• Title/Summary/Keyword: hematoxylin and eosin (H&E) staining

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Dioscorea japonica Thunb. Ethanolic Extract Attenuated Oxazolone-Induced Atopic Dermatitis-like Skin Lesions in BALB/c Mice

  • Jegal, Jonghwan;Park, No-June;Jo, Beom-Geun;Kim, Su-Nam;Yang, Min Hye
    • Natural Product Sciences
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    • v.25 no.3
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    • pp.261-267
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    • 2019
  • The rhizomes of Dioscorea japonica Thunb. are widely consumed as food and also used to treat diabetes and polyuria in Korea. This study was undertaken to study the anti-atopic dermatitis effects of a 95% ethanolic extract (DJE) of D. japonica in an oxazolone-stimulated murine model of atopic dermatitis (AD). The therapeutic effects of DJE on AD-like skin lesions were assessed on both ears. DJE (1%) or dexamethasone (0.5%; the positive control) were applied to skin lesions for three weeks. Serum levels of IgE and IL-4 were assessed by ELISA (enzyme-linked immunosorbent assay). Histopathological examinations were performed by hematoxylin and eosin (H&E) and toluidine blue staining and revealed DJE significantly reduced dermal thickness and inflammatory cell infiltration when applied to oxazolone-treated ear skin. DJE-treated AD mice also showed lower serum levels of IgE and IL-4 than oxazolone-stimulated controls. Our findings demonstrate DJE might be a useful safe, topical agent for the treatment of atopic diseases.

Comparative Research of Decalcification Methods for Quick Diagnosis on Bone Tissue (골조직의 신속한 진단을 위한 탈회방법의 비교 연구)

  • Kim, Sung-Chul;Back, Oun-Chul;Kim, Tai-Jeon;Bae, Hyung-Joon;Kang, Hee-Gyoo
    • Korean Journal of Clinical Laboratory Science
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    • v.37 no.1
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    • pp.47-55
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    • 2005
  • These studies were done to know decalcification methods to reduce the time of decalcification for quick bone tissue diagnosis. When bone tissue was decalcified with 10 % formic acid at room temperature, decalcification and hematoxylin & eosin (H&E) stains were complete and satisfactory after 12 hours, but some of the tissue sections fell off during staining. In this way, decalcification, H&E stains were complete and satisfactory after 24 hours, 36 hours and 48 hours, tissue sections didn't fall off during staining. When bone tissue was decalcified with 10 % formic acid in a $60^{\circ}C$ paraffin oven, decalcification and H&E stains were complete and satisfactory after 6 hours, but some tissue sections fell off during staining. In this way, decalcification and tissue sections were complete, with no falling off during staining after 8 hours, 10 hours, 12 hours, 14 hours, 24 hours, or H&E stains were satisfactory from 8 hours to 12 hours, but H&E stains appeared to reddish nucleus after 14 hours and 24 hours. Bone tissue was decalcified with 10 % formic acid for 6 hours, 12 hours and 24 hours at DECAL machine frequencies of 15 Hz and 45 Hz, and for 6 hours, 12 hours and 24 hours at a DECAL machine frequency of 90 Hz. Decalcification and H&E stains were complete and satisfactory after 6 hours at the 15 Hz and 45 Hz DECAL settings. Some of the tissue sections fell off during staining at the 15 Hz DECAL machine setting. At the 90 Hz setting, decalcification, H&E stains, and tissue sections were complete and satisfactory with no falling off during staining after 4 hours. In this way, decalcification, H&E stains, and tissue section were complete and satisfactory with no falling off during staining after 12 hours, 24 hours at all machine settings. Bone tissue was decalcified with 10 % formic acid for 6 hours, 12 hours and 24 hours at $37^{\circ}C$ 3 hours, 6 hours and 12 hours at $45^{\circ}C$ and 1 hours, 5 hours and 10 hours at $60^{\circ}C$ with the RHS-1 machine setting at 60Hz. At the temperatures of $37^{\circ}C$, $45^{\circ}C$, and $60^{\circ}C$ decalcification, H&E stains, and tissue sections were complete and satisfactory, with no falling off during staining except for after 6 hours at $37^{\circ}C$. 3 hours, 1 hours, or decalcification, H&E stains, and tissue sections were complete and satisfactory with no falling off during staining after 12 hours and 24 hours at $37^{\circ}C$, 6 hours and 12 hours at $45^{\circ}C$, and 5 hours at $60^{\circ}C$. But H&E stains appeared to reddish nucleus after 10 hours at $60^{\circ}C$. From the above reults, the authors were able to deduce that decalcification is accelerated by heat and frequency. We therefore think that it is necessary for machines which are similar to the RHS-1 machine to be maintained at the temperature evenly with agitation effect for quick decalcification.

