positively correlated with total cholesterol (r=0.217, p<0.05), LDL-cholesterol (r=0.271, p<0.01), triglyceride (r=0.233, p<0.05) and insulin (r=0.338, p<0.01), but was inversely correlated with HDL-cholesterol (r=-420, p<0.01). AdipoN levels were positively correlated with HDL-cholesterol (r=0.220, p<0.05) and were inversely correlated with total cholesterol (r=-0.196, p<0.05), LDL-cholesterol (r=-0.190, p<0.05), triglyceride (r=-0.199, p<0.05), SFT (r=-0.195, p<0.05) and VFT (r=-0.412, p<0.01). However, LeP levels showed a reverse trend to AdipoN. AdipoN level was significantly higher in non-obese participants (BMI<25 kg/m), but LeP concentration was significantly higher in obese participants (BMI>25 kg/m) than in non-obese. On multiple logistic regression analysis, obese were significantly associated with AdipoN (odds ratio=0.784) and LeP (odds ratio=1.494). These results suggested that AdipoN and LeP concentrations are affected abdominal fat and that dysfunction and/or declination in the production and secretion of neuropeptides might induced ultimately obese and CVD.

Antihepatotoxic effect of ethanol extracts from steam-dried ginseng berry on ᴅ-galactosamine/lipopolysaccharide-sensitized mice (ᴅ-galactosamine/lipopolysaccharide로 감작된 급성간독성 마우스 모델에서 인삼열매추출물의 간독성 개선 효과)

  • Jang, Su Kil;Park, Jun Sub;Ahn, Jeong Won;Jo, Boram;Kim, Hyun Soo;Kim, JeongHoon;Kim, Sang Yun;Park, Jung Youl;Lee, Do Ik;Park, Hee Yong;Joo, Seong Soo
    • Korean Journal of Food Science and Technology
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    • v.49 no.6
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    • pp.676-684
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    • 2017
  • The present study aimed to examine the hepatoprotective effects of ethanol extracts from steam-dried ginseng berry (SGBE) in both $\text\tiny{D}$-Galactosamine/Lipopolysaccharide ($\text\tiny{D}$-GalN/LPS)-sensitized mice and in vitro models. Our results clearly demonstrated that SGBE significantly reduced the level of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and lactate dehydrogenase in blood, and $TNF{\alpha}$ was normalized in 8 h after the treatment with $\text\tiny{D}$-GalN/LPS. Coincidently, major organs remained unimpaired when compared to $\text\tiny{D}$-GalN/LPS control group. Moreover, p38, which stimulates expression of NAFLD-associated cytokines, was markedly inhibited when treated with SGBE. In vitro analysis revealed that the main components of SGBE, linoleic acid and ginsenoside Re/Rd, may play a role in protecting liver from $\text\tiny{D}$-GalN/LPS-induced toxicity. Finally, we concluded that SGBE may be a promising therapeutic agent for preventing damage to the liver.

The Change of Serum Soluble E-selectin in Kawasaki Disease (가와사끼병에서의 혈청 Soluble E-selectin의 변화)

