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Neutrophil Chemotactic Activity in Bronchoalveolar Lavage Fluid of the Rats Exposed to Hyperoxia (고농도의 산소에 노출시킨 쥐의 기관지폐포세척액내 호중구 화학주성활성화도)

  • Song, Jeong Sup;Lee, Sook Young;Moon, Wha Sik;Park, Sung Hak
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.4
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    • pp.547-557
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    • 1996
  • Background : An excessive accumulation of neutrophils in lung tissue has been known to play an important role in mediating the tissue injury among the adult respiratory distress syndrome, idiopathic pulmonary fibrosis and cystic fibrosis by releasing toxic oxygen radicals and proteolytic enzymes. Therefore, it is important to understand a possible mechanism of neutrophil accumulation in lung tissue. In many species, exposure to hyperoxic stimuli can cause changes of lung tissues very similar to human adult respiratory distress syndrome and neutrophils are also functioning as the main effector cells in hyperoxic lung injury. The purpose of the present study was to examine whether neutrophils function as a key effector cell and to study the nature of possible neutrophil chemotactic factors found in bronchoalveolar lavage fluid from the hyperoxia exposed rats. Methods : We exposed the rats to the more than 95% oxygen for 24, 48, 60 arid 72 hours and bronchoalveolar lavage(BAL) was performed. Neutrophil chemotactic activity was measured from the BAT- fluid of each experimental groups. We also evaluated the molecular weight of neutrophil chemotactic tractors using fast performance liquid chromatography and characterized the substances by dialyzer membrane and heat treatment. Results : 1) The neutrophil proportions in bronchoalveolar lavage fluid began to rise from 48 hours after oxygen exposure, and continued to be significantly increased with exposure times. 2) chemotactic index for neutrophils in lung lavages from rats exposed to hyperoxia was significantly higher in 48 hours group than in control group, and was significantly increased with exposure time. 3) No deaths occured until after 48 hours of exposure. However, mortality rates were increased to 33.3 % in 60 hours group and 81.3 % in 72 fours group. 4) Gel filtration using fast performance liquid chromatography disclosed two peaks of neutrophil chemotactic activity in molecular weight of 104,000 and 12,000 daltons. 5) Chemotactic indices of bronchoalveolar lavage fluid were significantly deceased when bronchoalveolar lavage fluid was treated with heat ($56^{\circ}C$ for 30 min or $100^{\circ}C$ for 10 min) or dialyzed (dialyzer membrane molecular weight cut off : 12,000 daltons). Conclusion : These results suggested that the generation of neutrophil chemotactic factor and subsequent neutrophil influx into the lungs are playing an important roles in hyperoxia-induced acute lung injury. Neutrophil chemotactic factor in the lung lavage fluids consisted of several distinct components having different molecular weight and different physical characteristics.

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Effect of GnRH analogue on predicted adult height in girls with early puberty (조기사춘기 여아에서 성선자극호르몬 방출호르몬 효능약제가 예측성인신장에 미치는 효과)

  • Ahn, Byung-Hoon;Han, Heon-Seok
    • Clinical and Experimental Pediatrics
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    • v.49 no.5
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    • pp.552-557
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    • 2006
  • Purpose : The recent results observed in precocious puberty and the hope that interrupting puberty might increase adult height have led to an attempt to use GnRH agonist(GnRHa) in children with premature puberty and a poor growth prognosis. We aimed to analyze the growth promoting effect of GnRHa in girls with early puberty and low predicted adult height(PAH). Methods : Thirty six girls were recruited. They were grouped according to the GnRHa treatment period(group 1>6 mo, n=18; group 2<6 mo, n=18). The following variables were analyzed before and after GnRHa treatment : chronological age(CA), bone age(BA), ${\Delta}age$(CA-BA), height, target height (TH), PAH, serum IGF-1, IGFBP-3. Results : Duration of the GnRHa treatment was $0.89{\pm}0.81yr$($1.37{\pm}0.92yr$ in group 1, and $0.41{\pm}0.08yr$ in group 2). Before treatment, none of the variables were different between the two groups. There were no differences in the following variables the between two groups at the end of treatment : CA, BA, ${\Delta}age$, PAH, serum IGF-1, IGFBP-3. But, growth velocity(GV) and PAH increment during treatment were significantly reduced in group 1. Compared with initial PAH, PAH at the end of treatment was significantly increased($3.7{\pm}3.2cm$). The last serum levels of IGF-1 and IGFBP-3 were lower than those before treatment. Conclusion : Even though last PAH didn't approach TH, short term GnRHa administration in early puberty with low predicted PAH was somewhat effective. But, GnRHa administration suppressed the growth hormone-IGF-1 axis. Therefore, it is recommended that growth hormone(GH) should be used in combination with GnRHa.

