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Cytotoxic Mechanism of Docosahexaenoic Acid in Human Oral Cancer Cells (인체 구강암 세포주에서 Docosahexaenoic acid에 의한 세포독성 기전)

  • Hong, Tae-Hwa;Kim, Hoon;Shin, Soyeon;Jing, Kaipeng;Jeong, Soyeon;Lim, Hyun;Yun, Donghyuk;Jeong, Ki-Eun;Lee, Myung-Ryul;Park, Jong-Il;Kweon, Gi-Ryang;Park, Seung Kiel;Hwang, Byung-Doo;Lim, Kyu
    • Journal of Life Science
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    • v.23 no.5
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    • pp.689-697
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    • 2013
  • In the United States, about 40,000 new cases of oral cancer are diagnosed each year and nearly 7,800 patients died from it in 2012. Omega-3 polyunsaturated fatty acids have been found to have anticancer effects in a variety of cancer cell lines and animal models, but their effect in oral cancer remains unclear. This study was designed to examine the effect of docosahexaenoic acid (DHA, a kind of omega-3 fatty acid) on oral cancer cells and the molecular mechanism of its action. We found that exposure of squamous cell carcinoma-4 (SCC-4) and squamous cell carcinoma-9 (SCC-9) human oral cancer cells to DHA induced growth inhibition in a dose- and time-dependent manner. Meanwhile, in addition to the elevated levels of apoptotic markers, such as cleaved PARP, subG1 portion and TUNEL-positive nuclei, DHA led to autophagic vesicle formation and an increase in autophagic flux, indicating the involvement of both apoptosis and autophagy in the inhibitory effects of DHA on oral cancer cells. Further experiments revealed that the apoptosis and autophagy induced by DHA were linked to inhibition of mammalian target of rapamycin (mTOR) signaling by AKT inhibition and AMP-activated protein kinase (AMPK) activation in SCC-9 cells. Together, our results suggest that DHA induces apoptosis- and autophagy-associated cell death through the AMPK/AKT/mTOR signaling pathway in oral cancer cells. Thus, utilization of omega-3 fatty acids may represent a promising therapeutic approach for chemoprevention and treatment of human oral cancer.

Comparative Analysis on Anti-aging, Anti-adipogenesis, and Anti-tumor Effects of Green Tea Polyphenol Epigallocatechin-3-gallate (녹차의 폴리페놀류인 에피갈로카테킨-3-갈레이트에 의한 항노화, 항비만 및 항암효과에 대한 비교 분석)

  • Lim, Eun-Ji;Kim, Min-Jae;Kim, Hyeon-Ji;Lee, Sung-Ho;Jeon, Byeong-Gyun
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1201-1211
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    • 2018
  • The study compared the anti-aging, anti-adipogenesis, and anti-tumor effects of epigallocatechin-3- gallate (EGCG) in various cancer cell lines (SNU-601, MKN74, AGS, MCF-7, U87-MG, and A-549) and normal cell lines (MRC-5 fibroblasts, dental tissue-derived mesenchymal stem cells [DSC], and 3T3-L1 pro-adipocytes). Half inhibitory concentration ($IC_{50}$) values were significantly (p<0.05) higher in normal cell lines (~50 uM), when compared to that in cancer cell lines (~10 uM). For anti-aging effects, MRC-5 and DSC were exposed to 10 uM EGCG for up to five passages that did not display any growth arrest. Population doubling time and senescence-related ${\beta}-galactosidase$ ($SA-{\beta}-gal$) activity in treated cells were similar to untreated cells. For anti-adipogenic effects, mouse 3T3-L1 pre-adipocytes were induced to adipocytes in an adipogenic differentiation medium containing 10 uM EGCG, but adipogenesis in 3T3-L1 cells was not inhibited by EGCG treatment. For anti-tumor effects, the cancer cell lines were treated with 10 uM EGCG. PDT was significantly (p<0.05) increased in EGCG-treated SNU-601, AGS, MCF-7, and U87-MG cancer cell lines, except in MKN74 and A-549. The level of telomerase activity and cell migration capacity were significantly (p<0.05) reduced, while $SA-{\beta}-gal$ activity was highly up-regulated in EGCG treated-cancer cell lines, when compared to that in untreated cancer cell lines. Our results have demonstrated that EGCG treatment induces anti-tumor effects more efficiently as noted by decreased cell proliferation, cell migration, telomerase activity, and increased $SA-{\beta}-gal$ activity than inducing anti-aging and anti-adipogenesis. Therefore, EGCG at a specific concentration can be considered for a potential anti-tumor drug.

