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The role of $Na^+-Ca^{2+}$ exchange on calcium activated chloride current in single isolated cardiac myocyte in pulmonary vein of rabbit.

  • Kim, Won-Tae;Lee, Yoon-Jin;Ha, Jeong-Mi;Han Choe;Jang, Yeon-Jin;Park, Chun-Sik;Lee, Chae-Hun m
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.37-37
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    • 2003
  • We have shown the $Ca^{2+}$-activated chloride current is present in cardiac myocyte in rabbit pulmonary vein (Kim et al., 2002). This current amplitude was increased as [N $a^{+}$]$_{i}$ was increased and we suggested this chloride current may be involve in the spontaneous action potential frequency change. Since this current is activated by the increase of intracellular $Ca^{2+}$, we would like to test what is the inducer of the increase of [C $a^{2+}$]$_{i}$ between a L-type $Ca^{2+}$-current or a reverse mode of N $a^{+}$-C $a^{2+}$ exchange current. White rabbit (1.5 kg) was used and anesthetized with Ketamin (100 mg/kg). Pulmonary vein (PV) was isolated and sleeve area between left atrium and PV was dissected. Using collagenase (Worthington 0.7 mg/cc), single cardiac myocytes were isolated. In the presence of 15 mM of N $a^{+}$, three steps of voltage pulses were applied (holding potential : -40 ㎷, -80 ㎷ for 50 msec, 30 ㎷ for 5 msec, 10 ㎷ steps from -70 ㎷ to 60 ㎷). The inward and outward tail current was activated after brief 5 msec prepulse. The outward tail current was blocked by the removal of extracellular chloride substituted by glucuronic acid or by a chloride channel blocker, 5 mM 9-AC. But the inward tail current was still remained even though the amplitude was decreased. The reversal potentials were changed to the direction of the change of chloride equilibrium potential ( $E_{Cl}$ ) but the shift of equilibrium potential was not enough to match to the theoretical equilibrium potential shift. In the presence of L-type $Ca^{2+}$ channel blocker, nifedipine 1 uM, inward tail currents were greatly reduced but the outward current tail currents were still remained. In the presence of N $a^{+}$-C $a^{2+}$ exchange current blocker, 10 uM KB-R7943, the inward and outward tail currents were blocked almost completely. We tried to test the $Ca^{2+}$sensitivity of the chloride current with various [C $a^{2+}$]$_{i}$ in pipette solution from 100 nM to 1 uM but we failed to activate $Ca^{2+}$-activated chloride currents even though the cell became contracted in the presence of 1 uM $Ca^{2+}$. From these results, we could conclude that the increase of [C $a^{2+}$]$_{i}$ to activate the outward $Ca^{2+}$-activated chloride current was mainly induced by the activation of the reverse mode of N $a^{+}$-C $a^{2+}$ exchanger, But for the increase of [C $a^{2+}$]$_{i}$ to activate the inward tail current, L-type $Ca^{2+}$ current may be the major provoking current. Since the cytosolic increase of [C $a^{2+}$]$_{i}$ through pipette solution have failed to activate $Ca^{2+}$-activated chloride current, this chloride current may have very low $Ca^{2+}$ sensitivity or a comparmental increase $Ca^{2+}$ such as in subsarcolemmal space may activate the chloride current. Since there are several reports and models that the increase of $Ca^{2+}$ in subsarcolemmal space would be over several to tens of uM, both possibility may be valid together.uM, both possibility may be valid together.

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Immunocytochemical Study on the Change of the Activated T Cells in Peripheral Blood of the Pulmonary Tuberculosis Patients (폐결핵환자의 말초혈액에서 Activated T Cell의 변화)

