• Maxillofacial Plastic and Reconstructive Surgery
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• 제33권3호
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• pp.199-209
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• 2011

Purpose: Osteoporosis, is a major health problem for the elderly and post-menopausal women and shown to alter the properties of bone as well as impair bone healing around titanium implants in both human and animals. The objective of this study was to examine the effect of LIPUS with adipose-derived stem cells on the healing process around a titanium implant in rats with osteoporosis. Methods: Sixteen osteoporosis-induced rats were divided into two groups: an adipose-derived stem cell injected with Low Intensity Pulsed Ultrasound (LIPUS) application group and a control group. Titanium screw implants (diameter, 2.0 mm: length, 3.5 mm, Cowell Medi, Korea) were placed into both tibia of 16 rats, on 8 rats as the control group and the other 8 rats as the experimental group. Rats were sacrificed at different intervals from 1, 2, 4 and 8 weeks after implantation for histopathologic and immunohistochemical analyses. Results: Histopathological analysis revealed newly formed bone in experimental group earlier than that in control group. Especially at 1 week after implantation, more amounts of new bone matrix and collagen around the implant in the experimental group were seen compared with the control group. Immunohistochemical analysis showed that the levels of osteoprotegerin (OPG) expression in the experimental group were increased at early stages compared with that of control group until 2 weeks after implantation. But after 2 weeks, the expression level of OPG similar in both groups. The expression levels of receptor activator of nuclear factor kB ligand (RANKL) were stronger in the experimental group than the control group until 2 weeks after implantation. After 4 weeks, expression of RANKL in experimental group was similar to the control group. Conclusion: The results of this study suggest that LIPUS with Adipose-Derived Stem Cells in implantation could promote bone healing around titanium implants in rats with osteoporosis.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제31권2호
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• pp.116-129
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• 2005

Distraction osteogenesis is a new bone formation technique. There is a advantage of the environmental adaptation when distraction force is applied to the gap between osteotomy lines. But it has a disadvantage of long-term wearing of the appliance and long consolidation period. Therefore we make an effort to reduce it and repair normal function. Extracellular matrix proteins have a function to control the cellular growth, migration, shape and metabolism. In these, hyaluronic acid is a member of polysaccharide glycosaminoglycans (GAGs) and has a important function as bone formation and osteoinduction property. Purpose : In this experimental study in rabbit mandibular distraction osteogenesis, we investigated the bone enhancing property of hyaluronic acid and the expression of extracellular proteins such as osteocalcin and osteonectin. Materials and Methods : The experimental study was carried out on 24 Korean male white rabbits (both mandibular body, n=48). Distraction group was divided to distraction experimental (A, n=16) and distraction control (B, n=16) by the application of hyaluronic acid (Hyruan, LGCI, Seoul, Korea). Normal control group (C, n=16) was only osteotomized. After 5 days latency, distraction devices were activated at a rate of 1.4 mm per day (0.7 mm every 12hours) for 3.5 days. Animals were sacrificed at postoperative 3, 7, 14, and 28 days. H&E stain and immunohistochemical stain was done on decalcified section. Additionally RT-PCR analysis was done for the identification of the expression of osteocalcin and osteonectin. Results : The bone formation in distraction experimental group was much more than that in distraction and normal control group at postoperative 28 days. In immunohistochemical stain, osteocalcin was enhanced at only postoperative 14 days, but osteonectin was not different at each post-operation days. In RT-PCR analysis, osteocalcin was not different at each post-operation days, but osteonectin was strongly expressed in distraction experimental group at postoperative 7 days. The expression of osteocalcin and osteonectin was elevated during the healing period. Conclusion : We found the good bone formation ability of hyaluronic acid in distraction osteogenesis through the immunohistochemistry and RTPCR analysis to osteocalcin and osteonectin, known as a bone formation marker. The application of hyaluronic acid in distraction osteogenesis is a method to reduce the consolidation period.

• Archives of Plastic Surgery
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• 제41권6호
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• pp.654-660
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• 2014

