• 한국미생물·생명공학회지
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• 제39권4호
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• pp.364-373
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• 2011

인간배아줄기세포는 인간배아줄기세포가 가지는 전 분화능 등의 특이적 특성으로 인해 재생의학 분야에서 세포 치료제의 근원으로 널리 각광받고 있다. 그러나, 미분화 상태의 인간배아줄기세포를 세포치료제로 이용하기 위해서는 인간배아줄기세포 주 유래 기능성 세포를 확립이 반드시 요구된다. 본 연구에서는, 미분화 상태의 인간배아줄기세포주로부터 기능성 세포의 확립을 위해, 혈관계통의 세포로 분화를 유도하였으며, 분화 유도 후 세포의 크기 차이를 이용하여 특정 세포군 만을 분리하여 그 기능성을 비교 분석하였다. 그 결과, VEGF를 이용하여 분화 시킨 세포군에서 약 10%의 PECAM 양성 세포군을 확인할 수 있었으며, 분리 및 세포 이식을 위해 세포를 단일 세포군으로 만들었다. 단일 세포군의 형성 후, 유세포 분석기를 이용한 세포 분리 기법을 이용하여 FCS를 기준으로 한 세포 크기의 차이를 이용하여 특정 세포군 만을 분리하여, 하지 허혈 동물 모델로의 이식을 통해, 비 분리 세포군과 치료 효능을 비교 분석을 실시하였다. 세포 이식 4주 후, 혈류량 복구율이 FSC 기준 분리 군의 경우 54%, 비 분리군의 경우 17%를 보이는 것을 확인하였다. 이 결과는, 초기 분화 유도 후 세포 크기차이를 이용한 세포 분리법이 기능성 세포 획득에 이용될 수 있음을 시사한다. 이와 같은 방법을 통해 다양한 종류의 기능성 세포 분리에 이용될 수 있을 것이라 생각된다.

• Biomolecules & Therapeutics
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• 제26권3호
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• pp.298-305
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• 2018

Rhomboid family member 2 gene (Rhbdf2) is an inactive homologue lacking essential catalytic residues of rhomboid intramembrane serine proteases. The protein is necessary for maturation of tumor necrosis factor-alpha ($TNF-{\alpha}$) converting enzyme, which is the molecule responsible for the release of $TNF-{\alpha}$. In this study, Rhbdf2 knockout (KO) mice were produced by CRISPR/CAS9. To see the effects of the failure of $TNF-{\alpha}$ release induced by Rhbdf2 gene KO, collagen-induced arthritis (CIA), which is the representative $TNF-{\alpha}$ related disease, was induced in the Rhbdf2 mutant mouse using chicken collagen type II. The severity of the CIA was measured by traditional clinical scores and histopathological analysis of hind limb joints. A rota-rod test and grip strength test were employed to evaluate the severity of CIA based on losses of physical functions. The results indicated that Rhbdf2 mutant mice showed clear alleviation of the clinical severity of CIA as demonstrated by the significantly lower severity indexes. Moreover, a grip strength test was shown to be useful for the evaluation of physical functional losses by CIA. Overall, the results showed that the Rhbdf2 gene has a significant effect on the induction of CIA, which is related to $TNF-{\alpha}$.

• 한국임상수의학회지
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• 제29권1호
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• pp.82-86
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• 2012

5년령 중성화된 수컷 치와와가 수개월간의 오른쪽 후지 파행 증상으로 내원하였다. 신체 검사에서 통증을 동반한 4등급의 슬개골 내측 탈구를 확인 하였고 오른쪽 왼쪽 후지 파행등급 2 와 0을 각각 확인 하였다. 일반 방사선 사진에서 골증식체와 연골하 병변을 확인 하였고 오른쪽과 왼쪽 후지 골관절증 점수 20 과 12를 각각 확인하였다. 슬개골 탈구 정복술과 자가지방 유래 줄기세포 치료를 보호자의 동의 하에 결정 하였다. $1{\times}10^6$ 개의 줄기세포를 PBS, 인공 괄절액과 함께 슬개골 탈구 정복술 후에 주입 하였다. 주입 후 19개월에 오른쪽 왼쪽 후지 파행등급 0 와 0, 골관절증 점수 19 와 13을 각각 확인 하였고, 5년후에 파행등급 0 와 0, 골관절증 점수 14 와 15를 각각 확인 하였다. 본 증례를 통해 자가지방 유래 줄기 세포치료를 받은 관절에서 골관절증이 감소 된 것을 확인 할 수 있었다.

