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The Effect of Particulate Matter 10 from Asian Dust on the Production of Reactive Oxygen Species, TGF-β, NF-κB, PDGF-α and Fibronectin in MRC-5 Fibroblast Cells (폐 섬유모세포에서 황사의 미세먼지(Particulate Matter 10)가 활성산소족과 TGF-β, NF-κB, PDGF-α, Fibronectin의 생성에 미치는 영향)

  • Kim, Ah Hyun;Chon, Suyeon;Yoon, Jin Young;Kim, Yu Jin;Kyung, Sun Young;Lee, Sang Pyo;Park, Jeong Woong;Jeong, Sung Hwan
    • Tuberculosis and Respiratory Diseases
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    • v.67 no.6
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    • pp.528-535
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    • 2009
  • Background: Dust clouds blown by the wind from the arid deserts of Mongolia and Northeast China are known as Asian dust storms. Ambient particulate matter with a diameter <10 ${\mu}m$ ($PM_{10}$) is associated with the exacerbation of respiratory diseases and increased mortality of heart and lung disease patients. The fibrotic effects of $PM_{10}$ of Asian dust to pulmonary fibroblast cells are unknown. This study examined the production of reactive oxygen species (ROS), TGF-${\beta}$, NF-${\kappa}B$, PDGF-$\alpha$ and Fibronectin in fibroblasts exposed to Asian dust particles. Methods: Air samples were collected using a high volume air sampler (Sibata model HV500F) with an air flow of 500 L/min for at least 6 hours. The MRC-5 cells were exposed to 0, 50 and 100 ${\mu}g/mL$ of $PM_{10}$ for 24 hours. ROS was detected by measuring the level of oxidized DCF using FACS. TGF-$\beta$, NF-${\kappa}B$, PDGF-$\alpha$ and fibronectin were detected by western blotting. Results: There was no increase in the ROS, TGF-$\beta$ and PDGF-$\alpha$ levels in the MRC-5 cells exposed to $PM_{10}$. The NF-${\kappa}B$ level was higher in the MRC-5 cells exposed to 50 and 100 ${\mu}g/mL$ of $PM_{10}$ for 24 hours. The fibronectin level in the MRC-5 cells after 24 hours incubation with 50 ${\mu}g/mL$ $PM_{10}$ was significantly higher than the control group ($PM_{10}$ 50 ${\mu}g/mL$ 113.27${\pm}$8.65 of control, p=0.005). Conclusion: $PM_{10}$ from Asian dust increases the activation of NF-${\kappa}B$ and fibronectin expression in MRC-5 fibroblast cells.

The Effect of Saccharin on the Gene Expression of NF-κB and Inflammatory Cytokines in LPS-Stimulated SW480 Colon Cancer Cells (옥수수수염 추출물이 SW480 Colon Cancer Cell에서 NF-κB와 염증성 사이토카인 발현에 미치는 영향)

  • Choi, Hyunji;Kim, Sunlim;Kang, Hyeonjung;Kim, Myunghwan;Kim, Wookyoung
    • Journal of the Korean Dietetic Association
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    • v.25 no.3
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    • pp.217-228
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    • 2019
  • There have been no published studies concerning the anti-inflammatory effects of corn silk on colon cancer cells. Thus, this study was conducted to investigate the effect of corn silk extract containing high levels of maysin on inflammation and its mechanism of action in colon cancer cells. SW 480 human colon cancer cells were treated with $1{\mu}g/mL$ of lipopolysaccharide (LPS) to induce inflammation, and next they were treated with different concentrations of corn silk extract (0, 5, 10 and $15{\mu}g/mL$). The concentrations of nitric oxide (NO) were determined. The mRNA expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), interleukin-1beta ($IL-1{\beta}$) and interleukin-6 (IL-6), were determined. Western blot analysis was performed to determine the protein expressions of nuclear factor-kappa B ($NF-{\kappa}B$) and mitogen-activated protein kinases, and the latter consists of extracellular signal-related kinase (ERK), c-jun NH2-terminal kinase (JNK) and p38 MAP kinase (p38). The concentration of NO and the mRNA expression of iNOS were significantly and dose-dependently decreased in the corn silk-treated groups (P<0.05). The mRNA expression of $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 were significantly increased in the LPS-treated group (P<0.05), but these expressions were significantly and dose-dependently decreased in the corn silk treated groups (P<0.05). The protein expressions of $NF-{\kappa}B$ (in a dose-dependent fashion), ERK (at 10 and $15{\mu}g/mL$), JNK (at $15{\mu}g/mL$) and p38 (at 10 and $15{\mu}g/mL$) were significantly decreased with corn silk treatments (P<0.05). In conclusion, corn silk extract containing high levels of maysin seems to inhibit the LPS-induced inflammatory responses in SW480 colon cancer cells via the $NF-{\kappa}B$ pathway.

