• Applied Microscopy
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• v.41 no.3
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• pp.189-196
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• 2011

Histological study on egg envelope and oogenesis of Kichulchoia multifasciata (Pisces, Cobitidae) was carried out by light microscopes and scanning electron microscopes. Various developmental cells appeared in ovary of the specimen catched during November 2010. The cytoplasm of oogonia was acidic and many nucleoli were located at the inner side of nucleus membrane. The size of the oogonia was $103.9{\pm}24.7$ ${\mu}m$ with nucleus size $42.9{\pm}6.9$ (31.1~50.3) ${\mu}m$. Primary oocyte having $277.5{\pm}60.5$ (216.7~354.9) ${\mu}m$ in diameter began to accumulate yolk vesicles. As the developmental stages proceed, secondary oocyte grows larger to $617.6{\pm}85.1$ (503.4~723.6) ${\mu}m$, and eosinophilic yolk granules yolk granules appeared between the yolk vesicles occupying most cytoplasm, and there are some yolk mass formed already. There are some yolk mass formed already. Envelope of fertilized egg investigated by a scanning electron microscope had plenty of microvilli (2~3 ${\mu}m$ in length) over the entire egg surface and a micropyle. Especially, the microvilli surrounding the micropyle were longer than those of egg surface with $5.26{\pm}1.22$ ${\mu}m$.

• Journal of Periodontal and Implant Science
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• v.26 no.2
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• pp.511-521
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• 1996

Periodontal pocket is one of the most frequently developed clinical feature on the teeth with periodontal disease. In order to determine the gingival curettage effect of intrapocket irradiation of a pulsed Nd : YAG laser on periodontally involved teeth, bilateral 60 teeth with $4{\sim}6mm$ in probing pocket depth 1 week after supragingival scaling were selected. On half of them the intrapocket irradiation($300{\mu}m$ fiber optic, 1.5W power, for 2min.) of a pulsed-Nd : YAG laser(EL.EN.EN06O, Italy) was applied as the lased group. On the contralateral 30 teeth the subgingival curettage was accomplished by Gracey curettes as the curattage group. The periodontal pocket tissues were surgically excised by the modified Widman flap technique immediately after the intrapocket irradiation or subgingival curettage, subsequently fixed with 10% neutral formalin, sectioned in $4{\sim}6{\mu}m$ thickness, and stained with hematoxylin-eosin. Surface characteristics and incomplete removal of the pocket epithelium were evaluated under light microscope. And the difference between the lased group and the curettage group was statistically analyzed by Chi-square test in Microstat program. The results were as follows ; 1. The plane surface was observed more frequently in the curettage group(73.3%) than in the lased group(23.3%), and the rough surface was observed more frequently in the lased grOoup(63.3%) than in the curettage group(6.7%)(p<0.05). 2. The rate of incomplete removal of the pocket epithelium was relatively high in both the lased group(76.6%) and the curettage group(86.6%), and there was no significant difference between the lased group and the curettage group(p>0.l). The results suggest that the further studies including various power control of laser should be succeeded in order to obtain more favorable results by the intrapocket irradiation of a pulsed Nd:YAG laser than the subgingival curettage with Gracey curettes.

• Restorative Dentistry and Endodontics
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• v.34 no.2
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• pp.120-129
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• 2009

The purpose of this study was to investigate the pulpal response to direct pulp capping with dentin sialoprotein (DSP) -derived synthetic peptide in teeth of dogs, and to compare its efficacy to capping substances $Ca(OH)_2$ and white mineral trioxide aggregate (WMTA). A total of 72 teeth of 6 healthy male beagle dogs were used. The mechanically exposed pulps were capped with one of the following: (1) DSP-derived synthetic peptide (PEP group): (2) $Ca(OH)_2$ (CH group): (3) a mixture paste of peptide and $Ca(OH)_2$ (PEP+CH group): or (4) white MTA (WMTA group). The access cavity was restored with a reinforced glass ionomer cement. Two dogs were sacrificed at each pre-determined intervals (2 weeks, 1 month, and 3 months). After the specimens were prepared for standard histological processing, sections were stained with hematoxylin and eosin. Under a light microscope, inflammatory response and hard tissue formation were evaluated in a blind manner by 2 observers. In the PEP group, only 3 of 17 specimens showed hard tissue formation, indication that the DSP-derived synthetic peptide did not induce proper healing of the pulp. Compared with the CH group, the PEP group demonstrated an increased inflammatory response and poor hard tissue formation. The CH and WMTA groups showed similar results for direct pulp capping in mechanically exposed teeth of dogs.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.3
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• pp.183-190
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• 2001

