• Title/Summary/Keyword: 3T3 L1 cells

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Effect of Demecolcine-Assisted Enucleation and Recipient Cell Cycle Stage on the Development of Nuclear Transfer Bovine Embryos (Demecolcine 처리에 의한 탈핵과 수핵란 세포질의 세포 주기가 소 핵이식란의 발육에 미치는 영향)

  • Back J. J.;Park C. K.;Yang B. K.;Kim C. I.;Cheong H. T.
    • Reproductive and Developmental Biology
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    • v.29 no.3
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    • pp.175-180
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    • 2005
  • This study was conducted to examine the effects of demecolcine-assisted enucleation and recipient cell cycle stage on the development of bovine somatic cell nuclear transfer (NT) embryos. In vitro cultured oocytes for $16\~20$ h were classified by first polar body (1st PB) extrusion and cell cycle stage (MI and MII) and treated $0.4\;{\mu}L/mL$ demecolcine for 40 min before enucleation. Enucleated oocytes were fused electrically with bovine ear skin cells, activated by Ca-ionophore+DMAP, and cultured in vitro. Most of eggs ($86.2\%$) treated with demecolcine protruded a chromosome mass and enucleated efficiently ($98.8\%$, (P<0.05). Demecolcine did not have a deteriorative effect on the development of NT embryos. Developmental rate of NT embryos reconstituted with oocytes extruded 1st PB significantly higher than that of NT embryos produced by oocytes without 1st PB ($18.2\%\;vs.\;4.6\%\cdot$, P<0.05). Cleavage and blastocyst formation rate of embryos reconstituted with MI oocytes ($69.4\%\;and\;5.9\%$, respectively) were significantly lower than those of embryos reconstituted with MII oocytes ($96.7\%\;and\;23.9\%$, respectively, P<0.05). From the present result, it is suggested that domecolcine is useful for the enucleation of recipient oocytes in bovine NT procedures, and MII oocytes rather than MI oocytes are more appropriate for recipient cytoplasm. Although, the potential to develop into blastocysts of NT embryos produced by 1st PB-nonextruded and MI oocytes was very low, these oorytes could be used for NT.

Comparison of Distribution of Cadmium-109 and Gallium-67 in Sarcoma-Bearing Mice (Sarcoma-bearing Mice에서 Cadmium-109과 Gallium-67의 체내 분포의 비교)

  • Sohn, Myung-Hee;Chang, Sook-Kyeong;Chung, Kyung-Ho;Han, Young-Min;Kim, Chong-Soo;Choi, Ki-Chul;Yim, Chang-Yeol;Kang, Shin-Hwa
    • The Korean Journal of Nuclear Medicine
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    • v.28 no.1
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    • pp.98-105
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    • 1994
  • The search for tumor-avid agents for use in nuclear medicine imaging or therapy is a field of ongoing importance. Metallothionein (MT) is an intracellular protein that binds many metals with isotopes having imaging or radiotherapeutic potential. The purpose of the study was to determine whether uptake of radioisotopes that bind to MT is increased in tumor. We measured the uptake of Cd-109 and Ga-67 in tumor and normal tissues of sarcoma-bearing mice. Tumors were grown subcutaneously in female Balb/C mice from cultured Balb/3T3 cells transformed by the Moloney murine sarcoma virus (MMSV). When the tumors reached about 1 cm in diameter, mice were injected subcutaneously with Cd-109 and Ga-67. Eighteen and seventy-two hours later, the mice were sacrified. Organs and tissues were removed, weighed, and activity per mg tissue determined by gamma well-counting. Uptake of Cd-109 by MMSV tumors exceeded that by normal tissues examined, with the exception of liver and kidney (the organs known to be richest in MT). The tumor-to-tissue ratios of uptake for Cd-109 were far greater than those for Ga-67 for many normal tissues of great importance in terms of background activity (bone, intestine, fat, muscle, and blood). We concluded that metals that bind to MT may be useful for oncologic imaging or rediotherapy of cancer.

