• 대한핵의학회지
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• 제30권4호
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• pp.484-492
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• 1996

허혈성 뇌 병변에서 $^{99m}Tc$-glucarate의 섭취에 관한 연구를 하기 위하여 중뇌동맥 폐쇄 쥐 뇌허혈 모델을 재관류한 군과 하지 않은 군으로 나누어 만들었다. $^{99m}Tc$-glucarate와 [$^{18}F$]FDG를 연속적으로 투여하여 그 분포 양상을 이중 자가방사촬영법으로 관찰하였다. 조직의 괴사 여부를 알기 위하여 TTC 염색도 실시하여 동일한 뇌표본에 대하여 3가지의 영상을 동시에 얻을 수가 있었다. 얻은 영상으로 섭취 또는 비섭취정도를 보아 0점에서 3점까지 점수를 매겨 합하여 비교하였다. 수술한 쥐들 18마리 중 10마리가 신경학적 증상을 보이면서 살아남아 실험대상이 되었다. TTC 염색으로 확인한 경색크기는 재관류하지 않은 군이 컸다. [$^{18}F$]FDG 섭취양상은 TTC 염색과 거의 비슷하였다. 다만 일부에서 TTC 염색되는 곳에 [$^{18}F$]FDG 가 중간정도로 섭취되는 곳이 있었고 TTC 염색되지 않는 곳에 [$^{18}F$]FDG가 중간정도 섭취되는 곳이 있었다. TTC로 염색된 부위에는 $^{99m}Tc$-glucarate가 섭취되지 않았다. TTC로 염색되지 않는 곳에 일부분 $^{99m}Tc$-glucarate가 섭취되었다. TTC와 [$^{18}F$]FDG가 염색되거나 섭취되지 않는 곳에 $^{99m}Tc$-glucarate가 섭취되지 않는 곳이 있었다. 그러나 [$^{18}F$]FDG의 중간 정도의 섭취나 [$^{18}F$]FDG와 TTC사이에 부합하지 않는 곳 등과 $^{99m}Tc$-glucarate 섭취와 대응시키기 어려웠다 $^{99m}Tc$-glucarate가 재관류 군에서 더 넓고 많이 섭취되었다. 결론적으로 $^{99m}Tc$-glucarate는 비생존 허혈조직에만 섭취되는데 관류재개통에 따라 다양하게 섭취되었고 재관류모델에 더 많이 넓게 섭취되었다. 중뇌동맥폐색 및 재관류 모델로 $^{99m}Tc,\;^{18}F$ 및 TTC 동시영상을 얻어 생존능과 포도당대사의 부합 비부합 여부를 밝히고 그 의의를 조사한 후 $^{99m}Tc$-glucarate섭취와 상관을 조사하면 $^{99m}Tc$-glucarate섭취의 의의를 밝힐 수 있을 것으로 본다.

• 대한방사선기술학회지:방사선기술과학
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• 제24권2호
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• pp.19-22
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• 2001

The purpose of this study is to evaluate time course of signal enhancement on Gadomer-17 enhance MRI, and to correlate the size of enhanced area with that of the infarct area on 2'3'5'-triphenyl tetrazolium chloride(TTC) histochemical examination for the assessment of myocardial viability in reperfused Myocardial Infarction in a cat model. Tan cats(average weight: 3.8 kg) which had undergone 90 minutes of occlusion of the LAD followed by 90 minutes of reperfusion underwent MR T2-weighted imaging, and T1-weighted imaging, enhanced T1-weighted imaging. We used 1.5T Magneton Vision MRI system(Siemens, Erlangen, Germany). Signal intensities were measured in the enhanced and non-enhanced areas of enhanced T1-weighted imaging. and TTC histochemical staining the size of the abnormal signal area on each image was compared with that of the infarct area. Maximum enhancement was detected during a $40{\sim}60$ minute period with an average enhancement of $168{\pm}9.9%$ of normal myocardium. TTC staining revealed that the size of the high signal area on T2-weighted images and of the enhanced area on enhanced T1-weighted images was greater than that of the infarct area($T2=48.1%{\pm}3.7$, enhanced $T1=47.2%{\pm}2.6$, TTC $staining=38.7%{\pm}3.1$ ; p<0.05). In reperfused Myocardial Infarction in a cat model, enhanced MR imaging delineates reversibly and irreversibly damaged myocardium, with a strong enhancement and a broad temporal window. We may therefore expect that enhanced MR image is useful for demonstrating myocardial injury.

