• Parasites, Hosts and Diseases
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• 제26권1호
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• pp.27-32
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• 1988

뇌낭미충증의 면역혈청학적 진단에 있어서 간접 형광항체 법의 유용성을 평가하기 위하여 효소연결성 면역홉착 검사와 비교 검토하였다. 검사대상자는 확진된 뇌낭미충증 환자의 혈청 163예, 다른 뇌신경 증상 환자, 조층 및 흡충류 감염자 101예 및 건강인 대조군 100예로서 모두 364예이었다. 간점 형광항체 반응에는 인체 유구낭미충의 낭벽 항원을, 효소연결성 면역흡착 검사에는 낭액 항원을 사용하여 혈청내 특이 IgG 항체를 조사한 결과 두 방법간의 민감도 및 특이도에 큰 차이가 없었으며, 양성 및 음성의 동일한 힐청을 검사하였을 때 낭미충증 혈청의 90.8%가 서로 합치되어 밀접한 연관성을 나타내었다. 또한 장내 조충 감염증의 경우 두 방법 모두에서 높은 교차반응을 나타내었으나 간접 형광항체반응의 특이성이 더 좋았으며 특히 간접 형광항체 반응은 흡충류 감염자 혈청에서 교차반응을 나타내지 않았다. 이와 같은 결과는 혈청만을 사용하였을 경우 간접 형광항체반응의 민감도나 특이도가 효소연결성 면역홉착 검사와 차이가 없으며 뇌낭미충증의 진단에 매우 유용함을 나타내고 있었다.

• Archives of Plastic Surgery
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• 제40권6호
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• pp.748-753
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• 2013

Background Nasal reconstruction is one of the most difficult challenges for the head and neck surgeon, especially in the case of complex full thickness defects following malignant skin tumor resection. Full-thickness defects require demanding multi-step reconstruction. Methods Seven patients underwent surgical reconstruction of full-thickness nasal defects with a bi-pedicled forehead flap shaped appropriately to the defect. Patients were aged between 58 and 86 years, with a mean age of 63.4 years. All of the tumors were excised using traditional surgery, and in 4 of the patients, reconstruction was performed simultaneously following negativity of fresh frozen sections of the margins under general anesthesia. Results Nasal reconstruction was well accepted by all of the patients suffering non-melanoma skin tumors with acceptable cosmetic outcomes. The heart-shaped forehead flap was harvested in cases of subtotal involvement of the nasal pyramid, while smaller defects were reconstructed with a wing-shaped flap. No cartilaginous or osseous support was necessary. Conclusions This bi-pedicled forehead flap was a valid, versatile, and easy-to-implement alternative to microsurgery or multi-step reconstruction. The flap is the best indication for full-thickness nasal defects but can also be indicated for other complex facial defects in the orbital (exenteratio orbitae), zygomatic, and cheek area, for which the availability of a flap equipped with two thick and hairless lobes can be a valuable resource.

• Journal of Gastric Cancer
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• 제17권4호
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• pp.331-341
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• 2017

Purpose: This study aimed to evaluate the clinical significance of microscopic invasion to determine the adequate resection margin in early gastric cancer (EGC). Materials and Methods: A retrospective review was performed that included patients who underwent gastrectomy for clinical early gastric cancer (cEGC) at Seoul National University Hospital between January 2007 and December 2010. After subtracting the microscopic resection margin from the gross resection margin for each proximal or distal resection margin, microscopic invasion was represented by the larger value. Microscopic invasion and its risk factors were analyzed according to the clinicopathologic characteristics. Results: In total, 861 patients were enrolled in the study. Microscopic invasion of cEGC was $6.0{\pm}12.8mm$, and the proportion of patients with microscopic invasion ${\geq}0mm$ was 78.4%. In the risk group, tumor location, pT stage, and differentiation did not significantly discriminate the presence of microscopic invasion. The microscopic invasion of EGC-IIb was $13.9{\pm}16.8mm$, which was significantly greater than that of EGC-I. No linear correlation was observed between the overall tumor size and microscopic invasion (R=0.030). The independent risk factors for microscopic invasion ${\geq}20mm$ were EGC-IIb vs. EGC-I/IIa/IIc/III (odds ratio [OR], 3.103; 95% confidence interval [CI], 1.533-6.282; P=0.002) and male vs. female sex (OR, 1.655; 95% CI, 1.012-2.705; P=0.045). Conclusions: Male sex and EGC-IIb were independent risk factors for microscopic invasion ${\geq}20mm$. Examination of intraoperative frozen sections is highly recommended to avoid resection margin involvement, especially in cases of EGC-IIb.

