• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.31 no.1
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• pp.22-31
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• 2018

Objectives : In this study, we investigated whether Astragali Radix Pharmacocupuncture (ARP) has an effective on the full thickness defect wound healing process of mouse. Methods : A total of 50 mice (ICR mouse, 7 week-old male) were divided into control group and ARP group. A single full thickness skin defect was made on the dorsal side of the each mouse using an 8mm diameter biopsy punch. Control group were treated with 0.2㎖ saline and ARP group were treat with 0.2ml ARP at 8 points around the wound every three days total 4 times during the experimental period. The change in wound size, contraction rate, healing rate, and epithelization rate was measured by digital images taken on days 3, 6, 9, and 13, and evaluated using a digital image analysis program. Tissues were collected for histological analysis, RT-PCR, and Western blot on days 3, 6, 9, and 15. Results : The results are as follows. ARP group accelerated the rate of wound contraction, wound healing and epithelization compared to the control group. ARP group showed the decrease of inflammatory cells in early inflammatory phase compared to the control group. ARP group upregulated PECAM-1 mRNA and protein expression compared to the control group. ARP group inhibited the scar width and area compared to the control group. Conclusions: ARP showed positive effects on wound healing through the inhibition of inflammatory reaction and increase of PECAM-1 expression related to the wound healing process.

• Archives of Plastic Surgery
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• v.40 no.3
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• pp.181-186
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• 2013

Background Partial or complete necrosis of a skin flap is a common problem. Polydeoxyribonucleotide (PDRN) can be extracted from trout sperm and used as a tissue repair agent. The aim of this study was to investigate whether PDRN could improve the survival of random pattern skin flaps in rats. Methods Twenty-two male Sprague-Dawley rats were randomly divided into two groups: the PDRN treatment group (n=11) and the control group (n=11). Caudally pedicled random pattern skin flaps were elevated on their dorsal skin and resutured. The treatment group received daily intraperitoneal administration of PDRN (8 mg/kg/day), and the control group received fluid vehicle (NaCl 0.9%, 8 mg/kg/day) from day 0 to day 6. On day 7, the flap survival was evaluated and the harvested tissue surrounding the demarcation line of the necrotic area was stained with H&E, anti-rat vascular endothelial cell growth factor (VEGF) antibody, and PECAM-1/CD31 antibody. Results The average necrotic area of the flap in the PDRN group was significantly smaller when compared with that of the control group. Histologic and immunohistochemical evaluation showed that granulation thickness score and VEGF-positive staining cells were marked higher in the PDRN group than in the control group. PECAM-1/CD31-positive microvascular densities were significantly higher in the PDRN group when compared with the control group. Conclusions This study confirms that PDRN improves the survival of random pattern skin flaps in rats. These results may represent a new therapeutic approach to enhancing flap viability and achieving faster wound repair.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.221.2-222
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• 2003

Expressions of cellular adhesion molecules (CAMs) are closely related to the formation of early atherosclerosis, an age-dependent vascular disorder. However. previous research provided only limited and conflicted reports on age-related alterations of CAMs' expressions and even much less is known the modulation of CAMs by calorie restriction (CR), In this study, expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), E-selectin, P-selectin and platelet/endothelial cell adhesion molecule-1 (PECAM-1) in aorta and kidney were investigated by western blot and immuno-histochemical stain utilizing ad libitum (AL) and CR rat. (omitted)

• Biomedical Science Letters
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• v.12 no.2
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• pp.127-130
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• 2006

It is important to coordinated interaction among neurons, astrocytes and endothelial cells to maintain the function of brain. To study their regulatory mechanisms in vitro system, the co-culture system among the isolated cells from brain may be needed. However, the method for purifying brain microvascular endothelial cells (BMEC) far culture have not established yet. In this study, the proper culture methods of mice cells using two different strains, CD1 and C57BL6, to obtain the pure and plentiful endothelial cells were described. The flatted-round forms of CD1 endothelial cells grew on the collagen-IV coating plates, while the purified cells from C57 mice preferred type collagen-I dishes for their growth. Both cells displayed anti-PECAM-1 (CD31) and von Willebrand Factor immune-reactivity. These results indicated that different coating materials not only improve attachment of isolated cells but also promoting growth of cells, suggesting that this method of purifying murine Brain microvascular endothelial cells (BMEC) provides a suitable model to investigate blood-brain-barrier (BBB) properties within neurovascular unit in vitro.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.87-94
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• 2017

