• Journal of Nutrition and Health
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• 제13권3호
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• pp.117-125
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• 1980

섭취된 단백질이 성장하는 쥐의 체내에서 이용되는 효율을 알고, 지방 섭취량이 단백질 대사에 어떤 영향을 주는지 알기 위해서 생후 6주되고 평균 체중이 약 106g 정도되는 Sprague-Dawley 계 albino rat 숫컷 51마리를 사용하여 (3 마리는 개시시 도체분석에 사용하고, 48마리는 사양시험에 사용하였음) 시험 사료의 에너지 수준은 M.E.가(價)가 4000Kca1/kg으로 모두 같게 하되 단백질 공급 수준은 저단백질구(10%), 중단백질구(20%), 고단백질구(30%)로 하고, 지방 급여 수준은 각 단백질 수준마다 총 에너지의 20%(저지방구), 40%(고지방구)로 정하고 나머지 에너지는 전분과 포도당으로 보충한 6종의 사료처리구에 대한 사양시험, 대사시험 및 도체분석 시험을 실시했던바 얻어진 결과를 요약하면 다음과 같다. 1) 증체량은 사료의 단백질 수준을 증가시킬 경우 저지방구에서는 증가했으나 고지방구에서는 오히려 감소되있다. 사료 효율은 저지방구에서는 단백질 공급수준을 높일수록 증가되었으나 고지방구에서는 일정한 경향을 찾을 수 없었다. 단백질 효율에 있어서는 저단백질구의 그것이 높았는데 특히 저단백질 고지방구(LPHO)의 단백질 효율이 가장 높았으며 단백질 수준이 높아질수록 지방의 공급량에 관계없이 저하되었다. 2) 고형물, 단백질 총탄수화물의 소화율은 같은 수준의 단백질을 공급하는 경우 고지방구의 것보다 저지방구의 것이 높았으며, 단백질 급여 수준이 높을수록 고형물과 탄수화물의 소화율은 약간 감소되었으나, 단백질의 소화율은 오히려 약간 증가되었다. 지방의 소화율은 사료의 단백질이나 지방의 급여 수준에 큰 영향을 받지 않고 모두 높았다. 3) 질소 이용률에 있어서는 중단백질구(20%)인 경우 저지방구가 가장 높았으나, 고지방구는 가장 낮았다. 사료단백질 20%까지는 저지방 수준인 경우 질소이용률이 뚜렷하게 높은 것을 알 수 있었으나 고단백질구(30%)에서는 지방의 급여량에 따른 차이를 볼 수 없었다. 4) 에너지 섭취량은 사료의 단백질과 지방 수준에 따라서 영향을 받았다. 각 처리구별로 에너지의 대사율을 비교해 보면, 첫째로 사료의 단백질 수준이 높아질수록 에너지 대사율은 약간씩 낮아졌고, 둘째로 같은 단백질 수준에서는 저지방구의 에너지 이용률이 고지방구의 그것보다 훨씬 높았다. 5) 체성분은 시험 개시시 체조성에 미하여 체수분은 감소되었고 체단백질과 체지방은 모두 증가되었다. 단백질 공급 수준이 높아질수록 체수분은 높았고, 체단백질과 체지방은 모두 낮은 경향이 있었다. 단백질 수준이 같을 경우 고지방구의 체수분은 저지방의 그것보다 높았으며 체단백질과 체지방은 오히려 고지방구에서 낮은 경향을 보였다. 6) 이상에서 설명한 실험결과를 종합하여 볼 때 성장하는 흰쥐의 경우 단백질 20%, 지방 20%, 탄수화물 60%로 구성된 식이가 가장 이상적인 것으로 생각된다.