• 한국동물학회지
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• 제39권4호
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• pp.410-418
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• 1996

Gangiloside는 포유동물세포에 편재하는 막성분으로서, 이들은 세포상호간의 접착, 분화 및 정보전달과정에 과여하는 것으로 알려지고 있다. Rat난소는 주요한 Gangiloside로서 GM3를 함유하고 있으며, 본 연구에서는 폐쇄난포에서 이들의 분포여부와 난포의 발달 과정에서의 변화여부를 조사하기 위하여, Rat 난소의 동결절편을 이용해 GM3를 포함 11종류의 Gangilo-series Gangiloside에 대해 특이한 단일항체로서 염색시켰다. 폐쇄난포들에서 GM3는 난포발달이 진행되는 동안 시간적, 공간적으로 서로 다른 양식으로 발현하였다. 그러나 GM1을 포함한 다른 종류의 Gangiloside들은 면역조직화학적으로 검출되지 않았다. 일차난포에서 관찰되는 폐쇄난포들에서 GM3는 모든 교막세포와 난자에 인접한 과립막세포의 일부에서 발현하였다. 이차난포의 시기에서 이들 폐쇄난포의 GM3는 모든 교막세포와 과립막세포들에서 발현하였다. 이어서 발달하고 있는 그라프난포의 시기에서 관찰되는 폐쇄난포의 GM3발현은 이차난포에서의 분포패턴과 유사함을 보여 주었다.

• Archives of Pharmacal Research
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• 제24권4호
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• pp.360-366
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• 2001

Gangliosides are ubiquitous membrane components in mammalian cells and are suggested to play important roles in various functions such as cell-cell interaction, adhesion, cell differentiation, growth control and signaling. Among all ganglio-series gangliosides, GM3 has the simplest carbohydrate structure, and has been shown as a major gangliosides, in male reproductive system. To study GM3 distribution in the seminiferous tubule and epididymis, frozen sections were stained with specific monoclonal antibody (MAb) against ganglioside GM3. In the seminiferous tubule of testis, pachytene spermatocytes and spermmatids expressed ganglioside GM3, but not in spermatogonia and sertoli cells. Spermatogonia and sertoli cells near the basement membrane were negatively reacted to anti-GM3. In the epididymis, GM3 was expressed only in some interstitial cells. Taken togethers, these results suggest that the expression of ganglioside GM3 in rat seminiferous tubule and epididymis is spatio-temporally regu lated during spermatogenesis.

• Journal of Korean Neurosurgical Society
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• 제55권3호
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• pp.131-135
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• 2014

Objective : With the growing interests of bacteria as a targeting vector for cancer treatment, diverse genetically engineered Salmonella has been reported to be capable of targeting primary or metastatic tumor regions after intravenous injection into mouse tumor models. The purpose of this study was to investigate the capability of the genetically engineered Salmonella typhimurium (S. typhimurium) to access the glioma xenograft, which was monitored in mouse brain tumor models using optical bioluminescence imaging technique. Methods : U87 malignant glioma cells (U87-MG) stably transfected with firefly luciferase (Fluc) were implanted into BALB/cAnN nude mice by stereotactic injection into the striatum. After tumor formation, attenuated S. typhimurium expressing bacterial luciferase (Lux) was injected into the tail vein. Bioluminescence signals from transfected cells or bacteria were monitored using a cooled charge-coupled device camera to identify the tumor location or to trace the bacterial migration. Immunofluorescence staining was also performed in frozen sections of mouse glioma xenograft. Results : The injected S. typhimurium exclusively localized in the glioma xenograft region of U87-MG-bearing mouse. Immunofluorescence staining also demonstrated the accumulation of S. typhimurium in the brain tumors. Conclusion : The present study demonstrated that S. typhimurium can target glioma xenograft, and may provide a potentially therapeutic probe for glioma.

• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.100-115
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• 1999

