• Journal of Chest Surgery
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• v.35 no.6
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• pp.407-419
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• 2002

Atherosclerosis is a chronic inflammatory disease of the arterial wall characterized by progressive accumulation of lipids, cells, and extracellular matrix. Matrix metalloproteinases(MMPs) and tissue inhibitor of metalloproteinases(TIMPS) contribute to vascular matrix remodeling in atherosclerosis, and some cytokines may play role in the synthesis or activation of MMPs or TIMPs. Material and Method： We produced experimental atherosclerotic plaques in 9 rabbits by atherogenic hypercholesterol diet for 12 weeks, and 10 other rabbits were used as control group with standard laboratory chow, At that time, 19 rabbits were sacrificed and aorta, coronary arteries and blood specimens were prepared. The expressions of MMP-9, TIMP-2 and interleukin(IL)-18, and the bioactivity of IL-6 were investigated with H&E stain, immunohistochemical stain, immunoblotting(Western blot analysis), and bioassay. Result： Serum cholesterol in the experimental group increased up to 1258$\pm$262 mg/dL(control group： 41$\pm$7 mg/dL). All experimental group showed well-developed atherosclerotic plaques in aorta and coronary artery. The expression of MMP-9 in aorta and coronary artery of the experimental group showed significant increase than that of the control group by immunohistochemistry. Among the experimental group, complicated lesions with intimal rupture or complete luminal occlusion, demonstrated stronger expression of MMP-9. Interestingly, there was no difference in expression of TIMP-2 between the experimental and the control group. These findings were confirmed by Western blot analysis. The bioassay revealed significant up-regulation of serum bioactivity of IL-6 in the experimental group(4819.60$\pm$2021.25 IU/$m\ell$) compared to that of IL-6 in the control group(27.20 $\pm$ 12.19 IU/$m\ell$). IL-18 was expressed in all atherosclerotic plaques, whereas little or no expression was detected in the control group. Conclusion： The increased MMP-9 expression along with the unchanged TIMP-2 expression seem to be contributory factors in extracellular matrix degradation in atherosclerosis. Focal overexpression of MMP-9 may promote plaque destabilization and cause complications of atherosclerotic plaques such as thrombosis with/without acute coronary syndrome. Elevation of IL-6 and IL-18 may be more than just markers of atherosclerosis but actual participants in lesion development. Identification of critical regulatory pathway is important to improve the understanding of the cellular and molecular basis of atherosclerosis and may open the way for novel therapeutic strategies.

• Journal of Chest Surgery
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• v.37 no.6
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• pp.482-491
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• 2004

Background: The xenogenic or allogenic valves after in Vitro repopulation with autologous cells or in vivo repo-pulation after acellularization treatment to remove the antigenicity could used as an alternative to synthetic polymer scaffold. In the present study, we evaluated the process of repopulation by recipient cell to the acellu-larized xenograft treated with NaCl-SDS solution and grafted in the right ventricular outflow tract. Material and Method: Porcine pulmonary valved conduit were treated with. NaCl-SDS solution to make the grafts acellularized and implanted in the right ventricular outflow tract of the goats under cardiopulmonary bypass. After evaluating the functions of pulmonary valves by echocardiography, goats were sacrificed at 1 week, 1 month, 3 months, 6 months, and 12 months after implantation, respectively. After retrieving the implanted valved conduits, histopathologic examination with Hematoxylin-Eosin, Masson' trichrome staining and immunohistochemical staining was performed. Result: Among the six goats, which had been implanted with acellularized pulmonary valved conduits, five survived the expected time period. Echocardiographic examinations for pulmonary valves revealed good function except mild regurgitation and stenosis. Microscopic analysis of the leaflets showed progressive cellular in-growth, composed of fibroblasts, myofibroblasts, and endothelial cells, into the acellularized leaflets over time. Severe inflammatory respon-se was detected in early phase, though it gradually decreased afterwards. The extracellular matrices were regenerated by repopulated cells on the recellularized portion of the acellularized leaflet. Conclusion: The acellularized xenogenic pulmonary valved conuits were repopulated with fibroblasts, myofibroblasts, and endothelial cells of the recipient and extracellullar matrices were regenerated by repopulted cells 12 months after the implantation. The functional integrity of pulmonary valves was well preserved. This study showed that the acellularized porcine xenogenic valved conduits could be used as an ideal valve prosthesis with long term durability.

