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Effect of Trichostatin A on Anti HepG2 Liver Carcinoma Cells: Inhibition of HDAC Activity and Activation of Wnt/β-Catenin Signaling

  • Shi, Qing-Qiang;Zuo, Guo-Wei;Feng, Zi-Qiang;Zhao, Lv-Cui;Luo, Lian;You, Zhi-Mei;Li, Dang-Yang;Xia, Jing;Li, Jing;Chen, Di-Long
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7849-7855
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    • 2014
  • Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.

A study on Empowerment of the Clinical Dental Hygienists (임상치과위생사의 임파워먼트에 관한 연구)

  • Shin, Seon-haeng
    • Journal of dental hygiene science
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    • v.10 no.3
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    • pp.185-190
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    • 2010
  • This study was conducted to collect the basic data to provide for the purpose of the development of dental field. This research is to find the relationship between the level of recognized empowerment of hygienists working in dental clinics and the factors influencing on work achievement in the system of dental clinics. The population of the research is some randomely chosen dental clinics located in Seoul City during the period from March 22 to April 30, 2010, and the sample of 256 female dental hygienists working in the field were recruited as the analysis object group. The data on general characteristics and empowerment were collected by a questionnaire survey. The collected data were analyzed with the SPSS WIN 12.0 program. The following shows the results of this study. 1. The empowerment level of the analysis subject group was 3.63. It is significant statistically that the longer working career or higher annual salary, the higher empowerment level (p<0.05). 2. It shows the high level of empowerment in the group of hygienists who are in education counsel (p<0.01), and who have a permanent job (p<0.05) than others. 3. There is a significant correlation among capability, age, career, annual salary (p<0.01). 4. There is a significant correlation among self determination, age(p<0.05), career (p<0.01), annual salary (p<0.05). 5. In regression analysis, career is proved as one of the significant factors that is related to the empowerment in dental hygienists (p<0.05). This finding shows high level of empowerment in dental hygienists who hold high level of career, annual salary, education counsel jobs, or permanent jobs. Therefore, I suggest with the result of the study that the organization of dental clinic needs to improve its capability and efficiency with the efficient manpower management. Particularly, there need to be various kinds of leadership-trainning and education-program development to enhance empowerment in hygienists as professionals through independent self-decision making experiences and role do experiences.

Study on the Effect of Contact Angles of Elastic Rubber Impression Materials on the Surface of Working Cast (탄성 고무인상재의 접촉각이 작업모형 표면에 미치는 영향)

  • Kim, Joo-Won
    • Journal of dental hygiene science
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    • v.10 no.3
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    • pp.177-183
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    • 2010
  • This study was begun to search effect of contact angles of elastic rubber impression materials on the surface of working cast. Of elastic rubber impression materials with a Type III consistency, such as polysulfide, polyether and addition silicone, we selected one and then measured the contact angle after dripping a distilled water 3.3ml. Then, after pouring a dental anhydrite in three types of impression materials, we prepared a working cast and then examined its surface. Contact angle was measured using a full automatic contact angle measuring system (DM-700, KYOWA, Japan), and the surface of working cast was observed using a field emission scanning electron microscope (JSM-6700F, JEOL Ltd., JAPAN). The following results were obtained: 1) $Mean{\pm}SD$ (SD: standard deviation) of the initial contact angles were $91.3{\pm}20.5^{\circ}$ in the addition silicone materials, $90.0{\pm}2.2^{\circ}$ in the polyethers and $101.5{\pm}2.3^{\circ}$ in the polysulfides. These results indicate that mean values were similar but standard deviations of the three materials showed a great discrepancy. 2) As the time elapsed, addition silicone materials were found to have a contact angle decreased abruptly as compared with the remaining two types. That is, the initial contact angle was $91.3^{\circ}$ and it was abruptly decreased to $29.4^{\circ}$ after 25 seconds. 3) In the polyethers, the initial contact angle was $101.5^{\circ}$ and it was decreased to $90.7^{\circ}$ after 25 seconds. In the polysulfides, however, the initial contact angle was $90.0^{\circ}$ and it was $84.2^{\circ}$ after 25 seconds. This showed almost no changes in the initial contact angles. Moreover, its magnitude was greater than that seen in additional silicones. 4) There were significant differences in the contact angles between the three types of elastic rubber impression materials as the time elapsed (p<0.001). On an observation on the surface of working cast, addition silicone materials were found to have the most dense surface. This was followed by polysulfides and polyethers in a descending order.

