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Analysis of Stromal Cells Developed from Cord Blood CD34+ Cells (제대혈 CD34+ 세포에서 유래된 지지세포의 분석)

  • Ryu, Kyung-Ha;Park, Se-Jin;Kim, Kyung Hyo;Seoh, Ju-Young;Khan, Mohammad;Shin, Hee-Young;Ahn, Hyo-Seop
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.87-94
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    • 2001
  • Background: Cytokine-mediated ex vivo expansion has been proposed as a means of increasing the number of cord blood (CB) hematopoietic stem cells for transplantation. As well as stem cell number, stromal cells are necessary for functional maturation of hematopoiesis. The purpose of this study was to analyze the development of stromal cells during ex vivo expansion of CB $CD34^+$ cells. Methods : $CD34^+$ cells were purified from CB by magnetic bead selection. The levels of of interleukin-3, interleukin-$1{\beta}$, interleukin-6, granulocyte macrophagecolony stimulating factor and tumor necrosis factor-${\alpha}$ were measured in culture supernatants on 0, 1, 2, and 3 weeks, using ELISA techniques. CB $CD34^+$ cells were expanded in Iscoves modified Dulbeccos medium in the presence of several cytokines. The expression of E-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, platelet/endothelial cell adhesion molecule-1, von Willebrand factor, vimentin, and CD14 in newly developed stromal cells was examined by immunocytochemical method. Relevant extracellular matrix (ECM) proteins and proper cytokines were also assayed for the most suitable condition for expansion of stromal cells. Results: Several cytokines were found to have been produced by CB $CD34^+$ cells as well as bone marrow-derived $CD34^+$ cells. During ex vivo expansion of CB $CD34^+$ cells, stromal cells appeared in the culture by day 4 and expanded over the following 7-10 days before being confluent by day 2 1. These cells expressed surface markers characteristic of cells of endothelial lineage. Furthermore, these stroaml cells also expanded effectively when treated with thrombopoietin+flt-3 ligand+stem cell factor+leukemia inhibitory factor or 0.1% poly-L-lysine-coated wells. Conclusion: Stromal cells were developed during ex vivo expansion of CB $CD34^+$ cells and that this development could be enhanced further by treating the stromal cells with cytokines or ECM.

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External and Cranial Characters of the Tscherskia triton in Jeju Island, Korea (제주도에 서식하는 비단털쥐 (Tscherskia triton)의 외부형태 및 두개골 특징)

  • Park, Jun-Ho;Oh, Hong-Shik
    • Korean Journal of Environmental Biology
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    • v.35 no.3
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    • pp.389-397
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    • 2017
  • The purpose of this study was to investigate the characteristics of species by comparing the external shape traits and skull size of Tscherskia triton in Jeju Island from May 2014 to November 2016. Body weight, head-body length, ear length, and hind-foot length were not statistically significant (p>0.05), but tail length was statistically significant in females compared to males (p=0.003). To examine the secondary sexual dimorphism characteristics of the collected T. triton, B_HR, T_HR, E_HR, and H_HR were calculated based on the head-body length, and only T_HR was statistically significant (p=0.003). Because of comparing male and female averages, among the 21 traits, the greatest length of skull (GLS; p=0.020) and occipitonasal length (ONL; p=0.014) were statistically significant and length of incisive foramen (LIF) was statistically significant, when the significance level was 90%. However, the other 18 traits did not show statistically significant differences between males and females (p>0.05). Other than GLS, ONL, interorbital breadth (IB), and breadth of occipital foramen (BOF), there was female-biased SSD. The results of this study are the analysis of external morphology and skull characteristics of T. triton in Jeju Island and will be used as basic data to understand the morphological characteristics of T. triton. In addition, the data can be used as a basic information to compare characteristics of T. triton, which is known as a single species, inhabited in China and Russia. The data will play an important role in biogeography and ecology research of T. triton, based on geographical conditions, climate effects, and habitat environment.