• Journal of Yeungnam Medical Science
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• 제16권1호
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• pp.34-42
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• 1999

고혈당과 고지질혈증이 운동 중 골격근의 당원절약효과가 있는지 또는 골격근섬유의 특생에 따른 차이가 있는지를 규명하기 위해서 흰쥐(Sprague Dawley)를 대상으로 실험한 연구결과를 요약하면 다음과 같다. 먼저 고혈당에 의한 결과는 하지근 중 족척근의 운동부하 45분군의 당원농도가 고혈당군에서 대조군의 그것보다 약 13% 높아서 고혈당에 의한 족척근의 당원절약효과를 관찰할 수 있었으며 또한 백색 비복근의 운동부하 45분군에서 절약효과를 관찰할 수 있었다. 그러나 서근인 가자미근과 서근의 특성이 강한 적색 비복근에서는 모든 실험군에서 5-13%의 절약경향이 있었다. 이때 고혈당군의 혈당농도는 운동부하 전, 운동부하 30 및 45분후에 각각 $145.7{\pm}7.4$, $164{\pm}6.2$ 및 $156{\pm}6.8mg/dL$로서 대조각군의 혈당농도보다 유익하게 높았다. 고지질혈증은 복강내 heparin투여와 10% 인트라리포즈를 경구 투여하여 유도하였으며 대조군, 대조운동부하군 및 고지질혈증군에서 유리지방산 농도(uEq/L)가 $247{\pm}41.1$, $260{\pm}58.6$ 및 $444{\pm}23.1$로서 고지질혈증군이 다른 양 군보다 유의하게 높았다. 고지질혈증에 의한 근당원절약효과가 있는지 여부를 관찰한 결과 가자미근, 적색 비복근 및 족척근 등에서 통계적으로 유의한 절약효과를 관찰할 수 없었으나 백색 비복근에서 절약의 경향을 보였다. 이상의 결과로 보아 흰쥐에서 비교적 단시간의 운동부하 중 평균 155mg/dL의 고혈당을 유지하였을 때 족척근과 백색 비복근에서 근 당원이 절약되었으며 고지질혈증에 의해서는 근당원의 절약효과가 없었다. 또한 속근섬유의 근육에서 고혈당에 의한 당원절약효과가 서근보다 뚜렷하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권10호
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• pp.1407-1413
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• 2005

An experiment was conducted in order to assess the effect of level of RDP:UDP ratio and level of feeding concentrate on milk yield, milk composition and nutrient utilization in lactating crossbred cattle. Twenty four medium producing (-10 kg/d, 45 to 135 days postpartum) lactating crossbred cows were divided into four groups of six animals each in a 2${\times}$2 factorial completely randomized design. The cows in group 1 were fed concentrate mixture I containing 59:41 RDP:UDP ratio (low UDP) at normal plane (LUDP+NP), in group 2 were fed low UDP ration at 115% of NRC (1989) requirements (LUDP+HP), whereas cows in group 3 were fed concentrate mixture II containing 52:48 RDP:UDP ratio (high UDP) at normal plane (HUDP+NP) and in group 4 were fed high UDP ration at 115% of NRC (1989) requirements (HUDP+HP). Green jowar was fed ad libitum as the sole roughage to all the animals. The experimental feeding trial lasted for 105 days. The total dry matter intake (DMI), DMI/100 kg body weight, DMI/kg $W^{0.75}$, digestibilities of DM, OM, CP, CF, EE and NFE and intakes of TDN and DCP did not differ significantly among the different groups and also due to both UDP level and plane of nutrition and also due to their interaction. The total dry matter intake varied from 145 g in group 1 (LUDP +NP) to 152.57 g/kg $W^{0.75}$ in group 2 (LUDP+HP) diet. However, increase in milk yield with increased UDP level and also with increased plane of nutrition was observed consistently throughout the experimental period. The average milk yield was 7.66, 8.15, 8.64 and 9.35 kg in groups 1, 2, 3 and 4, respectively and there was no significant difference in milk yield among different groups of cows. The overall daily average milk yields in cows fed with low and high UDP diets were 7.91 and 8.99 kg, respectively and at normal and higher plane of feeding the milk yields were 8.15 and 8.75 kg/day, respectively. Thus, there was 13.65% increase in milk yield due to high UDP level and 7.36% due to higher plane of feeding. The daily 4% FCM yields were 9.20 kg for low UDP diet and 10.28 kg for high UDP diet, whereas it was 9.11 kg at normal plane of feeding and 10.37 kg at higher plane of feeding. Fat yields for the corresponding treatment groups were 0.37, 0.43, 0.41 and 0.48 kg, respectively. The 4% FCM yield and also fat yield did not differ significantly among different dietary treatments and also due to UDP level and plane of nutrition, however, 4% FCM yield was increased by 11.74% with high UDP level and 13.83% with higher plane of feeding. The values for total solids, fat, lactose, solids-not-fat and gross energy contents in milk differed significantly (p<0.05) among the different groups and were significantly (p<0.05) higher in milk of cows fed LUDP+HP diet followed by HUDP+HP diet. Total solids (14.65 and 13.83%), lactose (5.44 and 4.92%), solids-not-fat (9.44 and 8.83%) and gross energy (887 and 838 kcal/kg) of milk decreased significantly (p<0.05) with increased UDP level while total solids (13.84 and 14.64), fat (4.84 and 5.36%) and gross energy (832 and 894 kcal/kg) increased significantly (p<0.05) with increase in plane of feeding. Gross and net energetic efficiencies and also gross and net efficiencies of nitrogen utilization for milk production were not significantly different among different groups and also were not affected significantly due to either UDP levels or plane of feeding. Results of the present study suggest that, increasing UDP level from 41% to 48% of CP in concentrate mixture and also increasing plane of feeding from normal (100%) to 115% of NRC requirements maintain a consistently higher milk production.

