• Title/Summary/Keyword: Phase-contrast

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Increased Protein of the Secretory Leukocyte Pretense Inhibitor (SLPI) and the Expression of Growth Factors in NIH3T3 Cells by LPS Stimulation (NIH3T3 세포주에서 LPS자극에 의한 분비백혈구단백분해효소억제제 (SLPI)의 단백질증가와 성장인자들의 발현)

  • Lee, Sang-Hwa;Choi, Baik-Dong;Jeong, Soon-Jeong;Jang, Hyun-Seon;Kim, Byung-Ock;Lim, Do-Seon;Park, Joo-Cheol;Wang, Guan-Lin;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.36 no.3
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    • pp.165-172
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    • 2006
  • Secretory leukocyte protease inhibitor (SLPI) involves tissue protection against the destructive action of neutrophil elastase at the site of inflammation. Several studies on new functions of SLPI have demonstrated that SLPI may play a primary role in innate immunity than protease inhibitor, To identify the function of SLPI by lipopolysaccharide (LPS) stimulation in the embryonic fibroblast (NIH3T3) cells. we studied the expression of SLPI compared to other growth factors involving the LPS treatment. To address this, we performed the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of the SLPI and some growth factors such as VEGF. bFGF, and PDGF-BB after LPS stimulation. NIH3T3 cells were exposed 100 ng/mL Escherichia coli LPS for 30min, 60min, 90min, 24h, and 48h, respectively. The result of RT-PCR showed that SLPI and VEGF mRNA was expressed strongly in NIH3T3 without related to LPS stimulation. mRNA of bFGF was weakly expressed such as the expression of the control. PDGF mRNA expression gradually increased follows at time course. However, SLPI protein level was increased in lysates and culture medium by LPS stimulation. Phase contrast microscopic and scanning electron microscopic observation showed that the increased cell number and cytoplasmic enlargement of the NIH3T3 cells. Therefore, it suggests that the LPS upregulates SLPI expression in NIH3T3 cells. Moreover, secreted SLPI may stimulate cell proliferation and migration.

The effect of safflower seed fraction extract on periodontal ligament fibroblast and MC3T3-E1 cell in vitro (홍화씨 분획 추출물이 치주인대 섬유아세포와 MC3T3-E1 세포에 미치는 영향)

  • Huh, Ji-Sun;Kang, Jung-Hwa;Yoo, Yun-Jung;Kim, Chang-Sung;Cho, Kyoo-Sung;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.833-846
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    • 2001
  • Recently, use of natural medicine is getting more attention, and some of them are believed to be effective in the treatment of periodontitis. Among them, the seeds of safflower(Carthamus tinctrorius L.) have been proven to be effective through its use in bone diseases such as fracture and osteoporosis. During the last few years, studies using the seeds of safflower gown in Korea have been active, and it has been reported that safflower seed extract increase the proliferation and the alkaline phosphatase(ALP) activity of human periodontal ligament fibroblast(hPDLF), osteoblast, and that they promote the mineralization process. In animal studies, when safflower seed extract were administered orally new bone formation was promoted. Recently, in an effort to find out the most effective osteogenic components, among many components of the safflower seed, various safflower seed fraction extracts were obtained by multistep extraction of the safflower components using various solvents. Among these, saf-M-W fraction extracted by methanol and water was most effective in increasing osteogenic potential of osteoblasts. In this study, the effect of safflower seed fraction extract, saf-M-W, on the growth and differentiation of hPDLF and MC3T3-E1 cell was investigated. The toxicity of saf-M-W on both cells was measured using M'IT(3-(4,5dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide) test, and ALP activity was measured using the colorimetric assay of hPDLF. In addition, in MC3T3-El cells, the expression of ALP, bone sialoprotein(BSP) mRNA was observed using Northern blot, and the mineralized nodule formation Was observed using von Kossa stain and phase-contrast microscope. 1. In concentrations below $10{\mu}g/ml$, saf-M-W didn't show any toxicity on hPDLF and MC3T3-El cell. 2. The change in saf-M-W concentration had no effect on the ALP activity of hPDLF. 3. In MC3T-E1 cells, mRNA expressions of ALP and BSP were greater in the experimental group treated with $10{\mu}g/ml$ concentration of saf-M-W compared with the control group. 4. In MC3T3-El cells, abundance of mineralized nodules were formed in the experimental group treated with $10{\mu}g/ml$ Concentration of saf-M-W, while no mineralized nodule was formed in the control group. These results suggest that safflower seed fraction extract, saf-M-W. didn't show any toxicity on hPDLF and MC3T3-E1 cell at concentrations below $10{\mu}g/ml$ and effectively enhanced the differentiation and osteogenic potential of MC3T3-El cell.