Ethanol-induced Activiationof Transcription Factor NF-$\kappa$B and AP-1 in C6 Glial Cells

  • Park, Jae -Won;Shim, Young-Sup
    • Preventive Nutrition and Food Science
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    • v.4 no.3
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    • pp.209-214
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    • 1999
  • In this study, the effectof ethanol and acetaldehyde on DNA binding activities of NF-$textsc{k}$B and AP-1 were evaluated in C6 rat glial cells. Both NF-$textsc{k}$B and AP-1 are important transcription factors for the expression of various cytokines in glial cells. Our data showed that neither ethanol nor acetaldehyde induced conspicuous cell death of C6 cells at clinically realistic concentrations. When the DNA binding activities of nuclear NF-$textsc{k}$B and AP-1 were estimated using electrophoretic mobility shift assay (EMSA), ethanol(0.3%) or acetaldehyde(1mM) induced transient activation of these transcription factors, which attained peak levels at 4~8 hours and declined to basal levels at 12 hours after treatement . The supershift analysis showed that the increased activities of NF-$textsc{k}$B in ethanol/acetaldehyde-treated C6 cells were due to the preferential induction of p65/p50 heterodimer complex. The DNA binding activities of these transcriptional factors decreased below basal levels when cells were cultured with either ethanol or acetaldehyde for 24 hours, and showed the inhibitory effect of chronic ehtanol /acetaldehyde treatment on the activities of these transsriptional factors. Our data indicate that either ethanol or acetaldehyde can induce functional changes of glial cells throught bi-directional modulation of NF-$textsc{k}$B and AP-1 DNA binding activities.

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The Effect of Moutan Cortex on Pro-inflammatory Cytokines through NF-κB & MAPKs pathway in HMC-l (목단피의 NF-κB와 MAPKs 억제를 통한 Pro-inflammatory Cytokines 활성에 미치는 효과)

  • Park, Joo-Ho;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.22 no.2
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    • pp.1-18
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    • 2009
  • Objective : Moutan Cortex (the root bark of Paeonia suffruticosa Andr.) is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how MC(Moutan Cortex) affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted MC on the mast cell-mediated inflammatory responses. Method : We observed the effect of MC on compound 48/80-induced histamine release of rat peritoneal mast cells and the effect of administering MC on PCA in rat. We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of MC. The TNF-$\alpha$ protein levels were analysised by Western blot. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. NF-$\kappa$B, phospho-I$\kappa$B and MAPKs were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by luciferase assay. Result : 1. Enzyme immunoassay indicated that MC suppressed histamine secretion of rat peritoneal mast cells. 2. In PCA dependent on IgE, MC had anti-allergic effect of the internal surface of rat skin. 3. Western blot indicated that MC decreased TNF-$\alpha$ protein levels. 4. ELISA indicated that MC decreased TNF-$\alpha$, IL-6 but MC had no significant effect on IL-8 in HMC-1 cells. 5. RT-PCR indicated that MC decreased TNF-$\alpha$, IL-8 but MC had no significant effect on IL-6 in HMC-l cells. 6. Western blot indicated that MC suppressed the induction of MAPKs, NF-$\kappa$B & phospho-I$\kappa$B activity in HMC-1 cells. 7. Luciferase assay indicated that MC suppressed the PMA plus A23187-induced NF-$\kappa$B promoting activityin HMC-1 cells. Conclusion : In this study, we have found that MC is an inhibitor of NF-$\kappa$B, MAPKs & cytokines on the mast cell-mediated inflammatory responses.