Remission rate and remission predictors of Graves disease in children and adolescents (소아 및 청소년 그레이브스병 환자에서의 관해 예측 인자와 관해율)

  • Lee, Sun Hee;Lee, Seong Yong;Chung, Hye Rim;Kim, Jae Hyun;Kim, Ji Hyun;Lee, Young Ah;Yang, Sei Won;Shin, Choong Ho
    • Clinical and Experimental Pediatrics
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    • v.52 no.9
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    • pp.1021-1028
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    • 2009
  • Purpose:Medical therapy is the initial treatment for children with Graves disease to avoid complications of other treatments. However, optimal treatment for childhood Graves disease is controversial because most patients require relatively long periods of medical therapy and relapse is common after medication discontinuation. Therefore, this study aimed to search clinical or biochemical characteristics that could be used as remission predictors in Graves disease. Methods:We retrospectively studied children diagnosed with Graves disease, treated with anti-thyroid agents, and observed for at least 3 years. Patients were categorized into remission and non-remission groups, and the groups were compared to determine the variables that were predictive of achieving remission. Results:Sixty-four patients were enrolled, of which 37 (57.8%) achieved remission and 27 (42.2%) could not achieve remission until the last visit. Normalization of thyroid-stimulating hormone-binding inhibitory immunoglobulin (TBII) after treatment was faster in the remission group than in the non-remission group (remission group, $15.5{\pm}12.07$ vs. non-remission group, $41.69{\pm}35.70$ months). Thyrotropin-releasing hormone (TRH) stimulation tests were performed in 28 patients. Only 2 (8.3%) of 26 patients who showed normal or hyper-response in TRH stimulation test relapsed. Binary logistic regression analysis identified rapid achievement of TBII normalization after treatment as a significant predictor of remission. Six percent of patients achieved remission within 3 years and 55.8% achieved it within 6 years. Conclusion:Rapid achievement of TBII normalization can be a predictor of remission in childhood Graves disease. The TRH stimulation test can be a predictor of maintenance of remission.

GnRH Agonist Stimulation Test (GAST) for Prediction of Ovarian Response in Controlled Ovarian Stimulation (COH) (난소기능평가를 위한 Gonadotropin Releasing Hormone Agonist Stimulation Test (GAST)의 효용성에 관한 연구)

