• Korean Journal of Clinical Laboratory Science
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• v.50 no.3
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• pp.304-312
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• 2018

Regulatory T cells (Treg), known as immune-suppressors, may help modulate the immune response. In this study, we investigated the effect of Treg-derived $TGF-{\beta}1$ on pancreatic islet cell function in vitro and in vivo. One hundred eighty IEQ (islet equivalents) of pancreatic islets, the marginal amount to regulate blood glucose level after syngeneic islet transplantation in mouse type 1 diabetes (T1D) model, were co-cultured with $4{\times}10^6$ Treg cells for 48 hours. The changes in $TGF-{\beta}1$, interleukin-6 (IL-6), and insulin secretion levels were measured and analyzed among the Treg-only group, the islet-only group, and the Treg/islet co-cultured group. In the Treg/islet co-cultured group, IL-6 and insulin secretion levels were increased (P<0.0005, P<0.005) and islet viability was improved (P<0.005) compared with the islet-only group. Furthermore, after transplantation, the co-cultured islets regulated blood glucose levels efficiently in the T1D mouse model. These data suggest that Treg could improve islet functions and viability via the $TGF-{\beta}1$ secretion pathway (P<0.05~0.005), thus the use of Treg in islet transplantation should be explored further.

• Journal of Chest Surgery
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• v.32 no.2
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• pp.206-210
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• 1999

We report here 2 cases of deep-seated mediastinitis combined with sternal osteomyelitis after tracheal reconstruction which were successfully treated with sternectomy, in-situ or free omental transfer, and pectoralis major myocutaneous flap. In case I, an 8 year-old boy with deep seated mediastinitis and sternal osteomyelitis that developed after anterior tracheoplasty through a standard midline sternotomy. In case II, a 50 year-old female patient with mediastinal abcess and sternal osteomyelitis that developed after resection and end-to-end anastomosis of the trachea through an upper midline sternotomy. Treatments consisted of drainage and irrigation followed by wide resection of the infected sternum, placement of the viable omentum into the anterior mediastinal space, and chest wall reconstruction with a pectoralis major myocutaneous flap. The omentum was transferred as an in-situ pedicled graft in case I and a free graft in case II. Both patients have recovered smoothly wit out any events and have been doing well postoperatively.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.88-88
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• 2002

인공수정 및 수정란기술의 활성화에 따라 소에 있어서 인공수정은 90%이상이 실시되고 있으나, 수정란이식은 수정란의 생산이 안정적이지 않아 활성화에 많은 지장을 초래하고 있다. 이의 원인은 수정란이식에 의한 수태율의 저하가 가장 크며, 수태율 향상을 위하여 수란우에 progesterone, hCG 등의 주사가 실시되고 있다. 그러나 이는 수정란의 착상에 있어서 자궁의 환경을 개선한다고 하나, 착상의 정확한 기전의 구명은 미미한 상태이다. 한편 비장유래 macrophage가 황체를 자극하고 TGF-$\beta$의 생산을 유도하는 것으로 보고되고 있으며, IL-I $\alpha$와 $\beta$에 따라 TGF-$\beta$ 생산에 있어서 약간의 차이를 보이는 것으로 보고되고 있다. 따라서 본 연구는 비장유래 macrophage가 TGF-$\beta$의 생산시 임신관련 Cytokine인 IL-I$\alpha$와의 관계를 조사하기 위하여 실시하였다. 임신 및 비임신 도축 암소의 비장을 채취하여 얼음에 채워 실험실로 운반한 후 비장의 표면을 70%의 알콜로 세척하고, 표피를 벗겨 비장조직을 세절하여 10% FBS+DMEM에 넣어 조직을 눌러 짜면서 조직속의 세포를 분리하였다. 세척한 배양액은 4-5$m\ell$를 100mm 유리 petri dish에 넣고 39$^{\circ}C$, 5% $CO_2$, 95% 공기인 배양기에서 2시간이상 배양하였으며, 배양 후 냉장된 buffer A 용액으로 세척하여 유리 petri dish의 바닥에 부착된 macrophage만을 cell scraper로 분리하였다. 분리한 macrophage는 0.5-1 $\times$ $10^{6}$ cells/$m\ell$가 되게 조정하여, IL-I 을 0.001, 0.01, 0.1 또한 1 ng/$m\ell$를 첨가하여 농도에 따른 효과를 조사하였고, 각각 24, 48, 72, 96 또한 120시간을 배양하여 시간에 의한 효과도 실시하였다. 각 배치구에서 얻어진 배양액은 TGF-$\beta$를 조사하기 전까지 -2$0^{\circ}C$에 동결 보존하였다. TGF-$\beta$의 측정은 TGF-$\beta$ kit(promega, USA)를 이용하여 실시하였으며, 통계학적 분석은 Anova test를 Statview program을 이용하여 분석하였다. 시험의 결과 대조구에 비해 IL-I 첨가구는 2-3배의 TGF-$\beta$생산을 보였으며, 배양시간에 따른 생산은 시간이 지남에 따라 약간 상승하는 경향을 보였으나, 유의적인 차이를 보이지는 않았다. 또한 IL-I의 농도에 따른 생산의 변화는 IL-I의 농도에 따라 약간의 차이를 보였고 유의적인 차이는 인정되지 않았다. 임신 및 비임신의 경우 임신우의 비장 macrophage가 비임신보다는 약간 상승하는 거스로 나타났다. 이상의 결과로 볼 때 IL-I $\alpha$ 는$\beta$subunit 보다 TGF-$\beta$ 생산에 있어서 서로 다른 양상을 보일 것으로 추정되며, IL-I은 macrophage의 직접적인 영향을 주기보다는 황체세포를 매개로 한 자궁에 TGF-$\beta$의 생산을 유도하는 것으로 사료되며, 임신관련 cytokine에 대한 다양한 연구가 요구되고 있다.

