• Title/Summary/Keyword: Hematoxylin and eosin

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Histological Changes in Rat Testis by Injection of Hypertonic Saline (고장성 식염수 주사에 의한 흰쥐 정소의 해부학적 변화)

  • Kwak, Byung-Kuk;Lee, Chul-Sang;Lee, Sung-Ho
    • Development and Reproduction
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    • v.14 no.4
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    • pp.281-286
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    • 2010
  • Recent evidence has revealed that the intratesticular injection of hypertonic saline(20%) resulted in a chemically castrated state such as nadir testosterone levels in rats. To confirm the efficacy of this simple saline-injection method further, we investigated the changes in the gross and microscopic anatomy of testis. Our study comprised three groups; intact(control) group, orchidectomy group and saline-injection (experimental) group. Single dose of hypertonic saline (sterilized, $750{\mu}{\ell}/testis$) were directly administered into both testis of adult rats (about 300 g BW). Bilateral orchidectomy was performed at the same day of saline injection. Following 30 days post-injection, reproductive tissues were surgically removed, weighed and fixed for histological examination. The body weights were not changed in both orchidectomy group and saline-injection group when compared to those in intact group. The wet weights of testis were significantly decreased in saline-injection group when compared to those in intact group. The wet weights of epididymis and seminal vesicle and prostate were significantly decreased in orchidectomy group and saline-injection group when compared to those in intact group. Macroscopically, the testes exerted slight atrophy and the tunica albuginea seemed to be intact in saline injection group. Histologically, however, larger parts of testicular tissue underwent necrosis and were barely recognizable after hematoxylin-eosin staining. In the same section, only the opposite part of the injection site was stained showing abnormal state of cell layers mostly fibrosis and infiltrated leukocytes. Sloughing of immature germ cells from the basement membrane along with shedding cells in the intraluminal space was notable in most seminiferous tubules from the saline injected testis. The present study confirmed that the direct injection of hypertonic saline into testis can induce a castration-like, testosterone-depriving effects on accessory sex organs. Our findings suggest that the efficacy of this less expensive and minimally invasive method seems to be almost even with that of conventional orchidectomy and chemical castration, though more in-depth evaluation should be supported.

Neuroprotective effects of geneticin (G418) via apoptosis in perinatal hypoxic-ischemic brain injury (주산기 저산소성 허혈성 뇌손상에서 항고사를 통한 geneticin (G418)의 신경보호 효과)

  • Ju, Mi;Lee, Hyun Ju;Lee, Sun Ju;Seo, Eo Su;Park, Hye Jin;Lee, Kye Yang;Lee, Gyeong Hoon;Choi, Eun Jin;Kim, Jin Kyung;Lee, Jong Won;Chung, Hai Lee;Kim, Woo Taek
    • Clinical and Experimental Pediatrics
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    • v.51 no.2
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    • pp.170-180
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    • 2008
  • Purpose : Some antibiotics were known to exert neuroprotective effects in the animal model of hypoxic-ischemic (H-I) brain injury, but the mechanism is still unclear. A recent study reported that geneticin (G418), an aminoglycoside antibiotic, increased survival of human breast cancer cells by suppressing apoptosis. We investigated the neuroprotective effects of systemically administrated geneticin via anti-apoptosis following the H-I brain injury Methods : Seven-day-old Sprague-Dawley rat pups were subjected to unilateral (left) common carotid artery occlusion followed by 2.5 hours of hypoxic exposure and the cortical cell culture of rat brain was done under a hypoxic incubator. Apoptosis was measured in the injured hemispheres 7 days after H-I insult and in the injured cells from hypoxic chamber using morphologic analysis by Terminal dUTP Nick-end Labeling(TUNEL) assay and immunohistochemistry for caspase-3, and cytologic analysis by western blot and real time PCR for bax, bcl-2, and caspase-3. Results : The gross appearance and hematoxylin and eosin stain revealed increased brain volume in the geneticin-treated animal model of perinatal H-I brain injury. The TUNEL assay revealed decreased apoptotic cells after administration of geneticin in the cell culture model of anoxia. Immunohistochemistry showed decreased caspase-3 expression in geneticin-treated cortical cell culture. Western blot and real-time PCR showed decreased caspase-3 expression and decreased ratio of Bax/Bcl-2 expression in geneticin-treated animal model. Conclusion : Geneticin appears to exert a neuroprotective effect against perinatal H-I brain injury at least via anti-apoptosis. However, more experiments are needed in order to demonstrate the usefulness of geneticin as a preventive and rescue treatment for H-I brain injuries of neonatal brain.