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Improved Detection of Metastases by Step Sectioning and Immuno-Histochemical Staining of Axillary Sentinel Nodes in Patients with Breast Carcinoma

  • Ensani, Fereshteh;Enayati, Ladan;Rajabiani, Afsaneh;Omranipour, Ramesh;Alavi, Nasrinalsadat;Mosahebi, Sara
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5731-5734
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    • 2013
  • Background: The object of this study was to examine whether a new protocol including step-sectioning and immunohistochemistry (IHC) staining of axillary sentinel nodes (SN) would lead to detection of more metastases in patients with breast cancer. Materials and Methods: Sixty-nine tumor free sentinel lymph nodes were examined. Step frozen sectioning was performed on formalin fixed SN and stained both by hematoxylin and eosin (H and E) and cytokeratin markers using IHC. Any tumoral cell in IHC stained slides were considered as a positive result. Metastases up to 0.2 mm were considered as isolated tumor cells and 0.2 up to 2 mm as micrometastasis. Results: Mean age of the patients was $48.7{\pm}12.2$ years. Step sectioning of the SN revealed 11 involved by metastasis which was statistically significant (p<0.001). Furthermore, 15 (21.7%) of the patients revealed positive results in IHC staining for pan-CK marker and this was also statistically significant (p=0.001). Ten patients had tumoral involvement in lymph nodes harvested from axillary dissection and 4 out of 15 lymph nodes with positive result for CK marker were isolated tumor cells. However, 4 of 10 patients with tumor positive lymph nodes in axillary dissection were negative for CK marker and in contrast 6 of the pan-CK positive SN were in patients with tumor-free axillary lymph nodes. Conclusions: Both IHC and step sectioning improve the detection rate of metastases. Considering the similar power of these two methods, we recommend using either IHC staining or step sectioning for better evaluation of harvested SNs.

Effects of Gyejigabuja-tang on MIA-induced Osteoarthritis in Rats (계지가부자탕(桂枝加附子湯)이 MIA로 유도된 골관절염 Rat 모델에 미치는 영향)

  • Won, Je-Hoon;Woo, Chang-Hoon
    • Journal of Korean Medicine Rehabilitation
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    • v.25 no.2
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    • pp.51-64
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    • 2015
  • Objectives This study was carried out to investigate the effects of Gyejigabuja-tang extracts on the Monosodium iodoacetate (MIA) induced osteoarthritis in rats. Methods Osteoarthritis was induced by injection of MIA into knee joint cavity of rats. Rats are divided into 4 groups (normal, control, positive comparison group, GBT group, each n=5). Normal group was injected by normal saline into knee joint cavity only. Control group was induced for osteoarthritis by MIA and oral medicated with distilled water. Positive comparison group was injected with MIA and taken Joins tablet 25 mg/kg. GBT group was injected with MIA and taken Gyejigabuja-tang extracts 300 mg/kg. Positive comparison group and GBT group were oral medicated for each substance once a day for 4 weeks. ALT, AST and creatinine were evaluated for hepatotoxicity and nephrotoxicity. Hind paw weight bearing ability was examined and inflammatory cytokines (IL-$1{\beta}$, IL-6, TNF-${\alpha}$), $PGE_2$, $LTB_4$, osteocalcin and deoxypyridinoline (DPD) within serum were analysed. Knee joint structures were observed by Hematoxylin & Eosin (H&E), Safranin-O staining method. Results 1. Function of liver and kidney was not affected. 2. Hind paw weight bearing ability was significantly improved. 3. IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ in experimental group were significantly decreased compared with control group. 4. $PGE_2$, $LTB_4$, Osteocalcin and DPD in experimental group were decreased compared with control group. 5. In histopathologic observation, injury on synovial membrane and cartilage of experimental group was lesser than control group (H&E, Safranin-O staining). Conclusions Based on these results, it can be suggested that Gyejigabuja-tang has anti-inflammation effects on the MIA-induced osteoarthritis in rats.