ICM - Trophectoderm Cell Numbers of Mouse IVF/IVC Blastocysts (체외생산된 생쥐 배반포기배의 ICM과 Trophectoderm 세포수에 관한 연구)

  • Kim, E.Y.;Kim, S.E.;Uhm, S.J.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.1
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    • pp.25-32
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    • 1996
  • This work has been carried out to examine the number of Total, ICM and TE cells of F1 mouse blastcysts at day 4 after IVF by differential labelling of the nuclei with polynucleotide-specific fluorochromes and to obtain a fundamental information of preimplantation mouse embryo development. Blastocysts produced by superovulated B6CBA F1(C57BL/${\times}$CBA) eggs were inseminated with $1{\times}10^6$spermatozoa/ml and cultured in M16 medium at $37^{\circ}C$, 5% $CO_2$ incubator for 95hrs. Blastocysts were classified as early, middle, expanded and hatching stage according to the developmental morphology; blastocoel expansion and zona thickness. The results obtained in these experiments were summarized as follows; 1) The development rate of blastocysts at 95hrs after IVF was 86.7% and classified blastocysts to early, middle, expanded and hatching were 16.3%, 18.9%, 10.5% and 40.9%, respectively. 2) The numbers of total blastomere using bisbenzimide in the classified blastocysts to early, middle, expanded and hatching were 35.6${\pm}$1O.4, 49.4${\pm}$8.6, 60.8${\pm}$1O.7 and 62.7${\pm}$13.9, respectively. 3) In ICM and TE cell number by using differential labelling with polynucleotide-specific fluorochrome in the classified blastocysts to early, middle, expanded and hatching; ICM numbers were 9.6${\pm}$3.0, 13.6${\pm}$3.9, 16.0${\pm}$3.3 and 19.5${\pm}$4.6, respectively and TE cell numbers were 30.6${\pm}$5.1, 39.9${\pm}$5.8, 42.2${\pm}$8.1 and 43.7${\pm}$11.1, respectively. These results showed the same increase pattern according to development advance level. Also, when compared with the results of total count were obtained between bisbenzimide only and differential labelling, both of them showed the same increase pattern according to development level and at the same time their cell numbers were almost the same. So, rapid and simple cell count method using differential labelling can be used for the examination of later preimplantation development or as an indicator of embryo quality according to the variables of culture conditions.

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The Synthesis and MR Properties of New Macromolecular MR Contrast Agent (새로운 거대분자 MR 조영제의 합성 및 MR 특성에 관한 연구)

  • 장용민;장영환;황문정;박현정;전경녀;이종민;배경수;강봉석
    • Investigative Magnetic Resonance Imaging
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    • v.6 no.1
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    • pp.35-40
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    • 2002
  • Purpose : To evaluate the NMR relaxation properties and imaging characteristics of tissue-specificity for a newly developed macromolecular MR agent. Materials and methods : Phthalocyanine (PC) was chelated with paramagnetic ion, Mn.2.01g (5.2 mmol) of Phthalocyanine was mixed with 0.37g (1.4 mmol) of Mn chloride at $310^{\circ}C$ for 36 hours and then purified by chromatography (CHC13/CH3OH 98/2 v/v, Rf, 0.76) to obtain 1.04g (46%) of MnPC (molecular weight= 2000d). The $T1}T2$ relaxivity of MnPC was measured in 1.5T(64 MHz) MR using 0.1 mM MnPC. The MR image characteristics of MnPC was evaluated using spin-echo (TR/TE=500/14 msec) and gradient-echo (FLASH) (TR/TE=80/4 msec, flip angle=60) techniques in 1.57 MR scanner. The images of rabbit liver were obtained every 10 minutes up to 4 hours. To study the effect of concentration on image, 20 mM, 50 mM, 100 mM of MnPC were tested. Results : The relaxivities of MnPC at 1.5T(64MHz) were Rl=7.28 $mM^{-1}S^{-1},{\;}R2=55.56mM^{-1}S^{-1}$. Compared to the values of Gd-DTPA (Rl[=4.8 $mM^{-1}S^{-1})$], R2[=5.2 $mM^{-1}S^{-1}])$]), both T1/T2 relaxivities of MnPC were higher than those of Gd-DTPA. For both of SE and FLASH techniques, the contrast enhancement reached maximum at 10 minutes after bolus injection and the enhancement continued for more than 2 hours. When compared with small molecular weight liver agents such as Gd-EOB-DTPA, Gd-BOPTA and MnDPDP, MnPC was characterized by more prolonged enhancement time. The time course of MR images also revealed biliary excretion of MnPC. Conclusion : We developed a new macromolecular MR agent, MnPC. The relaxivities of MnPC were higher than those of small molecular weight Gd-chelate. Hepatic uptake and biliary excretion of MnPC suggests that this agent is a new liver-specific MR agent.