Background Reactive oxygen species (ROS) damages cell molecules, and modifies cell signaling. The nuclear factor E2-related factor (Nrf2) is a critical transcription regulator, which protects cells against oxidative damage. Nrf2 expression is increased in a large number of cancers. However, little information has been reported regarding the expression of Nrf2 in skin cancers. Hence, we explored the expression of Nrf2 protein in skin cancers. Methods The Nrf2 protein expression in 24 specimens, including 6 malignant melanomas (MM), 6 squamous cell carcinomas (SCC), 6 basal cell carcinomas (BCC), and 6 normal skin tissues, was evaluated by western blotting. Immunohistochemical staining was performed. The expression of Kelch-like ECH-associated protein 1 (Keap1), the key regulator of Nrf2, was also analyzed by western blotting. Results Small interfering RNA transfection to the melanoma cell line G361 confirmed that an approximately 66 kDa band was the true Nrf2 band. The western blot revealed that the Nrf2 protein was definitely expressed in normal skin tissues, but the Nrf2 expression was decreased in MM, SCC, and BCC. Immunohistochemical examination showed that expression of Nrf2 was decreased in all skin cancer tissues compared to the normal skin tissues. Keap1 was not expressed in all malignant skin tumors and normal skin tissues by western blot. Conclusions ROS was increased in various types of cancers which proteins were highly expressed or underexpressed. This study demonstrated that the expression of Nrf2 protein was down-regulated in human malignant skin tumors. We suggest that decreased expression of Nrf2 is related to skin cancers.

• 대한골관절종양학회지
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• 제14권2호
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• pp.106-118
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• 2008

목적: 세포주기의 조절곤란은 종양의 발생과 진행에 영향을 미친다. 몇가지 알려진 연부조직 육종에 대한 면역 조직화학적 예후인자 들이 있지만 상반된 연구도 있고 G1/S phase관계되는 인자에 대한 연구는 거의 없는 상태이다. 따라서 저자는 연부조직 육종의 재발 및 전이와 관계된 G1/S phase 세포조절주기 단백의 면역화학적 예후인자를 알아보고자 하였다. 대상 및 방법: 1998년 1월부터 2005년 12월까지 연부조직 육종으로 진단된 환자 중 최소한 1년이상 추시관찰이 가능하고 파라핀 블록의 보존상태가 비교적 양호한 43예의 환자를 대상으로 연구하였다. 지방육종 15예, 악성 섬유성조직구증 13예, 횡문근육종 5예, 활막육종 5예, 평활근육종 3예, 섬유육종이 2예였다. 모든 환자의 조직은 전 절제술로 제거된 조직을 대상으로 Cyclin D1, Cyclin E, CDK4, CDK, p16, p27, Rb, E2F-1, p53, Ki-67 등의 면역 조직화학적 발현을 조직 microarray 방법을 사용하여 측정하고 환자의 국소 재발 및 전이에 따른 예후를 비교 분석하였다. 결과: 국소 재발은 8예(19%)에서 일어났으며 이것은 Cyclin E(p=0.024), E2F-1(p=0.046)의 발현과 연관이 있었다. 전이는 16예(37%)에서 일어났으며 CDK4의 증가와 연관이 있었다(p=0.031). 결론: Cyclin E와 E2F-1이 연부조직 육종의 국소 재발과 관련있는 예후를 제공해주었고, CDK4가 전이를 예측할 수 있는 독립적인 예후를 제공해주었다. 따라서 조직 검사시 이들 표식자들의 적절한 사용이 환자의 예후를 예측하고 치료의 범위를 결정 짖는데 중요한 도움을 줄 수 있을 것이다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권5호
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• pp.418-425
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• 2006

Purpose: This study was aimed at evaluating the histological changes of new bone and expression of osteopontin (OPN) after mandibular distraction osteogenesis. Materials and Methods: Unilateral mandibular distraction (0.5 mm twice per day for 10 days) was performed in eight adult dogs. Two animals were sacrificed at 7, 14, 28 and 56 days after completion of distraction, respectively. The distracted bones and contralateral non-distracted control bones were harvested and processed for histological and immunohistochemical examinations. Results: The new bone was arranged to tension direction after distraction osteogenesis. 7 days after distraction, numerous osteoblasts lining the immature trabecular bone and fibroblast-like cells in the fibrous intrezone were observed. 14 days after distraction, the new formed trabecular bones were thickened compared with 7 days after distraction. 28 days after distraction, the fibrous interzone was almost filled with newly calcified bone, and it was more hardened at 56 days after distraction. Increased OPN signals detected in the osteoblasts lining the trabecular bone and fibroblast-like cells in the fibrous interzone at 7 and 14 days after distraction. At 28 days after distraction, the OPN was weakly expressed in the osteoblasts, and it was not detected in all cellular components of distracted bone at 56 days later of distraction. Conclusions: After distraction osteogenesis, the distracted zone was completely calcified during the 56 days of consolidation period. In this study, the staining intensity of OPN increased in the osteoblasts and fibroblast-like cells at 7 and 14 days after distraction. The expression pattern of this protein shown here suggested that OPN play an important role in the osteogenesis during the early consolidation period.