Potentials of Synbiotics for Pediatric Nutrition and Baby Food Applications: A Review (소아 영양 및 유아식 응용을 위한 신바이오틱스의 잠재력: 총설)

  • Jung, Hoo Kil;Kim, Sun Jin;Seok, Min Jeong;Cha, Hyun Ah;Yoon, Seul Ki;Lee, Nah Hyun;Kang, Kyung Jin
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.2
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    • pp.111-118
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    • 2015
  • Probiotic, prebiotic, and synbiotic substances as well as microorganisms were added to infant formula in an attempt to influence the intestinal microflora with an aim to stimulate the growth of lactic acid bacteria, especially bifidobacteria and lactobacilli. Over the last 10 years, new synbiotic infant formulas containing probiotics and prebiotics have been proposed in order to simulate the effect of breast-feeding on the intestinal microflora. Owing to their synergistic effect, the new synbiotics are expected to be more helpful than using probiotics and prebiotics individually. Maintenance of the viability of the probiotics during food processing and the passage through the gastrointestinal tract should be the most important consideration, since a sufficient number of bacteria ($10^8cfu/g$) should reach the intended location to have a positive effect on the host. Storage conditions and the processing technology used for the manufacture of products such as infant formula adversely affect the viability of the probiotics. When an appropriate and cost-effective microencapsulation methodology using the generally recognized as safe (GRAS) status and substances with high biological value are developed, the quality of infant formulas would improve. The effect of probiotics may be called a double-effect, where one is an immunomodulatory effect, induced by live probiotics that advantageously alter the gastrointestinal microflora, and the other comprises anti-inflammatory responses elicited by dead cells. At present, a new terminology is required to define the dead microorganisms or crude microbial fractions that positively affect health. The term "paraprobiotics" (or ghost probiotics) has been proposed to define dead microbial cells (not damaged or broken) or crude cell extracts (i.e., cell extracts with complex chemical composition) that are beneficial to humans and animals when a sufficient amount is orally or topically administered. The fecal microflora of bottle-fed infants is altered when the milk-based infant formula is supplemented with probiotics or prebiotics. Thus, by increasing the proportion of beneficial bacteria such as bifidobacteria and lactobacilli, prebiotics modify the fecal microbial composition and accordingly regulate the activity of the immune system. Therefore, considerable attention has been focused on the improvement of infant formula quality such that its beneficial effects are comparable to those of human milk, using prebiotics such as inulin and oligosaccharides and potential specific probiotics such as bifidobacteria, which selectively stimulate the proliferation of beneficial bacteria in the microflora and the indigenous intestinal metabolic activity of the microflora.

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The Influence of 5-Fluorouracil Administration Mode on the Expression of Phospholipase C and Ras Oncoprotein Associated with Regeneration of Rat Intestinal Mucosa Following Radiation (방사선 조사후 백서 공장 점막의 재생과정에서 5-fluorouracil 투여가 phospholipsse C 와 ras 암유전자단백의 발현에 미치는 영향)

