• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.446-454
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• 2017

Human papillomaviruses (HPVs) are major causes of cervical cancer. Sixteen high risk HPVs, including HPV 16, HPV 18, HPV31, HPV 33, HPV 35, HPV 39, HPV 45, HPV 51, HPV 52, HPV 53, HPV 56, HPV 58, HPV 59, HPV 66, HPV 68, and HPV 69 are found in cervical cancer. HPVs 16 and 18 are mainly presented in 70% of cervical cancer. Therefore, identifying the presence of these high-risk HPVs is crucial. The objective of this study is to establish the HPV ViroCheck for detecting 16 HR-HPVs and genotypes of HPVs 16 and 18, as well as to analyze the analytical performance of HPV ViroCheck. We performed the analytical sensitivity of HPV E6 / E7 genes of 16 high risk HPVs to confirm the limit of detection. Then, a cross reactivity of HPV ViroCheck with microorganisms and viruses related to the cervix were analyzed for analytical specificity. Analytical sensitivity of high risk HPV genotypes ranged from 1 to 100 copies when using cloned DNAs. The limit of detection was 10 cells for both SiHa and HeLa cells. Cervical-related microorganisms and viruses did not show cross-reactivity to HPV DNA. Moreover, the intra- and inter-assay coefficient variations (CVs) were below 5%. In conclusion, HPV Virocheck will be useful for the detection of 16 HR HPVs, as well as HPV 16 and HPV 18 genotypes rapidly.

• Korean Journal of Medicinal Crop Science
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• v.9 no.2
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• pp.156-165
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• 2001

Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

• Korean Journal of Microbiology
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• v.39 no.4
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• pp.260-266
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• 2003

In this study bacterial community was analyzed to evaluate the impacts of long-term use of agricultural chemicals on a soil ecosystem as well as to obtain fundamental data on the relationship. Sequences of 16S rRNA clones from a non-agricultural site and a tangerine orchard soil which has a history of long-term use of agricultural chemicals over 30 years were analyzed. This revealed that bacterial community containing 5 divisions and 18 genera was distributed in a tangerine orchard soil, while bacterial community containing 9 divisions and 44 genera was distributed. In a tangerine orchard soil site, the most abundant bacteria in subdivision level were placed into Proteobacteria γ group which occupied 56% of total clones. The other bacterial clones from the ocrhcard soil exposed to agricultural chemicals over 30 years were Acidobacteria group (25%), Fimicutes group (5%), Planctomycetes group (2%), Proteobacteria α (1%), δ group (1%), and Cyanobacteria group (1%). Whereas, the clones were from the non-agricultural site were distributed among the division or subdivision Acidobacteria group (14%), Planctomycetes group (13%), Proteobacteria α (10%), β (9%), δ (9%), Fimicutes group (8%), Verrucomicrobia group (8%), Actinobacteria group (6%), Proteobacteria γ group (3%), Bacteroidetes group (3%), Gemmatimonadetes group (3%), and Cyanobacteria group (1%). This finding suggests the possibility that long-term application of agricultural chemicals or fertilizers on a tangerine orchard might result in drastic reduction or alteration in the composition of the bacterial community in the contaminated soil site.

• Korean Journal of Head & Neck Oncology
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• v.14 no.2
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• pp.236-243
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• 1998

Objectives: Local invasion of the thyroid cancer that is invasion of the upper aerodigestive tract, neurovascular structures of the neck and superior mediastinum, is infrequent and comprises of 1-16% of well-differentiated thyroid cancer. However the proximity of the thyroid gland to these structures provides the means for an invasive cancer to gain ready access into theses structures and when invasion occurs, it is the source of significant morbidity and mortality. So locally invasive thyroid cancer should be removed as much as possible, but still much debates have been exist whether the surgical method should be radical or conservative. This study was desinged to evaluate the clinical characteristics and the surgical treatment of the locally invasive thyroid cancer. Material and Methods: At the department of otorhinolaryngology of Hallym university, 10 patients diagnosed as locally invasive thyroid cancer among the 81 patients treated for thyroid cancer between 1991 to 1997 were retrospectively evaluated. Results: Of the 10 patients, 3 patients had histories of previous surgical treatment with or without radiation or radioactive iodine therapy. The site of invasion of thyroid cancer were trachea(7 cases), recurrent laryngeal nerve(5 cases), mediastinal node(5 cases), esophagus(3cases), larynx(3cases), carotid artery(3 cases), pharynx(l case), and other sites(4 cases). The operation techniques included 1 partial laryngectomy and 1 partial cricoid resection, 2 shavings and 3 window resections of the trachea, 1 sleeve resection of the trachea with end-to-end anastomosis and 1 cricotracheoplasty for tracheal invasion, 2 shavings and 1 partial esophagectomies for esophageal invasion, and 1 wall shaving and 2 partial resections with $Gortex^{\circledR}$ tube reconstruction for carotid artery invasion, and so on. Conclusions: These data and review of literature suggest that the surgical method should be perfomed on the basis of individual condition and complete removal of all gross tumor with preservation of vital structures whenever possible will offer a good result.