Tissue engineering (TE) has been developed to create functional organs and tissue by combining 3D matrix and cells in vitro. Vascularization and angiogenesis are utmost important for supply of nutrients and oxygen in tissue engineered organs. The present study was performed to isolate and characterize primary endothelial cells (EC) from aorta of alpha 1, 3-enzyme galactosyltransferase knock out (GalT KO) pig, to minimize immune rejection and analyze body immune system for future xenotransplantation studies. Isolation of primary EC from aorta were performed by incubation with dispase for 8-10 min at $37^{\circ}C$. Primary EC were cultured in EC growth medium on different extra cellular matrix (ECM), either collagen or gelation. Primary EC exhibits morphological characteristics and showed positive expressions of EC specific marker proteins i.e. PECAM1, KDR and VWF despite of their ECM surface; however, on collagen based surface they showed increase in mRNA level analyzed by qPCR. Primary EC cultured on collagen were sorted by flow cytometer using KDR marker and cultured as KDR positive cells and KDR negative cells, respectively. KDR positive cells showed dramatically increased in PECAM1 and VWF level as compared to KDR negative cells. Based on the above results, primary EC derived from GalT KO are successfully isolated and survived continuously in culture without becoming overgrown by fibroblast. Therefore, they can be utilize for xeno organ transfer, tissue engineering, and immune rejection study in future.

• Journal of Life Science
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• v.23 no.12
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• pp.1532-1540
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• 2013

T-box transcription factor 2 (Tbr2) is a member of the T-box family of transcription factors and it plays an important role in brain development, progenitor cell proliferation, and the modulation of differentiation and function in immune cells, such as CD8+ T cells and natural killer cells. This study aims to elucidate the involvement of Tbr2 in the pathophysiological events following pilocarpine-induced status epilepticus in mice. Status epilepticus resulted in prominent neuronal cell death in discrete brain regions, such as CA3, the hilus, and the piriform cortex. Interestingly, when the immunoreactivity of Tbr2 was examined two days after status epilepticus, it was transiently increased in CA3 and in the piriform cortex. Tbr2-positive cells in CA3 and the piriform cortex were double-labeled with CD11b, a marker of microglia and a subset of white blood cells, such as monocytes, CD8+ T cells, and natural killer cells. Moreover, the double-labeled cells with Tbr2 and CD11b showed amoeboid morphology, and this data indicates that Tbr2-expressing cells may be reactive microglia or infiltrating white blood cells. Furthermore, clustered Tbr2-positive cells were observed in the platelet endothelial cell adhesion molecule-1 (PECAM-1)-positive blood vessels near the CA3 area, which suggests that Tbr2-positive cells may be infiltrating the white blood cells. Based on this data, this study is the first to indicate the involvement of Tbr2 in neuropathophysiology in status epilepticus.

• Archives of Plastic Surgery
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• v.35 no.2
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• pp.121-126
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• 2008

Purpose: Poly-N-acetyl glucosamine(PGlcNAc) nanofiber-based materials, produced by a marine microalga, have been characterized as effective hemostatic and angiogenic agents. The similarity between PGlcNAc patch and the natural extracellular matrix allows it to support new healthy tissue growth in an injured area and to encourage fluid absorption. In this study, we hypothesized that a poly-N-acetyl glucosamine fiber patch(PGlcNAc patch) may enhance wound healing in the db/db mouse. Methods: PGlcNAc patches were applied on one square centimeter, full-thickness, skin wounds in the db/db mouse model. Wounds(n=15 per group) were dressed with a PGlcNAc nanofiber patch for 1 hour(1 h), 24 hours(24 h) or left untreated(NT). After the application time, patches were removed and wounds were allowed to heal spontaneously. The rate of wound closure was evaluated by digital analysis of unclosed wound area in course of time. At day 10, wounds(n=7 per group) were harvested and quantified with immunohistochemical markers of proliferation(Ki-67) and vascularization (platelet endothelial cell adhesion molecule, PECAM-1). Results: Wounds dressed with PGlcNAc patches for 1 hour closed faster than control wounds, reaching 90% closure in 16.6 days, nine days faster than untreated wounds. Granulation tissue showed higher levels of proliferation and vascularization following 1 h treatment than the 24 h and NT groups. In addition to its hemostatic properties, the PGlcNAc material also appears to accelerate wound closure in healing-impaired genetically diabetic mice. Conclusion: This material, with its combination of hemostatic and wound healing properties, has the potential to be effective agent for the treatment of complicated wounds.