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Asbestos Exposure and Health Risk Assessment for the Residents Near the Abandoned Mining Area in Chungbuk, Korea (충북지역 폐석면광산 인근주민의 석면노출과 건강위해도 평가)

  • Shin, Jin-Ho;Oh, Seok-Ryul;Hwang, Soon-Yong;Chung, Sook-Nye;Kim, Ji-Hui;Nam, Eun-Jung;Lee, Jin-Hyo;Choi, Hee-Jin;Eom, Seok-Won;Chae, Young-Zoo;Park, Chul-Hwi
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.5
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    • pp.345-350
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    • 2012
  • To investigate the exposure and health risk assessment for the residents near the D-asbestos mine in Chungbuk, Korea. We analyzed asbestos in the 20 ambient air and 23 activity based samples near the mine. The airborne sample results are showed that 8 of 20 samples ranged between 0.0025 to 0.0029 f/cc (fiber per cubic centimeter) and the others were below the detection limit by phase contrast microscopy (PCM). In addition, asbestos fibers were under the detection limit or not being by transmission electron microscopy (TEM). Based on interview and survey targeting the local residents, we made the activity based sampling (ABS) scenarios fit to the conditions of field. At the same time, we calculated the excess lifetime cancer risk (ELCR) of these ABS scenarios according to the ELCR average value and 95% upper confidence limit (UCL). At the case of weed whacking, soil digging and sweeping yard scenario, 95% UCL of ELCR exceeded the $1{\times}10^{-4}$, acceptable risk range for exposure. Based on our study results, it is necessary safety measures such as risk communication, abatement or management of naturally occurring asbestos (NOA).

Study of Rat Mammary Epithelial Stem Cells In Vivo and In Vitro (생체 및 시험관에서 유선 상피 모세포의 분리와 동정)

  • Nam Deuk Kim;Kee-Joo Paik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.470-486
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    • 1995
  • Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

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A Study on the Chitin and Protein Contents in Shells of 5 Marine Crustaceans (5종(種)의 해산(海産) 갑각류각피(甲殼類殼皮)에서의 Chitin 및 단백질함량(蛋白質含量)에 관한 연구(硏究))

  • Lee, Mee-Sook;Seo, Jung-Sook;Mo, Su-Mi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.13 no.3
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    • pp.307-312
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    • 1984
  • The dried pure shells deprived of soft tissues were subjected to analysis of chitin-protein complexes from 5 species of marine crustaceans, including 2 species of crabs and 3 species of shrimps. The protein fractions were obtained from chitin-protein complexes under the varying conditions of extractions and the crude chitin was prepared from the shells by the sulfurous acid process. The crude chitin was purified through the extraction with several organic solvents such as dimethyl-acetamide, N-methylpyrrolidone. The purified chitin was also examined using the phase contrast microscope. Total protein contents of the shells were diverse, showing 9.6% for Portunus trituberculatus, 3.1%, Charybdis bimaculata, 9.4%, Penaeus japonicus, 10.9%, Metapenaeus intermedius and 5.8%, Squilla oratoria. Covalently bound protein varied with species from 2.1% for Charybdis bimaculata to 9.9% for Metapenaeus intermedius. The puified chitin contents of the shells were shown to 21.1% for Portunus tritube rculatus, 6.2%, Charybdis bimaculata, 20.2%, Penaeus japonicus, 27.1%, Metapenaeus intermedius and 25.5%, Squilla oratoria. Exceptionally low analytical value obtained with Charybdis bimaculata are supposed to be due to the very young subjects. The ratios of chitin to covalently bound protein in the shells were various such as 2.7 to 1 for Portunus trituberculatus, Penaeus japonicus and Metapenaeus intermedius, 3.1 to 1, Charybdis bimaculata and 6.1 to 1, Squilla oratoria. The microscope finding of the purified chitin showed the filamentous form in all the specimen.