  • Ryu, Gyeong-Ryeol;Park, Une-Sook;Park, Chong-Hwa;Jeoung, Pan-Joon;Hwang, Young-Sil;Lee, Yang-Jae;Chang, Ga-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.6
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    • pp.823-830
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    • 1995
  • Background: It has been found that Helper T cells in the peripheral blood are decreased in the cell mediated immunity in the pulmonary tuberculosis. But it has not been confirmed yet that only decrease in number of cells which has phenotype in the peripheral blood is defined to decrease in cell mediated immunity. The immunocytochemical study was performed to observe the change of the percentage of T-lymphocytes with their subsets and activated T cells in the peripheral blood of pulmonary tuberculosis and to know how many T cells would be activated, relative to resting cells in the peripheral blood. Methods: The peripheral blood obtained from twenty two patients and ten healthy controls were smeared on the gelatin coated slide glass prepared for of mononuclear cells. The double bridge technique of alkaline phosphatase-antialkaline phosphatase(APAAP) method was used. As the primary antibodies, $T_1$(anti-human T cell), $T_4$(anti-human helper/inducer T cells) and $T_8$(anti-human supressor/cytotoxic T cell) antibodies and interleukin-2 receptor (for early activated T cell), very late activation antigen (for activated cytotoxic T cell), T cell lineage specific activation antigen monoclonal actibodies were used. Results: 1) There were significantly decrease in the absolute number of $T_4$(+) cells but significantly increase of $T_8$(+) cells in the peripheral blood of pulmonary tuberculosis (p<0.05). 2) The percentage of $T_4$(+) cells showed significantly decrease in pulmonary tuberculosis but $T_8$(+)cells significantly increase(p<0.05). $T_4(+)/T_8(+)$ ratio showed significantly decrease in the peripheral blood of the pulmonary tuberculosis(p<0.05). 3) There were significantly increase in the absolute number of variable stages of activated T cells in the peripheral blood of the pulmonary tuberculosis(p<0.05). 4) The percentage of IL-2R, VLA-1, TLiSA were 6.45+1.56%, $7.64+1.34^*$, 10.45+1.16% in order which showed significantly increase in the peripheral blood of the pulmonary tuberculosis(p<0.05). Conclusion: We speculate that only a few percentage of T lymphocyte is activated in cell mediated immunity in pulmonary tuberculosis.

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Evaluations on Deodorization Effect and Anti-oral Microbial Activity of Essential Oil from Pinus koraiensis (잣나무 정유의 소취효과 및 구강균에 대한 항균활성 평가)

  • Hwang, Hyun Jung;Yu, Jung-Sik;Lee, Ha Yeon;Kwon, Dong-Joo;Han, Woong;Heo, Seong-Il;Kim, Sun Young
    • Korean Journal of Plant Resources
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    • v.27 no.1
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    • pp.1-10
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    • 2014
  • Essential oils of various plants have been known for potential biological effects such as antibacterial, antifungal, spasmolytic, antiplasmodial activities and insect-repellent property. Recently, the essential oils have attracted considerable interest in oral disease therapy. This essential oil has been known as being effective on easing sick house syndrome, giving forest aroma therapy effect and acting as repellent against pest. The essential oil of Pinus koraiensi, a native plant from Hongcheon-gun, Gangwon-do, was obtained by hydrodistillation. In light of its medicinal importance, in this study its composition, antibacterial activity and the reducing effect of offensive odor have been analyzed. The composition of essential oil was determined by GC and GC-MS. We have identified 14 compounds, of which 1R-${\alpha}$-pinene (19.38 %), 3-carene (10.21 %), camphene (9.82 %), limonene (9.00 %), bicyclo[2,2,1] heptan-2-ol (8.76 %) and ${\beta}$-phellandrene (7.98 %) were the main components. Essential oils from P. koraiensis, Chamaecyparis obtusa, Abies holophylla and Pinus densiflora were compared in terms of alleviating effect of malodors caused from formaldehyde, ammonia, trimethylamine and methylmercaptan. P. koraiensis essential oil was found to decrease the amounts of ammonia and trimethylamine by 75.17 % and 77.36 %, respectively. Antibacterial activity against Streptococcus mutans and Streptococcus sobrinus, which were known as oral cavity inducer, was investigated using the paper disc agar diffusion method. The inhibition zone was observed against S. mutans (5.97 mm) and S. sobrinus (1.40 mm), respectively. P. koraiensis essential oil shown effective deodorization and inhibitory activity against oral cavity in this study might be potential material in oral sanitary industry.

Association between the Human Surfactant Protein-A(SP-A) Gene Locus and Chronic Obstructive Pulmonary Disease in Korean Population (한국인에서 만성폐쇄성폐질환과 인체 폐 표면 활성제 단백-A 유전자 대립형질의 상관관계)

