• Title/Summary/Keyword: Hematoxylin-Eosin (H&E) stain

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The Effect of Genetically Modified Lactobacillus plantarum Carrying Bone Morphogenetic Protein 2 Gene on an Ovariectomized Rat

  • Jin, Eun-Sun;Kim, Ji Yeon;Yang, Jung-Mo;Kim, Jun-Sub;Min, JoongKee;Jeon, Sang Ryong;Choi, Kyoung Hyo;Moon, Gi-Seong;Jeong, Je Hoon
    • Journal of Korean Neurosurgical Society
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    • v.65 no.2
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    • pp.204-214
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    • 2022
  • Objective : Osteoporosis result from age-related decline in the number of osteoblast progenitors in the bone marrow. Probiotics have beneficial effects on the host, when administered in appropriate amounts. This study investigated the effects of probiotics expressing specific genes, especially the effects of genetically modified bone morphogenetic protein (BMP)-2-expressing Lactobacillus plantarum CJNU 3003 (LP) on ovariectomized rats. Methods : Twenty-eight female Wistar rats (250-300 g, 12 weeks old) were divided into four groups : the sham (control), the ovariectomy (OVX)-induced osteoporosis group (OVX), the OVX and LP (OVX/LP), OVX and genetically modified BMP-2-expressing LP (OVX/LP with BMP) groups. The three groups underwent bilateral OVX and two of these groups were administered two different types of LP via oral gavage daily. At 16 weeks post-OVX, blood was collected from the heart and the bilateral tibiae were extracted and were scanned by ex-vivo micro-computed tomography and stained with hematoxylin-and-eosin (H&E) and Masson's trichrome stain for pathological assessment. The serum levels of osteocalcin (OC), rat C-telopeptide of type I collagen (CTX-I), BMP-2, and receptor activator of nuclear factor-ĸB ligand (RANKL) were measured. Results : The 3D-micro-computed tomography images showed that the trabecular structure in the OVX/LP with BMP group was maintained compared with OVX and OVX/LP groups. No significant differences were detected in trabecular thickness (Tb.Th) between control and OVX/LP with BMP groups (p>0.05). Furthermore, a tendency toward increased BMD, trabecular bone volume, Tb.Th, and trabecular number and decreased trabecular separation was found in rats in the OVX/LP with BMP groups when compared with the OVX and OVX/LP groups (p>0.05). The H&E and Masson's trichrome stained sections showed a thicker trabecular bone in the OVX/LP with BMP group compared with the OVX and OVX/LP groups. There was no difference in serum levels of OC, CTX and RANKL control and OVX/LP with BMP groups (p>0.05). In contrast, significant differences were found in OC and CTX-1 levels between the OVX and OVX/LP with BMP groups (p<0.05). Conclusion : Our results showed that the expression of genetically modified BMP-2 showed inhibition effect for bone loss in a rat model of osteoporosis.

Effects of Danggwisayeokgaohsuyusaenggang-tang(當歸四逆加吳茱萸生薑湯) on Collagen-induced Arthritis in Mice (Collagen으로 유발된 생쥐의 관절염에 대한 당귀사역가오수유생강탕(當歸四逆加吳茱萸生薑湯)의 효과)

  • Kim, Min-Kyun;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.2
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    • pp.63-85
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    • 2011
  • Objectives : This study was carried out to know the effects of Danggwisayeokgaohsuyusaenggang-tang(hereinafter referred to DST) on arthritis induced by collagen on DBA/1 OlaHsd mice. Methods : For this purpose, DST was orally administered to mouse with arthritis induced by collagen II. Cytotoxicity, high performance liquid chromatograph(HPLC) analysis, arthritis index, value of immunocyte in draining lymph node and paw joint, cytokine were measured in vivo. Results : 1. The cytotoxicity against human fibroblast cells(hFCs) was not measured in any concentration. 2. In HPLC analysis, There are high peak patterns at 8 minute(min), 12 min, 35 min, 45 min. 3. The arthritis index was decreased significantly. 4. The degree of arthritis induced damage of joint of DST group is slight compared with control group in histopathologic observation(Hematoxylin and eosin stain(H&E), Masson's trichrome(M-T) staining). 5. In total cell counts of draining lymph node(DLN) and paw joint, the cells in DLN decreased significantly on DST 200 mg/kg and the cells in paw joint decreased significantly on 200 mg/kg and 50 mg/kg. 6. In DLN, $CD4^+/CD25^+$, $CD3^+/CD69^+$, major histocompatibility complex(MHC), class-II/$CD11c^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg $CD3^+/CD8^+$ cells decreased significantly on DST 200 200 mg/kg, $CD4^+$, $CD3^+/CD44^+$ cells decreased. 7. In paw joints, $CD4^+$, $CD11b^+/Gr-1^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg. 8. In joints, levels of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, cyclo-oxygenase-2(COX-2), NOS-II were decreased on DST 200 mg/kg and DST 50 mg/kg. 9. In analysing of cytokine in CD3/CD28 activated spleen, IL-17 was decreased significantly, IL-4 was increased significantly $INF-{\gamma}$ was decreased on DST 200 mg/kg. 10. In analysing of cytokine in collagen activated spleen, IL-17 were decreased significantly, IL-4 was increased significantly. Conclusions : This results demonstrated that DST suppressed the inflammatory progression of collagen-induced arthritis(CIA) mice and supported further studies are required to survey continuously in looking for the effective substance and mechanism in the future.