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Effects of water and ethanol extracts from Ojeok-san on inflammation and its related diseases (오적산의 추출 용매 차이에 따른 염증 및 염증 관련 질환에 대한 효능 비교)

  • Yoo, Sae-Rom;Jeong, Soo-Jin;Kim, Ye-Ji;Lim, Hye-Sun;Jin, Seong-Eun;Jeon, Woo-Young;Shin, In-Sik;Shin, Na-Ra;Kim, Seong-Sil;Kim, Jung-Hoon;Ha, Hye-Kyung;Lee, Mee-Young;Kim, Ohn-Soon;Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • The Journal of Internal Korean Medicine
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    • v.33 no.4
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    • pp.418-428
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    • 2012
  • Objectives : Ojeok-san, a traditional herbal formula, has been used for the treatment of cold illness and its related symptoms such as headache, nausea and indigestion. This study was performed to compare effects of water (OJSW) and 70% ethanol extracts (OJSE) of Ojeok-san on inflammation and its related diseases atopy, asthma and obesity in vitro. Methods : We performed HPLC to investigate contents of index components of OJSW and OJSE. We investigated the effects of OJSW and OJSE with an in vitro model, using 5 cell lines, specifically RAW 264. 7, HaCaT, MC/9, BEAS-2B and 3T3-L1. Results : HPLC analysis displayed that the contents of index components were higher in OJSE than OJSW. In lipopolysaccharide (LPS)-treated RAW 264.7 macrophages, OJSE significantly inhibited productions of interleukin (IL)-6, nitrite and prostaglandin $E_2$ ($PEG_2$). In TNF-${\alpha}$/IFN-${\gamma}$-treated HaCaT keratinocytes, OJSE significantly lowered levels of macrophage-derived chemokine (MDC) as well as regulated and normal T cell expressed and secreted (RANTES). OJSE also had a protective effect on inflammatory response by decreasing RANTES secretion in TNF-${\alpha}$-stimulated BEAS-2B cells. Conclusions : We conclude that OJSE could be more appropriate to enhance the biological activities against inflammation and its related diseases, and could be applied as a bioactive material for developing the potent anti-inflammatory agents.

Larvae Growth and Biochemical Composition Change of the Pacific Oyster Crassostra gigas, Larvae during Artificial Seed Production (참굴 Crassostrea gigas 인공종묘생산 시 유생의 성장과 체성분 변화)

  • Hur, Young-Baek;Min, Kwang-Sik;Kim, Tae-Eic;Lee, Seung-Ju;Hur, Sung-Bum
    • Journal of Aquaculture
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    • v.21 no.4
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    • pp.203-212
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    • 2008
  • A nutritional demand of oyster, Crassostrea gigas larva as part of research for improving of utilization of microalgae being used for the artificial oyster seed production. The change of body growth and biochemical compositions of larvae were investigated during larvae rearing in hatchery. The larvae were cultured in 60 M/T tank and fed mixture 6 different phytoplankton species, Isochrysis galbana (30%), Cheatoceros gracilis (20%), Pavlova lutheri (20%), Phaeodactylum triconutum (10%), Nannochryis oculata (10%) and Tetraselmis tetrathele (10%). The initial feeding amount was $0.3{\times}10^4cells/mL$ at three times a day to D-shaped larva and the feeding amount had been increased 30% gradually every two day since the larvae were raising. The larvae were developed from D shape to pediveliger stage for 12 days. The daily growth of shell length and hight were $5.8{\sim}30.8\;{\mu}m$ and $8.7{\sim}31.4\;{\mu}m$, respectively and weight gains were changed from D shape to pediveliger as follow: wet weight was $0.52{\sim}15.0\;{\mu}g/larva$, dry weight was $0.2{\sim}6.5\;{\mu}g/larva$, and ash free dry weight was $0.1{\sim}8.5\;{\mu}g/larva$. The larvae growth pattern shown a logarithmic phase from D shape to umbone stage but after that stage shown a exponential growth aspect. The crude protein, crude lipid and nitrogen free extract (NFE) of larvae during rearing periods were analyzed as $6.1{\sim}10.6%$, $0.6{\sim}1.1%$ and 1.0-2.7%, respectively. And the total amino acid contents of the larvae during rearing periods were in order as glutamic acid $1.26{\sim}2.24%$, aspartic acid $0.97{\sim}1.70%$, and methionine $0.12{\sim}0.33%$. Of the total fatty acid in the analyzed larvae, the saturated fatty acid (SSAFA) was decreased from 54.3% (D shaped larvae) to 17.1 % (pediveliger) as larvae development but the total mono-unsaturated fatty acid (${\Sigma}MOFA$) and Poly-unsaturated fatty acid (${\Sigma}PUFA$) were increased from 29.9% and 7.8% to 40.6% and 45.6%, respectively. By the way the each fatty acid of the larvae were composed of palmitic acid $9.89{\sim}36.95%$, oleic acid $12.17{\sim}32.29%$, linoleic acid $1.96{\sim}33.55%$, EPA $2.17{\sim}11.58%$ and DHA $1.95{\sim}4.51%$. As a result of this study, the larvae of oyster were demanded a various nutrients for healthy growth and the feeding control, expecially after umbone stage larvae are a rapidly growing time, is very important for success of artificial seed production.

