• The Korean Journal of Pain
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• 제9권2호
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• pp.326-335
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• 1996

Background: Though a number of studies have described the distribution of substance P(SP)-like immunoreactivity in the spinal cord, they have been focused on lamina I and II of the dorsal horn and there are little morphological studies on the topographic distribution and ultrastructure of the SP immunoreactive neurons especially in the ventral horn of the spinal cord. this study was conducted to identify distribution pattern of SP immunoreactive neurons and to difine the synaptic organization of their processes in ventral horn of the thoracic cord of the cat by preembbeding immunocytochemical method using SP antiserum. Methods: Five adults cats of either sex were used and deeply anesthetized by intramuscular injection of ketamine. After removal of the spinal cord, samples of thoracic cord were taken and placed in fresh fixative at $4^{\circ}C$ for 2 hours. Transverse sections $50{\mu}m$ thick were processed using the preembbeding immunocytochemical method and incubated consecutively in the specific primary antibody and the 10% normal goat serum, the rabbit anti-substance P antiserum, the biotin-labelled goat anti-rabbit IgG and finally the avidin-biotin-peroxidase complex. The processed tissue sections were throughly washed and stained in the black with 1% uranyl acetate. Section were examined on a electron microscope. Results: 1) SP immunoreactive neurons were observed in the gray matter around central canal. 2) In lamina I and II SP immunoreactivity was observed in both myelinated and unmyelinated nerve fibers, but in ventral horn only in the unmyelinated nerve fibers. 3) SP immunoreactive axon terminals with small round and large dense core vesicles made chemical synapses onto the dendrites of motor neurons in the ventral horn. Conclusion: SP immunoreactive neurons might play an important role in modulation of motor neurons in the ventral horn of the thoracic cord of the cat.

• Journal of Acupuncture Research
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• 제20권2호
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• pp.135-144
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• 2003

Objective : The purpose of this study is To investigate the effects of acupuncture and moxibustion at Zusanli(ST36) and Weishu(BL21) Methods : serum gastrin level by radioimmunoassay was measured at 5 days after acupuncture and moxibustion of those acupoints. Gastric endocrine cell(G cell and Histamine immunoreactive density) by avidin-biotinylated complex(ABC) technique, histological examinations(Alcian Blue-PAS Stain; Alcian blue-Periodic Acid Schiff reagent) of the gastric mucosa were also performed. Acupuncture applied to the ST36 acupoint increased gastrin level of serum, but moxibustion did not produced significant effect. All of acupuncture and moxibustion at BL21 acupoint increased gastrin level of serum significantly. In moxibustion at ST36 and BL21, the number of gastrin secreting cells in gastric mucosa, the density of immunoreactive histamin secreting cells and the density of body mucosa stained by PAS were decreased compare to acupuncture at ST36 and BL21. In acupuncture and moxibustion at BL21, the density of pylorus mucosa stained by PAS were increased compare to the groups applied to ST36. In the density of body mucosa stained by AB, moxibustion at BL21 and ST36 were increased compare to the other groups. Results : These data suggest that acupuncture and moxibustion at BL21 increased gastrin level of serum and those effects were more potent than acupuncture at ST36.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.321-340
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• 1995

The regeneration of destructed periodontal tissues is one of the ultimate objectives of periodontal therapy. Guided tissue regeneration technique was developed for the ideal regeneration of periodontal tissues. In order to investigate the role of fibronectin, laminin and tenascin in the regenerating process of periodontal tissues, the expanded PTFE barrier membranes(Gore Associates, USA) removed from the patients who had been treated by guided tissue regeneration(GTR) and guided bone regeneration(GBR) techniques were fixed in neutral formalin for 6-24 hours, embedded with paraffin, sectioned at $4-6{\mu}m$ in thickness, and immunohistochemically processed by Avidin-Biotin peroxidase complex method for detecting fibronectin, laminin and tenascin. Monoclonal mouse anti-human fibronectin antibody(Oncogene Science, USA., 1:100), monoclonal mouse anti-human laminin antibody(Oncogene Science, USA., 1:50) and mouse anti-human tenascin antibody(Oncogene Science, USA, 1:10) were used as primary antibodies. The light microscopic findings were as follows: (1) The distribution of fibronectin, laminin and tenascin was various according to the area of barrier membranes. (2) The distribution of fibronectin in case of GBR was extensive in the tissue on the outer surface of barrier membranes, and rare in the intervening space and on the inner surface. In case of GTR it was extensive on the outer surface and in the intervening space, and rare on the inner surface. (3) The distribution of laminin was rare in the tissue on the outer, the inner surface and intervening space of barrier membranes, regardless of GBR or GTR. (4) In case 'of GBR rare distribution of tenascin was observed on the outer surface only, except the inner surface and the intervening space of barrier membranes. In case of GTR the distribution of tenascin was extensive in the tissue on the outer surface, rare in intervening space and the inner surface. The results suggest that fibronectin, laminin and tenascin may play a important role in the regenerating process of periodontal tissue, and they may affect the outcome of healing.