• Journal of Chest Surgery
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• 제31권10호
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• pp.924-927
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• 1998

연구배경 : 심근의 허혈 또는 재관류에 의한 세포사에는 괴사 이외에 세포고사가 존재함이 알려져 있다. Bcl-2 단백은 세포질에 존재하는 단백으로 세포고사를 억제하는 기능을 하며 정상심근에서는 발현되지 않으나 심근경색의 급성기에서 발현됨이 보고되어 있다. 본 연구는 가토 허혈-재관류 심근에서 Bcl-2 단백의 발현 여부와 재관류의 시간에 따른 발현의 변화를 알아보고자 하였다. 방법: 평균 무게가 2.9Kg(1.5-4.8Kg)인 가토 39마리를 이용하였다. 허혈-재관류 모델의 각 실험동물에서 좌전하행지를 30분간 결찰한 다음 1, 4, 8, 12, 24시간, 3, 7일 동안 재관류시켰다. 이후 즉시 실험동물을 희생시킨 다음 심장을 적출하여 심근조직을 얻고 10% buffered formalin에 고정하였다. Bcl-2 단백의 발현은 파라핀에 포매된 조직에서 단일클론항체를 이용한 면역조직화학적 염색으로 확인하였다. 결과: 허혈-재관류 심근 중 12, 24시간, 3일 재관류군에서 Bcl-2 단백의 발현을 관찰할 수 있었으며, 특히 24시간 재관류 심근에서 잘 관찰되었다. Bcl-2 양성염색의 심근세포는 위험부위의 구제심근에서 관찰되었다. 결론: Bcl-2 단백은 심근의 허혈-재관류에서 급성기의 비교적 후기에 발현되며, 이는 재관류 초기에서 보다는 후기에서 세포고사를 억제하는데 일부 역할을 할 것으로 사료된다.

• Journal of Korean Neurosurgical Society
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• 제30권4호
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• pp.443-450
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• 2001

Objective : The cyclin-dependent kinase inhibitor $p27^{kip1}$ protein is a negative regulator of the cell cycle, and its degradation is required for entry into the S phase. Loss of $p27^{kip1}$ expression has been reported to be associated with aggressive behavior in a variety of tumors of epithelial and lymphoid origin. However, its association with various astrocytic tumors has not been clearly demonstrated. We studied to investigate the relationship of $p27^{kip1}$ expression with the biological behavior of astrocytic tumors in addition to study on the role of $p27^{kip1}$ in the tumorigenesis of these tumors. Patients and Methods : From 1990 to 1998, a total of 29 astrocytic tumor of all grades obtained by operative resection were included for evaluation. We studied the expression of $p27^{kip1}$ protein immunohistochemical assay in astrocytic tumors and compared the findings with the clinicopathologic parameters. Immunohistochemical staining was performed on formalin-fixed paraffin-embedded sections by the avidin-biotin-peroxidase complex method. According to WHO classification, all cases were divided into astrocytomas(4 cases), anaplastic astrocytomas(9 cases), and glioblastomas(16 cases) by 3 pathologists. Clinical information was obtained from medical records, and others such as location and size of tumors from imaging studies. Results : Mean $p27^{kip1}$ protein labeling indexes(LI, mean${\pm}$standard deviation) of astrocytomas, anaplastic astrocytomas, and glioblastomas were $80.6{\pm}9.1$, $63.6{\pm}21.0$, and $28.9{\pm}18.7$, respectively, and were inversely correlated with grade of glial tumors(p<0.0001). Mean $p27^{kip1}$ protein LI in the recurrent group was lower than that in the nonrecurrent group, but there was no significant difference statistically(p=0.464). Additionally, $p27^{kip1}$ protein expression did not show any significant relationship to other prognostic factors such as age(p=0.1643), tumor size(p=0.8), or location(p=0.8). Conclusion : These results suggested that reduced expression of $p27^{kip1}$ protein may play a important role in the malignant transformation process of astrocytic tumor cells.

• Clinical and Experimental Reproductive Medicine
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• 제35권1호
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• pp.61-67
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• 2008

목 적: Glucocorticoid가 황체용해에 미치는 효과의 기전의 일부를 면역 조직학적 연구를 통해 알아보고자 하였다. 연구방법: 8주령의 Rat 암컷 30마리 대조군, 덱사메타손 0.1 mg 투여군 및 덱사메타손 1mg 투여군의 3군으로 나누어 황체용해의 차이와 대식세포의 침투여부를 비교하기 위하여 광학현미경을 이용하여 계수하였다. 결 과: TUNEL 면역염색 결과 덱사메타손 0.1 mg 투여군과 1 mg 투여군 모두에서 대조군에 비해 황체용해의 비율이 현저히 감소하였고, ED1 면역염색 결과 덱사메타손 1 mg 투여군에서 대식세포의 황체 내 침투가 현저히 억제된 것으로 나타났다. ED1 면역염색 결과 덱사메타손 1 mg 투여군에서 대식세포의 면역반응성이 대조군에 비해 감소되어 나타났다. 결 론: 덱사메타손은 직접 황체세포에 작용하여 세포자멸사를 억제한다. 또한 대량 투여했을 때는 대식세포의 황체 내 침투를 억제하거나 황체 내에 이미 존재하는 대식세포의 활성화를 억제함으로써 황체용해를 억제한다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권2호
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• pp.277-290
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• 2008