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Temperature Sensitivity for Flowering of Bulblets in Lilium formolongi (신나팔나리(Lilium formolongi) 인편자구의 개화를 위한 온도 감응)

  • Goo, Dae-Hoe
    • Journal of Bio-Environment Control
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    • v.17 no.3
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    • pp.221-225
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    • 2008
  • Lilium formolongi 'Fl August' plantlets with scale-leaves and scale-bulb were treated at 10, 15, 20, $25^{\circ}C$ for 15, 30, or 45 days and planted in February, March, April and May. In the April planting, flowering percentage was below 10% and in the May planting no flowering occurred. Sprouting and flowering percentages were lower at the late planting times. Days to flowering of the April planting was 110.8 days compared to 128 days for the February planting. Plant height and numbers of leaves were reduced to 7.2 in May planting, compared to 40.5 leaves in February planting, and almost no flowers emerged in either the April or May plantings. Plantlets exposed to 10 or $15^{\circ}C$ flowered at 80 percent or higher at all treatment durations, but at 20 or $25^{\circ}C$ flowering percentages were lower, with 30% or less in the $25^{\circ}C$ treatment. In the $15^{\circ}C$ treatment days to flowering were less than 100 days, while the number of flowers and flower bud differentiation were greatest in the $15^{\circ}C$ treatment. Cytokinin and auxin were analyzed in bulblets grown at 15, 20 and $25^{\circ}C$. T-zeatin content was three times greater in the $15^{\circ}C$ treatment than at $25^{\circ}C$, but the content of indoleacetic acid (IAA) was less at $15^{\circ}C$ than at 20 and $25^{\circ}C$. In the $15^{\circ}C$ treatment, T-zeatin content was about twice the IAA content in the scale- bulblet. The auxin and cytokinin balance may affect flower bud differentiation. $15^{\circ}C$ with 30 days was most effective for flowering in scale-bulblet and planting of February and March were effective for flowering too.

Histopathological Changes in Cold-Stored Dog Lungs to the Preservation Solutions (냉한에서 보관된 황견의 폐에서 장기 보존액에 따른 조직 세포의 변화)

  • 김해균;이두연
    • Journal of Chest Surgery
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    • v.29 no.8
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    • pp.816-821
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    • 1996
  • Lung transplantation is the established treatment for the end stage lung disedse find preservation of the organ is a major obstacle In performing lung transplantation. For solving this problem, we evaluated the histopathologic changes for various preservation solutions. Male mongrel dogs of similar size and weight (15∼20 kg) were used. The dog lungs were flushed with 4fl normal saline(group 1 'n:5): Modified Euro-Collins solution(group 2 n:5) and University of Wisconsin solution (group 3 : n=6), 60m11kg through a catheter placed in the main pulmonary artery aft r flushing of PGE 1 (20ng1kg). The lungs were preserved for 60 hours and measured dry and wet weights. Histologic specimens were taken every 6 hours and %toed for light microscopic evaluation. The edema ratio of the lungs peaked in 12 hours although there was no difference between the groups. Histologically, alveolar septal changes developed in one case (20%) after 1 hour preservation with normal saline. In case of the University of Wisconsin solution, the alveolar septal distortions and swellings were seen in 1 cases (20%) after 6 hours preservation compared with 3 cases (60%) after 6 hours preservation with Modified Euro-Collins solution. Changes of the pneumocytes were observed after 24 hours preser- vation in group 1, after 48 hours preservation in group 2 and after 60 hours preservation in group 3. We conclude that University of Wisconsin solution might have a superior preservation effect compare to normal saline and Modified Euro-Collins solutions.