Neuroprotection of Recombinant Human Erythropoietin Via Modulation of N-methyl-D-aspartate Receptors in Neonatal Rats with Hypoxic-ischemic Brain Injury (신생 백서의 저산소성 허혈성 뇌손상에서 NMDA receptor 조절을 통한 유전자 재조합 인 에리스로포이에틴의 신경보호)

  • Jang, Yoon-Jung;Seo, Eok-Su;Kim, Woo-Taek
    • Neonatal Medicine
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    • v.16 no.2
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    • pp.221-233
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    • 2009
  • Purpose: Erythropoietin (EPO) has neuroprotective effects in many animal models of brain injury, including hypoxic-ischemic (HI) encephalopathy, trauma, and excitotoxicity. Current studies have demonstrated the neuroprotective effects of EPO, but limited data are available for the neonatal periods. Here in we investigated whether recombinant human EPO (rHuEPO) can protect the developing rat brain from HI injury via modulation of NMDA receptors. Methods: In an in vitro model, embryonic cortical neuronal cell cultures from Sprague-Dawley (SD) rats at 19-days gestation were established. The cultured cells were divided into five groups: normoxia (N), hypoxia (H), and 1, 10, and 100 IU/mL rHuEPO-treated (H+E1, H+ E10, and H+E100) groups. To estimate cell viability and growth, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was done. In an in vivo model, left carotid artery ligation was performed on 7-day-old SD rat pups. The animals were divided into six groups; normoxia control (NC), normoxia Sham-operated (NS), hypoxia-ischemia only (H), hypoxia-ischemia+vehicle (HV), hypoxia-ischemia+rHuEPO before a HI injury (HE-B), and hypoxia-ischemia+rHuEPO after a HI injury (HE-A). The morphologic changes following brain injuries were noted using hematoxylin and eosin (H/E) staining. Real-time PCR using primers of subunits of NMDA receptors (NR1, NR2A, NR2B, NR2C and NR2D) mRNA were performed. Results: Cell viability in the H group was decreased to less than 60% of that in the N group. In the H+E1 and H+E10 groups, cell viability was increased to >80% of the N group, but cell viability in the H+E100 group did not recover. The percentage of the left hemisphere area compared the to the right hemisphere area were 98.9% in the NC group, 99.1% in the NS group, 57.1% in the H group, 57.0% in the HV group, 87.6% in the HE-B group, and 91.6% in the HE-A group. Real-time PCR analysis of the expressions of subunits of NMDA receptors mRNAs in the in vitro and in vivo neonatal HI brain injuries generally revealed that the expression in the H group was decreased compared to the N group and the expressions in the rHuEPO-treated groups was increased compared to the H group. Conclusion: rHuEPO has neuroprotective property in perinatal HI brain injury via modulation of N-methyl-D-aspartate receptors.

Effect of One- or Two-Week Triple Therapy with Omeprazole, Amoxicillin, and Clarithromycin on Eradication of Helicobacter pylori Infection in Children (소아에서 Omeprazole, Amoxicillin, Clarithromycin 삼제 병합 요법의 치료기간별 Helicobacter pylori 제균율)