  • Jeong, Jae-Ho;Cho, Eun-Young;Lim, Jae-Woo;Cheon, Eun-Jeong;Ko, Kyong-Og;Lee, Kyung-Il
    • Clinical and Experimental Pediatrics
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    • v.48 no.5
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    • pp.539-544
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    • 2005
  • Purpose : The aim of this study was to investigate the pathophysiologic role of serum E-selectin, vascular endothelial growth factor(VEGF)-induced cell adhesion mollecule in Kawasaki disease(KD) and to look for the evidence of direct relationship between the plasma levels of soluble E-selectin and the incidence of coronary artery lesion(CAL). Methods : Changes in plasma levels of sE-selectin(n=98) over time were measured by enzyme-linked immunosorbent assay(ELISA) in 23 patients with acute KD and 25 age-matched febrile children. Results : Compared with control values, the peak levels of plasma sE-selectin were significantly elevated($mean{\pm}S.E$. : $22.89{\pm}12.53ng/mL$ vs $10.65{\pm}3.42ng/mL$, P=0.01) in KD. 5 patients with CAL, plasma sE-selectin levels before treatment were higher than in 18 patients without CAL($mean{\pm}S.E$. : $39.43{\pm}15.08ng/mL$ and $19.00{\pm}8.32ng/mL$, respectively; P=0.01). Plasma sE-selectin declined rapidly in the majority of KD patients regardless of the presence of CAL. Plasma sE-selectin levels after treatment and convalesent period were similar in KD patients with and without CAL. The plasma levels sE-selectin were correlated with those of white blood cell count(r=0.299, P<0.05), CRP(r=0.430, P<0.05), serum albumin(r=-0.483, P<0.05), serum protein(r=-0.502, P<0.05) and hemoglobin(r=-0.372, P<0.05) not with those of ESR, platelet, or duration of fever. There were significant differences in the initial level of serum sE-selectin between KD with and without CAL($mean{\pm}S.E$. : $39.44{\pm}15.08ng/mL$ vs. $19.00{\pm}17.18ng/mL$) in multivariated linear tests. Conclusion : Plasma sE-selectin levels were significantly higher in KD than in other febrile illness. Higher plamsa levels of sE-selectin may have potential as a predictor of CAL in patients with KD.

Ultrastructural analysis and quantification of autophagic vacuoles in wild-type and atg5 knockout mouse embryonic fibroblast cells (정상 및 atg5 유전자 제거 섬유아세포에서 자가포식체의 미세구조 및 이들의 정량적 분석)

  • Choi, Suin;Jeon, Pureum;Huh, Yang Hoon;Lee, Jin-A
    • Analytical Science and Technology
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    • v.31 no.5
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    • pp.208-218
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    • 2018
  • Autophagy is a cellular process whereby cytosolic materials or organelles are taken up in a double-membrane vesicle structure known as an autophagosome and transported into a lysosome for degradation. Although autophagy has been studied at the genetic, cellular, or biochemical level, systematic ultrastructural quantitative analysis of autophagosomes during the autophagy process by using transmission electron microscopy (TEM) has not yet been reported. In this study, we performed ultrastructural analysis of autophagosomes in wild-type (WT) mouse embryonic fibroblasts (MEFs) and autophagy essential gene (atg5) knockout (KO) MEFs. First, we performed ultrastructural analysis of autophagosomes in WT MEFs compared to atg5 KO MEFs in basal autophagy or starvation-induced autophagy. Although we observed phagopore, early, late autophagosomes, or autolysosomes in WT MEFs, atg5 KO MEFs had immature autophagosomes that showed incomplete closure. Upon starvation, late autophagosomes accumulated in WT MEFs while the number of immature autophagosomes significantly increased in atg5 KO MEF indicating that atg5 plays an important role in the maturation of autophagosomes. Next, we examined autophagosomes in the cell model expressing polyQ-expanded N-terminal fragment of huntingtin. Our TEM analysis indicates that the number of late autophagosomes was significantly increased in the cells expressing the mutant huntingtin, indicating that improving the fusion of autophagosome with lysosome may be effective to enhance autophagy for the treatment of Huntington's disease. Taken together, the results of our study indicate that ultrastructural and quantitative analysis of autophagosomes using TEM can be applied to various human cellular disease models, and that they will provide an important insight for cellular pathogenesis of human diseases associated with autophagy.