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Investigation of the Relationship between Interleukin-4 Promoter Polymorphism and Severity of Patients with Bronchial Asthma (천식 환자에서 증상의 정도에 따른 IL-4 유전자 다형에 관한 연구)

  • Kang, Sea-Yong;Shim, Jae-Jeong;Cho, Jae-Yun;Kwon, Young-Hwan;Lee, Seung-Yong;Kim, Je-Hyeong;Lee, Sang-Youb;Lee, So-Ra;Han, Seon-Ae;Kim, Han-Gyum;Kang, Kyung-Ho;Yoo, Se-Hwa;In, Kwang-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.3
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    • pp.529-535
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    • 1998
  • Background: Interleukin-4 plays an important role in pathogenesis of asthma, especially in developing atopy by means of switching B lymphocytes to produce IgE. It has been shown that there is polymorphism in the Interleukin-4 promoter region, transversion of cytosine to thymine at-598 from translation initiation site of IL-4 gene. There has also been quite a few works to reveal the role of the polymorphism of IL-4 gene in patients with asthma. We performed this investigation to determine the role of the polymorphism in the severity of symptoms of patients with asthma. We also examined the frequency and the type of the polymorphism in asthmatics compared with non-asthmatics as well. Method: The subjects enrolled in this study were 49 asthmatics and 33 non-asthmatics. All the asthmatics were classified as mild and moderate to severe by the NHLBI/WHO Workshop. DNA from both asthmatics and non-asthmatics was extracted, then performed ARMS(Amplification Refractory Mutation System) as well as RFLP using BsmFl restriction enzyme in order to confirm the polymorphism of Il-4 gene. Results: There was no significant difference in the occurrence of polymorphism of the IL-4 promoter sequence between asthm and non-asthma groups(P=0.7). Among those with polymorphisms, the number of C/C type was slightly more than C/T type in both asthmatics and non-asthmatics, 26 vs 21 in asthmatics and 18 vs 15 in non-asthmatics, which was, however, insignificant statistically. No significant relationship between the severity of asthma and the polymorphism was found(P=0.7). Conclusion: There was no significant difference between the severity of asthma and the IL-4 promoter polymorphism(P=0.709). Interestingly, the frequency of the polymorphism in both asthmatics as well as non-asthmatics was found to be even higher than that occurred in Caucasians. However, no significant difference in the frequency of the polymorphism was found in both groups.

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The Surgical Treatment of Atrial Fibrillation in Patients Undergoing Simultaneous Open Heart Surgery (심장세동의 수술요법)

  • Kim, Gi-Bong;Lee, Chang-Ha;Son, Dae-Won
    • Journal of Chest Surgery
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    • v.30 no.3
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    • pp.287-292
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    • 1997
  • .Itrial fibrillation is one of the most common cardiac arrhythmias requiring treatment. About 60% of patients with mitral valvular disease have atrial fibrillation and one third of patients with atrial fibrillation may have the past history of thromboembolic events. Between April 1994 and June 1995, 20 patients with organic heart diseases combined with atrial fibrillation underwent open heart surgery including Cox-maze 111 procedure. There were 6 men and 14 women with an average age of 48 years (range, 31 to 66 years). Nineteen patients had valvular heart diseases and 1 ventricular septal defEct (VSD). Mean duration of atrial fibrillation was 36 months (:42 months) (range, 1 to 132 months). T e past medical history of thromboembolic events was positive in 7 patients (35%) and left atrial thrombus was detected in 9 patients (45%). The concomitant procedures were mitral valve replacement (MVR) and aortic valve replacement (AVR) in 5 patients, MVR in 4, MVd and tricuspid annuloplasty(TAP) in 4, mitral valvuloplasty(Mln) in 3, Mln and Tln in 1, MIW and coronary artery bypass surgery in 1, AVR in 1, and patch closure of VSD in 1. Mean aortic cross-clamping time was 175 minutes (range, 116 to 270 minutes). Atrial fibrillation recurred in 16 patients (80%) during the early postoperative period, but, recurrent atrial fibrillation was converted to regular rhythm at postoperative forty-first day in average. There was no early or late death in this series of 20 patients and postoperative complications were inappropriate tachycardia in 5 patients (25%), low cardiac output syndrome in 3 (15%), aggravated hemiplegic in 1, and acute renal failure in 1. Mean follow-up interval of patient was 16.5 months (range, 10.5 to 24 months) and all patients are currently in regular rhythm. Seventeen patients (85%) are in sinus rhythm and 3 (15%) in junctional rhythm. Right atrial contraction was detected in 95% of patients and left atrial contraction in 63% on postoperative transthoracic echocardiogram. The surgical treatment of atrial fibrillation concomitant with open heart surgery is warranted in the recent clinical setting of improved myocardial protection technique, considering the untoward side-effects of atrial fibrillation.