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Development of Health Promotion Program through IUHPE - Possibilities of collaboration in East Asia - (IUHPE를 통한 건강 증진 프로그램의 발달-동아시아권의 공동연구의 가능성-)

  • Moriyama, Masaki
    • Proceedings of The Korean Society of Health Promotion Conference
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    • 2004.10a
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    • pp.1-16
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    • 2004
  • This paper considers the possibilities of health promotion from the following perspectives; (1) IUHPE, (2) socio-cultural similarities, (3) action research, and (4) learning from our past. 1. The IUHPE values decentralized activities through regions, and countries such as Japan, Korea, Hong Kong, Taiwan and China belong to NPWP region. Since IUHPE World Conference was held in Japan in 1995, Japan used to occupy more than 60% of NPWP membership. After 2001, membership is increasing rapidly in Chinese speaking sub-region. The transnational collaboration is still in its beginning phase. 2. Confucianism is one of key points. Confucian tradition should not be seen only as obstacles but as advantages to seek a form of health promotion more acceptable in East Asia. 3. Within the new public health framework, people are expected to create and live their health. However, especially in Japan, the tendency of 'lacking of face-to-face explicit interactions' is still common at health-promotion settings as well as academic settings. Therefore, the author tried participatory approaches such as asking WlFY (interactive questions designed for subjects to review their daily life and environment) and as introducing round table interactions. So far, majority of participants welcome new trials. 4. The following social phenomena are comparatively discussed after Japanese invasion and occupation of Korea ended in 1945; ·status of oriental medicine, ·separation of dispensary services, and ·health promotion specialist as a national license. In contrast to Japanese' tendency of maintaining the status quo and postponing of substantial social change, trend toward rapid and dynamic social changes are more commonly observed in Korea. Although all of above possibilities are still in their beginning stages, they are going to offer interesting directions waiting for further challenges and accompanying researches.

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Expression of Growth Factors and Secretory Leukocyte Protease Inhibitor (SLPI) in RAW264.7 Cells after Lipopolysaccharide (LPS) Stimulation (대식세포주 (RAW264.7)에서 박테리아성 지질다당류 (LPS) 자극에 의한 분비백혈구단백분해효소억제제 (SLPI)와 성장인자들의 발현)

  • Son, Wook-Hee;Choi, Baik-Dong;Jeong, Soon-Jeong;Wang, Guan-Lin;Hwang, Ho-Keel;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.37 no.2
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    • pp.93-102
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    • 2007
  • Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

High Glucose Induces Apoptosis through Caspase-3 Dependent Pathway in Human Retinal Endothelial Cell Line (인간망막 내피세포주에서 고농도 포도당이 caspase-3 경로를 통해 세포자연사 유도)