• Journal of Chest Surgery
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• v.40 no.4 s.273
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• pp.256-263
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• 2007

Background: Intimal hyperpiasia is characterized by a proliferation of vascular smooth muscle cells in the intimal layer Epigallocatechin-3-gallate (EGCG) is known to suppress smooth muscle cell proliferation. We propose that EGCG may have a protective effect against the development of intimal hyperplasia through the suppression of smooth muscle cell proliferation. Material and Method: Human umbilical vein endothelial cells (HUVEC) and rat aortic smooth muscle cells (RASMC) were cultured with different concentrations of EGCG, and proliferation and migration speed were measured. In 20 dogs, the autologous jugular veins were interposed into the carotid arteries. For the study group (n=10), the graft was stored for 30 minutes in EGCG solution and 300mM EGCG was applied to the perivascular space after grafting. After 6 weeks, the intimal and medial thickness was measured. Result: The proliferation of RASMC and HUVEC was suppressed with EGCG. The migration of RASMC was suppressed with EGCG, but that of HUVEC was not affected. In the in vivo study, the intimal thickness was thinner in EGCG group than in the control group (p<0.05), but the medial thickness did not show any difference. The intimal/medial thickness ratio was lower in the EGCG group (p<0.05). Conclusion: EGCG suppresses intimal hyperplasia after vascular grafting, and this may be mediated by prevention of migration and proliferation of vascular smooth muscle cells. The use of EGCG may offer new therapeutic modality to prevent intimal hyperplasia.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2001.11a
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• pp.71-73
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• 2001

In this study, we could improve transmission efficiency of germline chimeras by transfer of gonadal PGCs (gPGCs) cultured in vitro. Of hatched recipient chicks, 301 chickens (141 males and 160 females) were brought up to sexual maturity and these WLs (KOC) were mated with KOCs for testcross, resulting in 27 germline chimeras (15 males and 12 females) identified by black feather color of their progenies. The production efficiency of germline chimera production of experimental groups was observed (P=0.6831). The average transmission efficiency of proven germline chimeras was 0.6 ∼56.5% (15.0% on average). The transmission efficiency of experimental group which were transferred 10-days cultured gPGCs without Ficoll treatment was highest (49.7%) and that of experimental stock which transferred non-cultured gPGCs with Ficoll treatment was lowest (0.6%). The duration of in vitro culture before transferring was significantly important for the high efficiency of germline transmission. Transferring 10-days cultured gPGCs made the transmission efficiency higher rather than transferring non-cultured and 5-days cultured gPGCs, 50 times and 10 times, respectively (p<0.0001). However, Ficoll treatment for increasing the population ratio of gPGCs negatively affected the transmission efficiency and the effects of sexuality and the reciprocal interaction between treatments showed no significant differences. These findings demonstrated that the crucial factors for improving the germline transmission were the duration of in vitro culture prior to transfer. Thus, we developed the complete system for production of germline chimera using cultured gPGCs with highly improved efficiency and this system would be useful for genetic manipulation and obtaining the transgenic aves.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.97-104
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• 2005

