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Expression of Cell Cycle Related Genes in HL60 Cells Undergoing Apoptosis by X-irradiation (HL60 세포주에서 방사선 조사에 의한 Apoptosis와 세포 주기 관련 유전자의 발현 변화)

  • Kim, Jin-Hee;Park, In-Kyu
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.377-388
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    • 1998
  • Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.

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Immunogenicity of Synthetic Peptide Specific for Major Immunogenic Determinat of Hepatitis B Surface Antigen (B형간염(型肝炎) 표면항원(表面抗原)의 주면역원(主免疫原) 결정기(決定基)에 특이(特異)한 합성(合成) Peptide의 면역원성(免疫原性)에 관한 연구(硏究))

  • Shin, Kwang-soon;Han, Su-nam
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.7-17
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    • 1985
  • Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

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A Study for Crystal Growth Inhibition of Ettringite by Solution Synthesis Experiment (용액합성실험에 의한 에트린자이트 결정성장억제 연구)

  • Lee, Hyo-Min;Hwang, Jin-Yeon;Oh, Ji-Ho
    • Journal of the Mineralogical Society of Korea
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    • v.23 no.1
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    • pp.51-61
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    • 2010
  • Ettringite $(Ca_6[Al(OH)_6]_2(SO_4)_3{\cdot}26H_2O)$ is a sulfate mineral that shows a complicate property in concrete. It is often called as "a cancer of concrete" because secondary ettringite formation in hardened concrete often cause expansion and cracking of concrete due to its expansive crystal structure. In the present study, we tested the possibility for crystal growth inhibition of secondary ettringite by crystallization inhibitors that are commercially used for scaling inhibitors in Korea. For the test, we developed a method of ettringite solution synthesis. Three types of crystallization inhibitors were selected and examined the effects On ettringite growth inhibition. The experimental results of ettringite solution synthesis indicated that ettringite was successfully synthesized under condition that the mass balance between calcium hydroxide saturated solution and aluminum sulfate solution was attained. Monosulfate and semisulfate were synthesized when the ratio of $Ca^{2+}$ ions to ${SO_4}^{2+}$ ions was increased. The induction time of ettringite crystallization was less than 2 min. and crystallization was almost completed within an hour. The experimental results of ettringite crystallization inhibition showed that organic PBCT (2-Phosphonobutane-1,2,4-Tricarboxylic Acid) and inorganic SHMP (Sodium Hexametaphosphate) were relatively less effective on ettringite crystallization inhibition under experimental conditions. However, organic HEDP (1-Hydoxyethylidene-1,1-Diphosphonic Acid) effectively prevented ettringite growth with producing amorphous gel phase materials up to inhibitor concentration 0.1 vol.% of aluminum sulfate solution.

Cold Shock Response and Low Temperature Stable Transcript of DEAD-box RNA Helicase in Bacillus subtilis (DEAD-box RNA Helicase 유전자가 결핍된 Bacillus subtilis의 저온 충격 반응성과 저온 안정성 전사물)

  • Oh, Eun-Ha;Lee, Sang-Soo
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.289-294
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    • 2011
  • We investigated the cold shock sensitivity of DEAD-box RNA helicase gene deleted strains of in Bacillus subtilis CU1065. To understand cold shock effects, cells were cultivated at $37^{\circ}C$ to log phase ($O.D_{600}$=0.5-0.6) and then temperature was shifted to $15^{\circ}C$. Cold shock slow down the growth rate of wild type and deleted strains of DEAD-box RNA helicase gene (ydbR, yfmL, yqfR, deaD). The growth rate of ydbR deleted strain is 5 times severely reduced compared to that of wild type strain (CU1065). But the growth rate of other three (yfmL, yqfR, deaD) deleted strains is nearly equal to the growth rate of wild type. Compared to $37^{\circ}C$, the amount of ydbR and yqfR mRNA transcripts are increased at the growth temperature of $15^{\circ}C$. On the other hands the mRNA transcripts of yfmL and deaD are not changed at both conditions of $37^{\circ}C$ and $15^{\circ}C$. Upon cold shock treatment ydbR mRNA transcript is clearly increased. After treatment of rifampicin (bacteria transcription inhibitor) the amount of ydbR mRNA was measured. Temperature shift from $37^{\circ}C$ to $15^{\circ}C$ and rifampicin treatment showed slowly decay of ydbR mRNA. But at $37^{\circ}C$ and rifampicin treatment ydbR mRNA is rapidly reduced. These results showed that cold shock induction of ydbR mRNA resulted from the stability of ydbR mRNA and not from the transcription induction of ydbR. In relation to these results, we found the cold box element of csp (cold shock protein gene) in 5' untranslated region of ydbR gene. Cold shock induction of ydbR is caused by the stability of ydbR mRNA like the stability of csp mRNA.