• 대한한의학회지
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• 제20권4호
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• pp.82-92
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• 2000

The purpose of the present study was to explore the proper method for animal stroke model of Oriental medicine To this end, brain ischemia was induced by distal middle cerebral artery occlusion(dMCAO) and proximal middle cerebral artery occlusion(pMCAO) and evaluated with the method of Triphenyl Tetrazolium Chloride (TTC) staining and Swimming Behavior Test. Results demonstrated that first, infarct size and volume of pMCAO group were significantly bigger that those of dMCAO group. Second, analysis of swimming behavior test revealed that the percentage of left turning angles of pMCAO was significantly bigger than that of dMCAO. Third, during swimming behavior test, there were peculiar traces of small successive circles that represent motor dysfunction and conscious disturbance among dMCAO group. The results of the study thus indicate that non-invasive intraluminal method of pMCAO was the appropriate animal stroke model for Oriental medicine in the light of brain ischemia as hemiplesia and conscious disturbance.

• Korean Journal of Radiology
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• 제2권1호
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• pp.21-27
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• 2001

Objective: To identify and evaluate the lateral border zone by comparing the size and distribution of the abnormal signal area demonstrated by MR imaging with the infarct area revealed by pathological examination in a reperfused myocardial infarction cat model. Materials and Methods: In eight cats, the left anterior descending coronary artery was occluded for 90 minutes, and this was followed by 90 minutes of reperfusion. ECG-triggered breath-hold turbo spin-echo T2-weighted MR images were initially obtained along the short axis of the heart before the administration of contrast media. After the injection of Gadomer-17 and Gadophrin-2, contrast-enhanced T1-weighted MR images were obtained for three hours. The size of the abnormal signal area seen on each image was compared with that of the infarct area after TTC staining. To assess ultrastructural changes in the myocardium at the infarct area, lateral border zone and normal myocardium, electron microscopic examination was performed. Results: The high signal area seen on T2-weighted images and the enhanced area seen on Gadomer-17-enhanced T1WI were larger than the enhanced area on Gadophrin-2-enhanced T1WI and the infarct area revealed by TTC staining; the difference was expressed as a percentage of the size of the total left ventricle mass (T2= 39.2 %; Gadomer-17 =37.25 % vs Gadophrin-2 = 29.6 %; TTC staining = 28.2 %; p < 0.05). The ultrastructural changes seen at the lateral border zone were compatible with reversible myocardial damage. Conclusion: In a reperfused myocardial infarction cat model, the presence and size of the lateral border zone can be determined by means of Gadomer-17- and Gadophrin-2-enhanced MR imaging.

• Journal of Korean Neurosurgical Society
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• 제29권7호
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• pp.853-860
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• 2000

Objective : The purpose of our experimental study was to analysis the advantages and disadvantages in the reversible and irreversible cerebral ischemic models with rats by staining with Neutral Red(NR) solusion, 2% 2,3,5-triphenyltetrazolium chloride(TTC) and Hematoxylin & Eosin(H & E). Methods : We have measured the range of cerebral infarction in the rat to get a suitable ischemic model along the object of study with and without craniectomy. With craniectomy, 9 rats were sacrificed for irreversible cerebral ischemic model by means of ligation at proximal(group I) and distal(group II), and coagulation at proximal(group III) middle cerebral artery. Also, 6 rats were sacrificed for irreversible(group IV) and reversible(group V) cerebral ischemic model using nylon thread without craniectomy. The sizes of infarction were measured by staining the coronal sections of the brain with NR solusion, TTC and H & E. Results : There are no difference of physiological parameters comparing the each group. Cerebral infarction was not observed in group II, but it's volume was largest in group IV. Disadvantages of craniectomy group(I, II, III) are the long duration of operation and cortical damage by procedure. It's advantage is confirmation of the middle cerebral artery occlusion and cessation of blood flow through the operative microscope. In case of ischemic models using nylon thread (group IV, V), it is hard to identify the interruption or recirculation of blood flow through the middle cerebral artery, but the advantage is the simplicity of operative technique which reduces the operation time and minimizes the cerebral damage due to craniectomy. Therefore, it seems important to set up the reversible and irreversible ischemic models by carefully considering advantages and disadvantages listed above. Conclusion : TTC staining seems to be effective since it reflects the histological damage sufficiently and quickly. It is hoped that researches focused on ischemic penumbra, which became popular recently, will be further carried on with use of NR staining, optical microscope and electron microscope.