• Applied Microscopy
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• v.30 no.2
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• pp.193-204
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• 2000

Nowadays, mongolian gerbil is widely utilized in the research of brain and water deprivation because of congenitally incomplete Willis' circle, audiogenic seizure in low noise, and special cholesterol metabolism without water absorption for a long time. In this study, we intended to identify the time lapse changes in the general morphoogy and ultrastructure of the catecholaminergic neurons of mongolian gerbil brain in after long-term water deprivation. Fifteen mongolian gerbils were divided into 3 groups (5, 10, and 20-day water deprivation groups), each with 5 mongolian gerbils. Additional 5 mongolian gerbils which received water without limitation were used as a control. The brain sections were immunostained using tyrosine hysroxylase (TH), $ dopamine-\beta-hydroxylase$ (DBH), and phenylethanolamine-N-methyltrasferase (PMNT) antibodies. And immunoreactive cells were observed by electromicroscopy for the ultrastructural changes . The TH-immunoreactive (TH-IR) nerve cells were observed in the para- and peri-ventricular nucleus of the 3 rd ventricle in the hypothalamus and the substantia nigra. The number of TH-IR neurons in these areas was decreased from the 5th day of the water deprivation to the 10 th day and reincreased until 20 th day water deprivation. The shape and density of the dopamine-secreting cells identified by immunohistochemistry showed changes in the continuous water deprivation. Electron microscopy revealed a round nucleus in the neurons of control group but 5-day water deprivation group showed a dense and irregularly shaped nucleus. Also in the 5-day water-deprived group, mitochondria was decreased in number and junctins were disappered. Endoplasmic reticulum, Golgi complex did not show changes after water-deprivation. In this results, we can conclude that dopamine are involved in the water metabolism in mongolian gerbil, and mongolian gerbil could be used as an animal model for the researches of water deprivation.

• Radiation Oncology Journal
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• v.25 no.4
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• pp.193-200
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• 2007

Purpose: This study evaluated the pretreatment expression patterns of MDM2, p53, and pRb proteins to determine if the expression patterns could predict the outcome of concurrent chemoradiotherapy (CCRT) for esophageal squamous cell carcinoma and aid in the decisions for the selection of treatment modalities. Materials and Methods: Fifty-one patients that were treated with definitive chemoradiotherapy for stage $I{\sim}IVa$ esophageal squamous cell carcinoma were selected for this study. Radiotherapy was administered with daily $1.8{\sim}2\;Gy$ fractions up to a median dose of 54 Gy for primary tumors, and with four cycles of cisplatin/5-fluorouracil chemotherapy that was administered every 4 weeks, the first two cycles of which were administered concurrently with radiotherapy. Expression of MDM2, p53, and pRb was investigated by immunohistochemical analysis using pretreatment biopsy specimens. Results: MDM2, p53, and pRb were detected with high immunoreactivity in 19.6%, 27.5%, and 66.7% of the patients, respectively. However, there was no significant correlation between expression of these factors and clinical outcome. By the use of multivariate analysis with nine covariates-age, tumor location, tumor length, stage, pathological response, clinical response, MDM2 expression, p53 expression, and pRb expression, only pathological response and stage were significant factors for cause-specific survival. Conclusion: Expression of MDM2, p53, and pRb was not found to be clinically significant for predicting outcomes after CCRT in this study. Further studies with a larger patient population and longer follow-up periods are needed to re-evaluate the expression pattern and to identify new predictors for CCRT response.

• Journal of Chest Surgery
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• v.41 no.2
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• pp.149-159
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• 2008

Background: With increasing coronary bypass and peripheral vascular surgeries, the demand for homologous vascular or synthetic conduits has continued to grow, but wide-spread application has been limited by dismal patency rates. Although cryopreserved allograft valves may provide a suitable alternative, current viability or patency of implanted allograft vascular conduits has been unsatisfactory. Material and Method: We serially analyzed the outcomes of canine femoral artery and saphenous vein allograft implants after storage in either $4^{\circ}C\;or\;-170^{\circ}C$. Result: There were no differences in graft flow rate (patency) (p=0.264), rate of thrombosis (p=0.264), presence of endothelium (p=0.587), or immunohistochemical staining for thrombomodulin (p=0.657) were detected between grafts stored in $4^{\circ}C\;and\;-170^{\circ}C$. Greater flow occurred in the arterial grafts versus the venous grafts (p=0.030), irrespective of the preservation method, with a significantly lower incidence of thrombosis (p=0.030) in arterial allografts. There was a correlation coefficient of -0.654 between thrombosis and positive immunohistochemical staining for thrombomodulin (p=0.006) and a correlation coefficient of 0.520 (p=0.0049) between the endothelial presence and positive immunohistochemical staining for thrombomodulin. The relationship between the presence of endothelium and thrombomodulin expression failed to show any correlation within the first 2 weeks (p=0.306). However, a strong correlation was seen after 1 month (p=0.0008). Conclusion: Tissue storage in either $4^{\circ}C\;or\;-170^{\circ}C$ in 10% DMSO/RPMI-1640 preservation solution preserved grafts equally well. In terms of thrombosis and graft patency, arterial grafts were superior to venous grafts. Considering the poor correlation between thrombomodulin expression and the presence of an endothelium in the implanted graft within the first two weeks, grafts in this period would not be thromboresistant.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.62-62
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• 2003

