• Asian Pacific Journal of Cancer Prevention
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• v.14 no.9
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• pp.5467-5472
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• 2013

Objective: To construct short hairpin RNA (shRNA) eukaryotic expression vectors targeting Livin and Survivin genes, and to explore the impact of co-transfection of Livin and Survivin shRNA expression vectors on the biological behavior of HepG2 cells. Methods: shRNA eukaryotic expression vectors pSD11-Livin and pSD11-Survivin were designed and constructed then transfected into HepG2 cells separately or in combination. mRNA and protein expression in transfected cells was assessed by quantitative fluorescence PCR and Western blotting, respectively. Cell proliferation was measured by MTT assay and cell apoptosis by TUNEL assay. Results: The Livin and Survivin shRNA eukaryotic expression vectors were successfully constructed and transfected into HepG2 cells. The relative mRNA expression levels of Livin and Survivin in HepG2 cells co-transfected with pSD11-Livin and pSD11-Survivin were $0.12{\pm}0.02$ and $0.33{\pm}0.13$, respectively, which was significantly lower than levels in cells transfected with either pSD11-Livin or pSD11-Survivin (P<0.05). The relative protein expression levels of Livin and Survivin in the co-transfected cells were also significantly decreased compared to single-transfection (P<0.05). The inhibition rate of cell growth in the co-transfection group was higher than that in the single-transfection groups at 48 h, 60 h, or 72 h after transfection (P<0.01). The apoptotic rate increased to the greatest extent in the co-transfection group relative to any other group (P<0.05). Conclusions: Co-transfection with pSD11-Livin and pSD11-Survivin was more efficient than transfection with either vector alone in reducing the mRNA and protein expression of Livin and Survivin genes in HepG2 cells. Co-transfection also inhibited the proliferation of transfected cells more than the other groups, and induced cellular apoptosis more effectively.

• Proceedings of the Korean Vacuum Society Conference
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• 2014.02a
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• pp.218.1-218.1
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• 2014

In order to measure the absolute plasma density, various probes are proposed and investigated and microwave probes are widely used for its advantages (Insensitivity to thin non-conducting material deposited by processing plasmas, High reliability, Simple process for determination of plasma density, no complicate assumptions and so forth). There are representative microwave probes such as the cutoff probe, the hairpin probe, the impedance probe, the absorption probe and the plasma transmission probe. These probes utilize the microwave interactions with the plasma-sheath and inserted structure (probe), but frequency range used by each probe and specific mechanisms for determining the plasma density for each probe are different. In the recent studies, behaviors of each microwave probe with respect to the plasma parameters of the plasma density, the pressure (the collision frequency), and the sheath width is abundant and reasonably investigated, whereas relative diagnostic characteristics of the probes by a comparative study is insufficient in spite of importance for comprehensive applications of the probes. However, experimental comparative study suffers from spatially different plasma characteristics in the same discharge chamber, a low-reproducibility of ignited plasma for an uncertainty in external discharge parameters (the power, the pressure, the flow rate and so forth), impossibility of independently control of the density, the pressure, and the sheath width as well as expensive and complicate experimental setup. In this paper, various microwave probes are simulated by finite-different time-domain simulation and the error between the input plasma density in FDTD simulations and the measured that by the unique microwave spectrums of each probe is obtained under possible conditions of plasma density, pressure, and sheath width for general low-temperature plasmas. This result shows that the each probe has an optimum applicable plasma condition and reliability of plasma density measurement using the microwave probes can be improved by the complementary use of each probe.

• Molecules and Cells
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• v.39 no.9
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• pp.669-679
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• 2016

N-acetyl-D-glucosamine kinase (GlcNAc kinase or NAGK) primarily catalyzes phosphoryl transfer to GlcNAc during amino sugar metabolism. Recently, it was shown NAGK interacts with dynein light chain roadblock type 1 (DYNLRB1) and upregulates axo-dendritic growth, which is an enzyme activity-independent, non-canonical structural role. The authors examined the distributions of NAGK and NAGK-dynein complexes during the cell cycle in HEK293T cells. NAGK was expressed throughout different stages of cell division and immunocytochemistry (ICC) showed NAGK was localized at nuclear envelope, spindle microtubules (MTs), and kinetochores (KTs). A proximity ligation assay (PLA) for NAGK and DYNLRB1 revealed NAGK-dynein complex on nuclear envelopes in prophase cells and on chromosomes in metaphase cells. NAGK-DYNLRB1 PLA followed by Lis1/NudE1 immunostaining showed NAGK-dynein complexes were colocalized with Lis1 and NudE1 signals, and PLA for NAGK-Lis1 showed similar signal patterns, suggesting a functional link between NAGK and dynein-Lis1 complex. Subsequently, NAGK-dynein complexes were found in KTs and on nuclear membranes where KTs were marked with CENP-B ICC and nuclear membrane with lamin ICC. Furthermore, knockdown of NAGK by small hairpin (sh) RNA was found to delay cell division. These results indicate that the NAGK-dynein interaction with the involvements of Lis1 and NudE1 plays an important role in prophase nuclear envelope breakdown (NEB) and metaphase MT-KT attachment during eukaryotic cell division.

