• 한국임상수의학회지
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• 제34권5호
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• pp.377-380
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• 2017

An 8-year-old, shih-tzu female dog was referred due to neurological signs including paraparesis and back pain. On the complete blood count, hematologic analysis showed elevated leukocytosis. Serum biochemical analysis revealed elevated serum alkaline phosphatase concentration and C-reactive protein concentration. On the neurologic exam, the dog was suspected to have thoracolumbar myelopathy. On magnetic resonance imaging, there were masses within the spinal canal at L1-3 intervertebral disc space that were located dorsal to spinal cord. It was hyperintense on T1-, T2-weighted magnetic resonance images, Fluid-attenuated inversion recovery, and fat suppression images. The contrast-enhanced T1-weighted images showed no enhancement. The lesions were well circumscribed. The spinal cord was compressed and displaced ventrally by the mass. After removal of the masses via L1-L3 dorsal laminectomy, pyogranulomatous inflammation was confirmed by histopathological examination. Six months after surgery, the dog recovered uneventfully and remained fully ambulatory with no neurological deficits. This case demonstrates the utility of magnetic resonance imaging for the diagnosis of spinal canal pyogranulomatous inflammation.

• Archives of Reconstructive Microsurgery
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• 제19권1호
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• pp.12-20
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• 2010

Purpose: Industrial punch accidents involving fingers cause segmental injuries to tendons and neurovascular bundles. Although multiple-level segmental amputations are not replanted to regain function, most patients with an amputated finger want to undergo replantation for cosmetic as much as functional reason. The authors describe four cases of digital amputation by an industrial punch that involved the reinstatement of the amputated finger involving a joint and neurovascular bundle. Amputated segments were replanted to restore amputated surfaces and distal segments. Methods: A single institution retrospective review was performed. Inclusion criteria of punch injuries requiring replantation were applied to patients of all demographic background. Injury extent (size, tissue involvement), operative intervention, pre- and postoperative hand function were recorded. Result: Four cases of amputations were treated at our institute from 2004 to 2008 from industrial punch machine injury. Average patient age was 32.5 years (25~39 years) and there were three males and one female. Sizes of amputated segments ranged from $1.0{\times}1.0{\times}1.2\;cm^3$ to $3{\times}1.5{\times}1.6\;cm^3$. Tenorrhaphy was conducted after fixing fractured bone of the amputated segments with K-wire. Proximal and distal arteries and veins were repaired using the through & through method. The average follow-up period was thirteen months (2~26 months), and all replanted cases survived. Osteomyelitis occurred in one case, skin grafting after debridement was performed in two cases. Because joints were damaged in all four cases, active ranges of motion were much limited. However, a secondary tendon graft enhanced digit function in two cases. The two-point discrimination test showed normal values for both static and dynamic tests for three cases and 9 mm and 15 mm by dynamic and static testing, respectively, in one case. Conclusion: Though amputations from industrial punch machines are technically challenging to replant, our experience has shown it to be a valid therapy. In cases involving punch machine injury, if an amputated segment is available, the authors recommend that replantation be considered for preservation of finger length, joint mobility, and overall functional recovery of the hand.

• Journal of Korean Neurosurgical Society
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• 제55권1호
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• pp.43-47
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• 2014

Patients with neurofibromatosis 1 (NF1) are predisposed to develop central nervous system tumors, due to the loss of neurofibromin, an inactivator of proto-oncogene Ras. However, to our knowledge, only three cases of ependymomas with NF1 have been reported in the literature. The authors present a case of NF1 patient with a spinal cord ependymoma. She was referred for about half a year history of increasing numbness that progressed from her fingers to her entire body above the bellybutton. Magnetic resonance imaging revealed a relative-demarcated, heterogeneously enhanced mass lesion accompanied by perifocal edema in C5-7 level, a left-sided T11 spinous process heterogeneously enhanced mass in soft tissue, intervertebral disk hernia in L2-5 level, and widespread punctum enhancing lesion in her scalp and in T11-L5 level. The patient underwent C5-7 laminectomies and total excision of the tumor under operative microscope, and intraoperative ultrasonography and physiological monitoring were used during the surgery. Histopathologically, her tumor was found to be a ependymoma without malignant features (grade II in the World Health Organization classification). Therefore, no adjuvant therapy was applied. Following the operation, the patient showed an uneventful clinical recovery with no evidence of tumor recurrence after one year of follow-up.

