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Stem-leaf saponins from Panax notoginseng counteract aberrant autophagy and apoptosis in hippocampal neurons of mice with cognitive impairment induced by sleep deprivation

  • Cao, Yin;Yang, Yingbo;Wu, Hui;Lu, Yi;Wu, Shuang;Liu, Lulu;Wang, Changhong;Huang, Fei;Shi, Hailian;Zhang, Beibei;Wu, Xiaojun;Wang, Zhengtao
    • Journal of Ginseng Research
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    • v.44 no.3
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    • pp.442-452
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    • 2020
  • Backgroud: Sleep deprivation (SD) impairs learning and memory by inhibiting hippocampal functioning at molecular and cellular levels. Abnormal autophagy and apoptosis are closely associated with neurodegeneration in the central nervous system. This study is aimed to explore the alleviative effect and the underlying molecular mechanism of stem-leaf saponins of Panax notoginseng (SLSP) on the abnormal neuronal autophagy and apoptosis in hippocampus of mice with impaired learning and memory induced by SD. Methods: Mouse spatial learning and memory were assessed by Morris water maze test. Neuronal morphological changes were observed by Nissl staining. Autophagosome formation was examined by transmission electron microscopy, immunofluorescent staining, acridine orange staining, and transient transfection of the tf-LC3 plasmid. Apoptotic event was analyzed by flow cytometry after PI/annexin V staining. The expression or activation of autophagy and apoptosis-related proteins were detected by Western blotting assay. Results: SLSP was shown to improve the spatial learning and memory of mice after SD for 48 h, accomanied with restrained excessive autophage and apoptosis, whereas enhanced activation of phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway in hippocampal neurons. Meanwhile, it improved the aberrant autophagy and apoptosis induced by rapamycin and re-activated phosphoinositide 3-kinase/Akt/mammalian target of rapamycin signaling transduction in HT-22 cells, a hippocampal neuronal cell line. Conclusion: SLSP could alleviate cognitive impairment induced by SD, which was achieved probably through suppressing the abnormal autophagy and apoptosis of hippocampal neurons. The findings may contribute to the clinical application of SLSP in the prevention or therapy of neurological disorders associated with SD.

Sodium Salicylate(NaSaL) Induces Apoptosis of NCI-H1299 Lung Carcinoma Cells via Activation Caspase-3 Protease (NCI-H1299 폐암 세포주에서 Caspase-3 Protease 활성을 통한 Sodium Salicylate(NaSaL)의 세포고사)

  • Shim, Hyeok;Yang, Sei-Hoon;Bak, Sang-Myeon;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.5
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    • pp.485-496
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    • 2002
  • Background : Nonsteroidal anti-inflammatory drugs (NSAIDs) are useful in the chemoprevention of colon cancers. Continuous NSAID use results in a 40% to 50% reduction in the relative risk of colorectal cancer. The precise mechanism by which NSAIDs prevent and/or cause the regression of colorectal tumors is not known. Some investigators have reported that certain NSAIDs induce apoptosis and alter the expression of the cell cycle regulatory genes in some carcinoma cells when administered at a relatively high concentration. However, the possibility of NSAIDs application as chemopreventive agents in lung cancers remains to be elucidated. To address this question, the human lung cancer cell line NCI-H1299 was used to investigate whether or not NSAIDs might induce the apoptotic death of NCI-H1299 cells. Methods : A viability test was carried out using a MTT assay. Apoptosis was measured by flow cytometric analysis and unclear staining(DAPI). The talytic activity of the caspase family was measured by the fluirigenic cleavage of biosubstrates. To define the mechanical basis of apoptosis, western blot was performed to analyze the expression of the death substrates(PARP and ICAD). Results : NaSaL significantly decreased the viability of the NCI-H1299 cells, which was revealed as apoptosis characterized by an increase in the $subG_0/G_1$ population and unclear fragmentation. The catalytic activity of caspase-3 protease began to increase after 24 Hr and reached a peak 30 Hr after treatment with 10 mM NaSaL. In contrast, caspase-6, 8, and 9 proteases did not have a significantly altered enzymatic activity. Consistent with activation of caspase-3 protease, NaSaL induced the cleavage of the protease biosubstrate. Conclusion : NaSaL induces the apoptotic death of NCI-H1299 human lung cancer cells via the activation of caspase-3 protease.