Objective: Recently, microdissection of tissue sections has been used increasingly for the isolation of morphologically identified homogeneous cell populations, thus overcoming the obstacle of tissue complexity for the analysis cell-specific expression of macromolecules. The aim of the present study was to establish the minimal conditions required for the RNA extraction and amplification from the cells captured by the laser captured microdissection. Methods : Mouse ovaries were fixed and cut into serial sections (7 im thickness). Oocytes were captured by laser captured microdissection (LCM) method by using PixCell $II^{TM}$ system. The frozen sections were fixed in 70% ethanol and stained with hematoxylin and eosin, while the paraffin sections were stained with Multiple stain. Sections were dehydrated in graded alcohols followed by xylene and air-dried for 20 min prior to LCM. All reactions were performed in ribonuclease free solutions to prevent RNA degradation. After LCM, total RNA extraction from the captured oocytes was performed using the guanidinium isothiocyanate (GITC) solution, and subsequently evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) for glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). Results: With the frozen sections, detection of the GAPDH mRNA expression in the number of captured 25 oocytes were not repeatable, but the expression was always detectable from 50 oocytes. With 25 oocytes, at least 27 PCR cycles were required, whereas with 50 oocytes, 21 cycles were enough to detect GA PDH expression. Amount of the primary cDNA required for RT-PCR was reduced down to at least 0.25 $\grave{i}$ l with 50 oocytes, thus the resting 19.75 il cDNA can be used for the testing other interested gene expression. Tissue-to-slide, tissue-to-tissue forces were very high in the paraffin sections, thus the greater number of cell procurement was required than the frozen sections. Conclusion: We have described a method for analyzing gene expression at the RNA level with the homogeneously microdissected cells from the small amount of tissues with complexity. We found that LCM coupled with RT-PCR could detect housekeeping gene expression in 50 oocytes captured. This technique can be easily applied for the study of gene expression with the small amount of tissues.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.1
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• pp.92-97
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• 2005

Inhibition of inflammatory response, acceleration of basal cell growth, and balanced synthesis of extracellular matrix (ECM) are important in healing of cutaneous open wounds. In order to evaluate the healing effects of water extracts of Radix Astragali (the root of Astragalus membranaceus (Fisch.)) on open wound at early stage, the experimental open wounds were generated on the dorsal sides of SD rats under anesthesia. The boiled-water extracts of Radix Astragali $(100{\mu}l)$, soaked into an occlusive film dressing were applied once a day for eleven consecutive days. The healing process was assessed by measuring macroscopic appearance and wound areas of the open wounds. The molecular aspects of healing process by Radix Astragali extracts were also investigated by Hematoxylin-Eosin (H-E) double staining and immunohistological staining of collagen type I in the healed skin area, implying cell density and linear alignment of the granulation tissue, and ECM synthesis and its remodeling, respectively. The Astragali radix extracts were found to significantly accelerate the cutaneous wound healing by suppressing the inflammation and stimulating the basal cell growth in wounded area, as compared to epidermal growth factor (EGF).

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.2
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• pp.551-557
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• 2015