Histomorphological Comparison of the Coronary Artery and Available Graft Conduits (관상동맥 및 이식편으로 사용 가능한 혈관들의 조직형태학적 비교)

  • Kim, Yong-Ho;Lee, Seok-Ki;Yu, Jeong-Hwan;Kim, Si-Wook;Kang, Shin-Kwang;Lim, Seung-Pyung;Yu, Jae-Hyeon;Lee, Young
    • Journal of Chest Surgery
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    • v.41 no.5
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    • pp.563-567
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    • 2008
  • Background: The histomorphological properties of the left anterior desecending artery (LAD), the left internal thoracic arteries (LITA), the radial arteries (RA) and the intercostal arteries (ICA) were studied for their use as a conduit for coronary artery bypass grafting (CABG), and we compared them with each other. Material and Method: All the vessels were harvested from nineteen cadavers (17 males and 2 females). The mid-portion of the LAD, the mid-portion of the LITA, the distal RA and the mid-portion of the 5th ICA were obtained. All of them were stained with hematoxylin-eosin and with Van Gieson's elastin stain. The morphological characteristics were examined and the thicknesses of the intima and media (I/M ratio: the intima to media ratio) were compared using one-way ANOVA tests. Result: The mean age of the cadavers was $61.5{\pm}9.6$ years. The LITA and ICA were elastic arteries, and the LAD and RA were muscular arteries. The I/M ratio showed statistically significant differences: $0.07{\pm}0.03$ in the LITA, $0.16{\pm}0.11$ in the ICA, $0.45{\pm}0.29$ in the RA and $0.93{\pm}0.52$ in the LAD, respectively. Conclusion: This study showed that the degrees of intimal hyperplasia of the CA and the various conduits for CABG were different significantly. The ICA was found to have relatively favorable characteristics as a coronary by pass conduit, but its suitability for clinical use is a challenging issue.

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.

Airway Inflammation and Responses in the Bronchial Asthma Model in Sprague-Dawley Rats Sensitized by Ovalbumin (백서 기관지 천식 모텔에서 난황에 의한 기도염증과 기도반응)

  • Na, Moon-Jun;Lee, Byoung-Hoon;An, Chang-Hyeok;Kim, Jae-Yeol;Park, In-Won;Choi, Byung-Whui;Hue, Sung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.1
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    • pp.33-44
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    • 2000
  • Background: To evaluate airway responses and inflammation to antigen in Sprague-Dawley rat asthma model, we examined airway responses, serial histologic changes of the lung, and the relationship between airway responses and airway inflammation after antigen airway challenge. Methods: Sprague-Dawley rats were sensitized with subcutaneous injection of 10 ${\mu}g$ ovalbumin(OA). Antigen airway challenges were done 14~16 days after sensitization and the sensitized rats were sacrificed 1h($A_E$), 6~8h($A_L$) and 1day($A_D$) after airway challenge, to examine the histologic changes of the lung. Airway responses were measured by body plethysmograph and recorded by enhanced pause(Penh) as an index of airway obstruction 6~8h after antigen challenges. Nonsensitized controls(10 rats) were also challenged with antigen and sacrificed 1 day later. Histopathologic examination of two trachea, large bronchi, small bronchi, and vessels was performed to evaluate the severity of inflammation and eosinophilic infiltration with H&E stain. Results: In 17 of 20 rats(85%) in both groups, we observed airway responses. Among them, an early response(ER) in 15 rats(75%), an dual response in 5(25%), and an late response(LR) only in 2 rats(10%) displayed. There were no significant differences in the severity of inflammation among the trachea, large bronchi, small bronchi and vessels in all groups after antigen challenge(p>0.05) and between early and late responders. The significant eosinophil infiltration was observed in 5 rats(50%) of AL(p<0.05) compared with in AE and controls. Also, eosinophil infiltration was observed in higher trend in LR(57.1%) compared to ER(40%)(p>0.05). Conclusion: Sprague-Dawley rats sensitized with subcutaneous injection of OA showed a significant airway responses to antigen challenge. But antigen challenges caused a little eosinophil infiltration and no significant airway inflammation. Asthma model of Sprague-Dawley rats could be useful for antigen-induced airway responses, but this model has a limitation for the study of human asthma because of no significant pathologic change.