Annual Increase in Carbon and Nitrogen Stocks of Trees and Soils in a 'Niitaka' Pear Orchard Following Standard Fertilization Recommendations (표준 시비에 따른 '신고'배 수체 및 재배지 토양의 탄소 및 질소 저장량 변화)

  • Ro, Hee-Myong;Choi, Jin-Ho;Lee, Seo-Yeon;Lee, Tae-Kyu;Kim, Jong-Sung;Park, Ji-Suk;Choi, Jang-Jeon;Lee, Min-Jin
    • Horticultural Science & Technology
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    • v.33 no.4
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    • pp.591-597
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    • 2015
  • We determined the total C and N stocks in trees and soils after 1 year of fertilization in an experimental orchard with 16-year-old 'Niitaka' pear (Pyrus pyrifolia Nakai cv. Niitaka) trees planted at $5.0m{\times}3.0m$ spacing on a Tatura trellis system. Pear trees were fertilized at the rate of 200 kg N, 130 kg P and $180kg\;K\;ha^{-1}$. At the sampling time (August 2013), trees were uprooted, separated into six fractions [trunk, main branches, lateral branches (including shoots), leaves, fruit, and roots] and analyzed for their total C and N concentrations and dry masses. Soil samples were collected from 0 to 0.6 m in 0.1 m intervals at 0.5 m from the trunk, air-dried, passed through a 2-mm sieve, and analyzed for total C and N concentrations. Undisturbed soil core samples were also taken to determine the bulk density. Dry mass per tree was 5.6 kg for trunk, 12.0 kg f or m ain branches, 15.7 kg for lateral branches, 5.7 kg for leaves, 9.8 kg for fruits, and 10.5 kg for roots. Total amounts of C and N per tree were respectively 2.6 and 0.02 kg for trunk, 5.5 and 0.04 kg for main branches, 7.2 and 0.07 kg for lateral branches, 2.6 and 0.11 kg for leaves, 4.0 and 0.03 kg for fruit, and 4.8 and 0.05 kg for roots. Carbon and N stocks stored in the soil per hectare were 155.7 and 14.0 Mg, respectively, while those contained in pear trees were 17.8 and $0.2Mg{\cdot}ha^{-1}$ based on a tree density of 667 trees/ha. Overall, C and N stocks per hectare stored in the pear orchard were 173.6 and 14.2 Mg, respectively. Compared with results obtained in 2012, the amounts of C stocks have increased by $17.7Mg{\cdot}ha^{-1}$, while those of N stocks remained virtually unchanged ($0.66Mg{\cdot}ha^{-1}$).

Combined Effects of Biological Control Agent Two Native Shellfish on the Hibernal Diatom Bloom of Eutrophic Water (저온기 규조 발생억제를 위한 패류의 혼합적용)

  • Lee, Song-Hee;Hwang, Soon-Jin;Kim, Baik-Ho
    • Korean Journal of Ecology and Environment
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    • v.41 no.3
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    • pp.402-411
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    • 2008
  • Grazing effects of two freshwater shellfish, Unio douglasiae (UNIO) and Cipangoplaudina chinese malleata (CCM), on the hibernal diatom communities in eutrophic water was examined in a laboratory. Two animals having different feeding types used in the study were collected from Keum River (Boryeong, Korea), acclimatized in the artificial management system in a laboratory over at least one month, and starved in a laboratory for 2 days before the experiment. Experimental waters, which dominated by Synedra ulna-Scenedesmus sp. (ca. 80%), was collected from eutrophic lake, Lake Ilgam (Seoul) in Feb., 19, 2008 at AM 10:00, and used in the study after the filtration with 1 mm Nylon mesh. Feeding experiments were largely divided into three kinds of animal treatments; five densities of UNIO (U0, U1, U2, U3, U4) and CCM (C0, C1, C2, C3, C4), and four combined densities of two shellfish (M0, M1, M2, M3). U0 and C0 were control (no addition of UNIO or CCM), U1 or C1 (each animal density at 0.5 ind. $L^{-1}$), U2 or C2 (1.0 ind. $L^{-1}$), U3 or C3 (1.5 ind. $L^{-1}$), and U4 or C4 (2.0 ind. $L^{-1}$), respectively. Four combined treatments were consisted of M0 (no animals), M1 (U1+C3), M2 (U2+C2), and M3 (U4+C1), respectively. Under the presence of animal, the concentration of Chl-a and algal abundance were clearly decreased with the increase of UNIO density and the treated time, while in combined group a strong decrease of algal density showed with the increase of UNIO density. Total phytoplankton density shifted as the similar patterns to that of Chl-a concentration (r=0.6527, p<0.0001), however, there showed the differences following a species. There were strong decreases of dominant species Synedra ulna, Scenedesmus sp., Ankistrodesmus falcatus in UNIO treatment group, Diatoma vulgare in combined group, while Cryptomonas ovata in high density of CCM increased about 20% in algal density. Grazing rates (GRs) based on the concentration of Chl-a was depend on the kind of shellfish and treatment time; a strong feeding of CCM showed in the initial stage, and four hours later, UNIO and combined group with high UNIO density showed the high GRs. Interestingly, faeces production of shellfish was highest in combined group with high density of CCM, while their size over 60 ${\mu}m$ was much higher in production magnitude than that less 60 ${\mu}m$. Collectively, these results suggest that two domestic shellfish and its combined treatment have a strong potential as an effective biological controller of diatom bloom in cold eutrophic waters.