Organization and function of shoot apical meristem affecting growth and development in plants (식물의 생장과 발달에 영향을 미치는 슈트 정단분열조직의 체제와 기능)

  • Lee, Kyu Bae
    • Journal of Plant Biotechnology
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    • v.41 no.4
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    • pp.180-193
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    • 2014
  • In plants, a shoot apex has a small region known as the shoot apical meristem (SAM) having a group of dividing (initiating) cells. The SAM gives rise to all the groundabove structures of plants throughout their lifetime, and thus it plays important role in growth and development of plants. This review describes theories to explain the SAM organization and function developed over the last 250 years. Since in 1759 German botanist C. F. Wolff has described firstly the SAM, in 1858 Swiss botanist C. N${\ddot{a}}$geli proposed the apical cell theory from the observation of a large single apical cell in the SAM of seedless vascular plants: however, this view was recognized to be unsuitable to seed plants. In 1868, German botanist J. Hanstein suggested the histogen theory: this concept subdividing the SAM into dermatogen, periblem, and plerome was unable to generally apply to seed plants. In 1924, German botanist A. Schmidt proposed the tunica-corpus theory from the examination of angiosperm SAM in which two parts show different planes of cell division: this theory was proved to be not suitable to gymnosperm SAM, not have stable surface tunica layer. In 1938, American botanist A. Foster described zones in gymnosperm SAM based on the cytohistologic differentiation and thus called it a cytohistological zonation theory. With works by E. Gifford, in 1954, this zonation pattern was demonstrated to be also applicable to angiosperm SAM. As another theory, in 1952 French botanist R. Buvat proposed the m${\acute{e}}$rist${\grave{e}}$me d'attente (waiting meristem) theory: however, this concept was confuted because of its negation of function during vegetative growth phase to central initial cells. Rescent studies with Arabidopsis thaliana have found that formation and maintenance of the SAM are under the control of selected genes: SHOOTMERISTEMLESS (STM) gene forms the SAM, and WUSCHEL (WUS) and CLAVATA (CLV) genes function in maintaining the SAM; signaling between WUS and CLV genes act through a negative feedback loop.

The Impact of Service Level Management(SLM) Process Maturity on Information Systems Success in Total Outsourcing: An Analytical Case Study (토털 아웃소싱 환경 하에서 IT서비스 수준관리(Service Level Management) 프로세스 성숙도가 정보시스템 성공에 미치는 영향에 관한 분석적 사례연구)

  • Cho, Geun Su;An, Joon Mo;Min, Hyoung Jin
    • Asia pacific journal of information systems
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    • v.23 no.2
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    • pp.21-39
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    • 2013
  • As the utilization of information technology and the turbulence of technological change increase in organizations, the adoption of IT outsourcing also grows to manage IT resource more effectively and efficiently. In this new way of IT management technique, service level management(SLM) process becomes critical to derive success from the outsourcing in the view of end users in organization. Even though much of the research on service level management or agreement have been done during last decades, the performance of the service level management process have not been evaluated in terms of final objectives of the management efforts or success from the view of end-users. This study explores the relationship between SLM maturity and IT outsourcing success from the users' point of view by a analytical case study in four client organizations under an IT outsourcing vendor, which is a member company of a major Korean conglomerate. For setting up a model for the analysis, previous researches on service level management process maturity and information systems success are reviewed. In particular, information systems success from users' point of view are reviewed based the DeLone and McLean's study, which is argued and accepted as a comprehensively tested model of information systems success currently. The model proposed in this study argues that SLM process maturity influences information systems success, which is evaluated in terms of information quality, systems quality, service quality, and net effect proposed by DeLone and McLean. SLM process maturity can be measured in planning process, implementation process and operation and evaluation process. Instruments for measuring the factors in the proposed constructs of information systems success and SL management process maturity were collected from previous researches and evaluated for securing reliability and validity, utilizing appropriate statistical methods and pilot tests before exploring the case study. Four cases from four different companies under one vendor company were utilized for the analysis. All of the cases had been contracted in SLA(Service Level Agreement) and had implemented ITIL(IT Infrastructure Library), Six Sigma and BSC(Balanced Scored Card) methods since last several years, which means that all the client organizations pursued concerted efforts to acquire quality services from IT outsourcing from the organization and users' point of view. For comparing the differences among the four organizations in IT out-sourcing sucess, T-test and non-parametric analysis have been applied on the data set collected from the organization using survey instruments. The process maturities of planning and implementation phases of SLM are found not to influence on any dimensions of information systems success from users' point of view. It was found that the SLM maturity in the phase of operations and evaluation could influence systems quality only from users' view. This result seems to be quite against the arguments in IT outsourcing practices in the fields, which emphasize usually the importance of planning and implementation processes upfront in IT outsourcing projects. According to after-the-fact observation by an expert in an organization participating in the study, their needs and motivations for outsourcing contracts had been quite familiar already to the vendors as long-term partners under a same conglomerate, so that the maturity in the phases of planning and implementation seems not to be differentiating factors for the success of IT outsourcing. This study will be the foundation for the future research in the area of IT outsourcing management and success, in particular in the service level management. And also, it could guide managers in practice in IT outsourcing management to focus on service level management process in operation and evaluation stage especially for long-term outsourcing contracts under very unique context like Korean IT outsourcing projects. This study has some limitations in generalization because the sample size is small and the context itself is confined in an unique environment. For future exploration, survey based research could be designed and implemented.