• 대한수의학회지
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• 제35권2호
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• pp.317-326
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• 1995

The purpose of this study was to establish a method for in vitro culture of C parvum isolated in Korea by determination of suitable cell model to complete development of this parasite. The result obtained were summerized as follows: 1. To determine the most suitable cell line, six types of cell line were examined by microscopy. All cell lines were infected with C parvum and showed the highest infection score in HmLu cells. 2. The staining methods including DMSO-modified acid-fast(A-F) stain, hematoxylin-eosin(H & E) stain and immunofluorescence antibody(IFA) stain were applied to examine the infection of C parvum in cell culture. These staining methods were possible to examine the infection of C parvum in cell culture. The most sensitive one was IFA staining technique. 3. Developmental stages of C parvum in HmLu cell were observed. After the initial 8 hour incubation period, some trophozoites were observed. The meronts and gametes were appeared at 24-48 hour post inoculation(PI), and oocysts were observed firstly at 48-72 hour PI. 4. In H & E stain, the parasite appeared as basophilic within parasitophorous vacuole membrane(PVM) and lying in cytoplasm at near the nucleus of the host cells. It was able to distinguish the type I, type II meronts and gametes. 5. In DMSO-modified acid-fast stain, specific stained parasites were appeared firstly after 48 hour PI. The parasites were showed with different degrees of staining bright red color within PVM. 6. The endogenous stages of parasites in HmLu cell recovered at 48, 96, 120 and 144 hour after inoculation were reacted with rabbit immunized serum in immunofluorescence antibody and avidin-biotin complex peroxidase staining technique.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제15권3호
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• pp.217-228
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• 1993

The purpose or this study was to observe the histological alteration of Langerhans cells on wound healing process in applying low energy laser irradiation. For this study, 50 Spraque-Dewly rats, weighing 150Gm or more were devided into control, experimental control group(0), 47.5Hz(1), 190Hz(3), 380Hz(5), 760Hz(7), lased group. All the experimental animals were made excision wound on buccal mucosa, 2mm depth, and lased with stoma laser (904nm, semconductor type ASGaAI, Sedalac France) 47.5Hz, 380Hz, 960Hz, 3minutes one time respectively except experimental control group. After the experiment, experimental animals were sacrificed after 24hours, 48hours, 72hours on each. Taken specimens were embedden in paraffin, sectioned 6-8u in thickness. And the langerhans cell were detected using ant S-100 protein antibody, and histochemically processed with Avidin Biotin complex method. All the Langerhan cells were calculated under light microspe in 400 multiplication field and standard deviation, probability test between each group were evaluated using statistical analysis system(S.A.S)program. Following results were obtained. 1. Langerhan cells were increased in experimental control group compared to that in control group(P<0.01). 2. 24hour after experiments, Langerhans cell were decreased compare to that in control group and control experimental group 5, 1, 3. Probability test shows significance between control experimental and 5, 1, 3 group on a =0.05 range. 3. 48our after experiment, Langerhans cells were decreased compare to that on experimental control group, and probability test shows significance between control experimental and 3, 7, 5 group an a=0.05 range. 4. 72hour after experiments, Langerhans cells were decreased compare to that on experimental control group and probability test on group comparison shows significance between control experimental and 1, 5 and 1 between 3, 7 between 3, and 5, between 7, respeilively on a=0.05 range. 5. Langerhans cells number in experimental group were decreased compare to that on experimental control group in applying laser irradiation.