Background: Differentiating morphologic features based on hematoxylin-eosin (HE) staining is the most common method to classify pathological subtypes of non-small-cell lung cancer (NSCLC). However, its accuracy and inter-observer reproducibility in pathological diagnosis of poorly differentiated NSCLC remained to be improved. Materials and Methods: We attempted to explore the role of immunohistochemistry (IHC) staining in diagnosing pulmonary squamous cell carcinoma (SQCC) with poorly differentiated features by HE staining or with elevated serum adenocarcinoma-specific tumor markers (AD-TMs). We also compared the difference of epidermal growth factor receptor (EGFR) mutation rate between patients with confirmed SQCC and those with revised pathological subtype. Logistic regression analyses were used to test the association between different factors and diagnostic accuracy. Results: A total of 132 patients who met the eligible criteria and had adequate specimens for IHC confirmation were included. Pathological revised cases in poor differentiated subgroup, biopsy samples and high-level AD-TMs cases were more than those with high/moderate differentiation, surgical specimens and normal-level AD-TMs. Moreover, biopsy sample was a significant factor decreasing diagnostic accuracy of pathological subtype (OR, 4.037; 95% CI 1.446-11.267, p=0.008). Additionally, EGFR mutation rate was higher in patients with pathological diagnostic changes than those with confirmed SQCC (16.7% vs 4.4%, p=0.157). Conclusions: Diagnosis based on HE staining only might cause pathological misinterpretation in NSCLC patients with poor differentiation or high-level AD-TMs, especially those with biopsy samples. HE staining and IHC should be combined as pathological diagnostic standard. The occurrence of EGFR mutations in pulmonary SQCC might be overestimated.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.5
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• pp.1003-1011
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• 2009

This study was carried out to know the effects of Kwanjulbang-6(GJB-6) on the inhibition of arthritis. GJB-6 was orally administered to mouse with arthritis induced by collagen II . Cytotoxicity, hepatotoxicity, arthritis index, value of immunocyte in draining lymph node and paw joint, rheumatoid factor in serum were measured in vivo. The incidence of arthritis was significantly decreased. Total cell number of draining lymph node was significantly increased compared with control. Total cell number of paw joint was significantly decreased compared with control. The absolute number of $CD19^+$, $CD8^+$, $CD3^+/CD69^+$, $CD3^+/CD49b^+$, $CD4^+/CD44^+$, $CD3^+$, $CD4^+$, $CD4^+/CD25^+$ and $CD3^+/CD8^+$ cells in draining lymph node were significantly increased compared with control. The absolute number of $CD3^+$, $CD4^+$, $CD4^+/CD25^+$ and $CD11b^+/Gr-1^+$ cells in paw joint were significantly decreased compared with control. The absolute number of $B220^+/CD23^+$ and $MHCII^+/CD11c^+$ cell in draining lymph node were significantly decreased compared with control. The levels of IgG was decreased and The levels of IgM was significantly decreased compared with control. Anti-collagen II in serum was significantly decreased compared with control. With the hematoxylin and eosin stain, the cartilage destruction and synovial cell proliferation were decreased compared with control. With the Masson's trichrome stain, the expression of collagen fibers was decreased compared with control. Results showed that GJB-6 had immunomodulatory effects. So we expect that GJB-6 should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease.

• Journal of Periodontal and Implant Science
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• v.38 no.1
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• pp.59-66
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• 2008

Purpose: It has been shown that the inorganic polyphosphate is effective for the regeneration of bones through the preliminary animal test of rabbits. The most effective concentration of the polyphosphate, however, is not known yet. Moreover, the effectiveness of carriers inside human body is not confirmed.. Materials and Methods: In this study, we examined the effect of the concentration of the inorganic polyphosphate on the process of the bone regeneration using the 6 weeks old rabbits with the weight of 2.0 kg in average. We performed the experiment using TR-ePTFE membrane(membrane) filled with collagen immersed in 4%, 8% of inorganic polyphosphate, respectively, following removal of the proper sized cortical bones from the rabbit calvaria. The experimental results were compared with the one of the following four groups: The negative control group for membrane only, the positive control group for membrane filled with collagen, the first experimental group for membrane filled with collagen immersed in 4% of inorganic polyphosphate, and the second experimental group for membrane filled with collagen immerse in 8% of inorganic polyphosphate. The fragments of the tissue with membrane obtained from each group of the sacrificed rabbits for 8 or 16 weeks sustained after surgery were then prestained by the Hematoxylin-Eosin stain and coated by resin to form non-decalcified specimens for the histologic examination and analysis. New bone formation was assessed by histomorphometric and statistical analysis. Results: 1. All groups have shown better bone regeneration at 16weeks than 8weeks. 2. Negative control group has shown more bone regeneration relative to the other groups at 8 and 16 weeks. 3. All experimental groups have shown better bone regeneration relative to positive control group. 4. At 16 weeks, the first experimental group has shown more bone regeneration compared to the second experimental group. Exophytic bone formation is not good at the first and the second experimental groups compared with negative control group. But, the use of 4% inorganic polyphosphate was more effective to bone formation than the use of 8% inorganic polyphosphate. Conclusion: With above results, it is suggested the use of inorganic polyphosphate with vehicle under TR-ePTFE membrane.