Effects of Light Sources, Light Quality on the Growth Response of Leafy Vegetables in Closed-type Plant Factory System (완전제어형 식물공장에서 광원, 광질에 따른 엽채류 6종의 생육반응)

  • Kim, Sang Bum;Lee, Kyung Mi;Kim, Hae Ran;You, Young Han
    • Korean Journal of Ecology and Environment
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    • v.47 no.1
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    • pp.32-40
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    • 2014
  • This study was conducted to evaluate the growth response of economical six leafy vegetables that are crown daisy, pak-choi and four kinds of lettuce (Red leaf lettuce, Green leaf lettuce, Head lettuce, Romaine lettuce) by light treatment of LED in plant factory. The light treatments were composed of red, blue, red+farred, red+blue, red+blue+white LEDs, irradiation time ratio of the red and blue LED per minute (1 : 1, 2 : 1, 5 : 1, 10 : 1), and duty ratio of mixed light (100%, 99%, 97%). The following results were obtained in different LED light sources treatments: Shoot biomass and S/R ratio of romaine lettuce were the highest under mixed red+blue LEDs. S/R ratio of head lettuce was higher under mixed red+blue+white LEDs than red+blue LEDs. The others showed no difference in LED light treatment. Shoot biomass, total biomass and S/R ratio of green lettuce, head lettuce and pak-choi were highest in the higher red ratio (5 : 1) on irradiation time of red : blue LED ratios. By the different duty ratio (red+blue and red+blue+white LEDs), Under the mixed light of red+blue, shoot and root biomass of crown daisy and romaine lettuce were high in duty ratio of 100% and 99%, and S/R ratio was highest in all the 6 kinds in duty ratio of 97%. All the 6 kinds showed a fine growth state in low duty ratio (97%). Green lettuce, romaine lettuce and pak-choi showed relatively high shoot biomass and total biomass in low duty ratio of 97% under the mixed light of red+blue+white. S/R ratio of romaine lettuce and head lettuce were highest in the duty ratio of 97% with red+blue+white LEDs. Thus, we can cultivate stably without reference to external factors, if we use appropriate light sources and light quality in closed-type plant factory.

The Study on Consummer Behaviour of Poultry meat and Egg (닭고기와 계란의 소비에 대한 조사 연구)