Effect of Cassiae semen extract on ovalbumin-induced allergic asthma in mice (결명자 추출물의 난알부민 감작으로 천식이 유발된 마우스에서의 개선 효과)

  • Seo, Beom-Su;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.30 no.1
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    • pp.25-32
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    • 2015
  • Objectives : In this study, we investigated the effect of Cassia obtusifolia Linne (Cassiae Semen; CS) extract on ovalbumin (OVA)-induced allergic asthma in mice. Methods : CR was extracted with 70% ethanol. For in vitro study, HMC-1, human mast cells were treated with CS extract at 0.2 and $0.5mg/m{\ell}$ for 1 h, and then stimulated with compound (C) 48/80 for 30 min. Primary spleenocytes were isolated from the spleen of mice, treated with CS extract for 1 h, and then stimulated with ConA for 24 h. For in vivo study, mice were sensitized at day 0, 7 and 14 with 0.2% OVA and then airway challenged using neublizer at day 21, 23, 25, and 27 to induced allergic asthma. CS extract at doses of 100 and 300 mg/kg body weight was orally administered during OVA challenge once per a day. The levels of allergic mediators such as histamine, OVA-specific IgE, IL-4, and $IFN-{\gamma}$ were measured in the sera of mice or culture supernatants by EIA and ELISA, respectively. The expression of IL-4 and $IFN-{\gamma}$ gene was determined by RT-PCR. The histopathological change of lung tissues was observed with hematoxylin and eosin (H&E) and Periodic acid Schiff (PAS) staining. Results : The treatment of CS extract in HMC-1 cells significantly inhibited C48/80-induced degranulation, and histamine release. The treatment of CS extract in spleenocytes suppressed the expression of IL-4 and $IFN-{\gamma}$ mRNA. The administration of CS extract in OVA-induced asthmatic mice significantly decreased the levels of OVA-specific IgE, and IL-4 in a dose-dependent manner with OVA-control group. In addition, CS extract inhibited the infiltration of inflammatory cells and bronchiolar damage with epithelial thickening in lung tissues of OVA-induced asthma mice, and also mucin accumulation. Conclusions : These results indicate that CS extract prevents asthmatic damage through regulating the allergic immune response.

Development of Secretory Element in Murine Trachea and Larynx (백서 기관 및 후두에서 분비조직의 발생)

  • 조정일;박기현;김광문
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1993.05a
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    • pp.67-67
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    • 1993
  • Mucosal surface of the trachea and larynx is protected by several complex defense mechanism, namely mucociliary clearance, immunoglobulin, antibacterial secretory enzyme, which have also been demonstrated in the middle ear mucosa and eustachian tube. The morphologgy of secretory glands and cells of the trachea and larynx is well-known, but knowledge concerning their development related to secretory activity is still sketchy. The secretory element of the murine trachea and larynx, aging from gestational day 16 to postnatal day 21, was studied using hematoxylin & eosin and alcian blue/periodic acid-Schiff staining including lysozyme immunohistochemistry to investigate the development of secretory element of the murine trachea and larynx and to provide with basis of the future study for developmental morphology of the trachea and larynx. The results of this study suggest that the secretory activity starts to be established immediately after birth with the aeration of the lung.

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Evaluation of 20(S)-ginsenoside Rg3 loaded hydrogel for the treatment of perianal ulcer in a rat model

  • Jin, Longhai;Liu, Jinping;Wang, Shu;Zhao, Linxian;Li, Jiannan
    • Journal of Ginseng Research
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    • v.46 no.6
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    • pp.771-779
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    • 2022
  • Background: As a kind of common complication of the surgery of perianal diseases, perianal ulcer is known as a nuisance. This study aims to develop a kind of 20(S)-ginsenoside Rg3 (Rg3)-loaded hydrogel to treat perianal ulcers in a rat model. Methods: The copolymers PLGA1600-PEG1000-PLGA1600 were synthesized by ring-opening polymerization process and Rg3-loaded hydrogel was then developed. The perianal ulcer rat model was established to analyze the treatment efficacy of Rg3-loaded hydrogel for ulceration healing for 15 days. The animals were divided into control group, hydrogel group, free Rg3 group, Rg3-loaded hydrogel group, and Lidocaine Gel® group. The residual wound area rate was calculated and the blood concentrations of interleukin-1 (IL-1), interleukin-6 (IL-6), and vascular endothelial growth factor (VEGF) were recorded. Hematoxylin and eosin (H&E) staining, Masson's Trichrome (MT) staining, and tumor necrosis factor α (TNF-α), Ki-67, CD31, ERK1/2, and NF-κB immunohistochemical staining were performed. Results: The biodegradable and biocompatible hydrogel carries a homogenous interactive porous structure with 10 ㎛ pore size and five weeks in vivo degradation time. The loaded Rg3 can be released sustainably. The in vitro cytotoxicity study showed that the hydrogel had no effect on survival rate of murine skin fibroblasts L929. The Rg3-loaded hydrogel can facilitate perianal ulcer healing by inhibiting local and systematic inflammatory responses, swelling the proliferation of nuclear cells, collagen deposition, and vascularization, and activating ERK signal pathway. Conclusion: The Rg3-loaded hydrogel shows the best treatment efficacy of perianal ulcer and may be a candidate for perianal ulcer treatment.