• Korean Journal of Acupuncture
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• 제21권2호
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• pp.47-67
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• 2004

Objectives : In the present study, the effect of Scolopendrid Water-Alcohol Extract (SWAE) applied to acupuncture point BL23 (Shinsu) on the neuropathic pain was examined. A common source of persistent pain in humans is the neuropathic pain. Anti-convulsant drugs are used to treat the neuropathic pain. In the oriental medicine, Scolopendrid was used for long time to treat convulsant syndrome and back pain, etc. Methods : On the bases of the Scolopendrid clinical application, the effect of SWAE applied to the acupuncture point was tested in the rat model of neuropathic pain. Neuropathic pain was induced by tight ligation of L5 spinal nerve. When rats developed pain behaviors, One hundred microliter of SWAE was applied into the ipsilateral BL23 point at a dose of 10 mg/ml under enflurane anesthesia. The foot withdraw latency of the hind limb was measured for an indicator of pain level after each manipulation. Results : SWAE injection increased the mechanical threshold of the foot in the rat model of neuropathic pain significantly for the duration of 4h, suggesting a partial alleviation of pain. SWAE applied to BL23 point produced a significant improvement of mechanical sensitivity of the foot lasting for at least 4h. However, neither contralateral BL23 point, ST25 (Chonchu) point, nor LR3 (Taechung) point produce as much increase of mechanical sensitivity as ipsilateral BL23 point. And, this increase of mechanical sensitivity was dose-dependent. The improvement of mechanical threshold was interpreted as an analgesic effect. In addition, the analgesic effect of Scolopendrid 4 mg/kg injection is equivalent to that of gabapentin 50 mg/kg injection. The relations between SWAE-induced analgesia and endogenous nitric oxide(NO), inducible NO synthase (iNOS)/neuronal NO synthase (nNOS) were also examined. Results were turned out that both NO production and nNOS/iNOS protein expression which are increased by nerve injury were suppressed by SWAE injection applied to BL23 point. Conclusions : The data suggest 1) that SWAE produces a potent analgesic effect on the neuropathic pain model in the rat and 2) that SWAE-induced analgesia modulate endogenous NO through the suppression of nNOS/iNOS protein expression.

• Reproductive and Developmental Biology
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• 제34권2호
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• pp.89-94
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• 2010

Melatonin is induced by light information through the retina and leads to growth factor activation. Thus, we investigated the effects of melatonin by controlling the photoperiod of growing young rats. Male Sprague-Dawley rats (n=6; 4 weeks old) were divided into two experimental groups: the L/D group (normal photoperiod; light/dark: 12/12 h; lights on at 9:00 a.m.) and the L/L group (light/light: 24 h). Rat body weight and food consumption were measured daily for 8 weeks. After 8 weeks, the rats were anesthetized with a mixture of ketamine (50 mg/kg) and xylazine (10 mg/kg) and sacrificed. Tissue was then collected for RNA isolation (from brain, heart, liver, kidney, adrenal gland, testis, tibia, hind limb muscles). Also, serum was isolated from blood using a centrifugal separation. The L/L group had significantly lower body weight than the L/D group from 4 to 6 weeks (p<0.05). The L/D group had increased tissue mass, compared with the L/L group, but the difference was not statistically significant. The L/D group had a significantly higher melatonin concentration than the L/L group between the hours of midnight and 2:00 a.m (p<0.01). These results indicate that photoperiod length may affect the secretion of melatonin from the pineal gland. Also, the reduction of nocturnal melatonin secretion may retard the development of growing young rats. In future studies, we plan to compare exogenous melatonin administration with endogenous melatonin concentration induced by photoperiod control. Moreover, we will confirm whether the effects seen in pathological animal models can be reversed by controlling the photoperiod.