This study was conducted to investigate the effects of various factors such as recipient parity, delivery season, offspring number, pregnancy period, delivery type, midwifery type and dystocia, on the viability of calves derived from embryos produced in vitro. There was no difference in the abnormality of calves among treatments ($0\~25\%$, respectively). The incidence of a disease was significantly higher in delivered by multiparous $(40\%)$ than nulliparous$(9.9\%)$, in eutocia than dystocia group, in delivered on spring $(20.4\%)$ and winter $(22.7\%)$ than summer$(4.3\%)$ and autumn $(0\%)$, in single offspring $(18.4\%)$ than twin offsprings $(6.7\%)$, and in eutocia group $(17\%)$ than dystocia $(2.7\%)$, respectively (p<0.05). The rate of mortality was significantly higher when transferred into mulliparous $(22.3\%)$ than multiparous$(0\%)$, when were delivered within 270 day $(53.3\%)$ than over 270 day $(14.3\~16.1\%)$, when were dystocia $(41.7\%)$ than eutocia$(14.1\%)$ group, when were induced delivery $(44.4\%)$ than self-delivery $(18.1\%)$, when were non-midwifery $(34\%)$ than midwifery$(10.8\%)$, and when delayed midwifery $(31.6\%)$ than earlier midwifery$(11.5\%)$, respectively (p<0.05). The present study suggested that the proper treatment of parturition may be increased the viability of calves derived from in vitro.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.181-187
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• 2006

In recent years the number of patients waiting for organ transplantation has greatly outpaced the supply of human organs available, which leads to a renewed interest in pig-to-human xenotransplantation as an alternative. However, one of the biggest barriers in the xenotransplantation is presence of porcine endogenous retroviruses(PERV) that can infect human cells. In this study, to present a possible solution for this problem we tried to inhibit expression of PERVs using shRNAs(short hairpin RNA) at the level of RNA synthesis and virus release. The shRNA targeting the sequence of PERV A, B type was cloned into pSIREN-RetroQ vector under the control of polymerase-III U6-RNA gene promoter. Quantitative real-time PCR was performed to detect my alterations in mRNA production of PERV A, B targeted by the shRNA in each done. Depending on the target sequence of the shRNA, the transcription of PERV was decreased to as much as 4% and the number of progeny viruses was reduced to less than 1/200,000. Transgenic pigs producing such shRNAs may result in a highly reduced PERV expression in cells and organs, which is a prerequisite for safe xenotransplantations.

• Korean Journal of Environmental Agriculture
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• v.15 no.2
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• pp.198-206
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• 1996

Leaching of $^{54}Mn$, $^{60}Co$, $^{85}Sr$ and $^{137}Cs$ in paddy soil was studied using lysimeter cultures in a greenhouse. The radionuclides were applied to the water surfaces shortly before transplanting and five different times between transplanting and harvest. Fertilizer KCl and slaked lime were added simultaneously in the rate of 83kg and 200kg, respectively, per l0a following the first application after transplanting. Percolating water was sampled until two days before harvest. Concentrations of the radionuclides in percolating water decreased in the order of $^{85}Sr$ > $^{54}Mn$ > $^{60}Co$ > $^{137}Cs$ on the whole. Time taken to reach the maximum was the shortest for $^{137}Cs$(< one week) and the longest for $^{54}Mn$ and $^{85}Sr$. Six days' water dropping started 47 days after transplanting reduced the concentrations of $^{54}Mn$, $^{60}Co$, $^{85}Sr$ and $^{137}Cs$ by factors of 30-180, 3-75, 2-4 and 3-6, respectively, depending on the application time. After the significant decrease, $^{54}Mn$ concentration tended to gradually increase but $^{137}Cs$ did to the contrary Percent leaching varied 0.09-6.2% for $^{54}Mn$, 0.009-0.9% for $^{60}Co$, 1.4-14.4% for $^{85}Sr$ and 0.002-0.06% for $^{137}Cs$, with the application time. The highest leaching came from the application at 40 days after transplanting for all the radionuclides. The addition of KCl and lime increased percent leaching of the radionuclides by factors of 9, 85, 4 and 9, respectively.