β-elemene Induces Caspase-dependent Apoptosis in Human Glioma Cells in vitro through the Upregulation of Bax and Fas/FasL and Downregulation of Bcl-2

  • Li, Chen-Long;Chang, Liang;Guo, Lin;Zhao, Dan;Liu, Hui-Bin;Wang, Qiu-Shi;Zhang, Ping;Du, Wen-Zhong;Liu, Xing;Zhang, Hai-Tao;Liu, Yang;Zhang, Yao;Xie, Jing-Hong;Ming, Jian-Guang;Cui, Yu-Qiong;Sun, Ying;Zhang, Zhi-Ren;Jiang, Chuan-Lu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10407-10412
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    • 2015
  • Background: ${\beta}$-elemene, extracted from herb medicine Curcuma wenyujin has potent anti-tumor effects in various cancer cell lines. However, the activity of ${\beta}$-elemene against glioma cells remains unclear. In the present study, we assessed effects of ${\beta}$-elemene on human glioma cells and explored the underlying mechanism. Materials and Methods: Human glioma U87 cells were used. Cell proliferation was determined with MTT assay and colony formation assay to detect the effect of ${\beta}$-elemene at different doses and times. Fluorescence microscopy was used to observe cell apoptosis with Hoechst 33258 staining and change of glioma apoptosis and cell cycling were analyzed by flow cytometry. Real-time quantitative PCR and Western-blotting assay were performed to investigated the influence of ${\beta}$-elemene on expression levels of Fas/FasL, caspase-3, Bcl-2 and Bax. The experiment was divided into two groups: the blank control group and ${\beta}$-elemne treatment group. Results: With increase in the concentration of ${\beta}$-elemene, cytotoxic effects were enhanced in the glioma cell line and the concentration of inhibited cell viability ($IC_{50}$) was $48.5{\mu}g/mL$ for 24h. ${\beta}$-elemene could induce cell cycle arrest in the G0/G1 phase. With Hoechst 33258 staining, apoptotic nuclear morphological changes were observed. Activation of caspase-3,-8 and -9 was increased and the pro-apoptotic factors Fas/FasL and Bax were upregulated, while the anti-apoptotic Bcl-2 was downregulated after treatment with ${\beta}$-elemene at both mRNA and protein levels. Furthermore, proliferation and colony formation by U87 cells were inhibited by ${\beta}$-elemene in a time and does-dependent manner. Conclusions: Our results indicate that ${\beta}$-elemene inhibits growth and induces apoptosis of human glioma cells in vitro. The induction of apoptosis appears to be related with the upregulation of Fas/FasL and Bax, activation of caspase-3,-8 and -9 and downregulation of Bcl-2, which then trigger major apoptotic cascades.

BCR/ABL mRNA Targeting Small Interfering RNA Effects on Proliferation and Apoptosis in Chronic Myeloid Leukemia

