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Effect of Co-culture with Bovine and Porcine Oviductal Epithelial Cells on In Vitro Development of Mouse Embryos (마우스 수정란의 체외발달에 미치는 소와 돼지의 난관상피세포와의 공배양 효과)

  • Lee, S.;Hur, E.J.;Seok, H.B.
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.139-146
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    • 1997
  • This experiment was carried out to evaluate the effect of mouse early embryonic development in vitro by co-culture with bovine and porcine oviductal epithelial cells(BOEC and POEC). The 2-cell embryos were collected from the oviducts of the superovulated and mated does with D-PBS/15% FCS at 48 hours after hCG injection. The in vitro developmental rate of blastocyst formation and the number of nuclei in the embryos were examined. For a comparative study of in vi패 and in vitro development, the fresh blastocyst which developed in vivo for 120 hours after hCG injection was collected from the uterus, and their numbers of nuclei were also counted. The higher developmental rates of blastocyst formation was a, pp.ared from 91% to 97% when the embryos were co-cultured with a monolayer of bovine or porcine oviductal epithelial cells in TCM 199 or Ham's F-10 and MediCult IVF media. No significant difference in developmental rates was shown between bovine and porcine oviductal eptithelial cells. The number of nuclei in the embryos cultured for 72 hours under each conditions was significantly reduced it than blastocyst in vitro conditions. The number of nuclei in embryos cultured in TCM 199, Ham's F-10 and Medicult IVF medium were counted 68.1$\pm$6.00, 67.3$\pm$4.49, 66.4$\pm$5.64, and 94.3$\pm$8.61, 92.5$\pm$7.60, 92.1$\pm$6.10 with BOEC and 93.3$\pm$5.80, 92.9$\pm$6.53, 92.3$\pm$7.35 with POEC coculture, respectively. These numbers were lowered than 107.2$\pm$7.43 in vivo conditions. In conclusions, the coculture between the mouse early embryos, and oviductal epithelial cells of BOEC and POEC give to improve the developmental and hatching rates of blastocyst but in vivo culture systems for the growth of nuclei were ineligible than in vitro conditions.

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Inhibitory Mechanisms of Cell Cycle Regulation Induced by Indole-3-carbinol in Hepatocellular Carci-noma HepG2 Cells. (간암 세포주에서의 Indole-3-Carbinol에 의해 유도되는 세포주기 억제 기전)

  • 김동우;이광수;김민경;조율희;이철훈
    • Microbiology and Biotechnology Letters
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    • v.29 no.3
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    • pp.181-185
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    • 2001
  • The naturally occurring chemical indole-3-carbinol (13C), found in vegetables of the Brassica genus, is a promising anticancer agent that was shown previ- ously to induce a Gl cell cycle arrest of human breast cancer cell lines, independent of estrogen receptor signaling. The anticancer activity of 13C and the possible mechanisms of its action were explored in a human hepatocellular carcinoma cell line, HepG2. Treatment of HepG2 cells with 13C suppressed the growth of the cells. The growth sup- pression caused by 13C ($IC_{50}$/: 444$\mu$M) was found to be partially due to its ability to stop the cell cycle in HepG2 cells. Western blot analysis for the Gl phase artiest demonstrated that the expression-levels of cyclin-dependent kinase (Cdk4, Cdk6) and cyclic D were reduced strongly after treatment of Hep72 cells with 13C (4007M) for 24- 72 hrs. Furthermore, I3C selectively abolished the expression of Cdk6 in a dose- and time-dependent manner, and accordingly, inhibited the phosphorylation of retinoblastoma. Interestingly, after the HepG2 cells reached their max- imal growth arrest, the level of the p21, a well-known Cdk inhibitor, increased significantly. Therefore, it could be considered that the Gl arrest of HepG2 cells treated with 13C was due to the indirect inhibition of Cdk4/6 activities by p21 Western blot analysis for G2/M phase arrest of demonstrated the levels of Cdc2 and cyclin Bl werer reduced dramatically after the treatment of HepG2 cells with 13C ($40\mu$M) for 24-72 hrs. flow cytometry of propidium iodide-stained HepG2 cells revealed that 13C induces a Gl (53%,72hr incubation) and G2 (25%,24hr incubation) cell cycle arrest. Thus, our observations have uncovered a previously undefined antiproliferative pathway for r3C that implicates Cdk4/6 and Cdc2 as a target for cell cycle control in human HepG2 cells. However, the 13C-medi- ated cell cycle arrest and repression of Cdk4/6 production did not affect the apoptotic induction of HepG2 cell.