• Journal of the Korean Society of Clothing and Textiles
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• v.32 no.8
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• pp.1322-1331
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• 2008

The purpose of this study is to design an official costume of Joseon Ginyeo in the scholar’s banquet in 16th century, thereby making correct historical animation, drama costumes, and various costume contents. Every Ginyeo in and wore a long yellow coat with short front edge. 'Dan-ei', Ginyeo’s court costume, had very similar shape except color and collar, therefore the size of Dan-ei was adapted to design a yellow coat for drama actress. Ginyeo’s Jogori in these paintings was wide and long enough to cover undervelly under the edge of yellow coat, but It was designed more slim and long for better appearence and comfortable movement in my work. A skirt of Ginyeo was designed with circumference of about 360cm and length of about 105cm according as a skirt style of Joseon ladies in 16th century. Ginyeo's Jam(hairpin) was consist of a vase, a plum blossom, and a lotus pattern. And, a wide underpants with a single suspender, portable pouch, Un-hae(women’s shoes) with low heel are needed for drama actress in the role of Ginyeo. In this design, it is very important to save an estimate and time, select suitable mordern fabrics, reform discomportable points, express a charater’s role, and make a trendy appearence.

• Molecules and Cells
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• v.19 no.1
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• pp.104-113
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• 2005

Large inversions are well characterized in the chloroplast genomes of land plants. In contrast, reports of small inversions are rare and involve limited plant groups. In this study, we report the widespread occurrence of small inversions ranging from 5 to 50 bp in fully and partially sequenced chloroplast genomes of both monocots and dicots. We found that small inversions were much more common than large inversions. The small inversions were scattered over the chloroplast genome including the IR, SSC, and LSC regions. Several small inversions were uncovered in chloroplast genomes even though they shared the same overall gene order. The majority of these small inversions were located within 100 bp downstream of the 3' ends of genes. All had inverted repeat sequences, ranging from 11 to 24 bp, at their ends. Such small inversions form stem-loop hairpin structures that usually have the function of stabilizing the corresponding mRNA molecules. Intra-molecular recombination between the inverted sequences in the stem-forming regions are responsible for generating flip-flop orientations of the loops. The presence of two different orientations of the stem-loop in the trnL-F noncoding region of a single species of Jasminum elegans suggests that a short inversion can be generated within a short period of time. Small inversions of non-coding sequences may influence sequence alignment and character interpretation in phylogeny reconstructions, as shown in nine species of Jasminum. Many small inversions may have been generated by parallel or back mutation events during chloroplast genome evolution. Our data indicate that caution is needed when using chloroplast non-coding sequences for phylogenetic analysis.

• BMB Reports
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• v.41 no.11
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• pp.765-770
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• 2008

Undesirable hyperpigmentation that can arise from increased melanocyte activity may be alleviated by targeting active melanocytes for apoptosis. The role of Stathmin 1 as an important regulator of microtubule dynamics is well documented. The current study examined the potential of Stathmin 1-targeting strategies in eliminating active melanocytes. A vector to overexpress Stathmin 1 and vectors to express three distinct small hairpin RNAs to knockdown Stathmin 1 expression in normal melanocytes were produced and in cell cultures acted accordingly. Both overexpression and knockdown of Stathmin 1 led to a marked increase in melanocyte apoptosis, as indicated by the accumulation of apoptotic cells and increased levels of cleaved caspase-3. Both up- and down-regulation of Stathmin 1 expression inhibited the activity of differentiated melanocytes, as indicated by decreases in both melanin production and tyrosinase activity. Taken together, these results indicate that hyperactive melanocytes can be inhibited by altering Stathmin 1 expression.