• 대한자기공명의과학회:학술대회논문집
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• 대한자기공명의과학회 2002년도 제7차 학술대회 초록집
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• pp.114-114
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• 2002

Purpose： To know the fate of Gadophrin-2-enhanced areas in hearts with acute reperfused myocardial infarction. Method： The left anterior descending branches of coronary arteries were occluded for 90 min and reperfused for 60 min in 15 rabbits. Then, Gadophrin-2 (0.05 mmol/kg) was injected via ear veins. Short-axial T1-weighted spin echo images and fast cine images were obtained 24 hours after injection of Gadophrin-2. After four weeks, short-axial cine MRI was performed and T1-weighted spin echo Images were obtained 5 min and 15 min after injection of Gd-DTPA (0.2 mmol/kg). The animals were sacrificed and short-axial slices of the specimen were stained with 1.5% triphenyltetrazolium chloride (TTC) solution.

• 한국콘텐츠학회논문지
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• 제19권7호
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• pp.186-191
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• 2019

서론: 최근 노년층에서 뇌졸중 발병률이 증가하고 있다. 현재 뇌졸중 치료제 및 방법이 많이 개발되어 있으나 치료 후에도 후유증 등이 많이 남게 된다. 그래서 아직도 많은 과학자나 임상 의사들이 이를 치료하기 위한 약물 및 방법을 연구하고 있는 실정이다. 많은 연구 중 뇌졸중 치료 연구를 위한 표준화된 실험 동물연구는 드물며, 표준화된 Nylon thread를 이용한 중대뇌동맥 폐쇄모델(MCAO, middle cerebral artery occlusion)의 성공률에 대한 연구는 거의 없다 방법: 본연구는 $0.18{\pm}0.02mm$의 지름을 가진 5-0 Nylon thread를 중대뇌동맥에 삽입하였다. 60분 동안 삽입한 후에 봉합해 놓았던 부위를 다시 절개하여 Nylon thread를 빼내고, 막았던 혈관의 매듭을 풀어주어, 다시 혈액이 공급되게 하였다. 그로부터 23시간 후에 뇌를 내어 1mm 두께로 자른 후 1.5% TTC(2',3',5'-triphenyl-tetrazolium chloride)로 15분간 염색하고, 4% PFA(paraformaldehyde)로 15분 동안 고정하였다. 결과: Nylon thread를 삽입하여, MCA occlusion 50마리, ICA occlusion 14마리, 제대로 된 MCAO model보다 좀 더 깊게 들어간 distal MCAO model 36마리, 너무 깊은 MCA나 ACA까지 들어가서 상보적인 괴사를 나타내는 occlusion model 1마리, 그리고 경색이 일어나지 않은 마우스 50마리를 확인하였다. 결론: 이에 본 연구에서는 Nylon Thread를 생쥐의 무게에 따라 32~36g 인 생쥐는 9mm로 삽입하여주고, 37~40g인 생쥐는 9mm+0.5mm의 깊이로 삽입하여서 1hr의 occlusion과 23hr의 reperfusion을 주어 생쥐를 TTC 염색을 통하여 괴사가 일어난 부분을 확인하였고, 생쥐에서 가역적인 뇌혈관 경색으로 151말중 101마리에서 뇌경색을 유도 할 수 있었다(66.9%).