The purpose of this study was to investigate the distribution of calcitonin gene-related peptide containing nerve fibers in rat pulp after dentinl injury by means of immunohistochemistry and confocal laser scanning microscope. The Spague-Dawley rats weighing about 250-300gm were used. The animals were devided into normal control and experimental groups. Experimental animals were sacrified 1, 2, 4, 7, 10, 21days after dentinal injury (dentin cutting, and then acid etching with 35% phosphoric acid) on the maxillary molar teeth. The maxillary teeth and alveolar bone were removed and immersed in the 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), then were decalcified with 15% formic acid for 10 days. Serial frozen $50{\mu}m$ thick sections were cut on a cryostat. The rabbit CGRP antibody was used as a primary antibody with a dilution of 1:2000 in 0.01M PB. The sections were incubated for 48 hours at $4^{\circ}C$, and placed into biotinylated antirabbit Ig G as a secondary anti body with dilution of 1:200 in 0.01M PB and incubated in ABC(avidin-biotin complex). The peroxidase reaction was visualized by incubating the sections in 0.05% 3,3 diaminobenzidine tetrahydrochloride containing 0.02% $H_2O_2$. For the confocal laser scanning microscopic examination, Primary antibody reaction was same as immunoperoxidase stainning, but fluorescein isothiocyanate(FITC)-conjugate antirabbit IgG as a secondary antibody was used. The confocal laser scanning microscope was used for the examination. A series of images of optical sections was collected with a 20x objective at $3{\mu}m$ intervals throughout the depth of specimen. FITC fluerescence was registrated through a 488nm and 568nm excitation filter, and images were saved on optical disk. The stereoscopic images and three dimentionnal images were reconstructed by computer software, and then were analyzed. The results were as follows : 1. In normal control group, CGRP containing nerve fibers were coursed through the root with very little branching, and then formed a dense network of terminals in coronal pulp. 2. A slight increase in CGRP containing nerve fibers at 1 and 2day postinjury was noted subjacent to the injury site. In the 4day group, there were an extensive increase in the number of reactive fibers, followed by a partial return toward normal levels at 7~10 day postinjury, and return by 21days. 3. The sprouting of the CGRP containing nerve fibers was evident within 2day after dentinal injury, and by 4days there was a maximal increased, but was decreased at 7days and returned to normal 10~21 day postinjury. 4. In confocal laser scanning microscopic exammination, the distinct distribution pattern and sprouting reaction of CGRP containing nerve fibers were observed in stereoscopic images and three dimentional images. These results suggest that CGRP containing nerve fiber can be important role in the response to dental injury and pain regulation.

• Restorative Dentistry and Endodontics
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• 제31권3호
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• pp.192-202
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• 2006

본 연구의 목적은 흰쥐 상악 대구치를 발거 한 후 급속 냉동보존을 통해 치아를 보관하였을 때 in vivo MTT 검색법을 이용하여 치주인대세포의 활성도를 측정하고자 하였다. 실험 방법은 74마리 4주령의 암컷 Sprague-Dawley계의 흰쥐를 사용하여 각군당 10마리의 쥐에서 상악 좌우 제1, 2 대구치를 발거하여 모두 40개의 치아를 사용하였다. 대조군은 즉시 발치군이며 냉동군은 F medium에 5% Dimethylsulfoxide (DMSO) 6% Hydroxyethyl starch (HES)를 포함한 군(1군), 10% DMSO를 포함한 군(2군) 그리고 $Viaspan^(R)$에 5% DMSO 6% HES(3군), 10% DMSO를 포함한 군 (4군)으로 나누어 1주일간 액체질소에 냉동한 뒤 해동하였다. 냉장군은 F medium (5군)와 $Viaspan^(R)$ (6군)에 넣어 1주간 $4^{\circ}C$ 냉장고에서 보관하였다. 냉동 및 냉장보존한 후에는 in vivo MTT검색법을 시행하였다. 개개 치아의 치근면 단위면적으로 표준화하기 위해 치근을 1% eosin Y 용액에 12시간 담근 후 1% acid alcohol로 용해시켜 530 nm에서 측정한 흡광도 값을 in vivo MTT 측정값으로 나누었다. 통계 분석을 위해 Two way ANOVA와 Duncan's Multiple Range Test를 95% 신뢰 구간에서 시행하였다. 그리고 각 군당 2개의 치아를 같은 방법으로 처리한 후 $10{\mu}m$ 두께로 냉동 절단하여 광학 현미경과 편광 현미경하에서 관찰하였다. 1, 2군은 3, 4군보다. 높은 흡광도를 나타내었으며 6군은 5군보다. 높은 흡광도를 나타내었다(p<0.05). 광학 현미경 관찰에서 MTT 결정은 파란색을, 편광현미경에서는 밝은 오렌지색을 나타내었고 전체적으로 in vivo MTT검색법의 결과와 같은 양상을 나타내었다. 본 연구의 결과 5%, 10% DMSO를 사용한 F medium 배지로 냉동보존한 경우가 $Viaspan^(R)$을 배지로 냉동하였을 때나 냉장보존 하였을 때보다 통계적으로 유의차 있게 높은 세포 활성도를 나타내었다(p<0.05).