Establishment of I-131, Tc-99m Labeling Methods to In-house Anti-CEA Antibodies and Evaluation of the Immunological Characteristics (국산 항 CEA 항체의 I-131, Tc-99m 표지법 확립 및 면역학적 특성 분석)

  • Chung, June-Key;Lee, Dong-Soo;Lee, Myung-Chul;Chung, Hong-Keun;Koh, Chang-Soon;Hong, Mee-Kyoung;Choi, Seok-Rye;Seo, Il-Taek;Chung, Jun-Ho
    • The Korean Journal of Nuclear Medicine
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    • v.26 no.2
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    • pp.346-354
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    • 1992
  • Cancer cells have several tumor-associated antigens on the cell surfaces, and antibodies against these antigens have been developed by many investigators. Radiolabeled antibodies have been used as new methods to diagnose and treat malignant tumors. Especially anti-carcinoembryonic antigen (CEA) is the most popular antibody for these purposes. In this investigation, we tried to label $^{131}I$ and $^{99m}Tc $ to anti-CEA monoclonal antibodies which were developed in the Seoul National University College of Medicine. We found CEA-79 and CEA-92 antibodies had the better immunological characteristics among 8 anti-CEA monoclonal antibodies. And radioiodination of CEA-79 could be performed by chloramine-T method, while radioiodination of CEA-92 by iodogen method. To label these antibodies with $^{99m}Tc $, we used pretargeting transchelation as direct labeling method. At first, $^{99m}Tc $ was bound to glucaric acid, and monoclonal antibody was reduced by $\beta-mercaptoethanol$. When these were incubated together. $^{99m}Tc $ bound to glucarate was switched to monoclonal antibody because of higher affinity. We established conditions of several steps in this method. Anti-CEA monoclonal antibodies labeled with $^{131}I$ and $^{99m}Tc $ are expected to be used valuably in the detection and treatment of malignant tumors.

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Current Status of Korean Ginseng Research (한국인삼론(韓國人蔘論))

  • Han, Byung-Hoon
    • Korean Journal of Pharmacognosy
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    • v.3 no.3
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    • pp.151-160
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    • 1972
  • Recent achievements of scientific research on the pharmacologic activities and the chemical problems of dammalene glycosides, which are considered to be effective principles of Korean ginseng, are reviewed and analyzed in view of structure-activity relationship. 1) S. Shibata and his co-workers detected 12 glycoside spots of dammalene series on the two dimensional T.L.C. of total glycoside fraction from Japanese ginseng, and designated them Ginsenoside Rx(x=a, b, c, g, h, etc.) in the order of increasing Rf-value. The aglycones of those glycosides were characterized to be protopanaxadiol for the Ginsenoside $Rx(x=a,\;b_{1},\;b_{2},\;c,\;d,\;e,\;f)$ and protopanaxatriol for the Ginsenoside $Rx(x=g_{1},\;g_{2},\;g_{3},\;h_{1}\;'h_{2})$. Using Korean ginseng as the material for our study, the author and his coworkers isolated a new dammalene glycoside(Panax Saponin C), which comes under the category of protopanaxadiol glycosides based on the classification of S. Shibata et al., and characterized this saponin to be the glycoside of protopanaxatriol series. Furthermore, Panax Saponin C dissociated into $two\;components(C_{1}\;and\;C_{2}-acetate)$ by acetylation, both of which returned to original Panax Saponin C by deacetylation. Based on this result, more than 13 glycoside components of dammalene series will be expected in the Korean ginseng. 2) The structures of protopanaxadiol and protopanaxatriol, the genuine aglycones of dammalene glycosides, are fully established to be structural analogues by S. Shibata and his co-workers, therefore antagonistic and/or analogical activities will be expected for the pharmacologic activities of these glycoside series of structural analogues. K. Takaki and his co-workers found central nervous system (CNS) stimmulant activity from the glycosides of protopanaxatriol series and CNS-depressant activity from the glycosides of protopanaxadiol series. On the other hand, the author and his co-workers found stimmulating activity on the protein synthesis from both the series of dammalene glycosides with delayed and long-lasting characteristics. This delayed and long-lasting characteristics were also observed in the anti-inflammatory activity of glycosides of protopanaxatriol series on their time course tendency. For the convenience's sake of argument, pluralistic pharmacologic activities of dammalene glycosides, which were observed by many workers at various pharmacologic site, may be classified into two main categories; one is pan-cellular activity and the other is organ specific activity to the certain tissue which is a mass of cells differentiated to a certain direction for their special functions in the body. Based on the data of K. Takaki and those of the authors, following assumption will be probable; Pharmacologic activities of both series of glycosides of protopanaxadiol and protopanaxatriol aglycones may be antagonistic on their tissue-specific activities and analogic on their pan-cellular activities. Therefore, the mixture of these two series of glycosides in an appropriate ratio, as the case of total extract of Korean ginseng, will be probably beneficial to the host by increasing the synthesis of some functional proteins, due to the additive action of pan-cellular activity, and with the disappearance of any significant behavioral symptoms due to the antagonism of tissue specific activity. This fact will probably be the main reason why classical trials of pharmacologists failed in re-discovering the efficacy of Korean ginseng with their behavioral test. 3) The author and his co-workers achieved the synthesis of $C^{14}-labelled\;Panax\;Saponin\;A\;on\;C_{25}-C_{27}\;position\;of\;aglycone$ in the interest of tracer studies in vivo. The method will be applicable to other dammalene glycosides regardless of their chemical structure. 4) The author and his co-workers converted chemically betulafolienetriol, a triterpene component of Betula platyphylla, to the protopanaxadiol, one of genuine aglycone of dammalene glycosides.

