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노령 흰쥐의 뇌허혈 손상시 뇌대사활성의 변화 및 량격산화탕의 영향에 대한 [$^{14}C$2-Deoxyglucose Autoradiography 연구
손철훈,신정원,손영주,정혁상,원란,손낙원,Sohn, Cheol-Hoon,Shin, Jung-Won,Sohn, Young-Joo,Jung, Hyuk-Sang,Won, Ran,Sohn, Nak-Won 대한한의학회 2003 대한한의학회지 Vol.24 No.3
Objective : This study investigated the alteration of neural activity and effect of Yanggyuksanhwa-tang (Lianggesanhuo-tang) on cerebral ischemia of rats. Methods : Considering age-related impact on cerebral ischemia, aged rats (18 months old) were used for this study. Ischemic damage was induced by the transient occlusion of bilateral common carotid arteries (BCAO) with hypotension. Yanggyuksanhwa-tang (Lianggesanhuo-tang) was administered twice a day orally. Then alterations of neural activities in the brain of aged BCAO rats were measured by the [$^{14}C$]2-deoxyglucose autoradiography method. Results : The BCAO in aged rats led to significant decrease of neural activity in the whole brain. Treatment with Yanggyuksanhwa-tang (Lianggesanhuo-tang) significantly attenuated the decrease of neural activity in the whole brain following BCAO ischemia. Treatment significantly attenuated the decrease of neural activity in the CA1, CA2, CA3, dentate gyrus of the hippocampus, activated barrel, barrel cortex, somatosensory cortex, cingulate cortex, caudate putamen, and medial septal nucleus following BCAO in aged rats. Treatment with Yanggyuksanhwa-tang (Lianggesanhuo-tang) also significantly attenuated the decrease of neural activity in the anteroventral thalamic nucleus, ventral anterior thalamic nucleus, arcuate nucleus, posterior hypothalamic area, medial mammillary nucleus, lateral periaqueductal gray, dorsal raphe nucleus, interpeduncular nucleus, median raphe nucleus, and medial pontine nucleus. Conclusion : It can be suggested that Yanggyuksanhwa-tang (Lianggesanhuo-tang) has a neuroprotecuve effect on cerebral ischemia through the control of glucose metabolic rate and cerebral blood flow.
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최철원,정민호,박유수,손철훈,이홍래,고은경 대한암학회 2019 Cancer Research and Treatment Vol.51 No.2
Purpose The purpose of this study was to investigate the efficacy of stereotactic body radiation therapy (SBRT) as a tumor-associated antigen (TAA) presentation method for dendritic cell (DC) sensitization and evaluate its effect in combination with immunotherapy using an intratumoral injection of immature DCs (iDCs). Materials and Methods CT-26 colon carcinoma cell was used as a cancer cell line. Annexin V staining and phagocytosis assays were performed to determine the appropriate radiation dose and incubation time to generate TAAs. BALB/c mice were used for in vivo experiments. Cancer cells were injected into the right legs and left flanks to generate primary and metastatic tumors, respectively. The mice were subjected to radiation therapy (RT) alone, intradermal injection of electroporated DCs alone, or RT in combination with iDC intratumoral injection (RT/iDC). Tumor growth measurement and survival rate analysis were performed. Enzyme-linked immunospot and cytotoxicity assays were performed to observe the effect of different treatments on the immune system. Results Annexin V staining and phagocytosis assays showed that 15 Gy radiation dose and 48 hours of incubation was appropriate for subsequent experiments. Maximum DC sensitization and T-cell stimulation was observed with RT as compared to other TAA preparation methods. In vivo assays revealed statistically significant delay in the growth of both primary and metastatic tumors in the RT/iDC group. The overall survival rate was the highest in the RT/iDC group. Conclusion The combination of SBRT and iDC vaccination may enhance treatment effects. Clinical trials and further studies are warranted in the future.
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Jae-Ho Bae(배재호),Cheol-Hun Son(손철훈),You-Soo Park(박유수),Dong-Wan Kim(김동완),Sun-Hee Kim(김선희),Chi-Dug Kang(강치덕) 한국생명과학회 2009 생명과학회지 Vol.19 No.6
본 연구에서는, PBMC로부터 고순도 및 고수율의 단세포를 분리하기 위해 Histopaque 용액의 density 및 osmolarity를 PBS와 NaCl 용액으로 1.072 g/㎖ 및 335 mOsm로 조절하여 간단하고 경제적인 방법인 부유밀도구배원심분리법을 개발하였다. 위와 같이 조절된Histopaue용액을 사용하여 부유밀도구배 원심분리를 시행한 결과 단세포는 Top층에서 가장 많이 회수되었다. 회수된 단세포의 순도는 71.44~82.38%의 범위를 나타내었고 평균 74.75±3.84% 이었다. 또한, 수율은 21.02~53.63%의 범위를 나타내어 평균 32.62±11.16% 이었다. 이러한 방법의 부유밀도구배 원심분리에 의해 분리된 단세포를 수지상세포로 분화 유도시켜 그 형태, 표현형, 기능적인 면을 분석해 보았을 때 전형적인 수지상세포의 기능을 나타내었다. 결론적으로, PBMC로부터 단세포를 분리하기 위해 본 연구에서 사용된 부유밀도구배 원심분리 방법은 plastic adherence, elutriation 및 immunomagnetic selection보다 용이하고 경제적인 방법이다. 나아가 이러한 장점을 바탕으로 임상단계에서 수지상세포를 안전하고, 효율적으로 배양할 수 있는 closed system에도 적용이 가능할 것으로 기대된다. In this work, a simple, inexpensive and reproducible technique of flotation density gradient centrifugation was developed to isolate monocytes with high purity and yield from peripheral blood mononuclear cells (PBMC) using Histopaque solution, density and osmolarity of which were modified to 1.072 g/㎖ and 335 mOsm with phosphate-buffered saline (PBS) and sodium chloride (NaCl) solution, respectively. The average purity of monocytes was 74.75±3.84%, with the individual purity ranging from 71.44% to 82.38%. The average yield of monocytes was 32.62±11.16%, with the individual yields ranging from 21.02 to 53.63%. The monocytes isolated by floatation density gradient centrifugation could be successfully cultured into morphologically, phenotypically and functionally dendritic cells in vitro. In conclusion, the entire procedure seemed to be faster and more convenient, simple and cost-effective than other monocyte isolation methods, including plastic adherence and density gradient methods, and has the potential to be developed as a closed system for clinical scale generation of dendritic cells.
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