  • Park Kyung Ran;Lee Chung Sik;Kim Sung Sook;Lee Young Han;Ryu Sung Ho;Suh Pann-Ghill
    • Radiation Oncology Journal
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    • v.12 no.3
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    • pp.271-284
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    • 1994
  • Purpose : Phospholipase C(PLC) isozymes play significant roles in transmembrane signal transduction. PLC-${\gamma}1$ acts as the intracellular effector in signal transduction for cellular proliferation and differentiation. Ras oncoprotein is also involved in cell growth. We determined the biological significance of PLC and ras oncoprotein in regeneration following radiation and the effect of different modes of administration of 5-FU. Materials and Methods : To determine the effect of the administration mode of 5-FU on the regeneration of intestinal mucosa of rats following radiation, we compared the expression of PLC and ras oncoprotein in six groups. Group I had no treatment. Group II received radiation(8 Gy) only. Group III received radiation(8 Gy) and 5-FU(150mg/kg) continuous intravenous (iv) infusion for 12 hours. Group IV received radiation(8 Gy) and 5-FU(750mg/kg) iv bolus injection. Group V received only 5-FU(150mg/kg) continuous iv infusion for 12 hours, Group VI received only 5-FU (150mg/kg) iv bolus injection. Through immunoblotting and immunohistochemistry, we examined the expression of PLC and ras oncoprotein in rat jejunum at 96 hours after radiation or 5-FU administration and at 120 hours after radiation and 5-FU adminstration. We also investigated the histological findings using hematoxylin and eosin stain. Results : In the immunohistochemistry study, PLC-${\gamma}1$ expression was the highest in group III followed by groups II and VI in that order and was weakly positive in groups V and VI. PLC-${\gamma}1$ was hardly detected in the control group. The expression of ras oncoprotein was the same as the PLC-${\gamma}1$ expression for all groups. These results were confirmed by the histological findings regarding the mucosal regeneration. In the immunoblotting analysis, PLC-${\gamma}1$ expression was the highest in group III followed by group IV and II in that order. This difference between the immunoblotting and immunohistochemistry study was due to the high expression of PLC-${\gamma}1$ on the damaged surface epithelium rather than to its expression in the regeneration region as observed in the immunohistochemistry study for group IV. The expression of PLC-${\delta}1$ was positive only in group V and VI, which received both radiation and 5-FU, and the expression of PLC-${\beta}1$ was negligible for all groups. Conclusion : These results suggest that PLC-${\gamma}1$ mediated signal transduetion and ras oncoprotein may have a significant role in mucosal regeneration after radiation, and that continuous iv infusion of 5-FU may induce active regeneration in intestinal mucosa following radiation. In addition, the expression of PLC-${\delta}1$ in combined group of radiation and 5-FU implies that PLC-${\delta}1$ may be involved in signal transduction mediated by concerted action between radiation and 5-FU.

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The Measurements of Plasma Cytokines in Radiation-induced Pneumonitis in Lung Cancer Patients (폐암환자에서 방사선 폐렴 예측을 위한 혈장 Cytokine측정)

  • Hur Won Joo;Youn Seon Min;Lee Hyung Sik;Yang Kwang Mo;Sin Geun Ho;Son Choon Hee;Han Jin Yeong;Lee Ki Nam;Jeong Min Ho
    • Radiation Oncology Journal
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    • v.18 no.4
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    • pp.314-320
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    • 2000
  • Purpose :To investigate whether changes in plasma concentrations of transforming growth factor-$\beta$1(TGF-$\beta$1), tumor necrosis factor-alpha (TNF-$\alpha$) and interleukin-6 (IL-6) could be used to identify the development of radiation-induced pneumonitis in the lung cancer patients. Methods and Materials :Seventeen patients with lung cancer (11 NSCLC, 6 SCLC) were enrolled in a prospective study designed to evaluate clinical and molecular biologic correlation of radiation-induced pneumonitis. The study began in May 1998 and completed in July 1999. All patients were treated with radiotherapy with curative intent : 1.8 Gy per day, 5 fractions per week. Serial measurements of plasma TGF-$\beta$1, TNF-$\alpha$ and IL-6 were obtained in all patients before, weekly during radiotherapy and at each follow-up visits after completion of treatment. These measurements were quantified using enzyme linked immunosorbent assay (ELISA). All patients were evaluated for signs and symptoms of pneumonitis at each follow-up visit after completion of radiotherapy. High resolution CT (HRCT) scans were obtained when signs and symptoms of pneumonitis were developed after completion of radiotherapy. Results : Thirteen patients eventually developed signs and symptoms of clinical pneumonitis 씬file four patients did not. TGF-$\beta$ 1 levels were elevated in all 13 patients with pneumonitis, which showed characteristic pattern of elevation (38.45 ng/ml at pretreatment, 13.66 ng/ml during radiotherapy, then 60.63 ng/ml at 2-4 weeks after completion of radiotherapy). The levels of TNF- $\alpha$ and IL-6 were also elevated In the group of patients who developed pneumonitis but the pattern was not characteristic. Conclusions : Changes in plasma TGF$\beta$-1 levels before, during and after radiotherapy appears to be a useful means by which to identify patients at risk for the development of symptomatic pneumonitis. Other cytokines like TNF- $\alpha$ and IL-6 shows no meaningful changes in association with radiation pneumonitis.