Experimental Analysis in the Reversible and Irreversible Cerebral Ischemic Models in the Rat (백서의 가역성 및 비가역성 뇌허혈 모형의 실험적 고찰)

  • Song, Kwang Chul;Choi, Byung Yon;Kim, Seong Ho;Bae, Jang Ho;Kim, Oh Lyong;Cho, Soo Ho
    • Journal of Korean Neurosurgical Society
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    • v.29 no.7
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    • pp.853-860
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    • 2000
  • Objective : The purpose of our experimental study was to analysis the advantages and disadvantages in the reversible and irreversible cerebral ischemic models with rats by staining with Neutral Red(NR) solusion, 2% 2,3,5-triphenyltetrazolium chloride(TTC) and Hematoxylin & Eosin(H & E). Methods : We have measured the range of cerebral infarction in the rat to get a suitable ischemic model along the object of study with and without craniectomy. With craniectomy, 9 rats were sacrificed for irreversible cerebral ischemic model by means of ligation at proximal(group I) and distal(group II), and coagulation at proximal(group III) middle cerebral artery. Also, 6 rats were sacrificed for irreversible(group IV) and reversible(group V) cerebral ischemic model using nylon thread without craniectomy. The sizes of infarction were measured by staining the coronal sections of the brain with NR solusion, TTC and H & E. Results : There are no difference of physiological parameters comparing the each group. Cerebral infarction was not observed in group II, but it's volume was largest in group IV. Disadvantages of craniectomy group(I, II, III) are the long duration of operation and cortical damage by procedure. It's advantage is confirmation of the middle cerebral artery occlusion and cessation of blood flow through the operative microscope. In case of ischemic models using nylon thread (group IV, V), it is hard to identify the interruption or recirculation of blood flow through the middle cerebral artery, but the advantage is the simplicity of operative technique which reduces the operation time and minimizes the cerebral damage due to craniectomy. Therefore, it seems important to set up the reversible and irreversible ischemic models by carefully considering advantages and disadvantages listed above. Conclusion : TTC staining seems to be effective since it reflects the histological damage sufficiently and quickly. It is hoped that researches focused on ischemic penumbra, which became popular recently, will be further carried on with use of NR staining, optical microscope and electron microscope.

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Analysis of Anti-Allergic Activities by Extracts from Persimmon Sap-Stained Rayon and Cotton Fabrics (감물염색 직물의 추출물에 의한 항알러지 활성의 분석)

  • Lee, Sang-Han
    • Journal of Life Science
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    • v.20 no.5
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    • pp.794-798
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    • 2010
  • Allergic dermatitis is one of the most prevalent diseases in young/juvenile children worldwide. In this research, extracts with persimmon sap-stained fabrics (rayon and cotton) exhibited an elevation in $CD4^+$ cell numbers. MMP-2 and MMP-9 expressions by Hematoxylin-Eosin staining and immunohistochemistry revealed that the expressions were decreased by addition of the extracts. The present results collectively suggest that the active ingredients of persimmon sap-stained fabrics play an important role in inhibition of DNFB-induced-atopic symptoms in vivo.

Effect of dietary n-3 Polyunsaturated Fatty Acids on Bony Remodeling during Eruptive Tooth Movement

  • Kim, Hyun-Jin
    • International Journal of Oral Biology
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    • v.41 no.2
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    • pp.63-68
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    • 2016
  • The aim of this study was to investigate the effect of n-3 polyunsaturated fatty acids (PUFAs) on eruptive movement during tooth development. Sprague-Dawley (SD) rat pups were randomly divided into two groups; control group and experimental group. The experimental group was administered daily with n-3 PUFA by intraperitoneal (IP) injection. After 10 days postpartum, rat pups were sacrificed to evaluate the effect of n-3 PUFA on eruptive tooth movement. Histological analyses were by hematoxylin-eosin (H&E) staining. Tartrate-resistant acid phosphatase (TRAP) assay was performed to compare the osteoclast distribution in the bone matrix above the developing molar teeth. Incisor teeth eruptions were noticeably observed in IP group, as compared to control group. Rat pups in IP group showed faster tooth eruption on day 8 after birth. Through histological analyses, IP group showed thinner bone matrix and more osteoclasts above the $1^{st}$ molar teeth, as compared to control group. TRAP assay showed significantly stronger stained pattern that the osteoclast above the $1^{st}$ molar teeth in IP group, as compared to control group. The results suggested that n-3 PUFA could affect osteoclastic activity involved in bony remodeling during eruptive tooth movement.