Aquaporin (AQP) family protein은 일종의 수분 전달 통로 역할을 하는 단백질로 AQP를 통한 수분의 조절은 삼투압을 통한 물의 이동과 함께 조직내 정상적인 수분의 상성 유지에 필수적이다. 현재까지 11종의 AQP이 신장·뇌·정소·안구 등에서 발현이 확인되었다. AQP9은 물 뿐 아니라 carbamide, polyol, purine, pyrimidine, urea, glycerol 등의 이동에 관여한다. 본 연구에서는 생쥐에서 출생 후 성체에 이르는 동안 정소 내 AQP9의 발현, Leydig cell의 분화에 따른 AQP9의 발현을 조사하였다. 1, 2, 4, 8주령의 정소로부터 semiquantitative RT-PCR 및 real time PCR 법으로 AQP9의 발현을 분석한 결과 1주령에서는 발현되지 않았고 2주령에서는 미량이 발현되기 시작하였고, 4주령에서는 성체의 1/2수준으로 발현량이 급격히 증가하였고 성체에서는 다량으로 발현됨이 확인되었다. Semiquantitative RT-PCR 법과 real time PCR법을 비교할 때 주령별 발현 양상은 유사하였으나 4주령과 성체에서는 두 시험법 사이에 양적인 차이가 있었다. 면역조직화학염색 결과 주로 Leydig cell에서 AQP9의 발현이 확인되었다. 성체의 정소 균질액의 Western blot 상에서 분자량 80, 55, 35 및 23 kDa의 항원이 검출되어 dimer, trimer 형태로 존재할 가능성과 당쇄 결합에 의한 단백질의 변형이 있는 것으로 추정된다. 미성숙 개체의 정소에서는 23 form이 확인되는 반면 성체에서는 35 kDa form이 주로 발현되므로 정소에서 발현되는 AQP9의 경우 Post-translation 수준에서 AQP9의 변형이 수반되는 것으로 사료되며 AQP9의 기능과의 연관성은 추후 연구되어야 할 것이다. Leydig cell은 fetal 및 adult type 2종의 세포가 정소발달 과정에 출현, 사멸, 분화하며 이들은 각기 정소발달, 성숙과 정자형성에 필요한 steroidogenesis에 관여한다. 정소 내 AQP9의 발현은 17beta HSD의 발현 양상과 같게 나타나므로 성적 성숙에 따른 정소 내 AQP9의 발현의 증가는 adult type Leydig cell의 분화와 관련된 것으로 추측된다. 성체의 정소로부터 분리한 Leydig cell-enriched culture에 hCG를 처리한 결과 배양체의 AQP9의 발현이 증가하므로 AQP9은 LH 수용체 하위 신호전달과정을 통해 Leydig cell의 steroidogenesis 또는 생성된 steroids의 분비에 요구되는 수분 및 중성용질의 이동에 관여하는 것으로 사료된다.

• Journal of Life Science
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• v.28 no.10
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• pp.1220-1232
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• 2018

The purpose of this animal study was to evaluate, by histological analysis, bone regeneration in rabbit maxillary sinuses with an anorganic bovine graft (Bio-Oss) and a ${\beta}-tricalcium$ phosphate (${\beta}-TCP$) grafting. Bilateral sinus augmentation procedures were performed in 12 adult male rabbits. Rectangular replaceable bony windows were made with a piezoelectric thin saw insert. In the Bio-Oss group, Bio-Oss was grafted and in the ${\beta}-TCP$ group, ${\beta}-TCP$ was grafted and covered by replaceable bony windows. The animals were sacrificed at 2, 4, and 8 weeks after the surgical procedure. The augmented sinuses were evaluated by histomorphometric analysis using hematoxylin-eosin, Masson trichrome, and tartrate-resistant acid phosphatase stains and also by immunohistochemical analysis of proliferating cell nuclear antigen (PCNA), type I collagen, and osteocalcin content. Histologically, new bone formation was found on the surface of Bio-Oss and ${\beta}-TCP$ particles from 2 weeks and continued to 8 weeks. Significant higher new bone formation was revealed in the ${\beta}-TCP$ group than in the Bio-Oss group at 8 weeks. The amount of graft materials was significantly decreased in the ${\beta}-TCP$ group and the number of osteoclasts was significantly increased in the ${\beta}-TCP$ group from 4 to 8 weeks. Immunoreactivity to PCNA was reduced at 8 weeks. The expression of type I collagen was significantly increased in the ${\beta}-TCP$ group at 2 weeks, but was significantly increased in the Bio-Oss group at 8 weeks. Immunoreactivity to osteocalcin was increased from 2 to 8 weeks. These histological results can help in the selection of graft materials for implants. Both Bio-Oss and ${\beta}-TCP$ are proven graft materials, however, these results indicate that ${\beta}-TCP$ showed better bone regeneration results in rabbit maxillary sinus augmentation.