  • Na, Joo Ock;Oh, Myung Ho;Choi, Jae Sung;Seo, Ki Hyun;Kim, Yong Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.6
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    • pp.638-644
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    • 2006
  • Backgrounds: This study investigated whether or not a polymorphism in the gene encoding the surfactant protein A(SP-A) has any bearing on the individual susceptibility to the development of chronic obstructive pulmonary disease(COPD) in a genetically homogenous Korean population. Methods: The genotypes of 19 COPD patients and 20 healthy neonates as controls were tested using a polymerase chain reaction followed by restriction fragment length polymorphism analysis for the SP-A gene. Results: The specific frequencies of the 6A2 and 6A18 alleles of SP-A1 and the 1A2 allele of SP-A2 were much higher in the COPD group than control group (p<0.05). However, the frequencies of the 6A3 and 6A4 alleles of SP-A1 and the 1A0 allele of SP-A2 in the COPD group were significantly lower than the control group. In the COPD group, the frequencies of the +50 locus genotypes GG of SP-A1 and the +9 locus genotypes CC of SP-A2 were 85.0% and 60.6%, respectively, and 19.7% and 24.8% in the control group, respectively. The frequencies of the polymorphic genotypes or alleles showed a statistically significant difference between the COPD group and the control group (P<0.05). Conclusion: A genetic polymorphism in SP-A is associated with the development of COPD in the Korean population.

Presence of Leukemia-maintaining Cells in Differentiation-resistant Fraction of K562 Chronic Myelogenous Leukemia (만성 골수성 백혈병 K562세포의 분화 내성 분획에서 백혈병 유지 세포의 동정)

  • Lee, Hong-Rae;Kim, Mi-Ju;Ha, Gahee;Kim, So-Jung;Kim, Sun-Hee;Kang, Chi-Dug
    • Journal of Life Science
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    • v.23 no.2
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    • pp.197-206
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    • 2013
  • The present study investigated whether leukemia-maintaining cells reside in a differentiation-resistant fraction using a megakaryocytic differentiation model of K562 cells. Treatment with phorbol-12-myristate-13-acetate (PMA) significantly inhibited the colony-forming efficiency of the K562 cells. At a PMA concentration of 1 nM or higher, colony was not formed, but approximately 40% of K562 cells still survived in soft agar. Approximately 70% of colony-forming cells that were isolated following the removal of PMA after exposure to the agent were differentiated after treatment with 10 nM PMA for 3 days. The differentiation rate of the colony-forming cells was gradually increased and reached about 90% 6 weeks after colony isolation, which was comparable to the level of a PMA-treated K562 control. Meanwhile, imatinib-resistant variants from the K562 cells, including K562/R1, K562/R2, and K562/R3 cells, did not show any colony-forming activity, and most imatinib-resistant variants were CD44 positive. After 4 months of culture in drug-free medium, the surface level of CD44 was decreased in comparison with primary imatinib-resistant variants, and a few colonies were formed from K562/R3 cells. In these cells, Bcr-Abl, which was lost in the imatinib-resistant variants, was re-expressed, and the original phenotypes of the K562 cells were partially recovered. These results suggest that leukemia-maintaining cells might reside in a differentiation-resistant population. Differentiation therapy to eliminate leukemia-maintaining cells could be a successful treatment for leukemia if the leukemia-maintaining cells were exposed to a differentiation inducer for a long time and at a high dose.

Comparative Transcriptome Analysis of the Response of Two Lines of Rapeseed (Brassica napus L.) to Cold Stress (유채 두 계통에서 저온 스트레스에 반응하는 전사체 발현 비교 분석)

  • Lee, Ji-Eun;Kim, Kwang-Soo;Cha, Young-Lok;An, Da-Hee;Byun, Jong-Won;Kang, Yong-Ku
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.1
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    • pp.37-71
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    • 2021
  • Rapeseed is a typical winter crop, and its freezing stress tolerance is a major feature for winter survival. Therefore, it is important to comprehend clearly the physical and molecular mechanisms of rapeseed under freezing stress conditions. This study investigates the physical and transcriptome changes of two rapeseed lines, 'J8634-B-30' and 'EMS26', under cold acclimation and freezing temperature treatments. The proline content of 'J8634-B-30' at 5 ℃ increased 8.7-fold compared to that before treatment, and there was no significant change in that of 'EMS26' RNA-sequencing analysis revealed 5,083 differentially expressed genes (DEGs) of 'J8634-B-30' under cold acclimation condition. Among the genes, 2,784 (54.8%) were up-regulated and 2,299 (45.2%) were down-regulated. The DEGs of 'EMS26' under cold acclimation condition were 5,831 genes, and contained 2,199 up-regulated genes (37.7%) and 3,632 down-regulated genes (62.3%). Among them, only DEGs annotated in the cold response-related signaling pathways were selected, and their expression in the two rapeseed lines was compared. Comparative DEGs analysis indicated that cold response related signaling pathways are proline metabolism and ABA (Abscisic acid) signaling. And ICE (Inducer of CBF expression) - CBF (C-repeat-binding factor) - COR (Cold-regulated) signaling were the significantly differentially expressed transcripts in the two rapeseed lines. The major induced transcripts of 'J8634-B-30' induced P5CS (Δ'-pyrroline-5-carboxylate synthetase), which is related to proline biosynthesis, PYL (pyrabactin resistance-like protein, ABA receptor) and COR413 (cold-regulated 413 plasma membrane 1). In conclusion, these result provide a foundation for understanding the mechanisms of freezing stress tolerance in rapeseeds. Further functional studies should be performed on the freezing stress-related genes identified in this study, which can contribute to the transgenic and molecular breeding for freezing stress tolerance in rapeseed.