• Asian Pacific Journal of Cancer Prevention
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• 제15권1호
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• pp.467-474
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• 2014

Objective: To observe the effects of allogeneic and autologous transfusion on cellular immunity, humoral immunity and secretion of serum inflammatory factors and perforin during the perioperative period in patients with malignant tumors. Methods: A total of 80 patients (age: 38-69 years; body weight: 40-78 kg; ASA I - II) receiving radical operation for gastro-intestinal cancer under general anesthesia were selected. All the patients were divided into four groups based on the methods of infusion and blood transfusion: blank control group (Group C), allogeneic transfusion group (group A), hemodiluted autotransfusion Group (Group H) and hemodiluted autotransfusion + allogenic transfusion Group (A+H group). Venous blood was collected when entering into the surgery room ($T_0$), immediately after surgery ($T_1$) and 24h ($T_2$), 3d ($T_3$) and 7d ($T_4$) after surgery, respectively. Moreover, flow cytometry was applied to assess changes of peripheral blood T cell subpopulations and NK cells. Enzyme linked immunosorbent assays were performed to determine levels of IL-2, IL-10, TNF-${\alpha}$ and perforin. Immune turbidimetry was employed to determine the changes in serum immunoglobulin. Results: Both CD3+ and NK cells showed a decrease at $T_1$ and $T_2$ in each group, among which, in group A, CD3+ decreased significantly at $T_2$ (P<0.05) compared with other groups, and CD3+ and NK cell reduced obviously only in group A at $T_3$ and $T_4$ (P<0.05). CD4+ cells and the ratio of D4+/CD8+ were decreased in groups A, C and A+H at $T_1$ and $T_2$ (P<0.05). No significant intra- and inter-group differences were observed in CD8+ of the four groups (P<0.05). IL-2 declined in group C at $T_1$ and $T_2$ (P<0.05) and showed a decrease in group A at each time point (P<0.05). Moreover, IL-2 decreased in group A + H only at $T_1$. No significant difference was found in each group at $T_1$ (P<0.05). More significant decrease in group ?? at $T_2$, $T_3$ and $T_4$ compared with group A (P<0.05), and there were no significant differences among other groups (P>0.05). IL-10 increased at $T_1$ and $T_2$ in each group (P<0.05), in which it had an obvious increase in group A, and increase of IL-10 occurred only in group A at $T_3$ and $T_4$ (P<0.05). TNF-${\alpha}$ level rose at $T_1$ (P<0.05), no inter- and intra-group difference was found in perforin in all groups (P<0.05). Compared with the preoperation, both IgG and IgA level decreased at $T_1$ in each group (P<0.05), and they declined only in Group A at $T_2$ and $T_3$ (P<0.05), and these parameters were back to the preoperative levels in other groups. No significant differences were observed between preoperative and postoperative IgG and IgA levels in each group at $T_4$ (P>0.05). No obvious inter- and intra-group changes were found in IgM in the four groups (P>0.05). Conclusions: Allogeneic transfusion during the perioperative period could obviously decrease the number of T cell subpopulations and NK cells and the secretion of stimulating cytokines and increase the secretion of inhibiting cytokines in patients with malignant tumors, thus causing a Th1/Th2 imbalance and transient decreasing in the content of plasma immune globulin. Autologous transfusion has little impact and may even bring about some improvement oo postoperative immune function in patients with tumors. Therefore, cancer patients should receive active autologous transfusion during the perioperative period in place of allogeneic transfusion.

