• BMB Reports
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• v.40 no.5
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• pp.656-661
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• 2007

In this report, we describe an optimized method for generation of ephA8 BAC transgenic mice expressing the lacZ reporter gene under ephA8 regulatory sequences. First, we constructed a targeting vector that carries a 1.2 kb ephA8 DNA upstream of its first exon, a lacZ expression cassette, a kanamycin cassette, and a 0.7 kb ephA8 DNA downstream of its first exon. Second, the targeting vector was electroporated into cells containing the ephA8 BAC and pKOBEGA, in which recombinases induce a homologous recombination between the ephA8 BAC DNA and the targeting vector. Third, the FLP plasmid expressing the Flipase was electroporated into these bacteria to eliminate a kanamycin cassette from the recombinant BAC DNA. The appropriate structures of the modified ephA8 BAC DNA were confirmed by Southern analysis. Finally, BAC transgenic mouse embryos were generated by pronuclear injection of the recombinant BAC DNA. Whole mount X-gal staining revealed that the lacZ reporter expression is restricted to the anterior region of the developing midbrain in each transgenic embryo. These results indicate that the ephA8 BAC DNA contains most, if not all, regulatory sequences to direct temporal and spatial expression of the lacZ gene in vivo.

• The Journal of Internal Korean Medicine
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• v.32 no.2
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• pp.243-258
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• 2011

Objectives : This study investigated the hypolipidemic and antioxidant effects of Curcuma radix using a rat model induced by poloxamer 407 injection. Methods : Serum lipid parameters and oxidative stress-associated biomarkers were determined. Additionally, hepatic cholesterol and triglyceride as well as lipid metabolism-associated gene expressions were observed in hepatic tissue. Results : 1. Curcuma radix ameliorated elevation of serum cholesterol, triglyceride, LDL-cholesterol, MDA, hepatic cholesterol level, and reduction of serum TAC, SOD, GSH, GSH-reductase level. 2. Curcuma radix augmented up-regulated ACAT gene expression. 3. Curcuma radix almost completely ameliorated down-regulated CYP-7A1 but up-regulated HMG-CoA gene expression. Conclusions : The hypolipidemic and antioxidant properties of Curcuma radix were evidenced. This study provides a scientific basis for the clinical application of Curcuma radix and development of hypolipidemics using this herb in the future.

• Toxicological Research
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• v.25 no.2
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• pp.85-92
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• 2009

4,4'-Methylenedianiline (MDA) is an aromatic amine that is widely used in the industrial synthetic process. Genotoxic MDA forms DNA adducts in the liver and is known to induce liver damage in human and rats. To elucidate the molecular mechanisms associated with MDA-induced hepatotoxicity, we have identified genes differentially expressed by microarray approach. BALB/c male mice were treated once daily with MDA (20 mg/kg) up to 7 days via intraperitoneal injection (i.p.) and hepatic damages were revealed by histopathological observation and elevation of serum marker enzymes such as AST, ALT, ALP, cholesterol, DBIL, and TBIL. Microarray analysis showed that 952 genes were differentially expressed in the liver of MDA-treated mice and their biological functions and canonical pathways were further analyzed using Ingenuity Pathways Analysis (IPA). Toxicological functional analysis showed that genes related to hepatotoxicity such hyperplasia/hyperproliferation (Timp1), necrosis/cell death (Cd14, Mt1f, Timp1, and Pmaip1), hemorrhaging (Mt1f), cholestasis (Akr1c3, Hpx, and Slc10a2), and inflammation (Cd14 and Hpx) were differentially expressed in MDA-treated group. This gene expression profiling should be useful for elucidating the genetic events associated with aromatic amine-induced hepatotoxicity and for discovering the potential biomarkers for hepatotoxicity.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.12
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• pp.7375-7379
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• 2013