The process of wound repair involves an ordered sequence of events such as overlapping biochemical and cellular events that, in the best of circumstances, result in the restoration of both the structural and functional integrity of the damaged tissue. An important event during wound healing is the contraction of newly formed connective tissues by fibroblasts. The polypeptide growth factors, like transforming growth factor-${\beta}$(TGF-${\beta}$, insulin-like growth factor I (IGF- I) and epidermal growth factor (EGF), play very important mediator roles in the process of wound contraction. Deer antlers, as models of mammalian regeneration, are cranial appendages that develop after birth as extensions of a permanent protuberance (pedicle) on the frontal bone. Antlers contain various growth factors which stimulate dermal fibroblast growth. They are involved in digestion and respiration and are necessary for normal wound healing and skin health. In order to investigate and evaluate the effects of red deer antlers on skin wound site, the speed of full-thickness skin wound healing and the expression of IGF-I, TGF-${\beta}$ and EGF in skin wounds, three groups of skin full-thickness rat models with a high concentration of antler ointment, a low concentration of antler ointment and without antler ointment were compared. At post-injury days 0, 2, 4, 8, 16, 20, 32, 40 and 60, the skin wound area was measured, the expressions of IGF-I, TGF- ${\beta}$ and EGF mRNA were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and collagen formation by sirius red dye and the localization of IGF-I, TGF-${\beta}$ and EGF peptides were inspected by histological immunohistochemical techniques. Wound healing was significantly more rapid in antler treated skins. In addition, the wound treated with a high concentration antler ointment, a low concentration antler ointment, and the control closed completely at post-injury day 40, day 44 and day 60, respectively. Via RT-PCR, the expressions of IGF-I (day 8 and day 16), TGF-${\beta}$(day 8, day 16 and day 20) and EGF (day 4, day 8, day 16, and day 32) were obviously up-regulated in high concentration antler-treated skins compared to control skins. Similar results could be seen in the histological detection of collagen dye and immunohistochemical methods using the corresponding polyclone antibodies of IGF-I, TGF-${\beta}$ and EGF. These results illustrate that antlers stimulate and accelerate the repair of cutaneous wounds.

• 대한한의학회지
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• 제29권4호
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• pp.68-82
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• 2008

Objective: The purpose of our study was to show the effects of Hwalhyulmaksung-bang (Huoxiemoxing-fang, HHMSB) treatment on cBSA-induced in MN Mouse Model. Methods: We divided the 20 mice into 4 groups. One group, named NR, was not treated. The second group, named CT, was treated with cBSA (7mg/kg i.p) only. The third group, named HH-250, was treated with cBSA (7mg/kg i.p) and HHMSB extract (250mg/kg, p.o). The fourth group, named HH-500, was treated with cBSA (7mg/kg i.p) and HHMSB extract (500mg/kg, p.o). 4 weeks after cBSA, proteinuria, serum albumin, total cholesterol, serum creatinine, BUN, total cell number of spleen and kidney of all groups were measured. CD3e+/CD19+ and CD4+/CD8 cells ratio of peripheral blood, kidney and spleen of all groups were analyzed. $IL-1{\beta}$ and TNF-$\alpha$, IL-6, IgG, IgM, and IFN-$\gamma$ levels of all groups were gauged. Histological analysis of kidney tissue and immunohistochemical staining (CD4, CD8) of kidney were observed. Results: The level of proteinuria significantly decreased and serum albumin increased in the group treated with cBSA and HHMSB extract compared with the control. Total cholesterol decreased but not significantly. CD3e+/CD19cells ratio of peripheral blood is decreased, but CD4+/CD8cells ratio has no significancy. CD3e+/CD19+ and CD4+/CD8 cells percentage of kidney and spleen has no significancy. Level of $IL-1{\beta}$, IL-6 is significantly decreased, and IFN-$\gamma$ is significantly increased on HHMSB compared with control. Total IgG level significantly decreased on HHMSB compared with the control. Thickness of GBM decreased on histological analysis of kidney. Deposition of CD4 and CD8 decreased on immunohistochemical staining of kidney. Conclusions: We conclude that Hwalhyulmaksung-bang treatment may could be a useful remedy agents for treating Membranous Neuropathy(MN) induced by cationized bovine serum albumin.