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Kojic Acid, a Potential Inhibitor of NF-$textsc{k}$B Activation in Transfectant Human HaCaT and SCC-13 Cells

  • Moon, Ki-Young;Ahn, Kwang-Seok;Lee, Jin-seon;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • v.24 no.4
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    • pp.307-311
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    • 2001
  • The activation of NF-$\kappa$B induced by kojic Acid, an inhibitor of tyrosinase for biosynthesis of melanin in melanocytes, was investigated in human transfectant HaCaT and SCC-13 cells. These two keratinocyte cell lines transfected with pNF-$\kappa$B-SEAP-NPT plasmid were used to determine the activation of NF-$\kappa$B. Transfectant cells release the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the NF-$\kappa$B activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selective marker of geneticin resistance. NF-$\kappa$B activation was measured in the SEAP reporter gene assay using a fluorescence detection method. Kojic Acid showed the inhibition of cellular NF-$\kappa$B activity in both human keratinocyte transfectants. It could also downregulate the ultraviolet ray (UVR)-induced activation of NF-$\kappa$B expression in transfectant HaCaT cells. Moreover, the inhibitory activity of kojic Acid in transfectant HaCaT cells was found to be more potent than known antioxidants, e.g., vitamin C and N~acetyl-L-cysteine. These results indicate that kojic Acid is a potential inhibitor of NF-$\kappa$B activation in human keratinocytes, and suggest the hypothesis that NF-$\kappa$B activation may be involved in kojic Acid induced anti-melanogenic effect.

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Antioxidant and Anti-inflammatory Activities of Butanol Extract of Melaleuca leucadendron L.

  • Surh, Jeong-Hee;Yun, Jung-Mi
    • Preventive Nutrition and Food Science
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    • v.17 no.1
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    • pp.22-28
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    • 2012
  • Melaleuca leucadendron L. has been used as a tranquilizing, sedating, evil-dispelling and pain-relieving agent. We examined the effects of M. leucadendron L. extracts on oxidative stress and inflammation. M. leucadendron L. was extracted with methanol (MeOH) and then fractionated with chloroform ($CHCl_3$) and butanol (BuOH). Antioxidant activity of the MeOH extract and BuOH fraction were higher than that of both ${\alpha}$-tocopherol and butyrated hydroxytoluene (BHT). Total phenol content in the extracts of M. leucadendron L., especially the BuOH fraction, well correlated with the antioxidant activity. The anti-inflammatory activity of BuOH extracts were investigated by lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production, and cyclooxygenase-2 (COX-2) expression in RAW 264.7 macrophages. The BuOH fraction significantly inhibited LPS-induced NO and $PGE_2$ production. Furthermore, BuOH extract of M. leucadendron L. inhibited the expression of COX-2 and iNOS protein without an appreciable cytotoxic effect on RAW264.7 cells. The extract of M. leucadendron L. also suppressed the phosphorylation of inhibitor ${\kappa}B{\alpha}$ ($I{\kappa}B{\alpha}$) and its degradation associated with nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activation. Furthermore, BuOH fraction inhibited LPS-induced NF-${\kappa}B$ transcriptional activity in a dose-dependent manner. These results suggested that M. leucadendron L. could be useful as a natural antioxidant and anti-inflammatory resource.

A comparative study of the immuno-modulatory activities of ethanol extracts and crude polysaccharide fractions from Annona muricata L. (가시여지잎(Annona muricata L.) 에탄올 추출물과 조다당 분획분의 면역활성 비교)

  • Kim, Yi-Eun;Lee, Joeng-Hee;Sung, Nak-Yun;Ahn, Dong-Hyun;Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.49 no.4
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    • pp.453-458
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    • 2017
  • This study compared the immuno-modulatory effects of ethanol extracts (A. muricata L. ethanol extracts, ALE) and crude polysaccharide fraction (A. muricata L. crude polysaccharide fraction, ALP) from Annona muricata L. in macrophages. Immuno-modulatory activity was determined by assessing cell viability, nitric oxide (NO) production, inducible NO synthase (iNOS) expression and cytokine production in RAW 264.7 a macrophage cell line. Both ALE and ALP treatment did not affect cytotoxicity, and ALP treatment significantly increased NO production. Additionally, cytokine production [tumor necrosis factor ($TNF-{\alpha}$; $2909.04{\pm}4.1pg/mL$), interleukin (IL)-6; $662.84{\pm}5.3pg/mL$, and $IL-1{\beta}$; $852.37{\pm}2.2pg/mL$), was highly increased in the ALP ($250{\mu}g/mL$) treated group compared to the ALE ($250{\mu}g/mL$) treated group ($TNF-{\alpha}$; $1564.50{\pm}6.1pg/mL$, IL-6; $517.24{\pm}4.1pg/mL$ and $IL-1{\beta}$; $237.23{\pm}1.8pg/mL$). Moreover, ALP treatment considerably increased the expression of mitogen-activated protein kinase (MAPKs) and nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) in the macrophages. Therefore, ALP can induce macrophage activation through MAPK and $NF-{\kappa}B$ signaling and this can be a potential candidate for development of nutraceuticals.