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Traumatic Injuries of the Diaphragm (외상성 횡격막 손상)

  • 김덕실;허동명
    • Journal of Chest Surgery
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    • v.29 no.4
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    • pp.433-439
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    • 1996
  • From January 1980 to July 1995, 32 patients with blunt diaphragmatic trauma and 12 wish penetrating diaphragmatic trauma were treated at Kyungpook University Hospital. The mean age of patients with blunt trauma was 37.6 years, and with penetrating trauma 29.2 years. The mean age was older in patients with blunt trauma by 8.4 years (p< 0.05). When simple chest X-ray diagnosis was performed, 24cases (75%) of the diaphragmatic injuries were found in blunt trauma and 4 case(33%) in penetrating trauma. When opei.ations were done due to injuries of thoracic or abdominal organ, 7 diaphragmatic injuries (58%) were found in penetrating trauma. Herniation occurred in 24 patients (75%) in blunt trauma and 5 patients (42%) in penetrating truma. The size of injured diaphragms in 29 cases in which hernia was discovered was 10.9 $\pm$ 4.3cm. On the other hand, the size in 15 cases in which hernia did not occur was 3.5 $\pm$ 2.9cm. There was a remarkable difference in their size depending on the presence or absence of hernia (p< 0.051. That is, a close relation between the sire of injured diaphragms and hernia was shown. All diaphragmatic injuries were repaired primarily. In blunt trauma approach of repair were as follows : 20 (63%) thoracic, 9 abdominal, 2 thoracoabdominal, and 1 thoracic and in penetrating trauma 6 (50%) abdominal, 4 thoracic, and 2 thoracic and abdominal. The complication rate was 19% in blunt trauma and 25% in penetrating trauma. Two patients with blunt trauma died with a mortality rate of 6.3% . All patients with penetrating trauma recovered. This study suggests that diaphragmatic injury should be suspected in all patients w th blunt as well as penetrating injury of the chest and abdomen. The size of injured diaphragms was larger in blunt trauma than in penetrating trauma. For herniation, it appeared to be more common in blunt trauma. The morbidity and mortality were related primary to the severity of associated injuries.

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Growth and Morphological Characteristics of Introduced Sorghum Germplasm (도입 수수 유전자원의 생육 및 형태적 특성)

  • 강정훈;이호진
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.2
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    • pp.207-214
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    • 1996
  • This study was conducted to obtain fundamental information on forage sorghum breeding in forage crop field of Livestock Experiment Station at Suwon from 1986 to 1991. The charcterization of sorghum germplasm was performed through 1986 to 1987, and after parental lines were selected from diverse sorghum germplasm on the basis of flowering date, plant height and several morphological characters for forage sorghum Fl hybrids. The range of variation of 50% flowering date and plant height were greater in order of forage sorghum sudangrass and male sterile line of grain sorghum. The average flowering date was earlier in sudangrass and male sterile line of grain sorghum than forage sorghum lines from the tested sorghum germplasms. And the average plant height was tall in order of forage sorghum, sudangrass and male sterile lines of grain sorghum. There were remarkable morphological variations between sudangrass lines and male sterile lines of grain sorghum such as plant color, leaf midrib color, glume color, seed coat color, head compactness and shape, awns, grain covering and 100 seed weight.

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