  • Choi, In-Kyoung;Lee, Seung-Yun;Chung, Ki-Sup
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.5 no.1
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    • pp.19-25
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    • 2002
  • Purpose: The triple therapy with proton pump inhibitor (PPI) has been recognized as the treatment of choice in Helicobacter pylori (H. pylori) infection in adults. However, the effect of triple therapy with omeprazole, amoxicillin and clarithromycin (OAC) on eradication of H. pylori infection in children has not been established yet. This study was performed to evaluate the efficacy of OAC triple therapy and to compare the effect of one-week with two-week therapy on H. pylori eradication. Methods: From July 1998 to July 2000, 34 children with upper gastrointestinal symptoms, who underwent upper gastrointestinal endoscopy with biopsy at entry and 4 or more weeks after therapy, were enrolled in this study. H. pylori infection was assessed by CLO test and histologic examination (Hematoxylin-Eosin stain or Alcian yellow stain) with biopsy specimens. The regimen consisted of omeprazole (0.7 mg/kg/day), amoxicillin (50 mg/kg/day), and clarithromycin (25 mg/kg/day) for 1 week (n=21) or 2 weeks (n=13). Eradication of H. pylori was determined after the termination of treatment by the CLO test and histologic examination. Results: One-week treatment group consisted of 21 children (11 male, 10 female) with a mean age of $9.5{\pm}3.0$ years. Two-week group consisted of 13 children (4 male, 9 female) with a mean age of $9.9{\pm}4.0$ years. The endoscopic diagnoses included nodular gastritis in 19 cases, superficial gastritis in 7 cases, gastric ulcer in 4 cases, purpuric duodenitis in 2 cases, and normal in 2 cases. H. pylori was eradicated in 28 of total 34 children (82.4%). In 1-week group, H. pylori was eradicated in 17 of 21 children (81%). In 2-week group, H. pylori was eradicated in 11 of 13 children (84.6%). In remaining 6 cases in whom H. pylori had not been eradicated with OAC regimen, H. pylori infection persisted despite of the treatment with additional drugs such as colloidal bismuth subcitrate ($Denol^{(R)}$) and metronidazole. Conclusion: In this study, eradication rate of H. pylori with OAC regimen was 82.4%, and the triple therapy would be highly effective as primary treatment. However, there was no significant difference in the eradication rate between the 1-week and 2-week treatment group (P=0.785).

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THE BONE CHANGES OF THE MIDPALATAL SUTURE AFTER RAPID PALATAL EXPANSION IN YOUNG ADULT DOGS (유성견에서 상악골 급속확장 후 정중구개봉합부에서의 골조직 변화)

  • Park, Chang-Wook;Yoon, Young-Jooh;Kim, Kwang-Won
    • The korean journal of orthodontics
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    • v.27 no.3 s.62
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    • pp.391-399
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    • 1997
  • Rapid palatal expansion(RPE) is a method of inducing the new bone formation in the palate by separation of the midpalatal suture, which can be done conveniently by placing heavy force across the maxillary dental arch. This experiment was undertaken to examine the histologic changes after RPE and during retention period. Four young adult dogs(a control dog, three experimental dogs) aged 4 to 6 months old were used for this experiment. Expansion screw($Hyrax^{\circledR}$, Dentarum Inc.) was delevered to the palate and fumed 180 degrees every morning and evening for 8 days, giving a total expansion of 7.2mm. A control dog was sacrified at the starting point of this study without any treatment and three experimental dogs were sacrified after RPE, 14-day retention, and 28-day retention in each. Thereafter, those samples were observed with hematoxylin-eosin(H-E) stain, ground section(Villanueva stain), alkaline phosphatase(ALP) stain, tartrate-resistant acid phosphatase(TRA) stain. The results were as followings: 1. After RPE, collagen fiber bundles were stretched along the midpalatal suture and few osteoblasts were flattened-inactive state and also, a little osteoid tissues was observed. Few multinucleated osteoclasts which had TRAP-positive activity in their cytoplasm were seen in horizontal section, whereas a few osteoclasts were seen in frontal section, especially in the nasal floor side of palatal bone. 2. After 14-day retention, collagen fiber bundles were stretched along the midpalatal suture and few osteoblasts which had ALP-positive activity in their cytoplasm were seen. Few multinucleated osteoclasts which had TRAP-positive activity in their cytoplasm were seen in horizontal section, whereas a few osteoclasts were seen in frontal section, especially in the nasal floor side of palatal bone. 3. After 28-day retention, collagen fiber bundles were arranged like those of control dog and osteoblasts which showed a lot of immature bone formation were cuboidal shape and exhibited ALP-positive activity in their cytoplasm. Few multinucleated osteoclasts which had TRAP-positive activity in their cytoplasm were seen in horizontal section, whereas a few osteoclasts were seen in frontal section, especially in the nasal floor side of palatal bone. According to the above results, the new bone formation after rapid palatal expansion was examined after 14-day retention and significantly increased after 28-day retention.