• Microbiology and Biotechnology Letters
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• v.33 no.2
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• pp.96-105
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• 2005

For screening of autoregulator receptor gene from Streptomyces longwoodensis, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHIsite of pUC19 and transformed into the E. coli $DH5{\alpha}$. The isolated plasmid from transformant contained the fragment of 100 bp, which was detected on $2\%$ gel after BamHI treatment. The insert, 100 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridizations with the 100 bp fragment as a probe allowed to select a genomic clone of S. longwoodensis, pSLT harboring a 4.4 kb SphI fragment. Nucleotide sequencing analyses revealed a 651 bp open reading frame(ORF) were isolated protein showing moderate homology ($35{\sim}46\%$) with the ${\Gamma}$-butyrolactone autoregulator receptors from Streptomyces sp., and this ORF was named sltR The sltR/pET-17b plasmid was constructed to overexpress the recombinant SltR protein (rSltR) in E. coli BL21 (DE3)/pLysS, and the rSltR protein was purified to homogeneity by DEAE-Sephacel column chromatography, and DEAE-5PW chromatography (HPLC). The molecular mass of the purified rSltR protein was 55 kDa by HPLC gel-filtration chromatography and 28 kDa by SDS-PAGE, indicating that the rSltR protein is present as a dimer. A binding assay with tritium-labeled autoregulators revealed that the rSltR has clear binding activity with a A-factor type autoregulator as the most effective ligand.

• Journal of Life Science
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• v.16 no.6
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• pp.904-910
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• 2006

Dendritic cells (DCs) are the only antigen presenting cells (APCs) capable of initiating immune responses, which is crucial for priming the specific cytotoxic T lymphocyte (CTL) response and tumor immunity. Upon activation by DCs, CD4+ helper T cells can cross-prime CD8+ CTLs via IL-12. However, recently activated DCs were described to prime in vitro strong T helper cell type 1 $(Th_1)$ responses, whereas at later time points, they preferentially prime $Th_2$ cells. Therfore, we examined in this study the optimum kinetic state of DCs activation impacted on in vivo priming of tumor-specific CTLs by using ovalbumin (OVA) tumor antigen model. Bone-marrow-derived DCs showed an appropriate expression of surface MHC and costimulatory molecules after 6 or 7-day differentiation. The 6-day differentiated DCs pulsed with OVA antigen for 8 h (8-h DC) and followed by restimulation with LPS for 24 h maintained high interleukin (IL)-12 production potential, accompanying the decreased level in their secretion by delayed re-exposure time to LPS. Furthermore, immunization with 8-h DC induced higher intracellular $interferon(IFN)-{\gamma}+/CD8+T$ cells and elicited more powerful cytotoxicity of splenocytes to EG7 cells, a clone of EL4 cells transfected with an OVA cDNA, than immunization with 24-h DC. In the animal study for the evaluation of therapeutic or protective antitumor immunity, immunization with 8-h DC induced an effective antitumor immunity against tumor of EG7 cells and completely protected mice from tumor formation and prolonged survival, respectively. The most commonly used and clinically applied DC-based vaccine is based on in vitro antigen loading for 24 h. However, our data indicated that antigen stimulation over 8 h decreased antitumor immunity with functional exhaustion of DCs, and that the 8-h DC would be an optimum activation state impacted on in vivo priming of tumor-specific CTLs and subsequently lead to induction of strong antitumor immunity.