  • 남두희;오세정
    • Korean Journal of Poultry Science
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    • v.15 no.2
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    • pp.81-91
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    • 1988
  • The purpose of this research is to analyze the tendencies of poultry consumption in Korea. The information used is the data collected three times during the period from Sept. 1, 1985 to April 30, 1987 throughout the major cities. Those who participated in the survey are 2, 598 including housewives, nutritionists, cookers, group feeding institutions, woman's organizations, and the students of doing nutrition-related studies. Consumer preferences for poultry meat and eggs tend to move toward high quality and more strict sanitary standards. Following this line of consumer preference changes, the poultry product marketing supported by cold chain system is rapidly developing. Household consumption of poultry meat largely consists of hi-and semi-broilers but the household demand for these two broilers tends to decrease steadily over time. In general poultry meat consumption appears to be affected by consumer taste rather than market factors such as prices. In addition consumer choices are quite different depending on poultry meat parts which are preferred in order of drum sticks, wings, and breasts. In particular drum sticks are most preferred than any other parts. An important problem in poultry production is related to consumption seasonality since large part of poultry meat consumption is concentrated during the summer season. Another problem is associated with little development of cooking methods. At present there are two types of primary commercial cooking techniques, fries and samgaetang (boiled chicken with jinsang and rice). For promoting domestic poultry meat disposal and reducing the demand seasonality, new cooking methods should be developed and followed by more aggresive advertisements. In domestic egg trade, smaller packing units(i.e., 10 eggs per unit) tend to bi preferred to large ones (i.e., 30 eggs per unit). In consumers egg purchasing decisions nearness to the shops and convenience appear to be important factors. For egg shell colors consumers recognize that there is no difference in nutritional values. However, survey results show that consumers highest preference lies in eggs with brown color. Eggs are most popular among children and preferred in order of middle-and high-school students, 17-25 age people, and adults. Egg prices are concieved relatively cheap to its nutritional values. In house-holds eggs are consumed in the forms of fries, side dishes, and lunch basket dishes. However, high level of cholesterol content in eggs appears to be an important problem in promoting eggs consumption.

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Effects of Insamjungchuntang on the Respiratory System in Experimental Animals (인삼정천탕(人蔘定喘湯)이 호흡기계(呼吸器系)에 미치는 영향(影響))

  • Kim, Ho-Soon;Shin, Jo-Young
    • The Journal of Internal Korean Medicine
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    • v.19 no.1
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    • pp.409-427
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    • 1998
  • Insamjungchuntang has been used in Korea for many centuries as a treatment for respiratory disease. The effect of Insamjungchuntang on tracheal smooth muscle is not known. The purpose of the present study is to determine the effect of Insamjungchuntang on histamine and acetylcholine induced tracheal smooth muscle contraction in rats and guinea pigs. Guinea pig (500 g, male) and Sprague Dawley rats (200 g, male) were killed by $CO_2$ exposure and a segment (8-10 mm) of the thoracic trachea from each rat and guinea pig was cut into equal segments and mounted 'in pairs' in a tissue bath. Contractile force was measured with force displacement transducers under 0.5 g loading tension. The dose of histamine (His) and acetylcholine (Ach) which evoked 50% of maximal response $(ED_{50})$ was obtained from cumulative dose response curves for histamine and acetylcholine$(10^{-7}{\sim}10^{-4}\;M)$. Contractions evoked by His ($ED_{50}$) and Ach $(ED_{50})$ were inhibited significantly by Insamjungchuntang. In guinea pig tracheal smooth muscle, the mean percent inhibition of acetylcholine induced contraction was $38.58\(p<0.05)\;after\;10{\mu}l/ml$ Insamjungchuntang, $90.75\(p<0.01)\;after\;30{\mu}l/ml$. Insamjungchuntang and $133.17\(p<0.01)\;after\;100{\mu}l/ml$ Insamjungchuntang. In rat tracheal smooth muscle, the mean percent inhibition of acetylcholine induced contraction was $10.0\(p<0.05)\;after\;10{\mu}l/ml$ Insamjungchuntang, $80.71\(p<0.01)\;after\;30{\mu}/ml$ Insamjungchuntang and $118.29\(p<0.01)\;after\;100{\mu}l/ml$ Insamjungchuntang. Also, in guinea pig tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was $45.5\(p<0.01)\;after\;10{\mu}l/ml$ lnsamjungchuntang, and $93.17\(p<0.01)\;after\;30{\mu}l/ml$. lnsamjungchuntang $134.50\(p<0.01)\;after\;100{\mu}l/ml$ Insamjungchuntang. In rat tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was $37.83\(p<0.01)\;after\;10{\mu}l/ml$ lnsamjungchuntang, $90.5\(p<0.01)\;after\;30{\mu}l/ml$ Insamjungchuntang and $135.17\(p<0.01)\;after\;100{\mu}l/ml$ Insamjungchuntang. Propranolol $(10^{-7}\;M)$ slightly but significantly attenuated the inhibitory effects of Insamjungchuntang. Following treatment with propranolol, the mean percent inhibition caused by $100{\mu}l/ml$. Insamjungchuntang fell to 46.42% in guinea pig induced by acetylcholine contraction and by $100{\mu}l/ml$ Insamjungchuntang fell to 5.43% (p<0.05) in rat induced by acetylcholine contraction and the mean percent inhibition caused by $100{\mu}l/ml$ Insamjungchuntang fell to 49.0% in guinea pig induced by histamine contraction and by $100{\mu}l/ml$ Insamjungchuntang fell to 48.6% (p<0.05) in rat induced by histamine contraction. Indomethacin and methylene blue $(10^{-7}\;M)$ did not significantly alter the inhibitory effect of lnsamjungchuntang. Also, I could find the effects of lnsamjungchuntang and Insamjungchuntanggamorphine on the tracheal smooth muscle in guinea pig and rat did not change significantly. These results indicate that Insamjungchuntang can relax histamine and acetylcholine-induced contraction of guinea pig and rat tracheal smooth muscle, and that this inhibition involves sympathetic effects.