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Prevention of Photoaging and Wrinkle Formation in Hairless Mice Dorsal Skin by APB-03 (Hairless mice에서의 대두 홍삼 혼합 분말(APB-03)의 경구 반복 투여 시 피부 주름 생성 예방 효과)

  • Lee, Ji-Hae;Lee, Byoung-Seok;Yang, Mi-Suk;Byun, Bum-Sun;Kim, Wan-Gi;Kim, Bae-Hwan;Lee, Sang-Jun
    • Korean Journal of Food Science and Technology
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    • v.37 no.6
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    • pp.989-996
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    • 2005
  • Ultraviolet (UV) induces photo aging, erythema, sunburn, photo-toxicity, photo-allergy, and skin tumor, To investigate photo-protective effects of AmorePacific Beauty-03 (APB-03; mixture of red ginseng extract powder and soybean extract powder) on UV-induced damaged skins, 40 SKH hairless female mice were orally administered APB-03 or saline five times a week and irradiated with UV three times per week far up to 12 weeks. Visible skin changes and skin damage in dermis and epidermis by replica image analysis and histological analysis. In APB-03-treated group, better skin, negative replica appearance and less wrinkle formation were observed compared to the UV control group. These results demonstrate oral administration of APB-03 have photo-protective effects on UV-damaged hairless mouse skin.

A Study on the changes of periodontal tissue after orthodontic tooth movement in young adult dogs (유성견에서 교정적 치아이동에 따른 치주조직 변화에 관한 연구)

  • Kang, Nam-Yong;Yoon, Young-John;Kim, Kwang-Won
    • The korean journal of orthodontics
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    • v.27 no.4 s.63
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    • pp.559-568
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    • 1997
  • For orthodontic tooth movement, optimal orthodontic force should be maintained without periodontal breakdown and alveolar bone should be remodeled physiologically Therefore, To obtain proper occlusion through tooth movement within alveolar bone, we should know the biomechanics of teeth and supporting 4issues. The present study was performed to observe histologic changes of periodontal tissue immediately after application of orthodontic force and during the retention period in growing young adult dogs. In this study, experimental group contained between mandibular left canine and 1st molar and control group contained contralateral teeth of same animal. The .018'x.022' stainless steel closed coil spring(Dentaurum Co.) was ligated on the experimental teeth at initial 200gm-force from mandibular canine to 1st molar The animals(4 to 6 months aged young adult dogs) were sacrificed on 0, 14, 28 days after the finish of appliance activation, and then tissue samples were divided into hematoxylin-eosin(HE) staining section, ground section, alkaline phosphatase(ALP) staining section, and tartrate-resistant acid phosphatase(TRAP) staining section. Thereafter, the preparations were examined under light microscopy The following results were obtained: 1. Immediately after the finish of appliance activation, the periodontal space was increased in tension side, but decreased in pressure side compared to that of control. The hyalinized zone was also observed in the periodontium. 2. After the 14-day retention, peridontal space was decreased in tension side and slightly increased in pressure side compared to that of immediately after the finish of appliance activation. The hyalinized zone was repaired and a few osteoblasts showing slightly new bone formation were seen. Osteoblasts were scarcely observed along the alveolar bone. 3. Aftter the 28-day retention, the periodontal fibers are normally repaired. A lot of TRAP(+) osteoclasts md increased alveolar bone resorption were observed in pressure side, and AP(+) osteoblast and increased new bone formation were observed in tension side.

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Effects of BCG on the DNA Synthesis and Ultrastructure of Mouse Gastric Mucosal Epithelial Cells Inoculated with Ehrlich Carcinoma Cells (BCG가 Ehrlich 암세포를 이식한 생쥐의 위점막 상피세포의 DNA합성 및 미세구조에 미치는 영향)

  • Ko, Jeong-Sik;Ryoo, In-Sang;Park, Kyung-Ho;Park, Dae-Kyoon
    • Applied Microscopy
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    • v.39 no.3
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    • pp.205-218
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    • 2009
  • This experiment was performed to evaluate the morphological responses of the gastric epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (tumor control group and BCG-treated group). In the experimental groups, each mouse was inoculated with $1{\times}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of saline or BCG, each mouse was injected with a single dose of 0.7 ${\mu}Ci/g$ of methyl-$^3H$-thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and gastric tissues were taken and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab., England) in a dark room. The number of labeled epithelial cells in the gastric mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. And for electron microscopic observation, gastric tissues were prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. On the light microscopic study, gastric mucosae had no morphological changes following the injection of BCG. On the electron microscopic study, in the BCG-treated mice, myelin figures and multivesicular bodies within the gastric epithelial cells were observed more frequently than in those of the normal control ones. On the autoradiographic study, number of the labeled cells of normal control, tumor control and BCG-treated mice were 380.2 (${\pm}31.35$), 426.1 (${\pm}28.43$) and 301.8 (${\pm}34.63$), respectively. In the BCG-treated mice, poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently as compared in those of the normal control and tumor control ones. From the above results, BCG may suppress the DNA synthesis of the gastric epithelial cells, but does not results severe fine structural defect on the gastric epithelial cells. These results suggest that BCG is expected as one of the effective supplemental anticancer drugs.