The main aim of this study was to investigate the roles of GST-${\pi}$ and $pol{\beta}$ genes in the chemoresistance of esophageal carcinoma cells. Eukaryotic expression vectors containing each gene were constructed and transfected into EC9706 cells, and the biological effects of the two genes assessed based on a resistance index. We additionally investigated the in vitro and in vivo anti-resistance effects of GST-${\pi}$ and $pol{\beta}$ genes using recombinant lentiviruses carrying siRNAs against the two genes. Our results showed that upregulation of GST-${\pi}$ and $pol{\beta}$ genes suppresses chemosensitivity of esophageal carcinoma cells to cisplatin, while downregulation of these two genes with RNAi technology reverses this chemoresistance. Multi-site injection of recombinant lentivirus targeting the GST-${\pi}$ gene into transplanted cDDP tumors effectively reversed their chemoresistant phenotype. However, the same treatment against the $pol{\beta}$ gene did not lead to significant efficacy against chemoresistance.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.5
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• pp.355-359
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• 2000

Cocaine induces immediate early gene expression and behavioral changes by blocking dopamine transporters in the terminals of nigrostriatal neurons in the striatum. The pharmacological role of serotonin 2/1C (5HT2/1C) receptors in cocaine-induced expression of zif268 (NGFI-A, egr1 and Krox-24) mRNA, a member of the zinc finger, was investigated using quantitative in situ hybridization histochemistry in vivo. Behavioral alterations induced by cocaine were also monitored in relation with blockade of the receptors. Systemic injection of ritanserin (1 mg/kg, s.c.), a 5HT2/1C receptor antagonist, did not reverse behavioral alterations and zif268 mRNA gene expression induced by 15 mg/kg cocaine, i.p., in the dorsal and ventral striatum. These data indicate that ritanserin-sensitive 5HT2/1C receptors are not necessary for cocaine-induced behavioral alterations and zif268 mRNA gene expression in the striatum.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.5
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• pp.361-367
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• 2000

Cocaine functions as indirect dopamine and serotonin (5-hydroxytryptamine, 5HT) agonists and induces genomic and behavioral alterations in the striatum. Previously we demonstrated that ritanserin, a 5HT2/1C receptor antagonist, is not responsible for cocaine-induced behavioral alterations and zif268 mRNA gene expression in the striatum (see the previous paper in this issue). In this study, it was hypothesized that dopamine and 5HT2/1C receptors are required for cocaine-induced behavioral alterations and c-fos and zif268 mRNA expression. This hypothesis was addressed by infusing amperozide which antagonizes both 5HT2/1C and dopamine receptors and was analyzed using the quantitative in situ hybridization histochemistry in vivo. Systemic injection of amperozide (5 mg/kg, s.c.) significantly blocked increase in behavior, c-fos and zif268 mRNA expression induced by 15 mg/kg cocaine, i.p., in the dorsal striatum. These data suggest that dopamine and 5HT2/1C receptors are necessary for cocaine-induced behavioral alterations and immediate early gene expression in the dorsal striatum.

• Molecular & Cellular Toxicology
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• v.3 no.2
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• pp.119-126
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• 2007

1, 1-dichloroethylene (DCE) is well known hepatotoxicant as a model acute hepatotoxicity and selectively injure the bile canalicular membrane of centrilobular hepatocytes. In this study, we investigated hepatic gene expression and histopathological changes in response to DCE treatment. DCE was administered once daily at 20 mg/kg up to 14 days via intraperitoneal injection. Five mice were used in each test group and were sacrificed at 1, 7, and 14 days. Serum biochemical and histopathological analysis were performed for evaluation of hepatotoxicity level. Direct bilirubin and total bilirubin activities were slightly elevated in treated group at 7 days. DCE treatment for 7 days resulted in centrilobular hepatocyte hypertrophy and hepatocyte vacuolation, and mild hepatocyte vacuolation and high hepatocyte basophilia were observed in 14 days treated group. One hundred twenty three up-regulated genes and 445 down-regulated genes with over 2-fold changes between treated and control group at each time point were used for pathway analysis. These data may contribute in understanding the molecular mechanism DCE-induced hepatotoxicity.