Increased Carboxyhemoglobin and Serum Iron Concentration as an Indicator of increased Red Cell Turnover in Preeclampsia (임신성 고혈압 환자에서 적혈구교체의 지표로서 일산화탄소 혈색소와 혈청 철의 임상적 의의)

  • Kim, Sang-Heon;Lee, Kwang-Hee;Kim, Mi-Sook;Lee, Young-Gi;Park, Yoon-Kee;Lee, Tae-Hyung;Lee, Sung-Ho
    • Journal of Yeungnam Medical Science
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    • v.10 no.1
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    • pp.68-76
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    • 1993
  • Pregnancy induced hypertension is multifaceted syndrome with variable involvement of several key organ systems, so sensitive and specific laboratory tests for predicting severity and prognosis, and early diagnosis of this disease are required. Because heme catabolism results in equimolar production of carboxyhemoglobin, iron and bilirubin, a concomittant rise of these parameters would provide confirmation of increased heme catabolism. Microangiopathic hemolytic anemia may occurs in severe preeclampsia, but it is not known whether increased red cell turnover occurs with mild preeclampsia as complication. The purpose of this study was to confirm that increased heme catabolism also occurs in patients with mild preeclampsia. The analysis of data was done on 23 cases with mild preeclampsia and 35 normal pregnant women, who were admitted to Yeungnam University Hospital from October 1992 to March 1993. The results were as follows. 1. The mean antepartum serum iron concentration was significantly higher in the group with mild preeclampsia($86.5{\pm}6.1{\mu}g/dl$) than in the controls($53.2{\pm}5.3{\mu}g/dl$). 2. The mean antepartum and postpartum carboxyhemoglobin concentrations were significantly higher in the group with mild preeclampsia(antepartum : $2.55{\pm}0.42{\mu}g/dl$, postpartum : $1.21{\pm}0.4{\mu}g/dl$) than the controls(antepartum : $0.61{\pm}0.2{\mu}g/dl$, postpartum $0.53{\pm}0.2{\mu}g/dl$) 3. During postpartum, carboxyhemoglobin concentration in preeclampsia reduced significantly from antepartum level, but, there was no difference between antepartum and postpartum carboxyhemoglobin concentrations among controls. 4. Bilirubin concentrations were similar in both groups.

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A Clinical Study on Childhood Hemolytic Anemia According to Etiological Classification (소아 용혈성 빈혈 환자에서 원인에 따른 임상적 분석)

  • Kwon, Hae-Sik;Kang, Jung-Chul;Won, Sung-Chul;Oh, Seung-Hwan;Lyu, Chuhl-Joo
    • Clinical and Experimental Pediatrics
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    • v.46 no.9
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    • pp.883-888
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    • 2003
  • Purpose : The etiology of hemolytic anemia can be classified as either cellular or extracellular defects of red blood cells. The aim of this study was to investigate the clinical and laboratory findings of hemolytic anemia concerning its etiological classification. Methods : Clinical and laboratory findings of the patients with hemolytic anemia treated from January 1987 to May 2002 at Severance Hospital were analyzed retrospectively. They were divided into two groups based on the types of red cell defects(group I : erythrocytic defect, group II : extraerythrocytic defect). Results : Twenty one cases were included in group I, thirty four cases in group II, and three cases were unclassified. In group I, nineteen cases(90.5%) were diagnosed as hereditary spherocytosis and were proved to have red cell membrane disorders while two cases(9.5%) were shown to have red cell enzyme deficiencies. In group II, thirteen cases(38.2%) were noted as autoimmune hemolytic anemia, eleven cases(32.4%) as traumatic or microangiopathic hemolytic anemia, four cases(11.8%) as drug induced hemolytic anemia, two cases(5.9%) were related with systemic lupus erythematosus and one case(2.9%) with malignancy. Hemoglobin at the time of diagnosis(7.5 g/dL vs. 6.2 g/dL, P<0.05) and the incidence of splenomegaly(85.7% vs. 18.2%, P<0.05) were higher in group I though blood urea nitrogen(9.0/0.4 mg/dL vs. 27.8/1.6 mg/dL, P<0.05) was higher in group II. Conclusion : Comparing the clinical features of pediatric hemolytic anemia, we concluded as following : In cases associated with extraerythrocytic defect, blood tests revealed significant initial lower hematocrit with higher level of BUN and Cr while cases with erythrocytic defect, splenomegaly were more common noted.