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Limnological Characteristics and Influences of Free-floating Plants on the Woopo Wetland during the Summer (하계우포습지의 육수학적 특성 및 부유수생식물의 영향)

  • Joo, Gea-Jae;Kim, Gu-Yeon;Park, Sung-Bae;Lee, Chan-Woo;Choi, Sang-Ho
    • Korean Journal of Ecology and Environment
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    • v.35 no.4 s.100
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    • pp.273-284
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    • 2002
  • During January 1998-October 1999, the impact of free-floating plants (FFP) on limnology of the wetland ecosystem was evaluated through the investigation of physicochemical characteristics of the Woopo Wetland along with in situ manipulation experiments of aquatic plants. Flooding events occurred in the wetland during the summer period (Jun.-Aug.) and water levels rose to 2-3 m due to precipitation from the catchment and inflow from the main channel of the Nakdong River. Physicochemical parameters and plankton dynamics in the wetland during the summer were greatly influenced by floods and growth of free-floating plants. Dissolved oxygen (Jun.-Sept., 4.5${\pm}$2.5 mg/1; Oct.-May, 8,1 ${\pm}$4.0 mg/1) and pH (Jun.-Sept., 6.9${\pm}$0.4; Oct.-May,7.4${\pm}$0.8) levels were significantly lower during the summer than any other seasons. Three types of enclosure experiments (100 L, treatments with floating plants, screened and opened) were conducted under the presence and absence of sediment for 15 days in the 1999 summer. The treatments with sediment had higher levels of nutrient concentrations than those of the others. Among the treatments with sediment, nutrient concentrations in the treatments with free-floating plants were higher than the others. Zooplankton communities in each treatment showed a similar variation, although the scale of zooplankton densities differed. Rotifer community dominated the zooplankton at the initial phase of the experiment, but decreased drastically along with an increase of cladoceran and copepod communities. In conclusion, low levels of dissolved oxygen and pH in the Woopo Wetland during the summer seemed to be caused by a proliferation of free-floating plants and active decomposition process at the bottom of the sediment.

Improvement of an Simultaneous Determination for Clenbuterol and Ractopamine in Livestock Products using LC-MS/MS (LC-MS/MS를 이용한 축산물 중 clenbuterol과 ractopamine의 동시 분석법 개선)

  • Cho, Yoon-Jae;Chae, Young-Sik;Kim, Jae-Young;Kim, Hyung-Soo;Kang, Ilhyun;Do, Jung-Ah;Oh, Jae-Ho;Kwon, Kisung;Chang, Moon-Ik
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.25-33
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    • 2013
  • Clenbuterol and ractopamine, which are ${\beta}$-agonists, have been misused as a growth promoting agent in meat producing animals. Clenbuterol was banned for veterinary drug in Korea because of its problems regarding safety. Due to their adverse effects, such as cardiovascular and central nervous diseases on human health proper control and monitoring should be conducted. The existing analytical method of clenbuterol and ractopamine in the Food code was improved through our present study. The bovine muscle samples were subjected to enzymatic hydrolysis, extracted with ethyl acetate and defatted by hexane-methanol partitioning. A molecular imprinted polymer (MIP) solid phase extraction cartridge was used for clean-up and LC-MS/MS was operated in positive multiple reaction monitoring (MRM). Clenbuterol-$d_9$ and ractopamine-$d_3$ were used as an internal standard. The renewed method was validated according to the CODEX guideline. The limits of quantitation for clenbuterol and ractopamine were 0.2 and 0.5 ${\mu}g/kg$, respectively. The mean recoveries ranged in 104.2-113.5% for clenbuterol and in 107.6-118.1% for ractopamine. The improved method was able to save both time and expenses.