• Journal of Radiation Protection and Research
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• v.24 no.1
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• pp.17-22
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• 1999

This study was carried out to evaluate the biochemical and morphological effects of ionizing radiation on mouse ovarian follicles. Immature mice (ICR, 3 week-old) were irradiated with a dose of $LD_{80(30)}$ at KAERI. The ovaries were collected after 6 hours, 12 hours, 1 day, and 2 days post irradiation. With the morphological basis of the histological staining with hematoxylin-eosin, immunohistochemical preparation using in situ 3'-end labeling was evaluated. Flowcytometric evaluation of DNA extracted from the whole ovary was performed. The percentage of $A_0$ (subpopulation of cells with degraded DNA and with lower DNA fluorescence than $G_0/G_1$ cells), apoptotic, cells in the cell cycle was significantly higher in the irradiated group than in the control group. The number of in situ 3'-end labeled follicles increased at 6 hours post irradiation. All the analyses represented that the ionizing radiation-induced follicular atresia was taken place via an apoptotic degeneration. Such a degeneration underwent very fast and acutely. Therefore, it is concluded that the radiation-induced follicular degeneration is, like the spontaneous atresia, mediated by an acute apoptosis of follicular granulosa cells. Flowcytometric evaluation of cell cycles can make the role for quantifying the atretic follicles and understanding the mechanism of the radiation-induced cell death.

• Journal of Chest Surgery
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• v.37 no.3
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• pp.261-266
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• 2004

Although significant progress has been made in the surgical treatment of esophageal carcinoma as well as in the detection of early stage esophageal carcinoma by diagnostic techniques, the prognosis of the esophageal carcinoma patients remain poor. The p53 gene product is known to regulate cell growth and proliferation. And the nm23 gene was identified originally as an anti-metastatic influence whose expression was correlated inversely with tumor metastatic potential in murine melanoma cell lines. This experiment was intended to know the relationship among the p53 and nm23 gene expression versus clinicopahologic characteristics of the esophageal cancer. Total 40 cases were collected from patients who had undergone esophagectomy at St. Mary's Hospital, Catholic university of Korea. Immunohistochemical stain for p53 mutant-type protein and nm23 protein was graded as <10% positive tumor cells: negative; 10∼30% positive tumor cells: ＋ ; 30∼50% positive tumor cells: ＋＋, and >50% positive tumor cells: ＋＋＋. The tumor invasion was grades as none:－ ; mild:＋ ; moderate:＋＋ ; severe: ＋＋＋. Overexpression of p53 protein and nm23 was not associated with the survival and cliniocopathologic characteristics of the esophageal cancer. Moreover, the combination analysis of p53 and nm23 revealed that there was no relationship between the gene expression and the clinicopatholic characteristics of the esophageal cancer.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.2
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• pp.150-160
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• 2017

Atopic dermatitis (AD) is a chronic inflammatory skin disease. It is characterized by eczematous lesions, skin dryness, and pruritus. The existing treatment drugs for AD have side effects, especially if the drugs are taken for extended periods. Therefore, new alternative therapies are necessary. The aim of this study was to investigate the combined effects of curcumin administration and LED irradiation on AD. AD-like lesions were induced in BALB/c mice by repeated application of 1-chloro-2,4-dinitrobenzene (DNCB) to the shaved skin of the ear and neck. Thirty male BALB/c mice were divided into five groups: vehicle, DNCB, curcumin, LED, and curcumin+LED groups. Curcumin (0.1 g/kg/day) was administrated repeatedly during a period of 14 days (experimental period) and 630 nm LED irradiation ($5J/cm^2/day$) was performed in the acryl box once a day for 10 days, after inducing AD-like lesions via DNCB application. The severity of AD-like lesions was evaluated during the experimental period, using a modified SCORAD index. Both ear and neck skin tissues were examined histologically for epidermal thickness, mast cell, eosinophil counting, and dermal collagen density. Epidermal cell proliferation and apoptosis were detected using immunohistochemistry and TUNEL, respectively. These were all reduced in SCORAD index, epidermal thickness, collagen density, number of mast cell and eosinophil in dermis, and number of proliferating cell and apoptotic cell in epidermis by curcumin administration and 630 nm LED irradiation. Moreover, all parameters were significantly lower in the curcumin+LED group compared with the curcumin group and LED group. These results suggest that the combined therapy of curcumin and LED is more effective than a single treatment. We recommend that this can be a feasible alternative therapy to manage AD.