  • Zhu, Xi-Shan;Lin, Zi-Ying;Du, Jing;Cao, Guang-Xin;Liu, Gang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.4773-4780
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    • 2014
  • Background: To investigate the effects of small interference RNA (siRNA) targeting BCR/ABL mRNA on proliferation and apoptosis in the K562 human chronic myeloid leukemia (CML) cell line and to provide a theoretical rationale and experimental evidence for its potential clinical application for anti-CML treatment. Materials and Methods: The gene sequence for BCR/ABL mRNA was found from the GeneBank. The target gene site on the BCR/ABL mRNA were selected according to Max-Planck-Institute (MPI) and rational siRNA design rules, the secondary structure of the candidate targeted mRNA was predicted, the relevant thermodynamic parameters were analyzed, and the targeted gene sequences were compared with BLAST to eliminate any sequences with significant homology. Inhibition of proliferation was evaluated by MTT assay and colony-formation inhibiting test. Apoptosis was determined by flow cytometry (FCM) and the morphology of apoptotic cells was identified by Giemsa-Wright staining. Western blotting was used to analyze the expression of BCR/ABL fusion protein in K562 cells after siRNA treatment. Results: The mRNA local secondary structure calculated by RNA structure software, and the optimal design of specific siRNA were contributed by bioinformatics rules. Five sequences of BCR/ABL siRNAs were designed and synthesized in vitro. Three sequences, siRNA1384, siRNA1276 and siRNA1786, which showed the most effective inhibition of K562 cell growth, were identified among the five candidate siRNAs, with a cell proliferative inhibitory rate nearly 50% after exposure to 12.5nmol/L~50nmol/L siRNA1384 for 24,48 and 72 hours. The 50% inhibitory concentrations ($IC_{50}$) of siRNA1384, siRNA1276 and siRNA1786 for 24hours were 46.6 nmol/L, 59.3 nmol/L and 62.6 nmol/L, respectively, and 65.668 nmol/L, 76.6 nmol/L, 74.4 nmol/L for 72 hours. The colony-formation inhibiting test also indicated that, compared with control, cell growth of siRNA treated group was inhibited. FCM results showed that the rate of cell apoptosis increased 24 hours after transfecting siRNA. The results of annexinV/PI staining indicated that the rate of apoptosis imcreased (1.53%, 15.3%, 64.5%, 57.5% and 21.5%) following treamtne with siRNAs (siRNA34, siRNA372, siRNA1384, siRNA1276 and siRNA1786). Morphological analysis showed td typical morphologic changes of apoptosis such as shrunken, fragmentation nucleus as well as "apoptotic bodies" after K562 cell exposure to siRNA. Western blot analysis showed that BCR/ABL protein was reduced sharply after a single dose of 50nmol/L siRNA transfection. Conclusions: Proliferation of K562 cells was remarkbly inhibited by siRNAs (siRNA1384, siRNA1276 and siRNA1786) in a concentration-dependent manner in vitro, with effective induction of apoptosis at a concentration of 50 nmol/L. One anti-leukemia mechanism in K562 cells appeared that BCR/ABL targeted protein was highly down-regulated. The siRNAs (siRNA1384, siRNA1276 and siRNA1786) may prove valuable in the treatment of CML.

The Role of Heme Oxygenase-1 in Lung Cancer Cells (폐암세포주에서 Heme Oxygenase-1의 역할)

  • Jung, Jong-Hoon;Kim, Hak-Ryul;Kim, Eun-Jung;Hwang, Ki-Eun;Kim, So-Young;Park, Jung-Hyun;Kim, Hwi-Jung;Yang, Sei-Hoon;Jeong, Eun-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.3
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    • pp.304-313
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    • 2006
  • Background : Heme oxygenase-1 (HO-1) is an inducible enzyme that catalyzes the oxidative degradation of heme to form biliverdin, carbon monoxide (CO), and free iron. The current evidence has indicated a critical role of HO-1 in cytoprotection and also in other, more diverse biological functions. It is known that the high expression of HO-1 occurs in various tumors, and that HO-1 has an important role in rapid tumor growth because of its antioxidative and antiapoptotic effects. Therefore, the role of HO-1 was analyzed in human lung cancer cell lines, and especially in the A549 cell line. Material and Methods : Human lung cancer cell lines, i.e., A549, NCI-H23, NCI-H157 and NCI-H460, were used for this study. The expression of HO-1 in the untreated state was defined by Western blotting. ZnPP, which is the specific HO inhibitor we used, and the viability of cells were tested for by conducting MTT assaysy. The HO enzymatic activity, as determined via the bilirubin level, was also indirectly measured. Moreover, the generation of intracellular hydrogen peroxide (H2O2) was monitored fluorimetrically with using a scopoletin-horse radish peroxidase (HRP) assay and 2',7'-dichlorofluorescein diacetate (DCFH-DA). We have also transfected small HO-1 interfering RNA (siRNA) into A549 cells, and the apoptotic effects were evaluated by flow cytometric analysis and Western blotting. Results : The A549 cells had a greater expression of HO-1 than the other cell lines, whereas ZnPP significantly decreased the viability of the A549 cells more than the viability of the other lung cancer cells in a dose-dependant fashion. Consistent with the viability, the HO enzymatic activity also was decreased. Moreover, intracellular H2O2 generation via ZnPP was induced in a dose-dependent manner. Apoptotic events were, then induced in the HO-1 siRNA transfected A549 cells. Conclusion : HO-1 provides new important insights into the possible molecular mechanism of the antitumor therapy in lung cancer.