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Hazard Analysis of Tomato Farms at the Growing Stage for the Establishment of the Good Agricultural Practices (GAP) (토마토 농산물우수관리제도(GAP)모델 확립을 위한 재배단계 위해분석)

  • Park, Su-Hee;Kwon, Woo-Hyun;Heo, Rok-Won;Kim, Kyeong-Yeol;Shim, Won-Bo;Shim, Sang-In;Chung, Duck-Hwa
    • Journal of Food Hygiene and Safety
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    • v.27 no.2
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    • pp.152-160
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    • 2012
  • The objective of this study was to analyze hazards for the growing stage of 6 tomato farms (A, B, C; soli farms, D, E, F; Nutriculture farms) located in Gyeongsangnam-do to establish the good agricultural practices (GAP). A total of 144 samples for analyzing hazards collected from cultivation environments (irrigation water, soil, nutrient solution, and air) and personal hygiene (hands, gloves, and cloths) were assessed for biological (sanitary indications and major food borne pathogens) and chemical hazards (heavy metals). Total bacteria, coliform, and fungi were detected at levels of 0.2-7.2, 0.0-6.1, and 0.0-5.4 log CFU/g, mL, hand or 100 $cm^2$, respectively. Escherichia coli were only detected in the soil sample from B farm. In case of pathogens, Bacillus cereus was detected at levels of 0.0-4.4 log CFU/(g, mL, hand or 100 $cm^2$), whereas Staphylococuus aureus, Listeria monocytogenes, E. coli O157, and Salmonella spp. were not detected in all samples. Heavy metals as a chemical hazard were detected in soil and irrigation water, but levels of them were lower than the permit limit. In conclusion, chemical hazard levels complied with GAP criteria, but biological hazards at the growing stage of tomato farms were confirmed. Therefore a proper management to prevent microbial contamination is needed.

Agroenvironmental Characteristics and N.P Demand of Paddy Fields Irrigated with the Water of Nagdong River (낙동강 물 관개논의 농업환경 특성과 질소, 인산 요구량)

  • Kang, Ui-Gum;Lee, jae-Saeng;Ko, Ji-Yeon;Park, Chang-Young;Jung, Ki-Yeul
    • Korean Journal of Environmental Agriculture
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    • v.23 no.3
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    • pp.170-177
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    • 2004
  • Agroenvironmental characteristics in paddy fields irrigated with the water of Nagdong river were analyzed along the river watershed for two years from 1999. The sites monitored from upper reaches of the river were Andong, Sangju, Gumi, Goryeong, Changnyeong, Milyang and Pusan. In paddy soils, the contents of heavy metals such as Cd, Cr, Cu, Pb, Ni, Zn and As were around natural values showing the highest values in Pusan followed by Goryeong. In brown rice, the contents of heavy metals were lower than natural values. Soil chemical properties appeared higher values in the lower reaches including Goryeong than the upper ones. The highest parameters in Goryeong were pH ($5.9{\sim}6.1$), EC ($0.8{\sim}0.9\;dS/m$), $Av.P_2O_5$ ($155{\sim}201\;mg/kg$), exchangeable Ca ($6.7{\sim}7.4\;cmol^+/kg$), Mg ($1.92{\sim}2.50\;cmol^+/kg$), K ($0.18{\sim}0.21\;cmol^+/kg$) and those in Pusan were organic matter ($23.0{\sim}29.1\;g/kg$) and T-N ($1.6{\sim}1.8\;mg/kg$). In conclusion, the recommended rates of N fertilizer for rice cropping were 21.4%, 11.8% and 8.8% high for Andong, Sangju and Gumi, respectively and 14.9%, 4.6%, 4.5% and 11.5% low for Goryeong, Changnyeong, Milyang and Pusan, respectively reflecting the chemical properties of soils and the quality of irrigation water on the basis of 110 kgN/ha. In the case of phosphorous, the rates were 18.9% and 33.3% low for Changnyeong and others, respectively on the basis of $45\;kgP_2O_5/ha$. The populations of bacteria, fungi, actinomycetes, Bacillus, fluorescent Pseudomonas and Biomass C were high at the lower reaches including Goryeong, which showed relatively much nutrient contents of organic matter, total N and phosphorous etc.