  • Kim, Mee-Ran;Song, In-Ok;Yeon, Hye-Jeong;Choi, Bum-Chae;Paik, Eun-Chan;Koong, Mi-Kyoung;Song, Il-Pyo;Lee, Jin-Woo;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.163-170
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    • 1999
  • Objectives: The aims of this study are 1) to determine if GAST is a better indicator in predicting ovarian response to COH compared with patient's age or basal FSH level and 2) to evaluate its role in detecting abnormal ovarian response. Design: Prospective study in 118 patients undergoing IVF-ET using GnRH-a short protocol during May-September 1995. Materials and Methods: After blood sampling for basal FSH and estradiol $(E_2)$ on cycle day two, 0.5ml (0.525mg) GnRH agonist ($Suprefact^{(r)}$, Hoechst) was injected subcutaneously. Serum $E_2$ was measured 24 hours later. Initial $E_2$ difference $({\Delta}E_2)$ was defined as the change in $E_2$ on day 3 over the baseline day 2 value. Sixteen patients with ovarian cyst or single ovary or incorrect blood collection time were excluded from the analysis. The patients were divided into three groups by ${\Delta}E_2$; group A (n=30):${\Delta}E_2$<40 pg/ml, group B (n=52): 40 pg/ml${\leq}{\Delta}E_2$<100 pg/ml, group C (n=20): ${\Delta}E_2{\leq}100$ pg/ml. COH was done by GnRH agonist/HMG/hCG and IVF-ET was followed. Ratio of $E_2$ on day of hCG injection over the number of ampules of gonadotropins used ($E_2hCGday$/Amp) was regarded as ovarian responsiveness. Poor ovarian response and overstimulation were defined as $E_2$ hCGday less than 600 pg/ml and greater than 5000 pg/ml, respectively. Results: Mean age $({\pm}SEM)$ in group A, B and C were $33.7{\pm}0.8^*,\;31.5{\pm}0.6\;and\;30.6{\pm}0.5^*$, respectively ($^*$: p<0.05). Mean basal FSH level of group $A(11.1{\pm}1.1mlU/ml)$ was significantly higher than those of $B(7.4{\pm}0.2mIU/ml)$ and C $(6.8{\pm}0.4mIU/ml)$ (p<0.001). Mean $E_2hCGday$ of group A was significantly lower than those of group B or C, i.e., $1402.1{\pm}187.7pg/ml,\;3153.2{\pm}240.0pg/ml,\;4078.8{\pm}306.4pg/ml$ respectively (p<0.0001). The number of ampules of gonadotropins used in group A was significantly greater than those in group B or C: $38.6{\pm}2.3,\;24.2{\pm}1.1\;and\;18.5{\pm}1.0$ (p<0.0001). The number of oocytes retrieved in group A was significantly smaller than those in group B or C: $6.4{\pm}1.1,\;15.5{\pm}1.1\;and\;18.6{\pm}1.6$, respectively (p<0.0001). By stepwise multiple regression, only ${\Delta}E_2$ showed a significant correlation (r=0.68, p<0.0001) with $E_2HCGday$/Amp, while age or basal FSH level were not significant. Likewise, only ${\Delta}E_2$ correlated significantly with the number of oocytes retrieved (r=0.57, p<0.001). All four patients whose COH was canceled due to poor ovarian response belonged to group A only (Fisher's exact test, p<0.01). Whereas none of 30 patients in group A (0%) had overstimulation, 14 patients among 72 patients (19.4%) in group B and C had overstimulation (Fisher's exact test, p<0.01). Conclusions: These data suggest that initial $E_2$ difference after GAST may be a better prognostic indicator of ovarian response to COH than age or basal FSH level. Since initial $E_2$ difference demonstrates significant association with abnormal ovarian response such as poor ovarian response necessitating cycle cancellation or overstimulation, GAST may be helpful in monitoring and consultation of patients during COH in IVF-ET cycle.

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Repetition of Apoptosis Induced by Amiloride Derivatives in Human Umbilical Vein Endothelial Cells (제대정맥 내피세포에서 Amiloride 유도체에 의한 Apoptosis 반복)

  • Park, Kyu Chang;Park, Kyu Sang;Moon, Soo Jee
    • Clinical and Experimental Pediatrics
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    • v.46 no.1
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    • pp.56-66
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    • 2003
  • Purpose : Human umbilical vein endothelial cells(HUVECs) play an important role in regulating blood flow by releasing vasoactive substances. It has been reported that endothelial impairment and dysfunction might be a primary cause of placental vascular disease, which is manifested clinically as preeclampsia in mother and intrauterine growth restriction in fetus. Furthermore, the frequency of apoptotic changes is increased in umbilical and placental tissues from growth-restricted pregnancies. However, the various mechanisms of umbilical endothelial cell apoptosis have not been broadly proposed. We investigate the effects of amiloride derivatives on apoptotic death of HUVECs and identify their ionic mechanism. Methods : HUVECs were purchased from Clonetics, and cultured on endothelial cell growth medium. MTT assay and flow cytometry were used for assessing cytotoxic effect and confirming the apoptosis. Changes in intracellular ion concentrations were measured with specific fluorescent dyes and fluorescence imaging analysis system. Results : Amiloride derivatives elicited cytotoxic effects on HUVECs with dose-dependent manners and the rank order of potency is HMA($IC_{50}\;11.2{\mu}M$), MIA>EIPA>>amiloride. HMA-induced cytotoxicity is dependent on extra- and intracellular pH, that is, increase extra- and intracellular pH augmented the cytotoxic effects of HMA. HMA dose-dependently reduced intracellular major ions, such as $K^+$ and $Cl^-$. Interestingly, the depletion of intracellular ions induced by HMA was also significantly enhanced at alkaline extracellular pH. Conclusion : Amiloride derivatives induce apoptosis of HUVECs with dose and pH dependent manners. They reduce intracellular $K^+$ and $Cl^-$ concentration, which is also extracellular pH dependent.

