• Advances in Traditional Medicine
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• 제6권2호
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• pp.134-143
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• 2006

The capacities of bacterial DNA, extracted from Salmonella typhimurium, and lipopolysaccharide (LPS), extracted from Salmonella minnesota, to activate mouse peritoneal macrophages in vitro were compared. Activation was assessed by estimating e levels of 3 cytokines, IL-10, IL-12, and $IL-1{\beta}$, at time intervals of 3, 6, 9, and 24 h after addition of LPS and/or DNA to macrophage cultures. Cytokine levels in culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA) and cytokine mRNA levels were estimated based on band intensity in cultured cells by reverse transcriptase-polymerase chain reaction (RT-PCR). Results obtained demonstrated the ability of DNA and LPS to elicit increased production of all 3 cytokines as compared to controls. In the amount tested, LPS appeared to be a more potent inducer of IL-12, and $IL-1{\beta}$, whereas DNA induced higher levels of IL-10. DNA and LPS, used in combination, exhibited neither an additive nor a synergistic effect. Rather, an antagonist effect appeared to occur. RT-PCR results correlated well with ELISA.

• 한국동물위생학회지
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• 제36권2호
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• pp.147-150
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• 2013

This study was carried out to investigate the prevalence of honeybee (Apis mellifera) disease in cheonan and asan area. From September to November in 2012, 33 samples were collected from 33 apiculture farms in the regions and reverse transcriptase-polymerase chain reaction (RT-PCR) and polymerase chain reaction (PCR) was conducted. Among 33 samples, prevalence rate was 42% in Sac Brood Virus (SBV), 52% in Nosema, 21% in American foulbrood (AFB), 70% in European foulbrood (EFB), 97% in Stonebrood, 3% in Chalkbrood. The result indicate that stonebrood was most prevalent disease in apiculture farms in cheonan and asan area.

• Genomics & Informatics
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• 제3권4호
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• pp.172-174
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• 2005

Recent anti-cancer approaches have been based to target tumor-specifically associated and/or causative molecules such as RNAs or proteins. As this specifically targeted anti-cancer modulator, we have previously described a novel human cancer gene therapeutic agent that is Tetrahymena group I intron-based trans-splicing ribozyme which can reprogram and replace human telomerase reverse transcriptase (hTERT) RNA to selectively induce tumor-specific cytotoxicity in cancer cells expressing the target RNA. Moreover, the specific ribozyme has been shown to efficiently retard tumor tissues in xenograft mice which had been inoculated with hTERT-expressing human cancer cells. In this study, we assessed specificity of trans-splicing reaction in cells to evaluate the therapeutic feasibility of the specific ribozyme. In order to analyze the trans-spliced products by the specific ribozyme in hTERT-positive cells, RT, 5'-end RACE-PCR, and sequencing reactions of the spliced RNAs were employed. Then, whole analyzed products resulted from reactions only with the hTERT RNA. This study suggested that the developed ribozyme perform highly specific RNA replacement of the target RNA in cells, hence trans-splicing ribozyme will be one of specific agents for genetic approach to revert cancer.

• BMB Reports
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• 제41권12호
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• pp.863-867
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• 2008

CD320 has been recently discovered and reported as a follicular dendritic cell (FDC) protein. Although CD320 is known to enhance proliferation of germinal center (GC) B cells, little other information is available. In this study, we investigated its cellular distribution in the GC. Confocal microscopy of human tonsil sections revealed co-localization of CD320 with CD19 and CD38 but not with CD3 indicating that GC B cells expressed CD320 in addition to FDC. In purified GC B cells, CD320 expression was inhibited in the nucleus, membrane and cytoplasm. Reverse transcriptase-polymerase chain reaction confirmed CD320 mRNA expression in B cells. These finding indicate that CD320 is expressed in B cells in addition to FDC, and that its GC activity may be more complicated than previously thought.

• Journal of Ginseng Research
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• 제32권2호
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• pp.114-119
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• 2008

Terpene synthase plays a key role in biosynthesis of triterpene saponins (ginsenosides) and is intermediate in the biosynthesis of a number of secondary metabolites. A terpene synthase (PgTPS) cDNA was isolated and characterized from the root of Panax ginseng c.A. Meyer. The deduced amino acid sequence of PgTPS showed a similarity with A. deliciosa (AAX16121) 61%, V. vinifera (AAS66357) 61%, L. hirsutum (AAG41891) 55%, M. truncatula (AAV36464) 52%. And the segment of a terpene synthase gene was amplified by reverse transcriptase-polymerase chain reaction (RTPCR). We studied expression of terpene synthase under stressful conditions like chilling, salt, UV, and heavy metal stress treatment. Expression of PgTPS was increased gradually after exposure to stresses such as chilling, salt, and UV illumination. But its transcription seems to be reduced by cadmium and copper treatment.