• Journal of Chest Surgery
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• 제33권8호
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• pp.605-612
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• 2000

Background: Ischemic preconditioning enhances the tolerance of myocardium against ischemia/reperfusion injury, with the enhancement of the recovery of post-ischemic myocardial function. This study was disigned to assess whether the protective effect of ischemic preconditioning could provide one additional hour of myocardial preservation in four hour myocardial ischemia in a rate heart. Material and method: Fourty four Spargue-Dawley rats, weighing 300~450gm, were divided into four groups. Group 1(n=7) and group 3(n=12) were subjected to 30 minutes of aerobic Langendorff perfusion without ischemic preconditioning and then preserved in saline solution at 2~4$^{\circ}C$ for 4 hours and 5 respectively. Group 2(n=7) and group 4(n=18) were perfused in the same way for 20 minutes, followed by 3 minutes of global mormothermic ischemia and 10 minutes of perfusion and then preserved in the same cold saline solution for 4 hours and 5 hours respectively. Heart rate, left ventricular developed pressure(LVDP), and coronary flow were measured at 15 minutes during perfusion as baseline. Spontaneous defibrillation time was measured after reperfusion. Heart rate, LVDP, and coronary flow were also recorded at 15 minutes, 30 minutes, and 45 minutes during reperfusion. Samples of the apical left ventricular wall were studied using a transmission electron microscope. Result: Time of spontaneous defibrillation(TSD) was significantly longer in group 4 than in group 1(p<0.001), and TSD in group 1 was significantly longer in comparision to that of group 2(p<0.05). Heart rate at 45 minutes was significantly higher in group 1 than in group 4(p<0.05). Heart rate at 15 min was significantly higher in group 2 than in group 1(p<0.001) and in group 4 than in group 3(p<0.05). Left ventricular developed pressure(LVDP) at 30 minutes and 45 minutes was higher in group 1 than in group 4(p<0.01), LVDP at 45 minutes was higher in group 4 than in group 3(p<0.05). Rate-pressure product(RPP) at 30 minutes and 45 minutes was higher in group 1 than in group 4(p<0.05). RPP at 15 minutes was higher in group 2 than in group 1(p<0.01). RPP at 30 minutes and 45 minutes was higher in group 4 than in group 3(p<0.05). Group 2 showed relatively less sarcoplasmic edema and less nuclear chromatin clearance than group 1. Group 4 showed less myocardial cell damage than group 3, group 4 showed less myocardial cell damage than group 3, group 4 showed more myocardial cell edema than group 1. Conclusion: Ischemic preconditioning enhanced the recovery of postischemic myocardial function after 4 hours and 5 hours preservation. However, it was not demonstrated that ischemic preconditioning could definitely provide one additional hour of myocardial preservation in four hour myocardial ischemia in a rat heart.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제31권3호
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• pp.199-218
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• 2005

Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권5호
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• pp.375-395
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• 2006

Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

• Journal of Chest Surgery
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• 제33권7호
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• pp.531-540
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• 2000

연구배경; 심근세포내 에너지원인 단원 pool의 고갈이나 당대사의증가와 이로 인한 유산의 심근세포내 축적은 허혈 심근세포 손상의 중요한 원인으로 알려져 있다. 그러나 역설적으로 당원이 결핍된 용액으로 짧은 기간 동안 허혈-재관류를 반복(IP)한 경우와 유사한 결과를 가져올 수 있는 가능성을 조사하여 세포내 신호전달체계 중 PKC와의 관련성을 알아보고자 하였다. 대상 및 방법 ; Langendorff방법에 따라 관류하여 기준설 혈역학 값이 유지되면 전체허혈(5분)-재관류(10분) 1회 실시로 IP를 유도하고 45분 동안 전체 허혈후 120분 동안 재관류하였다. (IP군. n=13). 허혈 대조군(n=10)에서는 IPdjqt이 45분 동안 전체 허혈후 120 동안 재관류를 실시하였다. Glucose 결핍용액 투여 전처치군(n=12)에서는 기준선 혈역학 값이 유지되면 5분 동안 glucose를 포함하지 않은 관류액으로 관류한 후 10분 동안 표준 관류액으로 측정하였으며 실험 종료후 PKC활성도는 PKC-specific peptide와 32P-${\gamma}$-ATP incorporation으로 PKC활성도(nmol/g tissue)를 측정하였따. PKC 동종효소의 발현정도는 단클론항체($\alpha$,$\beta$,$\delta$,$\varepsilon$,ζ 등)를 사용하여 Western blot로 확인하였다. 심근경색 크기는 1% tetrazolium chloride로 염색하여 형태 계측하였다. 결과; 45분 동안 허혈LVDP(LV developed pressure), dP/dt 등은 다른 실험군에 비하여 IPrns에서 현저히 증가하였으나 glucose 결핍용액 투여 전처치군에서는 허혈 대조군과 큰 차이가 없었으며 관혈류량은 모든 실험군 사이에서 차이를 나타내지 않았다. 그러나 glucose 결핍용액 트여 저너치군(15$\pm$3.9%)과 IP군(19$\pm$1.2%)에서는 허혈 대조군(39$\pm$2.7%)에 비하여 심근경색 범위의 현저한 감소를 볼 수 있었다. (p<0.05). PKC 활성도는 기준선과 비교하여 허혈 대조군에서는 87% 정도를 감소하였으며 (p<0.05), IP 실시한 후와 IP후 45분 동안 허혈을 실시한 결우에는 각각 119, 145%로 현저히 증가하였다. (p<0.01). PKC 동종효소중 $\beta$, $\delta$, ζ 등에서는 발현정도에 유의한 변화가 없었던 반면 $\alpha$ 및 $\varepsilon$에서 양적인 변화를 관찰할 수 있었다. PKC-$\alpha$의 세포질분획의 발현은 기준선이나허혈 대조군과 비교하여 IPgn에 증가하는 경향을 나타내었으나, 이외의실험군에서는 큰 변화를 볼 수 없었다. PKC-$\alpha$의 세포막분획은 IP후롸, glucose 결핍용액 투여 전처치후, glucose 결핍용액 투여 전치치후 45분 동안 허혈후에 증가하는 경향을 나타내었다. PKC-$\varepsilon$의 세포질분획의 발현은 기준선이나 허혈 대조군과 비교하여 IPgn나 IPgn 45분 동안 허혈후, glucose 결핍용액 투여 전처치에 증가하는 경향을 나타내었으며 PKC-$\varepsilon$의 세포막분획은 IP후 45분 동안 허혈후, 또는 glucose 결핍용액 투여 전처치후에 발현이 증가하는 경향을 나타내었다. 결론 ; 이상으로 적출 관류 토끼 심장에서 glucose 결핍용액 투여로 전처치할 경우 후속된 장시간 동안의 허혈에 대하여 좌심실기능 회복 증가는 기대할 수 없으나 심근경색 범위가 감소되거나 한정되는 보호효과가 있음을 알 수 있었다.