• The Korean Journal of Zoology
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• v.34 no.3
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• pp.426-431
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• 1991

The present study examines the effect of naloxone, mu-opioid receptor antagonist, on prolactin (PRL) gene expression and secretion induced by estradiol (I) treahent in vivo. Adult rats were ovariectomized (OW) and implanted with Silastic capsules containing either vehicle (oil) or E. Three days later, NAL (2 mg/kg BW) or saline urere injected 30 min prior to sacrifice. To examine PRL secretion in vitro, the pituitaries were incubated in the superfusion system for 3 hrs. Superfusates were collected at 10 min intenrals on ice and subjected to PRL radioimmunoassay. Endogenous release of PRL in OU( + I rats was signifcantlv higher than that in OVX rats (mean $\pm$ SE; 24.5 $\pm$ 3.1 vs 14.5 $\pm$ 2.9 ns/10 min). A single injection of NAL clearly inhibited PRL release in Nitro from pituitaries derived from OW + I rats, but not from OW group. PRL myNA was determined by RNA-blot hybridisation assay with nicktranslated PRL CDNA. E stimulated PRL mRNA about 3 fold over that shown in OW group. Treahent of NAL suppressed the I-stimulated PRL myNA in OVX + I group, but not in OVX group. These data clearly showed that the NAL-induced inhibition of PRL secretion was well correlated with changes in PRL mRNA level and this inhibitory process appears to be mediated in I-dependent manner.

• Journal of fish pathology
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• v.23 no.3
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• pp.273-280
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• 2010

To determine whether rock bream Oplegnathus fasciatus can be protected from rock bream iridovirus (RBIV) infection by intramuscular injection of long double-stranded RNAs (dsRNAs), we compared protective effect of virus-specific dsRNAs corresponding to major capsid protein (MCP), ORF 084, ORF 086 genes, and virus non-specific green fluorescent protein (GFP) gene. Furthermore, to determine whether the non-specific type I interferon (IFN) response was associated with protective effect, we estimated the activation of type I IFN response in fish using expression level of IFN inducible Mx gene as a marker. As a result, mortality of fish injected with dsRNAs and challenged with RBIV was delayed for a few days when comparing with PBS injected control group. However, virus-specific dsRNA injected groups exhibited no significant differences in survival period when compared to the GFP dsRNA injected group. Semi-quantitative analysis indicated that the degree of antiviral response via type I IFN response is supposedly equal among dsRNA injected fish. These results suggest that type I IFN response rather than sequence-specific RNA interference might involve in the lengthened survival period of fish injected with virus-specific dsRNAs.

• Journal of Marine Life Science
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• v.2 no.1
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• pp.7-11
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• 2017

Sulfated polysaccharides are known to be immune-stimulators in mammals and can be used as food additives to enhance immunity. In this study, the immune-stimulating activity of water-soluble anionic macromolecules F₂ fractionation isolated from Codium fragile using ion-exchange chromatography was tested in olive flounder, Paralichythys olivaceus, in vitro and in vivo. The gene expression of interleukin (IL)-1β was adopted to check the immune-affection. As a result, in vitro study revealed that the expression of IL-1β was significantly upregulated in head kidney cells by 1 and 5 ㎍/ml of polysaccharides 4 h and by 5 ㎍/ml of polysaccharides at 24 h. In vivo, IL-1β gene expression in head kidney was significantly upregulated by 20 and 100 ㎍ of the polysaccharides at day 1 post-i.p. injection, while downregulated at day 3 but not significant. Meanwhile, in peritoneal cells, it was upregulated by 20 ㎍ of the polysaccharides at day 1 but the upregulation was sustained until day 3 though it was not significant. These results indicate that the sulfated polysaccharides from C. fragile are an immune-stimulator and might be potential feed additives for olive flounder.