  • Seo, Eun-Sun;Chae, Soo-Chul;Kho, Eun-Gyeong;Lee, Jong-Bin
    • Korean Journal of Environmental Biology
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    • v.27 no.1
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    • pp.66-72
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    • 2009
  • Diabetic Retinopathy (DR) is a leading cause of blindness among adults in the western countries. Hyperglycemia is a condition, that induces apoptotic cell death in a variety of cell types in diabetes, but the mechanism remains unclear. The aim of the study is to understand the effects of high Glucose on Human Retinal Endothelial Cells. Retinal endothelial cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) containing 5, 25 and 50 mM Glucose, incubated for 24, 36 and 48 hours in humidified 5 % CO$_2$ incubator at 37$^{\circ}C$. Human Retinal Endothelial Cell Line (HREC) were characterized for morphology with different treatment by phase contrast microscopic analysis. Number of dead and viable cells was counted by trypan blue exclusion and supported by MTT assay. The intracellular Hydrogen peroxide (H$_2$O$_2$), a Reactive Oxygen Species (ROS) generation in high glucose conditions was assessed by FOX II assay and apoptosis by caspase-3 assay. The high glucose treated cells undergoing DNA fragmentation was witnessed by Agarose gel electrophoresis. We found that the cells incubated with 25 and 50 mM glucose containing medium for 48 hours altered the morphology of the cell, induced apoptosis and DNA fragmentation. The dead cell number were high in 25 and 50 mM when compared to the cells incubated with 5 mM glucose for 24, 36, and 48 hours. Also, the H$_2$O$_2$ levels and the activity of caspase-3 were increased in high glucose treated cells. Conclusions/interpretation: Our results demonstrated that elevated glucose induces apoptosis in cultured HREC. The hyperglycemia-induced increase in apoptosis may be dependent on caspase activation. The association between ROS generation and caspase-3 activation on high glucose treated cells is yet to be investigated.

Characterization of Biopesticides (Bacillus thuringiensis) Produced in Korea (국내에서 생산된 Bacillus thuringiensis 살충제의 특성)

  • Kil, Mi-Ra;Kim, Da-A;Choi, Su-Yeon;Paek, Seung-Kyoung;Kim, Jin-Su;Jin, Da-Yong;Hwang, In-Chon;Yu, Yong-Man
    • The Korean Journal of Pesticide Science
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    • v.11 no.3
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    • pp.201-209
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    • 2007
  • Characteristics of the 5 biopesticides that included Bacillus thuringiensis and on the domestic markets were investigated. These products were contained different strains of B. thuringiensis, for examples; product A and E was B. thuringiensis subsp aizawai; product B was B. thuringiensis; product C was B. thuringiensis Berline var. kurstaki; product D was B. thuringiensis var. kurstaki. Number of active spores were counted because they could influence the bio-activity against target pests. Only product C are contained the fixed quantity as its label, however, product D and E were a tenth part, and product A and B were a hundredth part of their descriptions. The pHs of product A and B were measured 3.67 and 3.73, and C, D and E were 5, respectively. Typical bypyramidal crystals produced from B. thuringiensis was found in only product C under a phase contrast microscope. For the uniform formulation of products that conformed whether B. thuringiensis were equally spreaded on the crops, B. thuringiensis in the C, D and E were equally grown on the nutrient agar medium As a results, product A were more different from product C than any other products. When product A and C were bioassayed against different larval stages of diamondback moth, their mortalities with spraying application were showed 100% after 48 hours.

An integrated Method of New Casuistry and Specified Principlism as Nursing Ethics Methodology (새로운 간호윤리학 방법론;통합된 사례방법론)