Studies on Influence of Water-Proof Agents on the Properties of Mortar (방수제(防水劑)가 모르터의 제성질(諸性質)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Kim, Seong Wan;Sung, Chan Yong;Kim, Sun Young
    • Korean Journal of Agricultural Science
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    • v.14 no.2
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    • pp.358-372
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    • 1987
  • This study was performed to obtain data which can be applied to use of water-proof mortars. The data was based on the properties of water-proof mortars depending upon various mixing ratios to compare those of cement mortar. The water-proof agents used were retard and accelerate type which are being used as mortar structures. The water-proof agents, mixing ratios of cement to fine aggregate were 1:1, 1:2, 1:3 and 1:4. The results obtained were summarised as follows; 1. The results of flow test, water-cement ratio was increased with the increasing of mixing ratio. 2. The permeability were increased in poorer mixing ratio and higher water pressure. 3. The bulk density was decreased with the increasing of mixing ratio, and compressive and tensile strength were increased with increasing of the bulk density. 4. At 1:1 mixing ratio, the highest strengths were showed and strengths were decreased with the increasing of mixing ratio. 5. The absorption rates were increased in- poorer mixing ratio 6. The correlation between W/C, permeability, bulk density, compressive strength and absorption rate were highly significant as a straight line, respectively.

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Study on true nature of the Fung(風) and that of application to the medicine (풍(風)의 본질(本質)과 의학(醫學)에서의 운용(運用)에 관(關)한 고찰(考察))

  • Back, Sang Ryong;Park, Chan Kug
    • Journal of Korean Medical classics
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    • v.7
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    • pp.198-231
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    • 1994
  • Up to now, after I had examined the relation between the origin of Fung(風) and Gi(氣) and the mean of Fung in medical science, I obtained the conclusion being as follows. The first, Fung(風) means a flux of Gi(氣) and Gi shows the process by virtue of the form of Fung, namely, Fung means motion of Gi. In other words, it is flow of power. Accordingly, the process of all power can give a name Fung. The second, Samul(事物) ceaselessly interchange with the external world to sustain the existence and life of themselves. And they make a adequate confrontation against the pressure of the outside. This the motive power of life action(生命活動) is Gi and shows its the process on the strength of Fung. The third, Samul(事物) incessantly releases power which it has to the outside. Power released to the outside forms the territory of the established power in the environment of them and keep up their substance(實體) in the space time(時空). It can be name Fung because the field(場) of this power incessantly flows. The fourth, man operates life on the ground of the creation of his own vigor(生氣) for himself as the life body(生命體) of the independence and self-support. The occurence of this vigor and the adjustment process(調節作用) is supervised by Gan(肝). That is to say, Gan plays a role to regulate and manage the process of Fung or the action of vigor with Fung-Zang(風臟). The fifth, because the Gi-Gi adjustment process(氣機調節作用) of Gan is the same as the process of Fung, Fung that operates the cause of a disease is attributed to the disharmony of the process of the human body Gi-Gi. Therefore, the generating pathological change is attributed to the extraordinary of the function by the incongruity of Gi-Gi(氣機) or the disorder of the direct motion of Gi-Hyul(氣血). Because the incongruity of this Gi-Gi of the human body gives rise to the abnormal of Zung-Gi(正氣) in the human body properly cannot cope with the invasion of 'Oi-Sa(外邪). Furthermore, Fung serves as the mediation body of the invasion of other Sa-Gi(邪氣) because of its dynamics, By virtue of this reason, Fung is named the head of all disease. And because the incongruity of the Gi-Gi has each other form according to Zang-Bu(臟腑), Kyung-Lak(經絡), and a region, the symptoms of a disease appear differently in line with them as well. The sixth, Fung-byung(風病) is approximately separated Zung-Fung(中風) and Fung-byung(猍義의 風病). Zung-fung and Fung-byung is to be attributed to the major invasion of each Jung-gi and Fung-sa(正氣와 風邪). But these two kinds stir up the problem to the direct motion of Gi-hyul(氣血) and the harmony of Gi-Gi in the human body. When one cures it, therefore, Zung-fung has to rectify Gi-Gi and the circulation of Gi-hyul on the basis of the supplement of Jung-gi(正氣) and Fung-byung must make the harmony of Gi-Gi with the Gu-fung(驅風). -Go-gi(調氣), Sun-Gi(順氣). Hang-Gi(行氣) - All existing living things as well as man maintain life on the ground of the pertinent harmony between the soul(精神) and the body(肉體). As soon as the harmony falls down, simultaneously life disappears as well. And Fung which means the outside process between Gi(氣) and Gi(氣) makes the action of their life cooperative and unified, Accordingy, the understanding of Fung, first, has to start wi th the whole thought that not only all Samul(事物) but also the soul and the body are one.