• Pediatric Infection and Vaccine
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• v.21 no.2
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• pp.81-95
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• 2014

Purpose: Hematopoietic cell transplantation (HCT) recipients are vulnerable to invasive infection by Haemophilus influenzae type b (Hib) and Streptococcus pneumoniae (Sp). This study was performed to evaluate immune responses after Hib and Sp vaccination in Korean pediatric HCT recipients. Methods: Patients were prospectively enrolled at Samsung Medical Center during 2009-2011. ELISA tests to detect anti-PRP IgG antibody and antibodies to Sp serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F were performed at the Center for Vaccine Evaluation and Study, Ewha Medical Research Institute. Results: Ten patients (two allogeneic, eight autologous recipients) with median age 5.4 years (range 2.7-12.2 years) were enrolled. Before Hib vaccination, 60% of patients' anti-PRP IgG titers were below $0.15{\mu}g/mL$. After vaccination, 100% of patients' anti-PRP IgG titers increased above $0.15{\mu}g/mL$ (cut-off value for detection) and $1.0{\mu}g/mL$ (cut-off value for seroprotection). For pneumococcus, in 2-5 year-old patients, pre-vaccination geometric mean concentrations (GMCs) of IgG for six serotypes (4, 6B, 9V, 14, 18C, and 23F) were below $0.35{\mu}g/mL$ and at 5 months post-vaccination GMCs of IgG for all seven serotypes increased to above $0.35{\mu}g/mL$. In patients older than 5 years, pre-vaccination GMCs of IgG for four serotypes (4, 9V, 14, and 23F) were below $0.35{\mu}g/mL$ and at 3 months post-vaccination GMCs of IgG for all seven serotypes increased to above $0.35{\mu}g/mL$. Conclusion: Most HCT recipients had low or no protective antibodies to Hib and Sp before vaccination, but showed good immune responses to protective levels after vaccination.

• Journal of Korean Orthopaedic Sports Medicine
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• v.6 no.1
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• pp.38-44
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• 2007

Purpose: To evaluate status of reconstructed ACL and changes around graft through second-look arthroscopy after arthroscopic reconstruction of the ACL with autograft tendons. Material and Methods: Between Jun. 2003 and Feb. 2007, the second look arthroscopy was performed on 22 cases. Second-look arthroscopy was conducted on average 15.1 $(7\sim31)$ months after reconstruction. 15 cases received hamstring tendon autograft, 7 cases received bone-patellar tendonbone autograft. We measured graft tension using displacement by probing, synovial coverage by visual analysis at second-look arthroscopy The assay in Lysholm score, Lachmann test and KT-2000 arthrometer were evaluated status of reconstructed ACL. Results: The hamstring tendon group showed normal tension in 11 cases and lax tension in 4 cases. The patellar tendon group showed normal tension in 3 cases, lax tension in 2 cases and partial tear in 2 cases. In the hamstring tendon group, synovial coverage was good in 11 cases, half in 3 cases and pale in 1 case, and the bone-patellar tendon-bone group was good in 4 cases and half in 3 cases. The patellar tendon group was superior to the hamstring tendon group in Lysholm score (p<0.05), but insignificance difference in KT-2000 arthrometer results statistically. Conclusion: The hamstring tendon group was superior to the bone-patellar tendon-bone group in second-look arthroscopy after ACL reconstruction with autograft tendons results, but long term follow up will be necessary to further evaluated.