A Study on the Conversion to Feed Stuff from Cellulosic Biomass (섬유질자원(纖維質資源)의 사료(飼料) 전환(轉換))

  • Lee, Ke-Ho;Sung, Chang-Geun;Chung, Kyu-Ok
    • Applied Biological Chemistry
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    • v.27
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    • pp.29-46
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    • 1984
  • To utilize several species of hard wood as raw materials of feed products, fermentation characteristics of cellulosic substrates to single cell protein was investigated, and results were summarized as follows. Among the microorganisms investigated, Tricoderma viride was selected as one of the most cellulolytic. Mixed culture of fungi did not show a synergistic effect on cellulose degradation. When the fungi were cultured at $28^{\circ}C$ for 7 days in a medium containing wheat bran 25 g, cellulose 0.25 g, proteose peptone 0.025 g and tween 800.025 g, cellulotic activities on carboxy methyl cellulose and filter paper reached maximum at 12 hr. The alkali treatment resulted in increased degradation of substrate from 13 to 18% when treated with enzymes for 12h, and reducing sugar formation increased with decreased size of substrates. Glucose was a very good feedback inhibitor of the enzyme from T.viride than that of xylose. When the substrate was rehydrolyzed, hydrolysis rate was 31% to reducing sugars within 12 hr. Quantative anlysis with HPLC showed the ratio of glucose to xylose in sugar syrups as 1.77 to 1. For the purpose of producing cellulosic-single cell protein from the sawdust of mulberry tree, 15 strains of xylose-assimilating yeast were isolated from 42 samples of rotten woods and compost soils and examined for their ability to utilize xylose. Then three strains were selected by their strong xylose-assimilating activities. The cultivative condition, the growth characteristics, and protein and nucleic acid productivities of three strains were investigated. The results obtained were, 1. Wood hydrolysate of mulberry tree was assimilated by 5 strains of CHS-2, CHS-3, ST-40, CHS-12 and CHS-13. 2. The optimum initial pH and temperature for the growth of strain CHS-13 were 4.4 and $30^{\circ}C$. 3. The specific growth rate of strain CHS-13 was $0.23h^{-1}$ and generation time was 3.01 hrs at the optimum condition. 4. CHS-13 strain assimilated 81 % of sugar in wood hydrolysate. 5. CHS-13 strain was identified as Candida guilliermondii var. guilliermondii 6. When the CHS-13 strain was cultured in the wood hydrolysate containing yeast extract, L-protein content was increased with yeast extract concentration. 7. The L-protein and nucleic acid yields from wood hydrolysate were 0.73 mg/ml and $4.92{\times}10^{-2}\;mg/ml$ respectively. 8. An optimal nucleic acid content of CHS-13 strain was observed in the medium containing 0.2% of yeast extract.

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Protective Effect of Plantago asiatica L. Leaf Ethanolic Extract Against Ferric Nitrilotriacetate-Induced Prostate Oxidative Damage in Rats (랫드에서의 Fe-NTA 유발 산화스트레스에 대한 차전초 에탄올 추출물의 전립선보호 효과)