• Restorative Dentistry and Endodontics
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• 제19권2호
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• pp.372-383
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• 1994

Saliva plays an important role in modulating the oral microbial ecology. And it is suggested to influence the initiation and progression of the dental caries. To evaluate the correlations between the salivary antimicrobial agents and the caries susceptibility, the 51 subjects were divided into 3 groups according to caries experience ; caries resistant group, medium caries susceptible group, and high caries susceptible group. Stimulated whole saliva was collected, and the salivary levels were measured for lysozyme, lactoferrin, and secretory-IgA to Streptococcus mutans. The lysozyme level was estimated using Micrococcus diffusion plate, lactoferrin level was determined with a non-competitive avidin-biotin enzyme immunoassay, and the titer of secretory IgA to Streptococcus mutans was assayed with ELISA. The results were as follows: 1. Lysozyme levels of each group showed no significant difference statistically (p>0.05). 2. The caries resistant group and the medium caries susceptible group had significantly higher levels of lactoferrin than the high caries susceptible group (p<0.05). But no clear difference was observed between the caries resistant group and the medium caries susceptible group(p>0.05). 3. The caries resistant group and the medium caries susceptible group showed relatively higher levels of the secretory IgA to Streptococcus mutans than the pigh caries susceptible group, but no significant difference was observed statistically (p>0.05).

• Restorative Dentistry and Endodontics
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• 제15권2호
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• pp.58-76
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• 1990

This study was designed to elucidate the distribution of the immunoglobulins in the experimentally induced rat periapical lesions. The pulp exposure was performed in 80 molars from 40 rats and the animals were sacrificed at 15, 30, 60 and 90 days after the operation and examined and radiographed. Of the 80 samples, 56 samples were routinely sectioned ($4-6{\mu}$ in thickness) and stained with Hematoxylin-Eosin for the light microscopic examination and 50 samples were stained with toluidin blue for mast cells and 50 samples were stained using the Avidin-Biotin horseradish peroxidase for detecting the presence of Ig A, Ig E, Ig M and Ig G containing cells. The following results were obtained : 1. The periapical lesions could be observed in all of 80 teeth by radiogragh (100%) and the periapical lesions were detected in 50 samples of 51 samples by light microscopy (98%). The size of lesions increased with time lapse both by radiograph and by light microscopy(p<0.05). 2. Of the 50 samples, 19 samples were diagnosed as periapical abscesses, 18 as periapical granulomas, 10 as fibrous scar tissues and 3 cysts. 3. After pulp exposure, periapical granulomas were developed mostly in the 15 day group, with time lapse periapical abscesses and fibrous scar tissues increased. 4. In the 50 periapical lesions, the numbers of Ig G containing cell (57.2%) were prominent and the percentage of Ig A, Ig E and Ig M containing cells were 16.4%, 14.7% and 11.8% respectively. The numbers of all classes of immunoglobulin containing cell were highest in the periapical granulomas and lowest in the cysts(p<0.05). 5. The number of the mast cell and immunoglobulin containing cells decreased generally with time lapse after the pulp exposure and Ig A, Ig E, Ig M and Ig G containing cells and mast cells had the high correlation one another(>0.6).

• 한국가축번식학회지
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• 제22권1호
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• pp.19-27
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• 1998

The present study was performed to investigate the patterns of a, pp.arance of lectin in uterus of fetuses of 90 and 120 days old and neonates of Korean native goat. The carbohydrate markers were used in histochemistry for the determination of the lectin by staining of avidin-biotin-per-oxidase complex(ABC), rincinus communis agglutinin(RCA-I), ulexeuropalus communis agglutinin(UEA) and wheat germ agglutinin(WGA). 1. The effects of this study were as follows; 1. The binding reactions for Con-A were weak on the mucosal epithelium of endometrium in 90 and 120 days old fetuses, and neonates and moderate at the free surface of mucosal epithelia. 2. The binding reactions for DBA was partially moderate on the mucosal epithelium of endometrium and partially strong at the free surface of mucosal epithelia in 120 days old fetuses. In neonates, the reactions were strong on the mucosal epithelium and gland primordium of endometrium, and the secretions at the free surface showed strong reactions for DBA. But, in 90 days old fetuses, the reaction was negative. 3. The binding reactions for RCA-I were moderate on the mucosal epithelium of endometrium and at the free surface of mucosal epithelia in 90 days old fetuses. In 120 days old fetuses, the reactions were weak on the mucosal epithelium of endometrium and moderate at the free surface of mucosal epithelia. In neonates, the reactions were moderate on the mucosal epithelium of endometrium and strong at the free surface of mucosal epithelia and also strong in the uterine gland. 4. The binding reactions for UEA were negative in 90 and 120 days old fetuses and neonates. 5. In 90 days old fetuses, the binding reactions for WGA were generally weak on the mucosal epithelium of endometrium, but several epithelial cells showed moderate reaction for WGA. In 120 days old fetuses and neonates, the reactions were moderate on the mucosal epithelium and blood vessels of endometrium and strong at the free surface of mucosal epithelia.