• 한국어병학회지
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• 제22권1호
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• pp.67-73
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• 2009

양식산 복어에서는 tetrodotoxin(TTX)가 존재하지 않는다고 일반적으로 생각되고 있다. 본 연구에서는 실제로 양식산 황복(Takifugu obscurus)에는 TTX가 존재하지 않는지 확인하기 위해 생쥐를 이용한 bioassay로 여러 장기를 동시에 분석하였다. 자연산에서는 동물이 치사하는 수준의 독력을 함유한 시료가 있었으며 정소>간장>장>근육>피부의 순으로 독력이 확인되었다.양식산 시료에서는 TTX의 미량존재로 추정되는 졸림, 행동둔화, 후지마비 등의 증상이 나타났지만 치사동물은 발견되지 않았다. 이 결과는 양식 산 황복에서 TTX가 매우 낮은 농도로 존재할가능성을 제시하지만 기기분석법을 이용한 정확한 정량이 필요하다고 판단된다.

• Korean Journal of Acupuncture
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• 제21권3호
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• pp.59-76
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• 2004

Objectives : The usage of acupuncture has gained popularity for certain chronic pain conditions. However, the efficacy of acupuncture in various diseases has not been fully established and the underlying mechanism is not clearly understood. In the present study, the effect of electroacupuncture (EA) applied to yangno$(SI_6)$ on the neuropathic pain was examined. Methods : A common source of persistent pain in human is a neuropathic pain. Neuropathic pain was induced by tight ligation of L5 spinal nerve. When rats developed pain behaviors, EA was applied for 30 min. under enflurane anesthesia with repeated train stimuli at the intensity of 10X of muscle twitch threshold. The foot withdraw latency of the hind limb was measured for an indicator of pain level after each manipulation. Results : EA increased the mechanical threshold of the foot in the rat model of neuropathic pain significantly for the duration of 1 hr. suggesting a partial alleviation of pain. EA applied to SI6 point produced a significant improvement of mechanical sensitivity of the foot lasting for at least 1 h. However, $ST_{36}$ point did not produce any significant increase of mechanical sensitivity. The improvement of mechanical threshold was interpreted as an analgesic effect. The analgesic effort was specific to the acupuncture point since the analgesic effect on the neuropathic pain model could not be mimicked by EA applied to a point, $ST_{36}$. In addition, this analgesic effect of EA is mediated by a adrenergic mechanism of descending control of spinal cord from the brain. Conclusions : The data suggest that EA produces a potent analgesic effect on the neuropathic pain model in the rat; and 2) that EA-induced analgesia is mediated by a adrenergic mechanism of descending control in a point specific manner.

• 한국임상수의학회지
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• 제28권3호
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• pp.323-327
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• 2011

8개월령, 수컷 진도개가 오른쪽 후지 파행으로 의뢰되었다. 이 개는 방사선 검사를 통하여 오른쪽 대퇴골의 단순 골절로 진단되었다. 대퇴골절의 수술적 교정 이후, 술 후 진통제로 합성 아편계 유사 약물인 트라마돌이 정맥 내 주사로 투여되었다. 트라마돌의 투여 직후 발작이 시작되었다. 집중적인 환자 관리에도 불구하고 투여 약 17시간 후 이 개는 폐사하였다. 폐사를 하게 된 원인과 관련하여 트라마돌의 투여에 의한 발작이 연관되어 있을 것으로 추측할 수 있었다. 부검을 실시한 결과 조직병리학적 분석에 따라 본 환자는 괴사성 수막뇌염(NME)으로 진단되었다. 괴사성 수막뇌염을 가진 개에서 트라마돌의 투여가 폐사를 일으킨 발작의 유발과 관련되어 있을 것으로 생각되었다. 본 증례를 통해 트라마돌이 통증 관리에 있어 안전하고 효과적인 약물로 알려져 있으나 뇌염, 뇌수두증, 뇌병증 등의 신경계질환이 있는 환자에서는 주의 깊게 사용되어야 한다는 것을 알 수 있었다.