The Cross-talk Mechanisms of Constitutive Androstane Receptor (CAR) in the Regulation of its Activity, Energy Metabolism, Cellular Proliferation and Apoptosis (Constitutive Androstane Receptor (CAR)의 활성, 에너지 대사 및 세포의 증식과 사멸의 조절에 대한 CAR의 cross-talk 기전)

  • Min, Gyesik
    • Journal of Life Science
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    • v.30 no.2
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    • pp.211-220
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    • 2020
  • The activity of CAR can be regulated not only by ligand binding but also by phosphorylation of regulatory factors involved in extracellular signaling pathways, cross-talk interactions with transcription factors, and the recruitment, degradation, and expression of coactivators and corepressors. This regulation of CAR activity can in turn have effects on the control of diverse physiological homeostasis, including xenobiotic and energy metabolism, cellular proliferation, and apoptosis. CAR is phosphorylated by the ERK1/2 signaling pathway, which causes formation of a complex with Hsp-90 and CCRP, leading to its cytoplasmic retention, whereas phenobarbital inhibits ERK1/2, which causes dephosphorylation of the downstream signaling molecules, leading to the recruitment to CAR of the activated RACK-1/PP2A components for the dephosphorylation, nuclear translocation, and the transcriptional activation of CAR. Activated CAR cross-talks with FoxO1 to induce inhibition of its transcriptional activity and with PGC-1α to induce protein degradation by ubiquitination, resulting in the transcriptional suppression of PEPCK and G6Pase involved in gluconeogenesis. Regulation by CAR of lipid synthesis and oxidation is achieved by its functional cross-talks, respectively, with PPARγ through the degradation of PGC-1α to inhibit expression of the lipogenic genes and with PPARα through either the suppression of CPT-1 expression or the interaction with PGC-1α each to induce tissue-specific inhibition or stimulation of β-oxidation. Whereas CAR stimulates cellular proliferation by suppressing p21 expression through the inhibition of FoxO1 transcriptional activity and inducing cyclin D1 expression, it suppresses apoptosis by inhibiting the activities of MKK7 and JNK-1 through the expression of GADD45B. In conclusion, CAR is involved in the maintenance of homeostasis by regulating not only xenobiotic metabolism but also energy metabolism, cellular proliferation, and apoptosis through diverse cross-talk interactions with extracellular signaling pathways and intracellular regulatory factors.

Total Polyphenol Contents and Antioxidant Activities of Methanol Extracts from Vegetables produced in Ullung Island (울릉도산 산채류 추출물의 총 폴리페놀 함량 및 항산화 활성)

  • Lee, Syng-Ook;Lee, Hyo-Jung;Yu, Mi-Hee;Im, Hyo-Gwon;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.233-240
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    • 2005
  • To discover new functional materials using edible plants, antioxidant activities of methanol extracts from various parts of seven wild vegetables were investigated in vitro. Total polyphenol contents, determined by Folin-Denis method, varied from 16.74 to $130.22{\mu}g/mg$. Radical-scavenging activities of methanol extracts were examined using ${\alpha},\;{\alpha}-diphenyl-{\beta}-pirrylhydrazyl$ (DPPH) radicals and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) assay. Inhibition effects on peroxidation of linoleic acid determined by ferric thiocyanate (FTC) method and on oxidative degradation of 2-deoxy-D-ribose in Fenton-type reaction system were dose-dependent. Athyrium acutipinulum Kodama (leaf and rood), Achyranthes japonica (Miq.) Nakai (seed), and Solidago virga-aurea var. gigantea Nakai (root) showed relatively high antioxidant activities in various systems.

The Geochemistry of Copper-bearing Hydrothermal Vein Deposits in Goseong Mining District (Samsan Area), Gyeongsang Basin, Korea (경상분지내 삼산지역 열수동광상에 관한 지화학적 연구)