• Clinical and Experimental Pediatrics
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• v.49 no.2
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• pp.150-156
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• 2006

Purpose : The aims of this study included assessment of molecular-epidemiologic features during an outbreak of colonization of extended spectrum ${\beta}$-lactamase producing Klebsiella pneumoniae(ESBL-KPN) and re-evaluation of their colonized status one year later. Methods : Rectal swab cultures for ESBL-KPN from all hospitalized infants and newly admitted infants were obtained during the outbreak of colonization from July to December, 2000. The pattern of XbaI-digested chromosomal DNA of isolates were analyzed by pulsed-field gel electrophoresis. Weekly rectal swab cultures were obtained during the outbreak until patients were either discharged or decolonized. Patients discharged after being colonized had follow up stool cultures a year later. Results : A total of 80 patients(28.5 percent) were colonized. Of those, 53 whose pulsed-field gel electrophoresis(PFGE) was possible only once, were ESBL-KPN grouped into six cluster clones and 10 single clones : 28 patients(52.8 percent) were colonized with type A, the most common clone, followed by type B in 11 patients(20.8 percent). Of those 12 patients in whom serial PFGE was done more than twice, type A was predominant. Narrowed-down in strains occurred from types A, B, C, D and three single clones at initiation of the study into types A and type B after three months of strict infection control. Among 75 patients(93.7 percent) who were sent home after being colonized, 30 patients were re-called for stool cultures a year later : All of them were decolonized. Conclusion : This study demonstrates the importance of infection control as the diversity of ESBL-KPN strains could be narrowed into fewer strains. Colonization of ESBL-KPN could be reversed upon return to the community.

• Journal of Korean Society of Forest Science
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• v.104 no.2
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• pp.187-192
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• 2015

Individuals in the population under a particular environmental condition influencing recurrently for a long time could locally adapted and local adaptation is of a fundamental importance in a breeding program, conservation activities of genetic resources or evolutionary biology. Plants at northern range limits have higher probability of expressing an adaptative genetic trait. The natural community at the northern range limit of Ilex cornuta (Chinese holly) in Buan, Jeollanam-do in Korea was composed of adults of 744 and seedlings of 211 (hereafter Community) and is designated as the Korean Natural Monument (No. 122) by the law. At two adjacent areas to Community, 85 (hereafter Plantation I) and 27 hollies (hereafter Plantation II) were planted respectively for preparations of the next generation. Eighty-five trees were sampled for genetic analysis in the three groups. Fifty-two (36%) of the total 143 amplicons were polymorphic from four AFLP primer combinations. A total of thirteen genotypes was identified and just one genotype was for 52 trees of Community. Seven and five genotypes were shown for Plantation I and II, respectively. There was no identical genotype between Community and Plantation (I or II) or between two plantation groups. Number of private loci was 2 for Community, 6 for Plantation I and 4 for Plantation II. We presumed their genetic backgrounds were quite different with one another and the plantation groups were made independently because they were different not only the genetic compositions but also their ages. Considering the genetic monomorphism by AFLP markers, observations of only male trees and asexual propagation as layerage or cuttage, the hollies in Community might be a genet by root suckering from a single male tree, not the results of selective removal of female trees for ornamental use in the past.

• Asian Journal of Turfgrass Science
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• v.1 no.1
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• pp.7-17
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• 1987