Effect of the Hexane Extract of Saussurea lappa on the Growth of HT-29 Human Colon Cancer Cells (목향 헥산추출물이 대장암세포인 HT-29 세포의 증식에 미치는 영향)

  • Kim, Eun-Ji;Park, Hee-Sook;Lim, Soon-Sung;Kim, Jong-Sang;Shin, Hyun-Kyung;Yoon, Jung-Han
    • Korean Journal of Food Science and Technology
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    • v.40 no.2
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    • pp.207-214
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    • 2008
  • In Asia Saussurea lappa (SL) has been used as a traditional herbal medicine to treat abdominal pain and tenesmus. Recently, in vitro cell culture studies have shown that SL has anti-ulcer, anti-inflammatory, and anti-tumor properties. To explore its potential chemopreventive and chemotherapeutic effects in colon cancer, we examined whether the hexane extract of SL (HESL) could inhibit the growth of HT-29 human colon cancer cells, and investigated the mechanisms for this effect. The cells were cultured with various concentrations (0-5 ${\mu}g/mL$) of HESL. The results indicated that HESL markedly decreased the numbers of viable HT-29 cells; whereas at the concentration of 5 ${\mu}g/mL$, HESL slightly decreased the viable cell numbers of CCD 1108Sk human skin normal fibroblasts at 72 hr. HESL substantially increased the numbers of cells in the sub G1 phase, and dose-dependently increased apoptotic cell numbers. Western blot analysis of the total cell lysates revealed that HESL increased Bax protein levels, but did not affect Bcl-2 levels. HESL induced the cleavage of poly (ADP-ribose) polymerase and caspases 8, 9, 7, and 3. This study demonstrated that HESL inhibits cell growth and induces apoptosis in HT-29 cells, which may be mediated by its ability to increase Bax levels and activate the caspase pathway. These findings may lead to the development of new therapeutic strategies for colon cancer treatment.

Comparison of the Effect of Income-Redistribution before and after the Mergence of Medical Insurance Program for Self-employeds (지역의료보험 통합전후의 계층간 보험료 이전효과 비교)

  • 박재용;박재원
    • Health Policy and Management
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    • v.11 no.2
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    • pp.85-122
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    • 2001
  • This study compared and analyzed the effect of income-redistribution, collecting data on the basis of the estimated details of insurance contribution and individual money wage lists for each one year before and after the combination of medical insurance program for industrial workers, by systematic sampling, extracting 4,160 families(14,764 people) among people applied to medical insurance program for self employees in Taegu City on the basis of Oct. 1st in 1998 with 227 associations of medical insurance program for self employees and medical insurance program for government employees and private school teachers combined, comparing the effect of income redistribution of before and after the combination of medical insurance program for self employees. The insurance contribution by household after the combination of medical insurance program for self employees showed the increase rate of average 20.9%, among them households of 68.8% increased and 31.2% decreased. The effect of income-redistribution was more positive because the degree of inequality was more deepened from 0.64 of the before-combination to 0.45 of the after-one in decile distribution ratio, from 0.26 to 0.34 in Gini -coefficient. Decile distribution ratio on the basis of insurance benefits by household was from 0.09 in the before-combination to 0.14 in the after-one, Gini-coefficient from 0.16 in the before-combination to 0.57 in the after-one was a little lowered. And decile distribution ratio of insurance benefits on the basis of insurance contribution was higher from 1.08 in the before-combination to 1.23 in the after-one, concentration index was a little lowered from 0.14 to 0.11, the effect of income-redistribution was improved in the phase of insurance benefits. The income-transfer rate of medical insurance program for self employees (the occupied rate of insurance benefits/ the occupied rate of insurance contribution) showed a lower trend in all of the before and after-combination towards upper classes, it was known that the income-transfer rate was higher from 1st degree to 7th degree in the after-combination in comparison with the before-one, but the effect of income¬redistribution was high because the income-transfer rate was lowered from 8th degree to 10th degree. The rate of medical insurance benefits (insurance benefits/ insurance contribution) increased from 0.79 in the before-combination to 1.07 in the after-one, and showed over 1.0 under 3th degree before the combination, but all of it was higher than 1.0 under 7th degree after the combination, the after-combination was more improved than the before-one in view of the rate of insurance benefits. As the result of above, on the basis of Oct. 1st in 1998 that 227 associations of medical insurance program for self employees was combined into one, we could say that the equality of imposing medical insurance contribution was more re-considered in the after-combination than in the before-one. But this study analyzed with classes divided, anyway, on the basis of insurance contribution, we have limit in explaining the correct effect of income-redistribution, because it was not analyzed according to classes of income, though it helps to analogize the effect of income-redistribution. So there must be analysis about the effect of income-redistribution, on the basis of the system, building up the system to grasp the correct income of the insureds of medical insurance program for self employees.

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