• Journal of Chest Surgery
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• 제32권9호
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• pp.773-780
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• 1999

연구배경: 적출 흰쥐 심장에서 짧은 기간 동안 칼슘 파라독스를 반복하면(칼슘결핍 및 재투여에 의한 칼슘 전처치, calcium-free preconditioning-calcium depletion and repletion)하면 후속 되는 보다 긴 기간 동안의 허혈에 대하여 재관류시 심장기능 회복 증가, 괴사크기의 감소 등 허혈성 전처치에서 나타나는 것과 유사한 심근보호 효과가 있음이 최근 보고되어 있다. 본 실험에서는 적출 토끼 심장을 이용하여 칼슘결핍용액으로 전처치할 경우 ischemic preconditioning(이하 IP)와 유사한 심근보호 효과가 나타나는가의 유무를 기능 및 형태학적 측면에서 확인하고자 하였다. 대상 및 방법: 체중 1.5∼2.0 kg의 건강한 흰토끼(New Zealand White rabbit)의 적출 심장을 이용하여 Langendorff 방법에 따라 관류하여 일정한 기준치가 유지되면 전체 허혈(5분)-재관류(10분) 1회 실시로 IP를 유도하고 45분 동안 전체허혈후 120분 동안 재관류하였다(IP군, n=7). 대조군(n=7)에서는 IP없이 45분 동안 전체허혈후 120분 동안 재관류를 실시하였다. 칼슘결핍용액 투여 전처치군(n=7)에서는 5분 동안 칼슘결핍용액을 투여 후 10분 동안 칼슘이 포함된 관류액으로 관류하고 45분 동안 허혈을 실시한 후 120분 동안 재관류하였다. 허혈후 재관류 기간 동안 좌심실 기능, 관혈류 등을 측정하였으며 심근괴사 크기는 1% tetrazolium으로 염색하여 형태계측 하였다. 결과는 분산분석(ANOVA)을 실시하여 유의성이 있다고 판정되면 Tukey's post-hoc test로 검정하였다. 결과: 45분 동안 허혈후 LVDP, dP/dt, 관혈류 등은 다른 실험군에 비하여 IP군에서 현저히 증가하였으나 칼슘결핍용액 투여 전처치군에서는 오히려 허혈 대조군에 비하여 현저히 감소하였다. 그러나 칼슘결핍용액 투여 전처치군에서는 IP군에서와 같이 허혈 대조군에 비하여 심근괴사 범위가 현저히 감소되었다. 결론: 이상의 결과로 적출 관류 토끼 심장에서 칼슘결핍용액 투여로 전처치할 경우 후속된 장시간 동안의 허혈에 대하여 좌심실기능 회복 증가는 기대할 수 없으나 심근괴사 범위 한정 등의 보호효과가 있는 것으로 생각된다.