  • Choi, Sang Hoon;So, Chil Sup;Kweon, Soon Hag;Choi, Kwang Jun
    • Economic and Environmental Geology
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    • v.27 no.2
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    • pp.147-160
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    • 1994
  • Copper-bearing hydrothermal vein mineralization of the Samsan area was deposited in two stages (I and II) of quartz-calcite-sulfide veins which fill fissures in Cretaceous volcanic and sedimentary rocks of the Gyeongsang basin. The major ore minerals, chalcopyrite and sphalerite, together with pyrite, galena, hematite, and minor sulfosalts, occur with epidote and chlorite as gangue minerals in stage I quartz veins. Chlorite geothermometry, fluid inclusion and stable isotope data indicate that copper ore was deposited mainly at temperatures between $330^{\circ}C$ and $280^{\circ}C$ from fluids with salinities between 12 and 3 equiv. wt % NaCl. Evidence of fluid boiling indicates a range of pressures from ${\leq}100$ to 200 bars bars. Within ore stage I there was an apparent decrease in ${\delta}^{34}S$ values of $H_{2}S$ with paragenetic time, from 8.0 to 2.3 per mil. This pattern was likely achieved through progressive increases in activity of oxygen accompanying boiling and mixing. In the early part of the first stage, the high temperature, high salinity fluids gave way to progressively cooler and more dilute fluids of the late parts in the first stage and of the second stage. There is a systematic decrease in calculated ${\delta}^{18}O_{water}$ values with decreasing temperature in the Samsan hydrothermal system, from values of -86 per mil for early portion of stage I through -5.9 per mil for late portion of stage I to -6.3 per mil for stage II. The ${\delta}D$ values of fluid inclusion waters also decrease with paragenetic time from -76 per mil to -86 per mil. These trends combined with mineral paragenesis and fluid inclusion data are interpreted to indicate progressive cooler, more oxidizing meteoric water inundation of an early exchanged meteoric hydrothermal system.

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Effects of Donor Somatic Cell Conditions on In Vitro Development of Nuclear Transplanted Porcine Embryos (돼지 공여세포의 조건이 핵이식 수정란의 체외발달에 미치는 영향)

  • 홍승표;박준규;이명열;이지삼;정장용
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.213-221
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    • 2001
  • This study was conducted to examine in vitro developmental ability of porcine embryos after somatic cell nuclear transfer. The porcine ear fell was cultured in vitro for confluency in serum-starvation condition(TCM-199 + 0.5% FBS) far 3~6 days of cell confluency. The zona pellucida of IVM oocytes were partially drilled using laser system. Single somatic cell was individually transferred into enucleated oocytes. And the reconstructed embryos were electrically fused(single DC 1.9kv/cm, 30$\mu$ sec) with 0.3M mannitol. After electrofusion, embryos were activated(single AC 5v/mm, 5sec) and cultured in HCSU-23 medium containing 10% FBS at 39$^{\circ}C$, 5% $CO_2$ in air for 6 to 8 days. The fusion rate of donor cells was 45.6, 36.8 and 46.1% in 3~4, 5~6 days of serum starvation and non serum starvation(N-S), and were 52.7. 53.0 and 51.7% in 1~2. 5~6 and 13~14 passages of donor cell culture, respectively. No significant difference was found in the fusion rate of donor cells by the duration of serum starvation treatment or the number of donor cell passages. By the size of donor cells, however, the fusion rate was significantly higher(P<0.05) for reconstructed embryos derived from 25r $\mu$m $\geq$ site of donor cells (65.3%) than that of 25~30$\mu$ m(42.5%) or 30$\mu$ m(45.5%)$\leq$ cells. The cleavage rate was significantly (P<0.05) higher in 3~4 darts of serum starvation treatment(67.1%) than that in N-S (50.7%) or 5~6 days of starvation(57.1%). The activation rate by the size of donor cells in fused oocytes was 56.5, 68.8 and 58.5%, respectively, and was not significant.

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Factors Related to Poor School Performance of Elementary School Children (국민학교아동의 학습부진에 관련된 요인)