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Distribution of Nutrients and Chlorophyll α in the Surface Water of the East Sea (동해 표층수 중 영양염과 Chlorophyll α의 분포 특성)

  • Yoon, Sang Chol;Yoon, Yi Yong
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.19 no.2
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    • pp.87-98
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    • 2016
  • During the period between July 3 and 27 of 2009, water samples were collected from the Russian coast at a depth of 30m from 26 stations (including Ulleung and Japan basins) onboard the Russian survey vessel R/V Lavrentyev following 4 lines (D, R, E, and A). The samples were analyzed for nutrients and chlorophyll a contents. All parameters exhibited higher values in warm waters than in cold waters ($NH_4:1.8-fold$, $PO_4:1.8-fold$, $SiO_2:1.2-fold$, and chlorophyll-${\alpha}$:1.9-fold), except nitrates, which was 1.4-fold higher in cold waters than in warm waters. The horizontal distribution of ammonia, phosphate, and chlorophyll-${\alpha}$ was very similar to each other and showed the highest values in the waters near Russia, where a upwelling influence of cold current and bottom water prevails, while relatively low distribution was observed at the Ulleung Basin. On the other hand, nitrates showed the highest concentration at the Ulleung Basin, which is under the direct influence of the Tsushima warm water, and showed a gradual decrease northward. The N/P ratio showed the highest value in the Tsushima middle water, rather than in the North Korean Cold Water, the Tsushima Warm Water was the primary source of nitrate flow into the East Sea. However, the average concentration of phosphate in the warm waters was < $0.2{\mu}M$, thereby limiting phytoplankton growth, while a high concentration of phosphate in cold waters showed a direct correlation with chlorophyll-${\alpha}$. The results of principal component analysis for the identification of primary factors that influence the marine environment showed that principal component I was water temperature and principal component II was influenced chlorophyll-${\alpha}$ and nutrients. Therefore, Study area has greatest influenced by water temperature, and clearly distinct cold and warm water regions were observed in the East Sea.

The Design and Evaluation of a Diagonally Splitted Column to Improve Text Readability on a Small Screen (소형 스크린 상에서의 텍스트 가독성 향상을 위한 대각분할 칼럼 디자인과 평가)

