• The Journal of Internal Korean Medicine
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• v.29 no.2
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• pp.469-489
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• 2008

Objectives : The purpose of this study was to investigate the effects of Chungganhaeju-tang(Qingganjiejiu-tang) on alcoholic liver damaged by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment the rats were divided into the normal group, the control group(alcohol) and the sample group(CGHJT +alcohol). The ethanol was orally administered twice a day for 6 weeks in the control and sample groups. Water instead of ethanol was orally administered twice a day for 6 weeks in the normal group. CGHJT extract was orally administered once a day for 6 weeks in the sample group. The livers of each group were processed and assessed by histology, Western Blot, $Oxyblot^{TM}$, CBB and 2-dimensional electrophoresis. Results : In the histological findings of the liver, CGHJT inhibited hepatic fibrogenesis induced by alcohol. TIMP-1 decreased in the sample group assessed by western blot and statistical significance was noted by dot blotting(p<0.05). In the $Oxyblot^{TM}$, protein oxidation induced by alcohol treatment decreased with CGHJT. In the 2-dimensional electrophoresis finding, increased proteins alcohol such as HSP 60, 60kDa heat shock protein, 3-mercaptopyruvate sulfurtransferase were normalized by CGHJT. CGHJT was considered to normalize the anti-oxidation activity elevated by alcohol. In the 2-dimensional electrophoresis finding, increased oxidized proteins such as actin, prolyl 4-hydroxylase beta polypeptide, 94kDa glucose regulated protein(GRP94), heat shock protein 90-alpha(HSC86), calreticulin precursor(CRP55), ATP synthase beta chain mitochondrial precursor, caspase-8 precursor, and dihydrolipoamide succinyltransferase(E2) decreased with CGHJT. CGHJT was considered to reduce the oxidative stress of alcohol. Conclusion : Chungganhaeju-tang(Qingganjiejiu-tang) exerts an inhibitory effect against the fibrosis and protein oxidation induced by alcohol treatment of rat liver. CGHJT was considered to normalize the elevated anti-oxidation activity by alcohol and to reduce the level of oxidative stress due to alcohol.

• Fisheries and Aquatic Sciences
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• v.12 no.3
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• pp.179-184
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• 2009

Hemibarbus mylodon (Cypriniformes) is an endemic freshwater fish species in the Korean peninsula, for which urgent conservation efforts are needed. To understand their stress responses in relation to metal toxicity and thermal elevation, we performed a real-time RT-PCR-based expression assay of hepatic copper/zinc-superoxide dismutase (Cu/Zn-SOD), a key antioxidant enzyme, in response to experimental heavy metal exposure or heat treatment. The transcription of hepatic Cu/Zn-SOD was differentially modulated by acute exposure to Cu, cadmium (Cd), or Zn. Exposure to each metal at $5{\mu}M$ for 24 h revealed that Cu stimulated the mRNA expression of Cu/Zn-SOD to a greater extent than the other two heavy metals. The elevation in Cu/Zn-SOD transcripts in response to Cu exposure was dose-dependent (0.5 to $5{\mu}M$). Time course analysis of Cu/Zn-SOD expression in response to Cd exposure ($5{\mu}M$) revealed a transient pattern up to day 7. Exposure to thermal stress (an increase from 22 to $30^{\circ}C$ at a rate of $1^{\circ}C/h$ followed by $30^{\circ}C$ for 18 h) did not significantly alter SOD transcription, although heat shock protein 90 kDa (HSP90) transcription was positively correlated with an increase in temperature.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2003.10a
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• pp.177-177
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• 2003

Methylmercury (MeHg) is a well-known neurotoxicant that causes severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. In this study, suppressive subtractive hybridization (SSH) was performed to identify differentially expressed genes on human neuroblastoma cell line, SH-SY5Y treated with DMSO and MeHg (6.25 uM) for 6 hr. Differentially expressed cDNA clones were sequenced and were screened by dot blot to eliminate false positive clones. 13 of 35 screened genes were confirmed using real time RT-PCR. These genes include EB1,90-kDa heat-shock protein, chromosome condensation-related SMC-associated protein and brain peptide Al, etc. Analysis of these genes may provide an insight into the neurotoxic effects of MeHg in human neuronal cells and a possibility to develop more efficient and exact monitoring system of heavy metals as ubiquitous environmental pollutants.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.1
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• pp.241-246
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• 2003