• The Journal of Natural Sciences
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• v.14 no.1
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• pp.39-50
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• 2004

T7 RNA polymerase is a single subunit RNA polymerase able to accomplish whole transcription process without auxiliary factors. In order to study transcription elongation mechanism of phage T7 RNA polymerse, stepwise walking of RNA polymerase was established by immobilizing biotinylated DNA template with streptavidin bead, series of active and stable elongation complexes were obtained, Transcripts were radio isotope labeled at the 16thm 17th and 18th nucleotide residues so stable elongation transcription complex of T7 RNA polymerase containing 22-40 nucleotide residues could be identified. We identified the positions of stablely formed transcription elongation complexes of termination site in intrinsic hairpin-independent PTH terminator sequence through the established stepwise walking of wild-type of mutant R173C T7 RNA polymerases. The results suggest that stable elongation transcription complexes were at the site of passing PTH terminator signal by mutant R173C RNA polymerase.

• Molecules and Cells
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• v.38 no.10
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• pp.876-885
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• 2015

N-acetyl-D-glucosamine kinase (NAGK) plays an enzyme activity-independent, non-canonical role in the dendritogenesis of hippocampal neurons in culture. In this study, we investigated its role in axonal development. We found NAGK was distributed throughout neurons until developmental stage 3 (axonal outgrowth), and that its axonal expression remarkably decreased during stage 4 (dendritic outgrowth) and became negligible in stage 5 (mature). Immunocytochemistry (ICC) showed colocalization of NAGK with tubulin in hippocampal neurons and with Golgi in somata, dendrites, and nascent axons. A proximity ligation assay (PLA) for NAGK and Golgi marker protein followed by ICC for tubulin or dynein light chain roadblock type 1 (DYNLRB1) in stage 3 neurons showed NAGK-Golgi complex colocalized with DYNLRB1 at the tips of microtubule (MT) fibers in axonal growth cones and in somatodendritic areas. PLAs for NAGK-dynein combined with tubulin or Golgi ICC showed similar signal patterns, indicating a three way interaction between NAGK, dynein, and Golgi in growing axons. In addition, overexpression of the NAGK gene and of kinase mutant NAGK genes increased axonal lengths, and knockdown of NAGK by small hairpin (sh) RNA reduced axonal lengths; suggesting a structural role for NAGK in axonal growth. Finally, transfection of 'DYNLRB1 (74-96)', a small peptide derived from DYNLRB1's C-terminal, which binds with NAGK, resulted in neurons with shorter axons in culture. The authors suggest a NAGK-dynein-Golgi tripartite interaction in growing axons is instrumental during early axonal development.

• The KIPS Transactions:PartB
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• v.14B no.6
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• pp.461-472
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• 2007

The emphasis in genome projects has Been moving towards the sequence analysis in order to extract biological "meaning"(e.g., evolutionary history of particular molecules or their functions) from the sequence. Especially. palindromic or direct repeats that appear in a sequence have a biophysical meaning and the problem is to recognize interesting patterns and configurations of words(strings of characters) over complementary alphabets. In this paper, we propose an algorithm to identify variable length palindromic pairs(longer than a threshold), where we can allow gaps(distance between words). The algorithm is called palindrome algorithm(PA) and has O(N) time complexity. A palindromic pair consists of a hairpin structure. By composing collected palindromic pairs we build n-pair palindromic patterns. In addition, we dot some of the longest pairs in a circle to represent the structure of a DNA sequence. We run the algorithm over several selected genomes and the results of E.coli K12 are presented. There existed very long palindromic pair patterns in the genomes, which hardly occur in a random sequence.

• Experimental and Molecular Medicine
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• v.50 no.12
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• pp.4.1-4.19
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• 2018

Transforming growth factor $(TGF)-{\beta}$ signaling is increasingly recognized as a key driver in cancer. In progressive cancer tissues, $TGF-{\beta}$ promotes tumor formation, and its increased expression often correlates with cancer malignancy. In this study, we utilized adenoviruses expressing short hairpin RNAs against $TGF-{\beta}1$ and $TGF-{\beta}2$ to investigate the role of $TGF-{\beta}$ downregulation in cancer cell death. We found that the downregulation of $TGF-{\beta}$ increased the phosphorylation of several SAPKs, such as p38 and JNK. Moreover, reactive oxygen species (ROS) production was also increased by $TGF-{\beta}$ downregulation, which triggered Akt inactivation and NOX4 increase-derived ROS in a cancer cell-type-specific manner. We also revealed the possibility of substantial gene fluctuation in response to $TGF-{\beta}$ downregulation related to SAPKs. The expression levels of Trx and GSTM1, which encode inhibitory proteins that bind to ASK1, were reduced, likely a result of the altered translocation of Smad complex proteins rather than from ROS production. Instead, both ROS and ROS-mediated ER stress were responsible for the decrease in interactions between ASK1 and Trx or GSTM1. Through these pathways, ASK1 was activated and induced cytotoxic tumor cell death via p38/JNK activation and (or) induction of ER stress.