The Change of Antioxidant Enzyme (Superoxide Dismutase, Catalase, Glutathione Peroxidase) in the Endotoxin Infused Rat Lung (내독소 투여후 쥐의 폐조직내 Antioxidant (Superoxide Dismutase, Catalase, GSH-Peroxidase)의 변화에 대한 연구)

  • Song, Jeong-Sup;Kim, Chi-Hong;Kwon, Soon-Seog;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Han, Ki-Don;Moon, Hwa-Sik;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.2
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    • pp.104-111
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    • 1993
  • Background: Gram-negative bacterial endotoxin induced septicemia is known to be a leading cause in the development of adult respiratory distress syndrome(ARDS). The mechanism of endotoxin induced lung injury is mainly due to the activated neutrophils which injure the capillary endothelial cells by releasing oxidant radical and resulted in pulmonary edema. We studied the change of antioxidant enzyme in the case of large or small, intermittant dose of endotoxin infused rat lungs. Methods: Endotoxin was given to the rat through the peritoneal cavity in the dose of 7 mg/kg body weight in the large dose group and 1 mg/kg for 10 days in the small dose group. Bronchoalveolar lavage (BAL) was done and rats were killed at 6, 12, 24 hours after single endotoxin injection in the large dose group and 3, 7, 10 days after daily endotoxin injection for 10 days in the small dose group. The lungs were perfused with normal saline through the pulmonary artery to remove the blood and were homogenized in 5 volume of 50 mM potassium phosphate buffer containing 0.1 mM EDTA. After centrifuging at 100,000 g for 60 minute, the supernatent was removed and stored at $-70^{\circ}C$ until measuring for superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px) and protein. Results: We observed the following results. 1) The lung wet/dry weight ratio and albumin concentration in the BAL fluids were increased to peak at 12 hours and neutrophil number in the BAL fluids were peak at 6 hours after endotoxin injection in the large dose group. 2) Cu, Zn SOD (IU/mg protein) was significantly decreased after 6, 12 hours after endotoxin injection in the large dose group. 3) There were no singnificant change in the level of Mn SOD, catalase, GSH-Px after endotoxin injection in both groups. Conclusion: Endotoxin in the large dose group produced the acute pulmonary edema and decreased the Cu, Zn SOD in the lung tissue after injecting endotoxin at 6 and 12 hours. These phenomenon may be due to the cell membrane damage by endotoxin. Further research would be necessary whther giving SOD by intratracheal route or method to increase the synthesis of SOD may lessen the acute lung injury by endotoxin.

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Efficacy and Safety of Miniscalpel Acupuncture in Knee Degenerative Osteoarthritis Patients: A Study Protocol for a Randomized Controlled Pilot Trial (퇴행성 슬관절염 환자에 대한 도침요법의 효능 및 안전성 연구: 임상예비연구)

  • Jun, Seungah;Park, Mu Seob;Oh, Se Jung;Lee, Jung Hee;Gong, Han Mi;Choi, Seong Hun;Hwangbo, Min;Lee, Hyun-Jong;Kim, Jae Soo
    • Korean Journal of Acupuncture
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    • v.33 no.2
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    • pp.67-74
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    • 2016
  • Objectives : The Knee degenerative osteoarthritis patients are not satisfied with the conventional therapies of KDOA, which results in the use of alternative therapies. The miniscalpel acupuncture is effective in treating chronic soft tissue, releasing contractures. However, there is little scientific evidence supporting the use of miniscalpel acupuncture in knee degenerative osteoarthritis. This study was designed to obtain basic data for a further large-scale trial as well as provide information about the feasibility of miniscalpel acupuncture in knee degenerative osteoarthritis patients. Methods : We describe the protocol for a randomized controlled pilot clinical trial of 5 weeks duration. Twenty patients will be recruited and randomly allocated to two treatment groups: miniscalpel acupuncture treatment(experimental group); and acupuncture and electro-acupuncture treatment(control group). Miniscalpel acupuncture will be performed once with a 1-week interval for 3 weeks. Electro-acupuncture will be administered twice per week for a period of 3 weeks. The primary outcomes will be measured by visual analogue scale and range of motion. The secondary outcomes will be short-form McGill Pain Questionnaire and Western Ontario and McMaster Universities Osteoarthritis Index. Both primary and secondary outcomes will be measured at baseline and at 1, 2, 3 and 5 weeks(i.e. 2 weeks after treatment completion). Conclusions : This pilot study will provide a basic foundation for a future large-scale trial as well as information about the feasibility of miniscalpel acupuncture in knee degenerative osteoarthritis.