Bambusae Caulis in Liquamen(BCL) has been used to relieve the cough and asthma, and remove the phlegm in traditional Oriental medicine. In recent years, it was studied for its antiinflammatory, antiallergenic, immune-modulating, and anticarcinogenic capabilities. This experiment was performed to evaluate the microarray profiles of CD genes in human mast cells before and after BCL treatment. The results are as follows: The expression of 51 of the genes studied was up-regulated in the Bel-treated group; they include the genes coding L apoferritin, beta-2-microglobulin, ferritin light polypeptide, CD63, monocyte chemotactic and activating fact, heme oxygenase 1, CD140a, integrin alpha M, colony stimulating factor 2 receptor, eukaryotic translation elongation factor, CD37, interleukin 18, NADH dehydrogenase 1 beta, CD48, 5-lipoxygenase activating protein, interleukin 4, ribosomal protein L5, GABA(A) receptor-associated protein, beta-tubulin, integrin beta 1, CD162, CD32, lymphotoxin beta, alpha-tublin, integrin alpha L, CD2, CD151, CD331, 90 kDa heat shock protein, CD59, CD3Z, microsomal glutathione S-transferase 2, CD33, CD162R, cyclophilinA, CD84, interleukin 9 receptor, interleukin 11, CD117, CD39-Like 2, and so forth. The expression of 7 of the genes studied was down-regulated in the BCL-treated group; they include the genes coding con, CD238, SCF, CD160, CD231, CD24, and CD130. Consequently, the treatment of BCL on the human mast cells increased the expression of 51 genes and decreased the expression of 7 genes. These data would provide a fundamental basis to the traditional applications of Bambusae Caulis in Liquamen.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.542-549
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• 2010

In the present study, overexpression, purification, and characterization of Aeropyrum pernix K1 chaperonin B in E. coli were investigated. The chaperonin $\beta$-subunit gene (ApCpnB, 1,665 bp ORF) from the hyperthermophilic archaeon A. pernix K1 was amplified by PCR and subcloned into vector pET21a. The constructed pET21a-ApCpnB (6.9 kb) was transformed into E. coli BL21 Codonplus (DE3). The transformant cell successfully expressed ApCpnB, and the expression of ApCpnB (61.2 kDa) was identified through analysis of the fractions by SDS-PAGE (14% gel). The recombinant ApCpnB was purified to higher than 94% by using heat-shock treatment at $90^{\circ}C$ for 20 min and fast protein liquid chromatography on a HiTrap Q column step. The purified ApCpnB showed ATPase activity and its activity was dependent on temperature. In the presence of ATP, ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}$ and $50^{\circ}$, respectively. Specifically, the activity of malate dehydrogenase (MDH) at $85^{\circ}$ was greatly stabilized by the addition of ApCpnB and ATP. Coexpression of pro-carboxypeptidase B (pro-CPB) and ApCpnB in E. coli BL21 Codonplus (DE3) had a marked effect on the yield of pro-CPB as a soluble and active form, speculating that ApCpnB facilitates the correct folding of pro-CPB. These results suggest that ApCpnB has both foldase and holdase activities and can be used as a powerful molecular machinery for the production of recombinant proteins as soluble and active forms in E. coli.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.112-112
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• 2002

Under normal conditions, women produce a single dominant follicle that participates in a single ovuation each menstrual cycle. But Polycystic ovary syndrome(PCOS) conditions, folliculogenesis does not proceed normally. This condition leads to the accumlation of large numbers of small graffian follicles in which the theca interstitial cells (TIC) produce abnormally large amounts of androgen. PCOS is probably the most common endocrine disorder, affecting women of reprodutive age with 5-10% prevalence estimate. Chronic anovulation, hyperandrogenism, hirsutism, obesity, infertility and polycystic ovaries are clinical hallmarks of women with PCOS. Its etiology remains unknown. To investigate the gene expression pattern of ovary in PCO-induced rat, we used cDNA expression analysis. Total RNA was extracted from the ovary of PCO-induced rat and reverse-transcribed in the presence of［$\alpha$$^{32}$P］-dATP Which were hybridized to Atlas$^{TM}$ Rat Toxicology 1.2 array (Clontech) representing approximately 1176 rat genes. We compared gene expression between ovary of pco-induced immature female rats and control. Differential gene expression profiles were revealed (LIFR-alpha, ADRA1A, Heat shock 90-kDa protein A, PDGFRA). Reverse transcription-polymerase chain reaction(RT-PCR) was used to validate the relative expression pattern obtained by the cDNA array. The precise relationship between the altered expression of genes and PCO is a matter of further investigation. This study was supported by Korea Science and Engineering Foundation(KOSEF)