• 대한소아치과학회지
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• 제28권1호
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• pp.25-31
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• 2001

말초신경 손상에 의한 VIP의 변화를 연구하기 위해 흰쥐 하악대구치 치수제거 후 삼차신경절에서 VIP의 분포 및 반응강도를 공초점레이저주사현미경을 이용하여 관찰하였다. 체중 200g 내외의 Sprague-Dawley계 흰쥐를 대조군과 하악대구치 치수제거 후 14일군으로 분리하여 희생시켰다. 1차 항체로 rabbit anti-VIP, 2차 항체로 fluorescein isothiocyanate(FITC) conjugated anti-rabbit IgG를 사용하여 면역형광염색을 시행한 후 공초점레이저주사현미경으로 관찰하여 다음과 같은 결론을 얻었다. 1. 삼차신경절 하악부위에서 VIP 양성반응세포의 비율은 대조군에서 7.40%를, 실험군에서는 28.42%를 보였다. 대조군에 비해 실험군에서 양성반응세포의 증가를 보였다. 2. 삼차신경절 하악부위에서 VIP면역반응세포체에 대한 상대성 형광강도는 대조군에서 87.78을, 실험군에서는 138.65를 보였다. 대조군과 비교하였을 때 실험군에서 상대성 형광강도의 증가를 보였다. 3. 실험군의 광연속절편$(1{\mu}m)$ 관찰에서 VIP면역반응세포는 9개의 절편 대부분에서 강하게 나타났다. 축삭의 면역반응을 살펴보면, 대조군의 축삭에서는 약한 반응을 보였으며, 실험군의 축삭에서는 강한 면역반응을 보였다. 또한 양성 반응 세포체의 크기는 $20\sim25{\mu}m$의 중간 크기의 세포체에서 강한 면역반응을 보였다. 위의 결과로 보아 치수제거 후에 삼차신경절 하악부위에서 VIP 면역반응세포의 증가와 함께 상대성 형광강도가 높아졌음을 알 수 있었다.

• Applied Microscopy
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• 제24권4호
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• pp.61-77
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• 1994

The calcium-binding proteins (CaBP), parvalbumin (PV) and calbindin-D 28K (calbindin) are particularly abundant and specific in their distribution, and present in different subsets of neurons in many brain regions. Although their physiological roles in the neurons have not been elucidated, they are valuable markers of neuronal subpopulations for anatomical and developmental studies. This study is designed to characterize dorsal lateral geniculate nucleus (dLGN) neurons and axon terminals in terms of differential expression of immunoreactivity (IR) for two well-known CaBPs, PV and calbindin. The experiments were carried out on 6 adult monkeys. Monkeys were perfused under deep Nembutal anesthesia with 2% paraformaldehyde and 0.2% glutaraldehyde in 0.1M phosphate buffer. After removal, the brains were postfixed for 6-8 hr in 2% paraformaldehyde at $4^{\circ}C$ and infiltrated with 30% sucrose at $4^{\circ}C$. Thereafter, they were frozen in dry ice. Serial sections of the thalamus, at $20{\mu}m$, were made in the frontal plane with a sliding microtome. The sections were stained for PV and calbindin with indirect immunocytochemical methods. For electron microscopy, after infiltration with 30% sucrose the blocks of thalamus were serially sectioned at $50{\mu}m$ with a Vibratome in the coronal plane and stained immediately by indirect ABC methods without Triton X-100 in incubation medium. Stained sections were postfixed in 0.2% osmium tetroxide, dehydrated and flat-embedded in Spurr resin. The block was then trimmed to contain only a selected lamina or interlaminar space. The dLGN proper showed strong PV IR in fibers in all laminae and interlaminar zones. Particularly dense staining was noted in layers 1 and 2 that contain many stained fibers from optic tract. Neuronal cell body stained with PV was concentrated only in the laminae. In these laminae staining was moderate in cell bodies of all large and medium-sized neurons, and was strong in cell bodies of some small neurons together with their processes. Calbindin IR was marked in the neuronal cell body and neuropil in the S layers and interlaminar zones whereas moderate in the neuropil throughout the nucleus. Regional difference in distribution of PV and calbindin IR cell is distinct; the former is only in the laminae and the latter in both the S layer and interlaminar space. The CaBP-IR elements were confined to about $10{\mu}m$ in depth of Vibratome section. The IR product for CaBP was mainly associated with synaptic vesicle, pre- and post-synaptic membrane, and outer mitochondrial membrane and along microtubule. PV-IR was noted in various neuronal elements such as neuronal soma, dendrite, RLP, F, PSD and some myelinated or unmyelinated axons, and was not seen in the RSD and glial cells. Only a few neuronal components in dLGN was IR for calbindin and its reaction product was less dense than that of PV, and scattered throughout cytoplasm of soma of some relay neurons, and was also persent in some dendrite, myelinated axons and RLP. The RSD, F, PSD and glial elements were always non-IR for calbindin. Calbindin labelled RLP were presynaptic to unlabeled dendrite or dendritic spine and PSD. Calbindin-labeled dendrite of various sizes were always postsynaptic to unlabeled RSD, RLP or F. From this study it is suggested that dLGN cells of different functional systems and their differential projection to the visual cortex can be distinguished by differential expression of PV and calbindin.