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The Effect of Hydroxyproline and Pro-Hyp Dipeptide on UV-damaged Skin of Hairless Mice (자외선에 의해 피부가 손상된 hairless mouse에서의 hydroxyproline, Pro-Hyp 경구반복투여시 피부 상태 개선 효과)

  • Lee, Ji-Hae;Seo, Jeong-Hye;Park, Young-Ho;Kim, Wan-Gi;Lim, Kyung-Min;Lee, Sang-Jun
    • Korean Journal of Food Science and Technology
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    • v.40 no.4
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    • pp.436-442
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    • 2008
  • Hydroxyproline and Pro-Hyp dipeptide are the digestive products of collagen hydrolysate called collagen peptide. Some suggested that collagen peptides could improve aged or damaged skins, however, the effects of collagen peptides on the skin have not been known. In this study, we investigated the effects of digestive products of collagen peptides, hydroxyproline and Pro-Hyp dipeptide on skin quality using the UV-damaged dorsal skin of hairless mouse as a model system. Female SKH hairless mice were pre-irradiated with UV for 7 weeks, and then hydroxyproline, Pro-Hyp dipeptide were orally administered for 7 weeks with UV irradiation. Wrinkle formation (by replica image), skin elasticity, barrier status (by TEWL, transepidermal water loss), epidermis thickness, and biophysical changes in the stratum comeum (by hematoxylin & eosin staining) were examined. With the oral peptide treatment, effects such as skin barrier maintenance, anti-skin thickening, and recovery of the stratum corneum were observed. These results indicate that oral intake of collagen peptides may have beneficial effects on damaged skin cells.

Effect of Radiation Dose for Radiotherapy on Ovarian Follicle Atresia in Rat (치료 방사선량이 쥐의 난포 퇴축에 미치는 영향)

  • Lee, Won-Jeong;Seon, Jong-Ryul;Yoo, Se-Jong;Ahn, Bong-Seon
    • Journal of Radiation Protection and Research
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    • v.37 no.4
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    • pp.208-212
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    • 2012
  • In previous studies, ovarian follicle in rat has been used a higher radiation dose than that for cancer radiotherapy in clinical practice. The aim of this study was to evaluate the effect of radiation dose used for cancer radiotherapy on ovarian follicle atresia in rat. Mice of 4-week-old female were whole body irradiated with 2 cGy or 2 Gy (Mevatron 67, Siemens, Germany) and sacrificed by cervical dislocation. Ovaries were collected at 24 hours after irradiation to observe the degree of follicular atresia. Ovaries were fixed in neutral formaldehyde solution for 24 hours and embedded with paraffin. Cutted in $5{\mu}m$ thickness with microtome and stained with hematoxylin and eosin (H&E) and TUNEL immunohistochemical stain, and examined histologically under a light microscope. All data were presented as mean ${\pm}SD$, calculating the ratio of normal or atretic follicles to total ovarian follicles. Statistical analysis was performed by the Mann Whitney test using the SPSS ver 19.0. Ratio of atretic to total follicles of 2 Gy group was significantly higher than control or 2 cGy groups (p<0.05). Ratio of normal to total follicles of 2 Gy group was significantly lower than control group in preantral follicle (64.0 vs. 87.7, p=0.027). Ratio of normal to total follicles of 2 cGy group was significantly increased more than control or 2 Gy groups in antral follicle, and there were no significant difference between control and 2 Gy groups (p=0.522). Radiation dose of 2 Gy for cancer radiotherapy have a significant effect on ovarian follicle atresia in rat.

Histologic changes of tooth and periodontal tissues applying to contraction & intrusion force for the maxillary four incisors of dogs (성견 상악 4절치의 Contraction과 압하시 치아 및 주위 조직의 조직학적 변화에 관한 실험 연구)

  • Kim, Young-Kuk;Cha, Kyung-Suk;Lee, Jin-Woo
    • The korean journal of orthodontics
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    • v.29 no.5 s.76
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    • pp.535-549
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    • 1999
  • The Purposes of this study were to investigate the initial tissue changes on the teeth and surrounding tissues under contraction and intrusive force by contraction UTA. A control and experimental dogs, 10-months in age, were studied. Contraction and intrusive force(60gm) were applied at upper four incisors by contraction UTA. Experimental dogs were sacrificed at 2 weeks and 4 weeks after force application, respectively. In this study, 2 experimental groups were designed by the duration of force applied(E1, E2). The specimens were taken around the upper first and second incisor in each groups and were prepared for the H-E and MT stain for light microscopic observation. From the results of the study, the following conclusions may be drown. : 1. In control group, the periodontal ligament width was constant from apical third to cervical third of the root and periodontal fiber arrangement was horizontal or oblique in cervical third, oblique in middle third and apical third. In alveolar bond, smooth appearance was shown with osteoblast. 2. In experimental group 1, in proportion to force was concentrated at labial middle third and apical third of root of the upper first and second incisors, root of these tooth tipped labially and intruded at a time. 3. In experimental group 2, periodontal ligament width and arrangement was similar to control and observed strong calcified response at the labial middle third of root. But, alveolar bond resorption and cementum resorption were as before seen at labial middle third and apical third of root that force was concentrated.

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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.