• 기술사
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• 제28권4호
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• pp.54-75
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• 1995

본 보고서는 자동차 긴급 피난 차선에 관한 기술 자료, 문헌의 검토와 각 주별 운송 당국에 대한 설문 조사 결과를 기본으로 하여 작성되었다. 자동차 긴급 피난 차선(Truck Escape Ram-ps)이 미국의 고속도로에 채택된 것은 35년의 역사가 있으며 지금은 서부의 산악 지대로부터 에팔라치아까지, LA 교외는 물론 동북부의 조그마한 마을까지 약 27개 주에서 채택하고 있다 과거에는 자동차의 긴급 피난 차선의 위치는 통상 사고 발생 경위와 기술적인 판단에 따라 선정하였지만 지금은 대형 사고가 발생되기전에 자동차 긴급 피난 차선의 필요성과 위치를 선정할 수 있는 새로운 방범이 도입되고 있다. 방법중에는 경사 평가제(Grade Severity Ra-ting System : GSRS)가 유력한 방법으로 평가된다. 자동차 긴급 피난 차선의 설계법은 지금도 계속적으로 발전 단계에 있으며 현재는 (자갈 제동 노반식 피난 차선)의 선호도가 높은 것으로 되어 있다. 입자가 둥글고 균일한 입도 분포로서 입경이 13~18mm인 자갈이 회전 저항이 커서 피난 차선의 노반 재료로 많이 사용되고 있다. 긴급 피난 차선의 길이 결정을 위한 새로운 계산 방법은 공사비 절감에 초점을 두고 있으며 긴급 피난 차선의 목과 종점부의 처리에 관한 설계법에 대한 연구가 진행중에 있다. 각 주별 조사에 의하면 1주일에 I~2회 이상 이용되는 곳이 있으며 자동차의 이용이 적어 4륜 구동 차량의 운전자들이 차를 시험하는 등국도 운전 연습장으로 사용되는 곳도 있었다. 따라서 긴급 피난 차선의 목적 외 사용을 제한하는 안내 표지판의 설치와 규정 강화를 위한 노력이 요구되고 있다. 내리막길의 정상부에는 미리 안내판을 설치하고 브레이크 점검 구역을 설치해 두면 안전 운행에 도움을 줄 수 있을 것이며 공공 정펄 매체를 이용한 안내 광고를 함으로써 운전자가 한계 상태에 위치한 자동차의 긴급 피난 차선을 적극적으로 이용할 수 있도록 유도하는 방법이 사용되고 있다. 적절한 유지 관리는 긴급 피난 차선의 효율적인 운영에 필수적인 요소로서 사용 후 입도 조정 및 표면 처리(fluffing :부풀림)가 필요하며 설계시나 유지 관리시에 노반의 세립토 제거 작업은 긴급 피난 차선의 수명 연장은 물론 만족할 만한 기능 유지에 결정적인 요인이 될 것이다. 현행 시공법을 개선하기 위하여는 몇 개 분야의 추가 연구가 필요하단. 예를 들면 비용 편익 분석, 위치 선정 기법 확립, 진입 속도 및 피난 차선 연장의 결정 조건, 골재 기능 평가, 다수 진입로의 필요성, 효과적인 운전자 교육 정책 등이 그것이다. 자동차 긴급 피난 차선은 유용한 시설로 여러곳에서 그 기능이 발휘되고 있으므로 많은 고속도로 관계자들은 설계, 시공 및 유지 관리 측면에서 유의해야 할 것이다.