  • Um, Young-Rhan
    • Journal of Korean Academy of Nursing Administration
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    • v.3 no.1
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    • pp.51-64
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    • 1997
  • The purpose of the study was to introduce an integrated approach of new Casuistry and specified principlism in resolving ethical problems and studying nursing ethics. In studying clinical ethics and nursing ethics, there is no systematic research method. While nurses often experience ethical dilemmas in practice, much of previous research on nursing ethics has focused merely on describing the existing problems. In addition, ethists presented theoretical analysis and critics rather than providing the specific problems solving strategies. There is a need in clinical situations for an integrated method which can provide the objective description for existing problem situations as well as specific problem solving methods. We inherit two distinct ways of discussing ethical issues. One of these frames these issues in terms of principles, rules, and other general ideas; the other focuses on the specific features of particular kinds of moral cases. In the first way general ethical rules relate to specific moral cases in a theoretical manner, with universal rules serving as "axioms" from which particular moral judgments are deduced as theorems. In the seconds, this relation is frankly practical. with general moral rules serving as "maxims", which can be fully understood only in terms of the paradigmatic cases that define their meaning and force. Theoretical arguments are structured in ways that free them from any dependence on the circumstances of their presentation and ensure them a validity of a kind that is not affected by the practical context of use. In formal arguments particular conclusions are deduced from("entailed by") the initial axioms or universal principles that are the apex of the argument. So the truth or certainty that attaches to those axioms flows downward to the specific instances to be "proved". In the language of formal logic, the axioms are major premises, the facts that specify the present instance are minor premises, and the conclusion to be "proved" is deduced (follows necessarily) from the initial presises. Practical arguments, by contrast, involve a wider range of factors than formal deductions and are read with an eye to their occasion of use. Instead of aiming at strict entailments, they draw on the outcomes of previous experience, carrying over the procedures used to resolve earlier problems and reapply them in new problmatic situations. Practical arguments depend for their power on how closely the present circumstances resemble those of the earlier precedent cases for which this particular type of argument was originally devised. So. in practical arguments, the truths and certitudes established in the precedent cases pass sideways, so as to provide "resolutions" of later problems. In the language of rational analysis, the facts of the present case define the gounds on which any resolution must be based; the general considerations that carried wight in similar situations provide warrants that help settle future cases. So the resolution of any problem holds good presumptively; its strengh depends on the similarities between the present case and the prededents; and its soundness can be challenged (or rebutted) in situations that are recognized ans exceptional. Jonsen & Toulmin (1988), and Jonsen (1991) introduce New Casuistry as a practical method. The oxford English Dictionary defines casuistry quite accurately as "that part of ethics which resolves cases of conscience, applying the general rules of religion and morality to particular instances in which circumstances alter cases or in which there appears to be a conflict of duties." They modified the casuistry of the medieval ages to use in clinical situations which is characterized by "the typology of cases and the analogy as an inference method". A case is the unit of analysis. The structure of case was made with interaction of situation and moral rules. The situation is what surrounds or stands around. The moral rule is the essence of case. The analogy can be objective because "the grounds, the warrants, the theoretical backing, the modal qualifiers" are identified in the cases. The specified principlism was the method that Degrazia (1992) integrated the principlism and the specification introduced by Richardson (1990). In this method, the principle is specified by adding information about limitations of the scope and restricting the range of the principle. This should be substantive qualifications. The integrated method is an combination of the New Casuistry and the specified principlism. For example, the study was "Ethical problems experienced by nurses in the care of terminally ill patients"(Um, 1994). A semi-structured in-depth interview was conducted for fifteen nurses who mainly took care of terminally ill patients. The first stage, twenty one cases were identified as relevant to the topic, and then were classified to four types of problems. For instance, one of these types was the patient's refusal of care. The second stage, the ethical problems in the case were defined, and then the case was analyzed. This was to analyze the reasons, the ethical values, and the related ethical principles in the cases. Then the interpretation was synthetically done by integration of the result of analysis and the situation. The third stage was the ordering phase of the cases, which was done according to the result of the interpretation and the common principles in the cases. The first two stages describe the methodology of new casuistry, and the final stage was for the methodology of the specified principlism. The common principles were the principle of autonomy and the principle of caring. The principle of autonomy was specified; when competent patients refused care, nurse should discontinue the care to respect for the patients' decision. The principle of caring was also specified; when the competent patients refused care, nurses should continue to provide the care in spite of the patients' refusal to preserve their life. These specification may lead the opposite behavior, which emphasizes the importance of nurse's will and intentions to make their decision in the clinical situations.

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