The experiment with two levels of nitrogen (0. and 300kg / ha / year) and two levels of clipping height (1.5cm and 4cm) was conducted on the field during the period 3 June to 23 October 1985. Clonal lines of korean lawngrass ( Zoysia japonica Steud.) and manilagrass ( Zoysia matrella ( L.) Merr.)of Daejon origin were established in June, as individual clone in rows 30cm apart with a 40cm spacing between clones, actually 4 clones each plot. The results obtained were as follows : 1. When no nitrogen was applied to korean lawngrass, leaf blade which appeared during the August / early September period remained green for a period of about 10 weeks and even leaves emerged in late September lived for 42 days. However, leaf longevity did not exceed 8 weeks as nitrogen was applied. In contrast the leaf longevity of manilagrass which emerged during the mid - August / early September period was 11 weeks and, under the nitrogen applied, 9 weeks, indicating that the life - saen of individual leaf of manilagrass may be longer than that of korean lawngrass. Meanwhile, clipping height had no effect on the leaf longevity in both grasses. 2. During the July / August period, tiller number, green leaf number and DM weight of korean lawngrass were increased significantly with fertilizer nitrogen, but were not with two levels of clipping height. This trend was reversed after late September : no effect of nitrogen was appeared. Instead, lax clipping increased tiller number, green leaf number and DM weight. Green leaves stimulated by lax clipping resulted in the occurrance of more dead leaves in late October. 3. The increase of tiller number, green leaf number, and DM weight of korean lawngrass due to nitrogen application appeared to be of significance in early September. Unlike korean lawngrass, however, this significant increase was maintained to late October when new green leaves still emerge. Clipping height had little effect on the growth of manilagrass by early September, but since then, lax clipping stimulated leaf appearance, possibly resulting in a remained green color of manilagrass turf. 4. Among the stolons outgrown until early September, the primary stolon was not influenced by nitrogen and clipping treatments to produce only 2 - 3 stolons. However, 1st branch stolon as affected by nitrogen increased significantly, so most of stolons which occurred consisted of 1st branch stolon. 5. Until early September, stolon length obtained at nil nitrogen level was chiefly caused by lengthening the primary stolons. By applying nitrogen the primary stolons of korean lawngrass was longer than 1st branch stolons when severe clipping was involved and in turn, shorter than 1st branch stolons when lax clipping was concerned. In manilagrass, 1st branch stolons were much longer than the primary stolons when turf was clipped severely but in conditions of lax clipping, there was little difference in length between primary and 1st branch stolons. 6. Stolon nodes of both korean lawngrass and manilagrass were positively influenced by nitrogen, but no particular increases by imposing clipping height treatment was marked in manilagrass. Although the stolon of korean lawngrass was grown until late october, the growth stimulated by nitrogen was not so remarkable as to exceed that a by nil N. 7. The thickness of korean lawngrass and manilagrass was greatest in late September, but that of manilagrass did not differ significantly from that in late October. 8. The response of stolon length of korean lawngrass to lax clippings was not so great in late October as to that to severe clippings. On the other hand, the positive effect of lax clippings to stolon length in m anilagrass was confirmed even in late October.

• Journal of Gastric Cancer
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• v.8 no.4
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• pp.169-175
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• 2008

Purpose: The main target of 5-fluorouracil (5-FU) is thymidylate synthase (TS). A high TS expression has been identified as promoting resistance to 5-FU. For colorectal cancers, the response to 5-FU based adjuvant chemotherapy is different according to the microsatellite instability (MSI) status. The reports on the relationship between MSI and the TS expression in colorectal cancer have been inconsistent. No data is available for gastric cancer regarding the relationship between MSI and the TS expression. Therefore, we studied the relationship between MSI and the TS expression in gastric cancer. Materials and Methods: Ninety-nine consecutive patients who underwent radical gastrectomy for gastric cancer from January 2004 to May 2006 at Bundang CHA hospital were studied. MSI was assessed for five markers (BAT25, BAT26, D2S123, D5S346, and D17S250) by PCR and with using an ABI prism 3100 Genetic analyzer. The TS expression was analyzed by immunohistochemistry with using mouse anti-thymidylate synthase monoclonal antibody to the TS 106 clone. Results: Out of the ninety-nine patients, MSS/MSI-L was detected in 92 (92.1%) cases and 7 cases (7.1%) were MSI-H. A negative TS expression was found in 46 (46.5%) cases, a low TS expression was found in 33 (33.3%) and a high TS expression was found in 20 (20.2%). Out of 92 MSS/MSI-L patients, the number of patients with negative, low and high TS expressions were 46 (50%), 30 (32.6%) and 16 (17.4%), respectively. Out of the 7 MSI-H patients, the number of patients with negative, low and high TS expressions were 0 (0%), 3 (42.9%) and 4 (57.1%), respectively. The relationship between MSI-H and a high TS expression was statistically significant. Conclusion: Gastric cancer with MSI-H showed higher levels of a TS expression compared to the gastric cancer with MSS/MSI-L.