Comparison of TheraCal LC, Mineral trioxide aggregate, and Formocresolas pulpotomy agents in rat molar (백서에서 치수절단술에 사용하는 TheraCal LC, MTA 그리고 Formocresol의 비교)

  • Lee, Bin-Na;Song, Young-Sang;Lee, Go-Woon;Kim, Young-Hoon;Chang, Hoon-Sang;Hwang, Yun-Chan;Oh, Won-Mann;Hwang, In-Nam
    • Korean Journal of Dental Materials
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    • v.44 no.2
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    • pp.187-195
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    • 2017
  • TheraCal LC, a new light-cured, resin-modified calcium silicate-filled base/liner material, has been introduced as a pulpotomy agent. The aim of this study was to evaluate the capacity of hard tissue formation and pulpal response after pulpotomy with TheraCal LC. Twenty-two 9-week-old male rats were anesthetized, cavities were prepared in maxillary first molars and pulps were capped with formocresol (FC), mineral trioxide aggregate (MTA), and TheraCal LC. Specimens obtained from rats were scanned using a high-resolution micro CT system. The specimens were prepared and evaluated histologically, and immunofluorescence assay was performed to assess the dentin matrix protein-1 (DMP-1) expression. On micro CT analysis, the MTA and TheraCal LC groups showed thicker hard tissue formation than the FC group. On hematoxylin and eosin (H&E) staining, MTA and TheraCal LC groups showed dentine bridge formation with vital pulp beneath the materials. On immunofluorescence analysis, DMP-1 was highly expressed in the TheraCal LC group compared to the FC group. TheraCal LC showed similar capacity to form hard tissue as MTA when it was used as a pulpotomy agent. Because of its good manipulation and faster setting time compared to MTA, TheraCal LC could be considered as a good alternative to MTA.

Relationship Between Vascularity and Other Remodeling Parameters in Asthmatic Airway (기관지천식환자에서 기도내 혈관분포정도(vascularity) 및 다른 기도 재구성 소견과의 연관성)

  • Kim, Seung Joon;Lee, Sook Young;Kim, Myoung Sook;Lo, Dae Keun;Kwon, Soon Seog;Kim, Young Kyoon;Kim, Kwan Hyoung;Moon, Hwa Sik;Song, Jeong Sup;Park, Sung Hak
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.2
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    • pp.191-198
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    • 2003
  • Background : The pathological features in asthmatic airway remodeling are diverse. The aim of this study was to examine the degree of airway vascularity in relation to the other remodeling parameters in asthmatics. Methods : Bronchial biopsies were done in 34 asthmatic patients, and 6 control subjects. The basement membrane thickness and the subepithelial thickness were measured in the hematoxylin-eosin stained tissue, and the degree of vascularity was measured using type IV collagen immunostaining. Results : 1) Compared to the control subjects, the asthmatics showed a significant increase in the basement membrane thickness ($6.92{\pm}2.01{\mu}m$ vs $9.67{\pm}2.84{\mu}m$, p<0.05) and the subepithelial thickness ($44.49{\pm}31.92{\mu}m$ vs $121.22{\pm}72.79{\mu}m$, p<0.05). 2) Compared to the control subjects, the asthmatics showed a significant increase in the vascular area per unit submucosal area ($4.51{\pm}2.13%$ vs $10.32{\pm}6.08%$, p<0.05). In addition, the number of vessels per unit submucosal area showed an increased tendency without statistical significance. 3) In the asthmatics, the number of vessels and the vascular area per unit submucosal area showed no correlation with the basement membrane thickness, the subepithelial thickness, the severity, the forced expiratory volume in 1 second($FEV_1$), and the methacholine provocative concentration 20($PC_{20}$). Conclusion : This study showed that vascularity was an important parameter in asthmatic airway remodeling but it was not related to the other remodeling parameters such as the basement membrane thickness and the subepithelial thickness. Each of these asthmatic remodeling parameters may have a different clinical significance. Therefore, further studies will be needed.