  • Hong, Seung-Taek;Hong, Chung-Oui;Nam, Mi-Hyun;Ma, Yuan-Yuan;Hong, Yun-Jin;Son, Da-Hee;Chun, Su-Hyun;Lee, Kwang-Won
    • Journal of Food Hygiene and Safety
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    • v.26 no.3
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    • pp.260-265
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    • 2011
  • Plantago asiatica L. (P. asiatica) has been used as one of the popular folk medicines in Asia for human health care practices. Various activities of P. asiatica have been reported, such as anti-oxidant, anti-glycation, anti-inflammatory and hepatoprotective activity. Therefore, the potential of P. asiatica to reduce oxidative stress has been studied in several ways for over 20 years, especially at liver and kidney. However no investigation has been reported revealing its protective effect on prostate. Method: Treatment of P. asiatica leaf ethanolic extract (PLE) (1 g/kg body weight (b.w.), 2 g/kg b.w., or 4 g/kg b.w.) were given separately to animals for pretreatment once per day for 7 days, and on the seventh day ferric nitrilotriacetate (Fe-NTA; 0.24 mmol Fe/kg b.w.), which is known as an oxidative stress-inducer at prostate, was administrated by i.p to negative control group. At the end of the study period, dissection was carried out for detecting the prostate protective effect of PLE. Result: Fe-NTA-treated animals produced reactive oxygen species (ROS) resulting in depletion of antioxidant biomaker, such as glutathione (GSH), glutathione reductase (GR), and glutathione s-transferase (GST) and increase of lipid peroxidation in prostate. However, PLE pretreatment resulted in an increase in the GSH, GST and GR levels concentration dependent manner and in an significant decrease in the levels of lipid peroxidation. Conclusion: Our data suggest that PLE may be effective in protecting oxidative stress-induced damage of prostate, and PLE may be an chemopreventive agent against Fe-NTA-mediated prostate oxidative damage.

The Effects of Isopropyl 2-(1,3-dithioetane-2-ylidene)-2-[N-(4-methyl-thiazol-2-yl)carbamoyl]acetate (YH439) on Potentiated Carbon Tetrachloride Hepatotoxicity (상승적 화학적 간독성에 미치는 YH439의 영향)

  • Kim, Sang-Geon;Cho, Joo-Youn
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.407-416
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    • 1996
  • The reactive intermediates formed during the metabolism of therapeutic agents, toxicants and carcinogens by cytochromes P450 are frequently capable of covalently binding to tissue macromolecules and causing tissue damage. It has been shown that YH439, a congener of malotilate, is effective in suppressing hepatic P450 2E1 expression. The present study was designed to further establish the mechanistic basis of YH439 protection against toxicant by assessing its effects against chemical-mediated potentiated hepatotoxicity. Retinoyl palmitate (Vit-A) pretreatment of rats for 7 days substantially enhanced carbon tetrachloride hepatotoxicity, as supported by an ${\sim}5-fold$ increase in serum alanine aminotransferase (ALT) activity, as compared to $CCl_4$ treatment alone. The elevation of ALT activity due to Vit-A was completely blocked by the treatment of $GdCl_3$ a selective inhibitor of Kupffer cell activity. Concomitant pretreatment of rats with both YH439 and Vit-A resulted in a 94% decrease in Vit-A-potentiated $CCl_4$ hepatotoxicity. YH439 was also effective against propyl sulfide-potentiated $CCl_4-induced$ hepatotoxicity. Whereas propyl sulfide (50 mg/kg, 7d) enhanced $CCl_4-induced$ hepatotoxicity by >5-fold, relative to $CCl_4$ treatment alone, concomitant treatment of animals with both propyl sulfide and YH439 at the doses of 100 and 200 mg/kg prevented propyl sulfide-potentiated $CCl_4$ hepatotoxicity by 35% and 90%, respectively. Allyl sulfide, a suppressant of hepatic P450 2E1 expression, completely blocked the propyl sulfide-enhanced hepatotoxicity, indicating that propyl sulfide potentiation of $CCl_4$ hepatotoxicity was highly associated with the expression of P450 2E1 and that YH439 blocked the propyl sulfide-enhanced hepatotoxicity through modulation of P450 2E1 levels. Propyl sulfide- and $CCl_4-induced$ stimulation of lipid peroxidation was also suppressed by YH439 in a dose-related manner, as supported by decreases in malonedialdehyde production. The role of P450 2E1 induction in the potentiation of $CCl_4$ toxicity and the effects of YH439 were further evaluated using pyridine as a P450 2E1 inducer. Pyridine pretreatment substantially enhanced the $CCl_4$ hepatotoicity by 23-fold, relative to $CCl_4$ alone. YH439, however, failed to reduce the pyridine-potentiated toxicity, suggesting that the other form(s) of cytochroms P450 inducible by pyridine, but not suppressible by YH439 treatment, may play a role in potentiating $CCl_4-induced$ hepatotoxicity. YH439 was capable of blocking cadmium chloride-induced liver toxicity in mice. These results demonstrated that YH439 efficiently blocks Vit-A-enhanced hepatotoxiciy through Kupffer cell inactivation and that the suppression of P450 2E1 expression by YH439 is highly associated with blocking of propyl sulfide-mediated hepatotoxicity.

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