• Korean Journal of Acupuncture
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• 제17권1호
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• pp.101-121
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• 2000

The purpose of this morphological studies was to investigate the relation to the meridian, acupoint and nerve. The common locations of the spinal cord and brain projecting to the the gallbladder, GB34 and common peroneal nerve were observed following injection of transsynaptic neurotropic virus, pseudorabies virus(PRV), into the gallbladder, GB34 and common peroneal nerve of the rabbit. After survival times of 96 hours following injection of PRV, the thirty rabbits were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by PRV immunohistochemical staining method, and observed with light microscope. The results were as follows: 1. In spinal cord, PRV labeled neurons projecting to the gallbladder, GB34 and common peroneal nerve were founded in thoracic, lumbar and sacral spinal segments. Densely labeled areas of each spinal cord segment were founded in lamina V, VII, X, intermediolateral nucleus and dorsal nucleus. 2. In medulla oblongata, The PRV labeled neurons projecting to the gallbladder, GB34 and common peroneal nerve were founded in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nucleus, medullary reticular nucleus, dorsal motor nucleus of vagus nerve, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus, gigantocellular nucleus, lateral paragigantocellular nucleus, principal sensory trigeminal nucleus and spinal trigeminal nucleus. 3. In Pons, PRV labeled neurons were parabrachial nucleus, Kolliker-Fuse nucleus and cochlear nucleus. 4. In midbrain, PRV labeled neurons were founded in central gray matter and substantia nigra. 5. In diencephalon, PRV labeled neurons were founded in lateral hypothalamic nucleus, suprachiasmatic nucleus and paraventricular hypothalamic nucleus. 6. In cerebral cortex, PRV labeled neuron were founded in hind limb area.This results suggest that PRV labeled common areas of the spinal cord projecting to the gallbladder, GB34 and common peroneal nerve may be first-order neurons related to the somatic sensory, viscero-somatic sensory and symapathetic preganglionic neurons, and PRV labeled common area of the brain may be first, second and third-order neurons response to the movement of smooth muscle in gallbladder and blood vessels.These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory system monitoring the internal environment, including both visceral sensation and various chemical and physical qualities of the bloodstream. The present morphological results provide that gallbladder meridian and acupoint may be related to the central autonomic pathways.

• Journal of Korean Neurosurgical Society
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• 제30권7호
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• pp.831-841
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• 2001

Objective : Somatosensory evoked potentials(SSEPs) have been used widely both experimentally and clinically to monitor the function of central nervous system and peripheral nervous system. Studies of SSEPs have reported the various recording techniques and patterns of SSEP. The previous SSEP studies used scalp recording electrodes, showed mean vector potentials which included relatively constant brainstem potentials(far-field potentials) and unstable thalamocortical pathway potentials(near-field potentials). Even in invasive SSEP recording methods, thalamocortical potentials were variable according to the kinds, depths, and distance of two electrodes. So they were regarded improper method for monitoring of upper level of brainstem. The present study was conducted to investigate the characteristics of somatosensory evoked field potentials(SSEFPs) of the cerebral cortex that evoked by hindlimb stimulation using ball electrode and the pathways of SSEFP by recording the potentials simultaneously in the cortex, VPL nucleus of thalamus, and nucleus gracilis. Methods : In the first experiment, a specially designed recording electrode was inserted into the cerebral cortex perpendicular to the cortical surface in order to recording the constant cortical field potentials and SSEFPs mapped from different areas of somatosensory cortex were analyzed. In the second experiment, SSEPs were recorded in the ipsilateral nucleus gracilis, the contralateral ventroposterolateral thalamic nucleus(VPL), and the cerebral cortex along the conduction pathway of somatosensory information. Results : In the first experiment, we could constantly obtain the SSEFPs in cerebral cortex following the transcutaneous electrical stimulation of the hind limb, and it revealed that the first large positive and following negative waves were largest at the 2mm posterior and 2mm lateral to the bregma in the contralateral somatosensory cortex. The second experiment showed that the SSEPs were conducted by way of posterior column somatosensory pathway and thalamocortical pathway and that specific patterns of the SSEPs were recorded from the nucleus gracilis, VPL, and cerebral cortex. Conclusion : The specially designed recording electrode was found to be very useful in recording the localized SSEFPs and the transcutaneous electrical stimulation using ball electrode was effective in evoking SSEPs. The characteristic shapes, latencies, and conduction velocities of each potentials are expected to be used the fundamental data for the future study of brain functions, including the hydrocephalus model, middle cerebral artery ischemia model, and so forth.