  • Park, Jung-Han;Kim, Gui-Yeon;Her, Kyu-Sook;Lee, Ju-Young;Kim, Doo-Hie
    • Journal of Preventive Medicine and Public Health
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    • v.26 no.4 s.44
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    • pp.628-649
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    • 1993
  • This study was conducted to investigate the factors related to the poor school performance of the elementary school children. Two schools in Taegu, one in the affluent area and the other in the poor area, were selected and a total of 175 children whose school performance was within low 10 percentile (poor performers) and 97 children whose school performance were within high 5 percentile (good performers) in each class of 2nd, 4th and 6th grades were tested for the physical health, behavioral problem and family background. Each child had gone through a battery of tests including visual and hearing acuity, anthropometry (body weight, height, head circumference), intelligence (Kodae Stanford-Binet test), test anxiety (TAI-K), neurologic examination by a developmental pediatrician and heavy metal content (Pb, Cd, Zn) in hair by atomic absorption spectrophotometry. A questionnaire was administered to the mothers for prenatal and prenatal courses of the child, family environment, child's developmental history, and child's behavioral and learning problems. Another questionnaire was administered to the teachers of the children for the child's family background, arithmatic & language abilities and behavioral problem. The poor school performance had a significant correlation with male gender, high birth order, broken home, low educational and occupational levels of parents, visual problem, high test anxiety score, attention deficit hyperactivity disorder (ADHD), poor physical growth (weight, height, head circumference) and low I.Q. score. The factors that had a significant correlation with the poor school performance in multiple logistic regression analysis were child's birth order (odds ratio=2.06), male gender(odds ratio=5.91), broken home(odds ratio=9.29), test anxiety score(odds ratio=1.07), ADHD (odds ratio=9.67), I.Q. score (odds ratio=0.85) and height less than Korean standard mean-1S.D.(odds ratio=11.12). The heavy metal contents in hair did not show any significant correlation with poor school performance. However the lead and cadmium contents were high in males than in females. The lead content was negatively correlated with child's grade(P<0.05) and zinc was positively correlated with grade (P<0.05). among the factors that showed a significant correlation with the poor school performance, high birth order, short stature and ADHD may be modified by a good family planning, good feeding practice for infant and child, and early detection and treatment of ADHD. Also, teacher and parents should restrain themselves from inducing excessive test anxiety by forcing the child to study and over-expecting beyond the child's intellectual capability.

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Geochemistry and Genesis of Hydrothermal Cu Deposits in the Gyeongsang Basin, Korea : Hwacheon-ri Mineralized Area (경상분지내 열수동광상의 지화학 및 성인연구 : 화천리지역 광화대)

  • So, Chil-Sup;Choi, Sang-Hoon;Yun, Seong-Taek
    • Economic and Environmental Geology
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    • v.28 no.4
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    • pp.337-350
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    • 1995
  • The Hwacheon-ri mineralized area is located within the Cretaceous Gyeongsang Basin of the Korean peninsula. The mineralized area includes the Hwacheon, Daeweon, Kuryong and Cheongryong mines. Each of these mines occurs along copper-bearing hydrothermal quartz veins that crosscut late Cretaceous volcanic rocks, although some disseminated ores in host rocks also exist locally. Mineralization can be separated into three distinct stages (I, II, and III) which developed along preexisting fracture zones. Stage I is ore-bearing, whereas stages II and III are barren. The main phase of ore mineralization, stage I, can be classified into three substages (Ia, Ib and Ic) based on ore mineral assemblages and textures. Substage Ia is characterized by pyrite-arsenopyrite-molybdenite-pyrrhotite assemblage and is most common at the Hwacheon deposit. Substage Ib is represented by main precipitation of Cu, Zn, and Pb minerals. Substage Ic is characteristic of hematite occurrence and is shown only at the Kuryong and Cheongryong deposits. Some differences in the ore mineralization at each mine in the area suggest that the evolution of hydrothermal fluids in the area varied in space (both vertically and horizontally) with respect to igneous rocks relating the ore mineralization. Fluid inclusion data show that stage I ore mineralization mainly occurred at temperatures between ${\approx}350^{\circ}$ and ${\approx}200^{\circ}C$ from fluids with salinities between 9.2 and 0.5 wt.% eq. NaCl. In the waning period of substage Ia, the high temperature and salinity fluid gave way to progressively cooler, more dilute fluids of later substage Ib and Ic (down to $200^{\circ}C$, 0 wt.% NaCl). There is a systematic decrease in the calculated ${\delta}^{18}O_{H2O}$ values with paragenetic time in the Hwacheon-ri hydrothermal system from values of ${\approx}2.7$‰ for substage Ia, through ${\approx}-2.8$‰ for substage Ib, to ${\approx}-9.9$‰ for substage Ic. The ${\delta}D$ values of fluid inclusion water also decrease with decreasing temperature (except for the Daeweon deposit) from -62‰ (substage Ia) to -80‰ (substage Ic and stage III). These trends are interpreted to indicate the progressive cooler, more oxidizing unexchanged meteoric water inundation of an initial hydrothermal system which is composed of highly exchanged meteoric water. Equilibrium thermodynamic interpretation of the mineral assemblages with the variation in amounts of chalcopyrite through the paragenetic time, and the evolution of the Hwacheon-ri hydrothermal fluids indicate that the solubility of copper chloride complexes in the hydrothermal system was mainly controlled by the variation of temperature and $fo_2$ conditions.

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