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Relation of Interleukin-10 in Bronchoalveolar Lavage Fluid and Airway Inflammation in Bronchial Asthma (기관지천식에서 기관지폐포세척액내 IL-10과 기도염증정도의 연관성)

  • Lee, Sook-Young;Youn, Hung-Gue;Shin, Youn;Lee, Sang-Haak;Kim, Seok-Chan;Kim, Kan-Hyoung;Moon, Hwa-Sik;Song, Jeong-Sup;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.1
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    • pp.44-52
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    • 1999
  • Background : Airway infiltration by inflammatory cells, particularly of eosinophils, is one of the characteristic features of asthma. Several mechanisms for the recruitment of eosinophil is focused on the CD4+ T lymphocyte for the preferential production of Th2-c1erived cytokines. Interleukin-10(IL-10) is identified cytokine with potent antiinflammatory activity. This molecule has been shown to inhibit the release of cytokine from inflammatory cells including Th2 cell, and also to inhibit eosinophil survival. We therefore attempted to determine whether decreased synthesis of IL-10 in the lung of bronchial asthma may contribute to inflammation that is characteristics of this dease. Method: Subjects were patients with bronchial asthma(n=23) and normal controls(n=11). IL-10 produced from peripheral mononuclear cell(PBMC) and in bronchoalveolar lavage(BAL) fluid was measured by ELISA method. Degree of bronchial inflammation was assessed by total cell counts and eosinophil percents in BAL fluid, eosinophil infiltration on bronchial biopsy tissue and $PC_{20}$ for methacholine. Results: The IL-10 level produced by PBMC and in BAL fluid from patient with bronchial asthma were not different with normal controls(respectively, $901.6\pm220.4$ pg/ml, $810.9\pm290.8$ pg/ml for PBMC, $24.5\pm9.5$ pg/mL $30.5\pm13.5$ pg/ml for BAL fluid p>0.05). There were significant negative correlation between IL-10 in BAL fluid and eosinophil percents in BAL fluid or degree of eosinophil infiltration in bronchial biopsy (respectively r=-0.522, r=-0.4486 p<0.05). However there was no difference of IL-10 level according to $PC_{20}$ for methacholine. There were no correlation between IL-10 production by PBMC and peripheral blood eosinophil counts or serum eosinophilic cationic protein levels(respectively r=0.1146, r=0.0769 p>0.05). Conclusion: These observation suggest that IL-10 may participate but not acts the crucial role in regulation of the airway inflammation in bronchial asthma.

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Studies on the Glucose Isomerizing Enzyme -Part II. Some Properties of the Glucose Isomerizing Enzyme of K-17 strain- (포도당(葡萄糖) 이성화(異性化) 효소(酵素)에 관(關)한 연구(硏究) -제2보(第二報); 분리선정(分離選定)된 균주효소(菌株酵素)의 성질(性質)에 대(對)해서-)

  • Seu, J.H.;Kim, C.K.;Ki, W.K.;Rhee, I.K.;Kwon, T.J.;Woo, D.L.
    • Applied Biological Chemistry
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    • v.11
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    • pp.49-53
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    • 1969
  • Some properties of glucose isomerizing enzyme which produced by the strain K-17 in xylose containing nutrient broth medium were investigated. The optimal pH for enzyme reaction was indicated about 7.2 and optimal temperature was about $75^{\circ}C$. The same optimal temprature was indicated by both cell free extract and acetone dried cells using as enzyme. The glucose isomerizing enzyme from strain K-17 was not inhibited by the high concentration of substrate even in a suturated glucose solution, but most enzyme was inactivated by the heat treatment at $80^{\circ}C$. The maximum fructose forming ratio from glucose was about 50 percents.