• Journal of Korean Medicine Rehabilitation
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• v.18 no.2
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• pp.45-60
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• 2008

Objectives : Sopung-tang(Shufeng-tang) is a famous herbal prescription that treated ischemic brain injury. This study was designed to evaluate the effects of Sopung-tang(Shufeng-tang) on congnition and motor function recovery after ischemic brain injury in rats. Methods : Male rats were divided into 4 groups. Those rats caused ischemic brain injury by occlusion of MCA as Longa method. Control group I was per os normal saline for 7 days after ischemic brain injury. Control group II was per os normal saline for 14 days after ischemic brain injury. Experimental group I(Ex I) was taken with Sopung-tang(Shufeng-tang) for 7 days after ischemic brain injury. Experimental group II(Ex II) was taken with Sopung-tang(Shufeng-tang) for 14 days after ischemic brain injury. The author carried out neurological, cognitive motor behavior tests and histological assessment. Neurological motor behavior tests consist of limb placement test, beam-walking test and horizontal wire test. Morris water maze test was used for cognitive motor behavior test. In the histological assessment test, TTC(2,3,5-triphenylteterazolium chloride) staining, Hematoxylin & Eosin staining and immunohistochemical staining were experimented. Results : 1. In neurological motor behavior tests, motor function recovery was significantly increased in the experimental groups as compared with control groups(p<0.05). Especially Ex II was significantly increased as compared with Ex I(p<0.05). 2. In Morris water maze test, congnitive motor function recovery was significantly increased in the experimental groups as compared with control group(p<0.05). Especially Ex II was significantly increased as compared with Ex I(p<0.05). 3. In the immunohistochemical staining for the expression of BDNF in hippocampus, more immune reaction was investigated in the experimental groups as compared with control groups. Especially most immune reaction was experimented in the EX II. Conclusions : According to the above results, Sopung-tang(Shufeng-tang) can treat on the congnition and motor function recovery after ischemic brain injury in rats. And it is effective method in expression of BDNF in hippocampus.

• The Korea Journal of Herbology
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• v.28 no.4
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• pp.77-81
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• 2013

Objectives : Horse oil (HO) has been used long time as the folk medicine of many Asian countries such as Korea, Mongol, China, India and Japan. HO has been used for anti-bacterial, anti-inflammatory, and anti-pruritic purposes in skin. However, it is still largely unknown whether HO modulates the skin condition. In this study, we attempted to evaluate the anti-inflammatory effect of HO on the 1 % of 2, 4-dinitro-1-chlorobenzene (DNCB)-induced contact hypersensitivity in Balb/c mice. Methods : To find the anti-inflammatory effect of HO, contact hypersensitivity, a local inflammatory response of skin, was induced on the back of Balb/c mice by sensitization and repeated application by 1% DNCB and HO treated 2 weeks on the 1% of DNCB-treated Balb/c mice. Excised mice skins were stained with hematoxylin and eosin and serum IgE level was measured by mouse IgE ELISA kit. Results : In this study, we found that HO reduced erythema by 1% of DNCB treated Balb/c mice. Also, HO recovered histopathological features such as the thickening of epidermis, hyperkeratosis and the infiltration of inflammatory cells in 1% of DNCB treated Balb/c mice. In addition, HO reduced IgE level on the serum obtained from blood of 1% of DNCB-treated Balb/c mice. Conclusion : Taken together, these results showed that HO could be used as a pharmaceutical material with anti-inflammatory effects by reducing of erythema, IgE level and recovering of histopathological features skin on DNCB-induced contact hypersensitivity in Balb/c mice model.