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Quality Characteristics and Antioxidant Activity of Spirulina added Yogurt (스피루리나 첨가 요구르트의 품질 특성 및 항산화능)

  • Shin, Yu-Mi;Son, Chan-Wok;Sim, Hyun-Jung;Kim, Min-Hee;Kim, Mi-Yeon;Kwon, Oh-Yun;Kim, Mee-Ree
    • Korean journal of food and cookery science
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    • v.24 no.1
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    • pp.68-75
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    • 2008
  • The aim of this study was to evaluate the quality characteristics and antioxidant activity of yogurt containing spirulina. Yogurt base was prepared from skim milk added with $0.25{\sim}1%(w/v)$ spirulina powder and fermented with lactic acid bacteria (S. thermophilus : L. bulgaricus = 1 : 1) at $40^{\circ}C$ for 12 hr. Kiwi puree and oligosaccharides were then added. The addition of 1% spirulina powder stimulated the growth of lactic acid bacteria, which showed the highest viable cell count ($3.4{\times}10^9$ CFU/mL), and increased the titratable acidity (1.10%). The viscosity range of the yogurt was 6,000 to 9,000 cP, and the sugar content of the yogurt was around 18 $^{\circ}Brix$. The antioxidant activities were determined using the DPPH method, and the hydroxyl radical scavenging activity of the yogurt containing spirulina was higher than that of the control. The sensory evaluation scores for appearance, odor, taste, overall acceptability and buying intention were higher in the yogurt containing 0.25% spirulina than in the other groups. The amount of macronutrients in the yogurt containing spirulina was higher than that in the control. In addition, the amounts of micronutrients in the yogurt containing spirulina was significantly increased. According to these results, the optimum concentration of spirulina powder is around 0.25%.

Bioactive Utility of the Extracts from Rhus verniciflua Stokes (RVS) : Biological Function of the Extracts from RVS (옻나무 추출물의 생리활성 이용에 대한 연구 : 옻나무 추출물의 생물학적 기능)

  • Lim, Kye-Taek;Lee, Jeong-Chae
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.238-245
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    • 1999
  • Antioxidative effects of the water or ethanol extracts from Rhus verniciflua Stokes (RVS) were measured by protection against hydroxyl radicals in mouse brain tissue culture. In the water extracts from RVS, cell viabilities were estimated 60.0, 66.0, 72.0, 84.0 and 90.0% at addition of 1, 2, 4, 7 and $10{\mu}L$, respectively, compared with GO (20 mU/mL) alone. The cell viability in the ethanol extracts was similarly with water extracts. In the antitumor effects, the results showed that percentages of the HeLa cell death were approximately 24% for 12 hrs, 57% for 48 hrs at addition of 10%/well ethanol extracts respectively. To know inhibition of tumor growth, in vivo, mice (BALB/c) were inoculated with 0.25 mL CT-26 $(1{\times}10^6\;cells/mL)$ subcutaneously. After the generation of tumor, the results of RVS extracts (ethanol, water) injection showed generally that the tumor size in BALB/c was reduced. For physicochemical characterization of the RVS extracts, purified substances of water or ethanol extracts were analized with SDS-PAGE and ICP spectrometer. In electrophoresis, gel showed 2 bands (210, 230 KDa). The results of ICP verified that RVS extracts contain $Cu^{2+}$ in both samples. Conclusively, this substance might be a laccase which has a biological effective function, as a natural bioactive substance.

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Influences of Sowing Time and Nursery Period on Growth and Yield of Perilla frutescens BRITTON var. acuta KUDO (파종기(播種期) 및 육묘기간(育苗期間)이 자소(紫蘇)의 생육(生育) 및 수량(收量)에 미치는 영향(影響))

  • Park, Hi-Jin;Chung, Dong-Hee;Kim, Sang-Gon;Kwon, Byung-Sun
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.1
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    • pp.1-4
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    • 1995
  • To determine the optimum sowing time and nursery period in Perilla frutescens in the southern areas of Korea, perilla frutescens cv. red Perilla local cultivar was grown under three different sowing dates and nursery periods. The blooming period of the area which was sown on the seedbed in Apr.10 and carried out the growing seedling in 30days is Aug. 12 and it is two days earlier than that of the area, Aug.14, sown on the seedbed in Apr. 20 and carried out the growing seedling in 30days of the same month and it is also six days earlier than that of the area, Aug.18, sown on the seedbed in April 30 and conducted the growing seedling in the same date. The stem lengths are 135cm,131cm and 125cm respectively and the number of branches are 26.4, 25.3 and 23.6 respectively. The fresh weight of stem and leaf at the area sown on the seedbed in Apr.10 and conducted the growing seedling in 30days with the width of leaf over 5cm in the middle of Aug. and at the beginning of Sep. is 2,476kg/10a and it shows more increase of 172kg than that of fresh weight of stem and leaf with 2,304kg/10a which was sown on the seedbed in Apr.20 and earned out the growing seedling in 30days, and it also shows more increase of 411kg than that of fresh weight of stem and leaf with 2,065kg/10a at the area sown on the seedbed in April 30 and con­ducted the growing seedling in the same date, The fresh weight of seeds are 609.5kg/10a,509.3kg/10a and 463.2kg/10a respectively and $100.2{\sim}146.3kg$ is more increased. Therefore, the seedling period of the proper seedbed for high yield bumper crop of perilla frutescens for exporting to Japan is April 10 and the number of days for seedling is 30 days.