• 한국토양비료학회지
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• 제11권4호
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• pp.283-295
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• 1979

농업(農業)의 기대성장(期待成長)과 현실성장을 비교(比較)하기 위하여 1953-77. 기간(期間)을 3단계(段階)로 구분하여 역사적접근법(歷史的接近法)을 시도(試圖)하였으며 농경지(農耕地)의 타목적전환과 전작물(田作物)의 수요(需要)를 전망(展望)하였다. 야산개발의 필요성(必要性)을 검토(檢討)하기 위하여는 농경지(農耕地)의 외연적확대가 가능(可能)한 가를 살펴보았으며 개간지(開墾地)의 작부유형(作付類型)을 분류(分類)하였다. 야산개발의 지역농업(地域農業)에 미친 효과로는 개간농가(開墾農家)의 경영규모확대와 경지규모별호수분포가 대농화경향(大農化傾向)을 보이는 것 같고 개간지(開墾地)의 지가가 기경지(旣耕地)보다 빠른 상승율(上昇率)을 보였다. 야산개발지의 생산성(生産性)은 6년차(年次)만 되면 기경지수준에 가깝게 되고 따라서 소득지수도 이와 유사(類似)한 경향(傾向)을 보이고 있다. 야산개발사업의 수익성은 다른 토지개발사업보다 높고 작목별(作目別)로 보면 식량작물(食糧作物)보다는 경제작물이 높고 단작(單作)보다는 이모작(二毛作)의 경우가 유리(有利)하게 나타났다.

• Radiation Oncology Journal
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• 제17권1호
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• pp.57-64
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• 1999

목적 : Paclitaxel(Taxol)은 미소관의 집합을 촉진시키고 분해를 방지하여 세포주기 중 유사분열을 정지시킴으로써 방사선조사와 병용할 경우 방사선감작제로서의 가능성이 있다. 흰쥐의 대장점막에서 paclitaxel이 방사선의 효과에 미치는 영향을 파악하기 위하여 본 실험을 시도하였다. 대상 및 방법 :실험군은 세군으로 나누어 paclitaxel 단독군은 paciltaxel 10mg/kg을 복강내 1회 주입하였고, 방사조사 단독군은 8Gy를 전복부에 단일조사하였으며, paclitaxel과 방사선 병용군은 paclitaxel(10mg/kg)을 복강내 주입 후 24시간에 방사선조사 단독군과 동일한 방법으로 조사하였다. 실험완료 후 대장점막에서 유사분열수, apoptosis와 기타 점막의 변화를 시간별로(6시간$\~$5일) 비교관찰하였다. 결과 : Paclitaxel 주입시 대장점막에서 유사분열의 빈도는 증가되지 않았고 apoptosis는 주입 후 24시간에 관찰할 수 있었으며, 소수포형성, 비정형성 및 배상세포의 감소는 paclitaxel 주입 후 6시간부터 3일까지 심하게 보였다. 방사선 조사 단독시 apoptosis는 6시간과 24시간에 관찰할 수 있었으며, 대장점막의 소수포형성, 비정형성 잊 배상세포의 감소는 24시간에 보이기 시작하여 3일에 심하게 보였다. Paclltaxel 주입 후 24시간에 방사선조사하여 apoptosls는 3일에 나타났으며 소수포형성, 비정형성 및 배상세포의 감소는 6시간부터 3일까지 나타났다. Paclitaxel과 방사선의 병용군에서 방사선조사 단독군에 비하여 apoptosis는 증가되지 않았으나, 소수포형성, 비정형성 및 배상세포의 감소는 6시간과 24시간에 증가되었으며(P<0.05), 이것은 방사선과 paclitaxel의 각각의 세포독성의 첨가효과가 있었다. 결론 : 흰쥐 대장에서 paclitaxel은 유사분열에 영향을 미치지 않았으나, 방사선조사에 의한 세포손상과 동일한 변화로 apoptosis, 소수포형성, 비정형성 및 배상세포의 감소를 유발하였다. Paclitaxel과 방사선조사 병용군에서 대장점막에 소수포형성, 비정형성 및 배상세포 감소가 방사선조사 단독군에 비하여 현저히 증가되어 paclitaxel은 방사선에 대하여 첨가효과가 있었다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권2호
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• pp.95-110
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• 2006