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Transforming growth factor-β promoted vascular endothelial growth factor release by human lung fibroblasts (인간 폐섬유아세포에서 TGF-β 자극에 의한 VEGF 분비)

  • Park, Sang-Uk;Shin, Joo-Hwa;Shim, Jae-Won;Kim, Deok-Soo;Jung, Hye-Lim;Park, Moon-Soo;Shim, Jung-Yeon
    • Clinical and Experimental Pediatrics
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    • v.51 no.8
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    • pp.879-885
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    • 2008
  • Purpose : The human lung fibroblast may act as an immunomodulatory cell by providing pro-inflammatory cytokines and chemokines, which are important in airway remodeling. Vascular endothelial growth factor (VEGF) induces mucosal edema and angiogenesis. Thymus and activation regulated chemokine (TARC) induces selective migration of T helper 2 cells. We investigated whether human lung fibroblasts produced VEGF and TARC, and the effects were augmented with the co-culture of fibroblasts and human bronchial smooth muscle cells (HBSMC), and whether dexamethasone can inhibit the proliferation and the release of VEGF in lung fibroblasts. Methods : Human lung fibroblasts were cultured with and without HBSMC, growth-arrested in serum-deprived medium, and pretreated with dexamethasone for 16 hours. After 24-hour stimulation with platelet derived growth factor-BB (PDGF-BB) and/or transforming growth factor-${\beta}$ (TGF-${\beta}$), culture supernatant was harvested for assays of VEGF and TARC. Cell proliferation was assayed using BrdU cell proliferation ELISA kit. Results : 1) The release of VEGF was significantly increased after stimulation with TGF-${\beta}$, and its release was augmented when co-stimulated with PDGF and TGF-${\beta}$. 2) VEGF release induced by PDGF or TGF-${\beta}$ was inhibited by dexamethasone. 3) There was no synergistic effect on the release of VEGF when human lung fibroblasts were co-cultured with HBSMC. 4) Dexamethasone did not suppress human lung fibroblasts proliferations. 5) Neither TGF-${\beta}$ nor PDGF induced TARC release from lung fibroblasts. Conclusion : Human lung fibroblasts may modulate airway remodeling by release of VEGF, but they have no synergistic effects when co-cultured with HBSMC. Dexamethasone suppresses VEGF release, not proliferation of lung fibroblast.

Coagulation and Fibrinolysis in Exudative Pleural Effusions (삼출성 흉막액에서 응고 및 섬유소 용해계에 관한 연구)

  • Ryu, Jeong-Seon;Lee, Hong-Lyeol
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1214-1222
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    • 1998
  • Background : The intrapleural hypofibrinolysis is caused by mainly excessive concentration of pleural plasminogen activator inhibitor-1 antigen(PAI-1 Ag), which binds tissue type plasminogen activator. In pleural inflammation induced by sclerosing agents for pleurodesis, levels of pleural PAI-1 antigen increase in relation to decreasing D-dimer levels. It has been known that the pleural mesothelial cells have the capability of secreting PAI-1 Ag in response to inflammation in vivo. Therefore, we estimated whether pleural inflammation changes the balance between fibrinolytic and coagulative properties in exudative pleural effusions. Method : The thirty cases was included in our study. We determined the pleural levels of glucose, lactic dehydrogenase(LDH), pH and the counts of white blood cell(WBC), polymorpho leukocyte(PMN), lymphocyte as the parameters of pleural inflammation and cellular components of pleural fluid. The plasma level of fibrinogen in fluid and the neutrophil count in blood were determined. The levels of D-dimer, PAI-1 Ag and thrombinantithrombin III complex(TAT) were determined by ELISA(Behring, Marburg, Germany). Result : The causes of pleural effusion were as following : tuberculous in 14 cases, malignant in 10 cases and parapneumonic in 6 cases. The levels of pleural D-dimer, PAI-1 Ag and TAT was significantly higher than that of plasma(p<0..001). The severity of pleural inflammation did not correlated with pleural D-dimer, PAI-1 Ag, TAT and their plasma levels. But the level of pleural TAT correlated with pleural WBC and lymphocyte count. Conclusion : We found that the severity of pleural inflammations did not correlated with pleural D-dimer, PAI-1 Ag, TAT and the possibility of local production of PAI-1 antigen is present.

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