• Journal of Acupuncture Research
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• 제36권3호
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• pp.140-146
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• 2019

Background: This study investigated the effects of Vipera lebetina turanica snake venom (SV) on cerebral infarction induced by middle cerebral artery occlusion in mice. Methods: Following cerebral infarction, SV was injected intravenously or added to BV2 cell culture. Tissue injury was detected using triphenyltetrazolium chloride (TTC) staining, neurological deficit score, NO, ROS, and GSH/GSSG assays, qPCR, Western blot, and cell viability. Results: Cerebral infarction caused by middle cerebral artery occlusion as observed by TTC staining, showed SV inhibited cell death, reducing the number of brain cells injured due to infarction. SV treatment for cerebral infarction showed a significant decrease in abnormal behavior, as determined by the neurological deficit score. The oxidation and inflammation of the cells that had cerebral infarction caused by middle cerebral artery occlusion (NO assay, ROS, GSH/GSSG assay, and qPCR), showed significant protection by SV. Western blot of brain infarction cells showed the expression of iNOS, COX-2, p-IkB-${\alpha}$, P38, p-JNK, p-ERK to be lower in the SV group. In addition, the expression of IkB increased. BV2 cells were viable when treated with SV at $20{\mu}g/mL$ or less. Western blot of BV2 cells, treated with 0.625, 1.5, $2.5{\mu}g/mL$ of SV, showed a significant decrease in the expression of p-IkB-${\alpha}$, p-JNK, iNOS, and COX-2 on BV2 cells induced by LPS. Conclusion: SV showed anti-inflammatory and anti-oxidant effects against cerebral infarction and inflammation.

• Journal of Korean Neurosurgical Society
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• 제67권3호
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• pp.333-344
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• 2024

Objective : Markers of neuroinflammation during ischemic stroke are well characterized, but additional markers of neural damage are lacking. The study identified associations of behavioral disorders after stroke with histologic neural damage and molecular biological change. Methods : Eight-week-old, 25 g male mice of the C57BL/6J strain were subjected to middle cerebral artery occlusion (MCAO) to induce ischemic stroke. The control group was a healthy wild type (WT), and the experimental group were designed as a low severity MCAO1 and a high severity MCAO2 based on post-stroke neurological scoring. All groups underwent behavioral tests, realtime polymerase chain reaction, triphenyltetrazolium chloride (TTC) staining and Hematoxylin and Eosin staining. One-way analysis of variance was used to analyze statistical significance between groups. Results : In TTC staining, MCAO1 showed 29.02% and MCAO2 showed 38.94% infarct volume (p<0.0001). The pro-inflammatory cytokine interleukin (IL)-1β was most highly expressed in MCAO2 (WT 0.44 vs. MCAO1 2.69 vs. MCAO2 5.02, p<0.0001). From the distance to target in the Barnes maze test, WT had a distance of 178 cm, MCAO1 had a distance of 276 cm, and MCAO2 had a distance of 1051 (p=0.0015). The latency to target was 13.3 seconds for WT, 27.9 seconds for MCAO1, and 87.9 seconds for MCAO2 (p=0.0007). Prospero homeobox 1 (Prox1) was most highly expressed in MCAO2 (p=0.0004). Doublecortin (Dcx) was most highly expressed in MCAO2 (p<0.0001). Conclusion : The study demonstrated that histological damage to neural cells and changes in brain mRNA expression were associated with behavioral impairment after ischemic stroke. Prox1 and Dcx may be biomarkers of neural damage associated with long-term cognitive decline, and increased expression at the mRNA level was consistent with neural damage and long-term cognitive dysfunction.

• Journal of Acupuncture Research
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• 제27권4호
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• pp.29-38
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• 2010

Objectives : The purpose of the study is to determine the neuroprotective effect of modified Boyanghwano-tang(mBHT), a traditional Korean medicine, on the transient focal cerebral ischemia in rats. Methods : Focal ischemia and reperfusion were induced by middle cerebral artery occlusion(MCAO) for 90 min, followed by 144 h reperfusion in rats. mBHT(200mg/kg body weight, p.o.) was administrated in rats once a day during reperfusion. At the end of treatment, brain infarction was measured by TTC staining, and histological change was observed by H&E staining. The expressions of Bax, Bcl-2 and cytochrome c in ischemic brains were determined by immunofluorescent analysis. Results : mBHT significantly reduced the cerebral infarct volumes of the MCAO rats. mBHT also attenuated the neuronal cell death and the expressions of pro-apoptotic molecules, bax and cytochrome c in ischemic brains. Further, mBHT significantly increased the survival time of ischemeic rats and the expression of anti-apoptotic molecule, Bcl-2 in ischemic brains. Conclusions : Our results suggest that mBHT is neuroprotective and may prove to be useful adjunct in the treatment of ischemic stroke.