Role of Oxygen Free Radical in the Expression of Interleukin-8 and Interleukin-$1{\beta}$ Gene in Mononuclear Phagocytic Cells (내독소에 의한 말초혈액 단핵구의 IL-8 및 IL-$1{\beta}$ 유전자 발현에서 산소기 역할에 관한 연구)

  • Kang, Min-Jong;Kim, Jae-Yeol;Park, Jae-Seok;Lee, Seung-Joon;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.6
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    • pp.862-870
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    • 1995
  • Background: Oxygen free radicals have generally been considered as cytotoxic agents. On the other hand, recent results suggest that small nontoxic amounts of these radicals may act a role in intracellular signal transduction pathway and many efforts to reveal the role of these radicals as secondary messengers have been made. It is evident that the oxygen radicals are released by various cell types in response to extracellular stimuli including LPS, TNF, IL-1 and phorbol esters, all of which translocate the transcription factor $NF{\kappa}B$ from cytoplasm to nucleus by releasing an inhibitory protein subunit, $I{\kappa}B$. Activation of $NF{\kappa}B$ is mimicked by exposure to mild oxidant stress, and inhibited by agents that remove oxygen radicals. It means the cytoplasmic form of the inducible tanscription factor $NF{\kappa}B$ might provide a physiologically important target for oxygen radicals. At the same time, it is well known that LPS induces the release of oxygen radicals in neutrophil with the activation of $NF{\kappa}B$. From above facts, we can assume the expression of IL-8 and IL-$1{\beta}$ gene by LPS stimulation may occur through the activation of $NF{\kappa}B$, which is mediated through the release of $I{\kappa}B$ by increasing amounts of oxygen radicals. But definitive evidence is lacking about the role of oxygen free radicals in the expression of IL-8 and IL-$1{\beta}$ gene in mononuclear phagocytic cells. We conducted a study to determine whether oxygen radicals act a role in the expression of IL-8 and IL-$1{\beta}$ gene in mononuclear phagocytic cells. Method: Human peripheral blood monocytes were isolated from healthy volunteers. Time and dose relationship of $H_2O_2$-induced IL-8 and IL-$1{\beta}$ mRNA expression was observed by Northern blot analysis. To evaluate the role of oxygen radicals in the expression of IL-8 and IL-$1{\beta}$ mRNA by LPS stimulation, pretreatment of various antioxiants including PDTC, TMTU, NAC, ME, Desferrioxamine were done and Northern blot analysis for IL-8 and IL-$1{\beta}$ mRNA was performed. Results: In PBMC, dose and time dependent expression of IL-8 and IL-$1{\beta}$ mRNA by exogenous $H_2O_2$ was not observed. But various antioxidants suppressed the expression of LPS-induced IL-8 and IL-$1{\beta}$ mRNA expression of PBMC and the suppressive activity was most prominant when the pretreatment was done with TMTU. Conclusion: Oxygen free radical may have some role in the expression of IL-8 and IL-$1{\beta}$ mRNA of PBMC but that radical might not be $H_2O_2$.