Research on Archive Opening and Sharing Projects of Korean Terrestrial Broadcasters and External Users of Shared Archives : Focusing on the Case of the 5.18 Footage Video Sharing Project 〈May Story(Owol-Iyagi)〉 Contest Organized by KBS (국내 지상파 방송사의 아카이브 개방·공유 사업과 아카이브 이용자 연구 KBS 5.18 아카이브 시민공유 프로젝트 <5월이야기> 공모전 사례를 중심으로)

  • Choi, Hyojin
    • The Korean Journal of Archival Studies
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    • no.78
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    • pp.197-249
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    • 2023
  • This paper focus on the demand for broadcast and video archive contents by users outside broadcasters as the archive openness and sharing projects of terrestrial broadcasters have become more active in recent years. In the process of creating works using broadcasters' released video footage, the study examined the criteria by which video footage is selected and the methods and processes utilized for editing. To this end, the study analyzed the the case of the 5.18 footage video sharing project 〈May Story(Owol-Iyagi)〉 contest organized by KBS in 2022, in which KBS released its footage about the May 18 Democratic Uprising and invited external users to create new content using them. Analyzing the works that were selected as the winners of the contest, the research conducts in-depth interviews with the creators of each work. As a result, the following points are identified. Among the submitted works, many works deal with the direct or indirect experience of the May 18 Democratic Uprising and focus on the impact of this historical event on individuals and our current society. The study also examined the ways in which broadcasters' footage is used in secondary works. We found ways to use video as a means to share historical events, or to present video as evidence or metaphor. It is found that the need for broadcasters to provide a wider range of public video materials such as the May 18 Democratic Uprising, describing more metadata including copyright information before releasing selected footage, ensuring high-definition and high-fidelity videos that can be used for editing, and strengthening streaming or downloading functions for user friendliness. Through this, the study explores the future direction of broadcasters' video data openness and sharing business, and confirms that broadcasters' archival projects can be an alternative to fulfill public responsibilities such as strengthening social integration between regions, generations, and classes through moving images.

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EFFECT OF INDUCTION CHEMOTHERAPY ON FLAP SURVIVAL RATE IN MICROSURGERY (종양수술전 화학요법이 미세수술시 피판생존율에 미치는 영향)

  • Kim, Uk-Kyu;Kim, Yong-Deok;Byun, June-Ho;Shin, Sang-Hun;Chung, In-Kyo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.29 no.6
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    • pp.421-429
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    • 2003
  • Purpose : Neoadjuvant chemotherapy is commonly used to treat cancer patients as adjunct treatment, but if the microvascular tissue transfer is performed simulataneously with cancer resection surgery, the induction chemotherapy might affect the survival rate of vascularized free flap. Our study will focus on the effect of induction chemotherapy on the free flaps which were made on white rat abdomen after injection of 5-FU. Materials and Methods: The experimental rat groups were divided into three groups (total 24 rats) as a normal control group, 24 hrs group after 5-FU injection, 3 days group after 5-FU injection. Inferior abdominal island flaps of 8 Sprague Dawley rats on each group were made and immediately were induced into an ischemic state by clamping the supplying inferior epigastric artery and vein with microvascular clamp for a hour to induce a similiar free flap circumstance, then the inferior abdominal skin flaps were reperfused by releasing the clamps. The flaps on abdomen were repositioned and sutured. The experimental data for flap survival rate was collected by digital photo taking, analysed by computer image program to compare with the flap luminosity. The rats were sacrificed at 3 days, 5 days, 7 days after flap preparation and specimens of the flap were taken and stained with H-E staining. The microscopic finding was made under magnification of 200 and 400. Results: 1. Gross findings on each groups showed the healing condition was good as following sequences; normal, 24 hrs group after chemotherapy, 3 days group after chemotherpy. 2. The values of flap luminosity for evaluation of flap survival rate also showed the same sequences as gross findings of healing state. 3. The microscopic findings of epidermis necrosis, inflammation state, dermis fibrosis, vessel change, fatty tissue layer thinning were compared with each group. The 3 days group after chemotherapy showed remarkably poor healing condition compared to other groups. Conclusion: Chemotherapy agents affected the healing process of free flap, but healing condition was recovered spontaneously as post-injection periods passed out. In opposite to our expectation, 3 days group showed the bad flap condition in comparing with 24 hours group which was considered as immatured body circulation state of chemotherapy agent. It showed that 3 weeks in human being after chemotherapy was not proper as timing of microvascular tissue transfer if 3 days group in rat was considered as same healing period of 3 weeks in human being. More delayed healing timing than 3 weeks might be required in clinical application of free tissue transfer.