• Molecules and Cells
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• v.45 no.12
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• pp.950-962
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• 2022

Aging is a major risk factor for common neurodegenerative diseases. Although multiple molecular, cellular, structural, and functional changes occur in the brain during aging, the involvement of caveolin-2 (Cav-2) in brain ageing remains unknown. We investigated Cav-2 expression in brains of aged mice and its effects on endothelial cells. The human umbilical vein endothelial cells (HUVECs) showed decreased THP-1 adhesion and infiltration when treated with Cav-2 siRNA compared to control siRNA. In contrast, Cav-2 overexpression increased THP-1 adhesion and infiltration in HUVECs. Increased expression of Cav-2 and iba-1 was observed in brains of old mice. Moreover, there were fewer iba-1-positive cells in the brains of aged Cav-2 knockout (KO) mice than of wild-type aged mice. The levels of several chemokines were higher in brains of aged wild-type mice than in young wild-type mice; moreover, chemokine levels were significantly lower in brains of young mice as well as aged Cav-2 KO mice than in their wild-type counterparts. Expression of PECAM1 and VE-cadherin proteins increased in brains of old wild-type mice but was barely detected in brains of young wild-type and Cav-2 KO mice. Collectively, our results suggest that Cav-2 expression increases in the endothelial cells of aged brain, and promotes leukocyte infiltration and age-associated neuroinflammation.

• Molecular & Cellular Toxicology
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• v.2 no.4
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• pp.266-272
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• 2006

Carpal tunnel syndrome (CTS) is one of the most common disorders by under pressure of the median nerve at the wrist in these days. However, pathological mechanism of CTS is unknown. We carried out this study to identify the changes of gene expression and to evaluate possible mechanism in CTS. 120 CTS patients and 30 control patients were included in this study. Patients with a history of diabetes, hypertension, thyroid diseases, and arthritis were excluded. CTS patients were divided to three experimental groups-Mild, Moderate, and Severe group-according to elecrodiagnosis. Radioactive cDNA microarrays (Nylon membrane including 1,152 genes) were used to examine the difference of gene expression profile in CTS. We identified up-regulated genes by more than 2.0 value of z-ratio, and down-regulated genes by less than-2.0 value of z-ratio. 20 genes such as the ITGAL, ITGAM, PECAM1, VIL2, TGFBR2, RAB7, RNF5 and NFKB1 were up-regulated, and 28 genes such as PRG5, CASP8, CDH1, IGFBP5, CBX3, HREV107, PIN, and WINT2 were down-regulated. These genes were related with TGF beta signaling pathway, NF-Kb signaling pathway, antiapoptotic pathway and T cell receptor signaling pathway. However, there were no differences in gene expression profiles according to severities of symptoms. We suggest that CTS could be related with proinflammatory mechanism and antiapoptotic mechanism.

• Tuberculosis and Respiratory Diseases
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• v.48 no.5
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• pp.757-765
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• 2000

Background : Angiogenesis plays a critical role in human tumor growth and metastasis. Microvessel count as a measure of tumor angiogenesis, has been significantly correlated with invasive and metastatic patterns in breast. prostate and cutaneous carcinomas. Materials and Methods : Fifty patients with curatively resected non-small cell lung cancer were evaluated. Tumor tissues embedded in paraffin block were stained by anti CD 31 (PECAM, platelet endothelial cellular adhesion molecule) using immunohistochemical method to assess microvessel count. Microvessels were counted in the most active areas of neovascularization(microscopy, 200$\times$). Results: 1) Mean microvessel count was 47.1$\pm$17.7(per 200$\times$field) in total 50 cases. 2) Mean microvessel count of adenocarcinoma (54.4$\pm$19.9) was significantly higher than that of squamous cancer (43.9$\pm$16.2) (p<0.05), but there were no relationship between microvessel count and TNM stages. 3) Median survival time, 2-year and 5-year survival rates of the low microvascular group (microvessel count<45, 22 cases) were 61 months, 80% and 40%, respectively, and those of the high microvascular group(microvessel count$\geq$45, 28 cases) were 46 months, 75% and 12%, respectively. As results, prognosis of low microvascular group is statistically significantly superior to that of the high microvascular group (p=0.0162, Kaplan-Meier, log-rank). Conclusion : Angiogenesis assessed by microvessel count can be used as one of the significant prognostic factors in non-small cell lung cancer.