• 한국동물위생학회지
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• 제41권1호
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• pp.57-61
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• 2018

Canine peripheral nerve sheath tumors (PNSTs) are spindle cell tumors that arise from Schwann cells, perineural cells, fibroblasts or all of them. Based on the morphology and biologic behavior, PNSTs are divided into benign PNST (BPNST) and malignant PNST (MPNST) forms. The aim of this study is to diagnose the two cases of neoplastic tissue samples with features of PNSTs by the histopathology and immunohistochemistry. The study was performed using two specimens from small animal clinic. The first case, A was a mass, 3~4 cm in diameter, extruded from vaginal mucosa of 10-year-old spayed female mixed-breed dog. And the second case, B was a subcutaneous mass, 1.5 cm in diameter, which is originated from right hind leg of 9-year-old castrated male mixed-breed dog. Two cases were stained with hematoxylin and eosin (H&E) for histopathological examination. And also immunohistochemistry (IHC) was performed by the avidin-biotin peroxidase complex (ABC) method with antibodies specific for the following proteins: S-100 protein, smooth muscle actin (SMA) and epidermal growth factor receptor (EGFR). In results, Antoni B schwannoma pattern characterized by pleomorphic, round and fusiform polygonal cells was seen in A. In B, Antoni A pattern, densely packed spindle cells arranged in interlacing bundles was seen in addition to Antoni B pattern. In IHC, cytoplasms of neoplastic cells were diffusely labeled for S-100 expression in A and B. For SMA, both A and B show negative expression. And for EGFR, A shows negative expression but B shows partially positive expression in areas of Antoni B schwannoma pattern. The histopathologic features of two cases coupled with the S-100 immunoreactivity led to a diagnosis of PNST. For SMA, both A and B show negative expression. The diagnosis of A will be a BPNST with the negative result and B will be a MPNST with the positive result for EGFR.

• 대한수의학회지
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• 제39권3호
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• pp.407-416
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• 1999

This study was carried out to investigate the NPY-immunohistochemical characteristics of the olfactory bulb in the striped field mouse(Apodemus agrarius). The animals were anesthesized with thiopental sodium and perfused with 4% paraformaldehyde through left ventricle and aorta. Brains were removed and tranfered 10%, 20% and 30% sucrose. Sections were then cut on a cryostat into $40{\mu}m$-thick. The tissue immunostained with avidin-biotinylated complex method. The main olfactory bulb consisted of seven circumferential laminae : an olfactory nerve fiber layer, a glomerular layer with glomeruli surrounding by periglomerular cells, an external plexiform layer having granule and tufted cells, a mitral cell layer, a narrow internal plexiform layer, a granule cell layer forming several cell rows and a layer of white matter. The accessory olfactory bulb had four layers : an olfactory or vomeronasal nerve fiber layer, a glomerular layer consisting of small glomeruli, a mixed layer not distinguishing the external plexiform/mitral cell/granule cell layers and a granule cell layer. Most of NPY-immunoreactive(NPY-IR) neurons in main olfactory bulb were localized in the deeper portion of granule cell layer, white matter and anterior olfactory nucleus. In addition, some NPY-IR neurons were identified in the external plexiform layer. The shape of NPY-IR neurons of all olfactory bulb were predominant round or oval, sometime multipolar in shape. And most NPY-IR processes were parallel to long axis of white matter. In accessory olfactory bulb, NPY-IR neurons were not found in all region.