Lipopolysaccharide-induced Synthesis of IL-1beta, IL-6, TNF-alpha and TGF-beta by Peripheral Blood Mononuclear Cells (내독소에 의한 말초혈액 단핵구의 IL-1beta, IL-6, TNF-alpha와 TGF-beta 생성에 관한 연구)

  • Jung, Sung-Hwan;Park, Choon-Sik;Kim, Mi-Ho;Kim, Eun-Young;Chang, Hun-Soo;Ki, Shin-Young;Uh, Soo-Taek;Moon, Seung-Hyuk;Kim, Yang-Hoon;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.846-860
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    • 1998
  • Background: Endotoxin (LPS : lipopolysaccharide), a potent activator of immune system, can induce acute and chronic inflammation through the production of cytokines by a variety of cells, such as monocytes, endothelial cells, lymphocytes, eosinophils, neutrophils and fibroblasts. LPS stimulate the mononucelar cells by two different pathway, the CD14 dependent and independent way, of which the former has been well documented, but not the latter. LPS binds to the LPS-binding protein (LBP), in serum, to make the LPS-LBP complex which interacts with CD14 molecules on the mononuclear cell surface in peripheral blood or is transported to the tissues. In case of high concentration of LPS, LPS can stimulate directly the macrophages without LBP. We investigated to detect the generation of proinflammatory cytokines such as interleukin 1 (IL-1), IL-6 and TNF-$\alpha$ and fibrogenic cytokine, TGF-$\beta$, by peripheral blood mononuclear cells (PBMC) after LPS stimulation under serum-free conditions, which lacks LBPs. Methods : PBMC were obtained by centrifugation on Ficoll Hypaque solution of peripheral venous bloods from healthy normal subjects, then stimulated in the presence of LPS (0.1 ${\mu}g/mL$ to 100 ${\mu}g/mL$ ). The activities of IL-1, IL-6, TNF, and TGF-$\beta$ were measured by bioassaies using cytokines - dependent proliferating or inhibiting cell lines. The cellular sources producing the cytokines was investigated by immunohistochemical stains and in situ hybridization. Results : PBMC started to produce IL-6, TNF-$\alpha$ and TGF-$\beta$ in 1 hr, 4 hrs and 8hrs, respectively, after LPS stimulation. The production of IL-6, TNF-$\alpha$ and TGF-$\beta$ continuously increased 96 hrs after stimulation of LPS. The amount of production was 19.8 ng/ml of IL-6 by $10^5$ PBMC, 4.1 ng/mL of TNF by $10^6$ PBMC and 34.4 pg/mL of TGF-$\beta$ by $2{\times}10^6$ PBMC. The immunoreactivity to IL-6, TNF-$\alpha$ and TGF-$\beta$ were detected on monocytes in LPS-stimulated PBMC. Some of lymphocytes showed positive immunoreactivity to TGF-$\beta$. Double immunohistochemical stain showed that IL-1$\beta$, IL-6, TNF-$\alpha$ expression was not associated with CD14 postivity on monocytes. IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$mRNA expression were same as observed in immunoreactivity for each cytokines. Conclusion: When monocytes are stimulated with LPS under serum-free conditions, IL-6 and TNF-$\alpha$ are secreted in early stage of inflammation. In contrast, the secretion of TGF-$\beta$ arise in the late stages and that is maintained after 96 hrs. The main cells releasing IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ are monocytes, but also lymphocytes can secret TGF-$\beta$.

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