  • Kim Yeon-Ji;Lee Woo-Hun
    • Archives of design research
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    • v.19 no.4 s.66
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    • pp.51-60
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    • 2006
  • Nowadays, reading text from screens is prevailing in everyday life. The advent of mobile information devices such as a cellular phone, PDA, and e-book reader facilitates us to enjoy various text-based contents any time and anywhere. Most studies comparing screen and paper readability show that screens are less readable than paper. Furthermore, the decrease of line length and number of lines that can be displayed on the screen of mobile information devices deteriorate text readability. This study investigated parameters affecting text readability on small screens and designed a new text layout to improve readability. We suggested a diagonally splitted layout of rectangular column, which is supposed to facilitate eye movement to trace text flow with ease. The experiment comparing readability between a traditional rectangular column and a diagonally splitted column was conducted. The result of experiment revealed that there is no significant difference between the two text layouts in terms of subjective satisfaction of reading task and a level of comprehension. However, in the screen size of $4000mm^2\;and\;8000mm^2$, reading speed was increased 18.9% and 34.0% respectively from a traditional rectangular column to a diagonally splitted column. We conducted a consecutive experiment to scrutinize the cause that improved the performance in readability task remarkably. The readability of text in a traditional rectangular column was compared with a left triangular column and a right triangular column in the condition of $4000mm^2/3:1$ ratio screen. The performance measurements revealed that participants read 21.1% and 67.6% faster respectively with the left triangular column and right triangular column than with the rectangular column. In consequence, the improvement of readability in the diagonally splitted column was attributed mainly to the increase of reading speed in the right triangular column. This research verified that the diagonally splitted column improve text readability on a small screen and this result is expected to make a contribution to designing an efficient text layout for mobile information devices

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Hydrochemical and Isotopic Characteristics of Major Streams in the Daejeon Area (대전지역 도심하천의 수리화학적 및 동위원소적 특성)

  • Jeong, Chan-Ho;Moon, Byung-Jin
    • Economic and Environmental Geology
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    • v.42 no.4
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    • pp.315-333
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    • 2009
  • In this study, the hydrochemical and the isotopic characteristics of major streams in the Daejeon area were investigated during rainy and dry seasons. The stream water shows the electrical conductivity of the range of $37{\sim}527{\mu}s$/cm, and pH $6.21{\sim}9.83$. The chemical composition of stream waters can be grouped as three types: the upper streams of Ca(Mg)-$HCO_3$ type, Ca(Mg)-$SO_4(Cl)$ type of middle streams flowing through urban area, and Na(Ca)-$HCO_3$(Cl, $SO_4$) type of the down streams. Based on in-situ investigation, the high pH of stream waters flowing through urban area is likely to be caused by the inflow of a synthetic detergent discharging from the apartment complex. The electrical conductivity of stream waters at a dry season is higher than those of at a rainy season. We suggest that the hydro-chemical composition of stream waters in the Daejeon area was affected by the discharging water from the sewage treatment facilities and anthropogenic contaminants as well as the interaction with soil and rocks. ${\delta}D$ and ${\delta}^{18}O$ values of the stream waters show the relationship of ${\delta}D=6.45{\delta}^{18}O-7.4$, which is plotted at a lower area than global meteoric water line(GMWL) of Craig(1961). It is likely that this isotopic range results from the evaporation effect of stram waters and the change of an air mass. The isotope value shows an increasing trend from upper stream to lower stream, that reflects the isotopic altitude effect. The relationship between ${\delta}^{13}C$ and $EpCO_2$ indicates that the carbon as bicarbonate in stream water is mainly originated from $CO_2$ in the air and organic materials. The increasing trend of ${\delta}^{13}C$ value from upper stream waters to lower stream waters can be attributed to the following reasons: (1) an increasing dissolution of $CO_2$ gas from a contaminated air in downtown area of the Daejeon, and (2) the increment of an inorganic carbon of groundwater inflowed into stream by base flow. Based on the relationship between ${\delta}^{34}S$ and $SO_4$ of stream waters, the stream waters can be divided into four groups. $SO_4$ content increases as a following order: upper and middle Gab stream${\delta}^{34}S$ value decreases as above order. ${\delta}^{34}S$ value indicates that sulfur of stream waters is mainly originated from atmosphere, and is additionally supplied by pyrite source according to the increase of sulfate content. The sulfur isotope analysis of a synthetic detergent and sewage water as a potential source of the sulfur in stream waters is furtherly needed.