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Core Loss Analysis of Non-oriented Electrical Steel Under Magnetic Induction Including Higher Harmonics

  • Cho, Chuhyun;Son, Derac;Cho, Youk
    • Journal of Magnetics
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    • v.6 no.2
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    • pp.66-69
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    • 2001
  • The actual magnetic induction waveform of cores in electrical machines is not sinusoidal i.e. higher harmonics are always included. Thus the core loss in actual electrical machines is different from the core loss which is measured by the standard method, because the waveform of magnetic induction should be sinusoidal in the standard testing method. Core loss analysis under higher harmonic induction is always important in electric machine design. In this works we measured the core loss when a hysteresis loop has only one period of an ac minor loop of higher harmonic frequency, depending on the position of the ac minor loop of relative to the fundamental harmonic frequency. From this experiment, the core loss P(B/sub 0/f/sub 0/, B/sub h/, nf/sub 0/)) under a higher harmonic magnetic induction B/sub h/ could be expressed by the linear combination the core loss at fundamental harmonic frequency P/sub c/(B/sub 0/, f/sub 0/), the core loss of ac minor loop at zero induction region of the major hysteresis loop P/sub cL/ (B/sub h/, nf/sub 0/), and the core loss of an ac minor loop in the high induction region of the major hysteresis loop P/sub cH/ (B/sub h/, nf/sub 0/) i.e., P/sub c/, (B/sub 0/, f/sub 0/, B/sub h/, nf/sub 0/)=P/sub c/ (B/sub 0/, f/sub 0/,)+(n-1)[k₁(B/sub 0/) P/sub cL/ (B/sub h/, nf/sub 0/)+(1-k₁(B/sub 0/)) P/sub cH/ (B/sub h/, nf/sub 0/)]. This will be useful formula for electrical machine designers and one of effective methods to predict core loss including higher harmonic induction.

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The Hardness Water Production By RO/NF/ED Linking Process From Deep Seawater (RO/NF/ED 연계 공정에 의한 고경도 담수 제조)

  • Moon, Deok-Soo;Kim, Kwang Soo;Gi, Ho;Choi, Mi Yeon;Jung, Hyun Ji;Kim, Hyun Ju
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.16 no.4
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    • pp.227-238
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    • 2013
  • The purpose of this study is to develop a process technology to produce high hardness drinking water which meet drinking water standard, remaining useful minerals like magnesium and calcium in the seawater desalination process while removing the sulfate ions and chloride ions. Seawater have been separated the concentrated seawater and desalted seawater by passing on Reverse Osmosis membrane (RO). Using Nano-filtration membrane (NF), We were prepared primary mineral concentrated water that sodium chloride were not removed. By the operation of electro-dialysis (ED) having ion exchange membrane, we were prepared concentrated mineral water (Mineral enriched desalted water) which the sodium chloride is removed. We have produced the high hardness water to meet the drinking water quality standards by diluting the mineral enriched desalted water with deionized water by RO. Reverse osmosis membranes (RO) can separate dissolved material and freshwater from seawater (deep seawater). The desalination water throughout the second reverse osmosis membrane was completely removed dissolved substances, which dissolved components was removed more than 99.9%, its the hardness concentration was 1 mg/L or less and its chloride concentration was 2.3 mg/L. Since the nano-filtration membrane pore size is $10^{-9}$ m, 50% of magnesium ions and calcium ions can not pass through the nano-filtration membrane, while more than 95% of sodium ions and chloride ions can pass through NF membrane. Nano-filtration membrane could be separated salt components like sodium ion and chloride ions and hardness ingredients like magnesium ions and calcium ions, but their separation was not perfect. Electric dialysis membrane system can be separated single charged ions (like sodium and chloride ions) and double charged ions (like magnesium and calcium ions) depending on its electrical conductivity. Above electrical conductivity 20mS/cm, hardness components (like magnesium and calcium ions) did not removed, on the other hand salt ingredients like sodium and chloride ions was removed continuously. Thus, we were able to concentrate hardness components (like magnesium and calcium ions) using nano-filtration membrane, also could be separated salts ingredients from the hardness concentration water using electrical dialysis membrane system. Finally, we were able to produce a highly concentrated mineral water removed chloride ions, which hardness concentration was 12,600 mg/L and chloride concentration was 2,446 mg/L. By diluting 10 times these high mineral water with secondary RO (Reverse Osmosis) desalination water, we could produce high mineral water suitable for drinking water standards, which chloride concentration was 244 mg/L at the same time hardness concentration 1,260 mg/L. Using the linked process with reverse osmosis (RO)/nano filteration (NF)/electric dialysis (ED), it could be concentrated hardness components like magnesium ions and calcium ions while at the same time removing salt ingredients like chloride ions and sodium ion without heating seawater. Thus, using only membrane as RO, NF and ED without heating seawater, it was possible to produce drinking water containing high hardness suitable for drinking water standard while reducing the energy required to evaporation.