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Activation Mechanism of Protein Kinase B by DNA-dependent Protein Kinase Involved in the DNA Repair System

  • Li, Yuwen;Piao, Longzhen;Yang, Keum-Jin;Shin, Sang-Hee;Shin, Eul-Soon;Park, Kyung-Ah;Byun, Hee-Sun;Won, Min-Ho;Choi, Byung-Lyul;Lee, Hyun-Ji;Kim, Young-Rae;Hong, Jang-Hee;Hur, Gang-Min;Kim, Jeong-Lan;Cho, Jae-Youl;Seok, Jeong-Ho;Park, Jong-Sun
    • Toxicological Research
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    • v.24 no.3
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    • pp.175-182
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    • 2008
  • DNA-dependent protein kinase(DNA-PK) is involved in joining DNA double-strand breaks induced by ionizing radiation or V(D)J recombination and is activated by DNA ends and composed of a DNA binding subunit, Ku, and a catalytic subunit, DNA-PKcs. It has been suggested that DNA-PK might be $2^{nd}$ upstream kinase for protein kinase B(PKB). In this report, we showed that Ser473 phosphorylation in the hydrophobic-motif of PKB is blocked in DNA-PK knockout mouse embryonic fibroblast cells(MEFs) following insulin stimulation, while there is no effect on Ser473 phosphorylation in DNA-PK wild type MEF cells. The observation is further confirmed in human glioblastoma cells expressing a mutant form of DNA-PK(M059J) and a wild-type of DNA-PK(M059K), indicating that DNA-PK is indeed important for PKB activation. Furthermore, the treatment of cells with doxorubicin, DNA-damage inducing agent, leads to PKB phosphorylation on Ser473 in control MEF cells while there is no response in DNA-PK knockout MEF cells. Together, these results proposed that DNA-PK has a potential role in insulin signaling as well as DNA-repair signaling pathway.

A Study on Factors Affecting Youth Employee's Labor Mobility and Employment Status Transition (청년취업자의 노동이동 및 고용형태 전환에 영향을 미치는 요인에 관한 연구)

  • Ban, Jung-Ho;Kim, Kyung-Hee;Kim, Kyung-Huy
    • Korean Journal of Social Welfare
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    • v.57 no.3
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    • pp.73-103
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    • 2005
  • This study takes of youth panel data(2002-2003), there is the purpose to know to youth employed's labor mobility conditions of employment status conversion and examine on factors affecting conversion of employment status. Main analysis result and policy imply, is as following. First, although youth employed's non-standard employment shows some decrease, employment youth hierarchy was construed that youth of our country is consisting very changefully because appear great turnover that is converted by unemployment or Not economically activity population same period. Specially, non-standard employment phenomenon of woman or low in scholarship person appeared notedly, and because phenomenon that is converted by unemployment or Not economically activity population is expose that is deepened, discriminating policy of government dimension is required for employment stabilization of these class. Second, show result that danger to escape to non-standard job risk trap which seeking employment activity of youth class is arranged case or company which is formed by official path is suitable becomes low, must formulate path of employment about youth class and improve qualitative level of employment through suitable job placement education of youth class or function (technology) level. Third, when was construed, but take into account that the although large enterprise have low risk in non-standard job, recently employment of youth class consists very limited, rather small scale business or smaller enterprise's competitive power preferably need to be plan. Finally, danger to non-standard job youth employed's company form is government connection wonder was expose that high, Such result can do that it is difficult by limited research period, but reflect actuality that youth unemployment policy of our country is enforced laying stress on public labor or unregular job employment such as internship system. Therefore, current youth unemployment policy may have to change by employment policy that can secure stable work record by youth class or act as bridge-building that promote conversion by full-time job.

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