• 한국동물학회지
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• 제35권1호
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• pp.80-86
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• 1992

The nucleotide sequences of 185 rRNAs of the five Korean decapods were partially determined by the direct sequencing method using the reverse transcriptase. ne average GC content of five species was 51.1% which is higher than that of yeast(45.0%) and lower than those of frog (53.0%) and rat (55.6%). This result follows the general patterns of the GC content in the nucleotides of the nucleic acid shown among the various phylogenetic groups. The average ratio of transrional/transversional nucleotide substitution of pairwise comparison among six species (including Anemia salina) was 1.200 $\pm$ 0.310 when whole region alas examined. However, the ratio showed some differences when the conservative regions and variable regions frere separatelv examined. The molecular phylogenies of the five species were constructed by using two different tree making methods. In general the results support the previously reported molecular phylogeny of the decapod crustaceans. However, our results indicate thats in the analysis of the sequence dat3, the UPGMA clustering method of the distance matrix method should be carefully employed after considering the rate of nucneotide substitution in the different regions of the molecule.

• Archives of Pharmacal Research
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• 제19권6호
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• pp.486-489
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• 1996

A simple, rapid, specific and sensitive method for the detection of serum hepatitis C virus (HCV) RNA using the reverse transcription-polymerase chain reaction (RT-PCR) technique without conventional RNA extraction was developed. HCV template RNA from serum was obtained by boiling the serum at $95^{\circ}C$ for 2 min, cooling rapidly in ice and removing the proteins by cetrifugation. RT-PCR amplifications including the reverse transcription and first PCR amplification were performed in one vessel containing both of reverse transcriptase and Taq DNA polymerase. The detection of HCV RNA from $10^{-3}{\mu}l$. serum was possible with this method. The suitability of this method for clinical analysis was evaluated by assaying HCV RNA in 225 patient samples including anti-HCV antibody negatives (13 samples) and positives (212 samples) by enzyme-linked immunosorbent assay test (ELISA). Detections of HCV RNA with this method were in 4 of 13 anti-HCV antibody negative samples (30.8%) and 95 of 212 positive samples (44.8%). The present method can be completed in 1 hr and has a wide range of application for the clinical utilities to determine the viral RNAS.

• 생약학회지
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• 제44권3호
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• pp.275-280
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• 2013

This study was carried out to investigate the effect of Origanum vulgare extracts on cell proliferation of human hair dermal papilla cell (HHDPC) using sulforhodamine B (SRB) assay, antioxidant activity by 1,1-diphenyl-2-picryl hydrazyl (DPPH) method, expression of insulin-like growth factor-1 (IGF-1) by analyzing reverse transcriptase polymerase chain reaction (RT-PCR) and hair growth in a shaving animal model of C57BL/6 mice topically applying with an amount of 0.1 mL once a day for 3 weeks. The mice were divided into 4 groups including normal group (saline, N), negative control group (dimethyl sulfoxide, NC), positive control group (5 mg/mL minoxidil, PC), and experimental group (Origanum vulgare extracts, OV). Treatment of OV didn't show cytotoxicity in HHDPC up to 10 ${\mu}g/mL$ and exhibited antioxidant activity with $IC_{50}$ of 31.0 ${\mu}g/mL$. IGF-1 expression in the skin was significantly (p<0.05) increased in the PC and OV compared to the N or NC. PC and OV also showed a prominently promoted hair regrowth compared to the N or NC in hair growth observation. The hair regrowth of OV was significantly higher than that of PC (p<0.05). Therefore, these results indicate that O. vulgare extracts effectively stimulated hair growth in an animal model.

• Parasites, Hosts and Diseases
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• 제41권4호
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• pp.209-219
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• 2003

The evolutionary course of the CsRn1 long-terminal-repeat (LTR) retrotransposon was predicted by conducting a phylogenetic analysis with its paralog LTR sequences. Based on the clustering patterns in the phylogenetic tree, multiple CsRn1 copies could be grouped into four subsets, which were shown to have different integration times. Their differential sequence divergences and heterogeneous integration patterns strongly suggested that these subsets appeared sequentially in the genome of C. sinensis. Members of recently expanding subset showed the lowest level of divergence in their L TR and reverse transcriptase gene sequences. They were also shown to be highly polymorphic among individual genomes of the trematode. The CsRn1 element exhibited a preference for repetitive, agenic chromosomal regions in terms of selecting integration targets. Our results suggested that CsRn1 might induce a considerable degree of intergenomic variation and, thereby, have influenced the evolution of the C. sinensis genome.

• 한국초지조사료학회지
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• 제37권3호
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• pp.237-241
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• 2017

Drought is one of the detrimental factors that impair plant growth and productivity. In this study, we applied annealing control primer (ACP)-based reverse transcriptase PCR (polymerase chain reaction) technique to identify differentially expressed genes (DEGs) in maize leaves in response to drought stress. Two-week-old maize seedlings were exposed to drought (DT) by suspending water supply. DEGs were screened after 3 days of DT-treated samples using the ACP-based technique. Several DEGs encoding 16.9 protein, antimicrobial protein, hypothetical protein NCLIV_068840, thioredoxin M-type were identified in maize leaves under drought stress. These genes have putative functions in plant defense response, growth and development. These identified genes would be useful for predictive markers of plant defense, and growth responses under drought stress in plants.