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Growth and electrical properties of $Sr_2$$({Ta_{1-x}},{Nb_x})_2$)$O_7$ thin films by RF sputtering (RF Sputtering을 이용한 $Sr_2$$({Ta_{1-x}},{Nb_x})_2$)$O_7$ 박막의 성장 및 전기적 특성)

  • In, Seung-Jin;Choi, Hoon-Sang;Lee, Kwan;Choi, In-Hoon
    • Korean Journal of Materials Research
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    • v.11 no.5
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    • pp.367-371
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    • 2001
  • In this paper, theS $r_2$(T $a_{1-x}$ , N $b_{x}$)$_2$ $O_{7}$(STNO) films among ferroelectric materials having a low dielectric constant for metal-ferroelectric-semiconductor field effect transistor(MFS-FET) were discussed. The STNO thin films were deposited on p-type Si(100) at room temperature by co-sputtering with S $r_2$N $b_2$ $O_{7(SNO)}$ ceramic target and T $a_2$ $O_{5}$ ceramic target. The composition of STNO thin films was varied by adjusting the power ratios of SNO target and T $a_2$ $O_{5}$ target. The STNO films were annealed at 8$50^{\circ}C$, 90$0^{\circ}C$ and 9$50^{\circ}C$ temperature in oxygen ambient for 1 hour. The value of x has significantly influenced the structure and electrical properties of the STNO films. In the case of x= 0.4, the crystallinity of the STNO films annealed at 9$50^{\circ}C$ was observed well and the memory windows of the Pt/STNO/Si structure were 0.5-8.3 V at applied voltage of 3-9 V and leakage current density was 7.9$\times$10$_{08}$A/$\textrm{cm}^2$ at applied voltage of -5V.of -5V.V.V.

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Mutational Analysis of MECP2 Gene in 34 Rett Syndrome (Rett 증후군 34례의 MECP2 유전자 변이에 관한 연구)

  • Park, Sang Jo;Hwang, Tae Gyu;Son, Byeong Hee;Kim, Chul Min
    • Clinical and Experimental Pediatrics
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    • v.45 no.10
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    • pp.1263-1272
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    • 2002
  • Purpose : Rett syndrome(RTT) is an X-linked dominant neurodevelopmental disorder affecting 1 per 10,000-15,000 female births worldwide. It was initially described by Andreas Rett in 1966. RTT involves developmental regression characterized stereotypic hand movements, tremors, gait apraxia, seizures, deceleration of head growth after the age of 6-18 months. The disease-causing gene was identified as MECP2 on chromosome Xq28. We carried out mutational analysis of MECP2 genes in RTT patients. Methods : Whole blood(5 cc) of 34 sporadic RTT patients was collected in EDTA-anticoagulated tubes. Genomic DNA was extracted from peripheral blood using the E.Z.N.A. blood DNA kit. Four exons of the MECP2 gene were amplified by PCR in 34 Korean with RTT. We carried out PCR divided the exon three into two parts and the exon four into five parts. Primer sequences designed by Amir et al. in 1999 were almost used(AF030876). Sequencing primers used were the same as PCR. DNA sequencing reactions were performed using an ABI 377 DNA sequencer and ABI PRISM dye terminator cycle sequencing reaction kit(Perkin-elmer). The results were compared with the normal DNA sequence(X99686). To confirm the change of sequence on novel mutations, RFLP analysis was performed. Results : The MECP2 mutations were detected in 23(67.6%) of the 34 patients. The mutations consisted of 12 different types including nine missense and three nonsense mutations. Of these, three (L100V, G161E and T311M) mutations were newly identified. Most of the mutations discovered are located within MBD(39.1%) and TRD(39.1%). In this study, three(T158M, R270X, R306C) mutations were identified high frequency. Conclusion : MECP2 gene was also an important cause of Korean RTT patients. MECP2 gene study is an important tool for diagnosis of Korean RTT patients.