The treatment for oral and maxillofacial carcinoma with chemotherapeutic agents is evaluated by many effective methods to reduce the tumor mass and cancer cell proliferation. However these chemotherapy have many serious side effects, such as bone marrow suppression, renal toxicity, G-I troubles. Therefore a possible approach to develop a clinically applicable chemotherapeutic agent is to screen anticancer activity of Taxol which is known to have very little side effect and have been used to breast cancer and ovarian carcinoma. Taxol is a new anti-microtubular anti-cancer agent extracted from the bark of the Pacific yew, Taxus brevifolia. Paclitaxel(Taxol) acts by promoting tubulin polymerization and over stabilizing microtubules agianst depolymerization. Despite the constant improvements of methods of the cancer treatment especially chemotherapy, the rate of cancer metastasis and recurrent are not decreased. Thus the investigation of new drug which have very little side effect and a possible clinically application continues to be a high priority. Considering that the Taxol have shown very effective chemotherapeutic agent with relatively low toxicity in many solid tumors, it deserves to evaluate its efficacy in oral squamous cell carcinoma. In this study, to investigate the in-vivo and in-vitro anti-cancer efficacy of Taxol in oral squamous cell carcinoma and lastly, the potency of Paclitaxel in the clinical application for oral cancer was evaluated. In vivo study, after HN22 cell line were xenografted in nude mice, the growth of tumor mass was observed, 3 mg/Kg taxol was injected intraperitoneally into nude mice containing tumor mass. The methods of these study were measurement of total volume of tumor mass, histopathologic study, immunohistochemical study, drug resistance assay, growth curve, MTT assay, flow cytometry, cDNA microarray in vivo and in vitro. The results were obtained as following. 1. The visual inspection of the experimental group showed that the volume of the tumor mass was slightly decreased but no significant difference with control group. 2. Ki-67 index was decreased at weeks 4 in experimental group. 3. Microscopic view of the xenografted tumor mass showed well differentiated squamous cell carcinoma and after Taxol injection, some necrotic tissue was seen weeks 4. 4. The growth curve of the tumor cells were decreased after 1day Taxol treatment. 5. According to the MTT assay, HN22 cell line showed relative drug resistancy above $5\;{\mu}g/ml$ concentrations of Taxol. 6. In drug resistance assay, the decrease of cell counts was seen relatively according to concentration. 7. In Flow cytometry, G2M phase cell arrests were seen in low concentration of the Taxol, while S phase cell arrests were seen in high concentration of the Taxol. 8. Using cDNA microarray technique, variable gene expression of ANGPTL4, TXNRD1, FAS, RRAGA, CTGF, CYCLINEA, P19, DUSP5, CEBPG, BTG1 were detacted in the oral squamous cell carcinoma cell after taxol treatment. In this study paclitaxel is effective against oral squamous cell carcinoma cell lines in vitro, but week effect was observed in vivo. So we need continuous study about anticancer effect of taxol in vivo in oral squamous cell carcinoma.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권4호
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• pp.299-305
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• 2010

Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.

• Journal of Periodontal and Implant Science
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• 제31권1호
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• pp.1-23
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• 2001

Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.