• Title/Summary/Keyword: Cell Observation

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FORMATION OF EXTRACELLULAR MATRIX COMPONENTS DURING DEVELOPMENT AND REPAIR OF PERFORATION OF THE RAT DENTIN AND PULP (흰쥐 대구치의 치수강 노출 후 치유 및 형성과정에서 치수와 상아질 기질내의 교원질과 당단백의 분포에 관한 면역조직화학적 연구)

  • Kim, Byung-Wooh;Min, Byung-Soon
    • Restorative Dentistry and Endodontics
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    • v.21 no.1
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    • pp.35-53
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    • 1996
  • The development and repair requires the formation of new tissues comprised of various extracellular matrix components. The present study investigated the formation and distribution of the major ECM components such as type I collagen, type III collagen, fibronection, bone sialoprotein, and osteonection during development and repair. For developing observation. Sprague-Dawley rats weighing $27{\pm}1gm$ were sacrificed. For repair observation, Sprague-Dawley rats weighing $110{\pm}5gm$ were used. The pulp perforation were prepared on mesial surface of the maxillary first molar by using 1/2round bur. At 5 days after perforation, rats were sacrificed by perfusion with 3 % paroformaldehyde. The maxillary first molar region were cut, demineralized, dehydrated and embedded in paraffin. Immunostaining the ECM components was achieved by the avidin-biotin complex method. The results as follows : 1. Bright immunoreaction for fibronectin was present in the basement membrane at the inner epithelial-mesenchymal interface, especially concentrated in the blood vessel walls, cell membrane of odontoblasts, and initial predentin. 2. Type I and III collagen was observed in the newly formed pulp tissue, predentin, and its intensity increased as more of these components during repair. 3. Strong immunostaining for bone sialoprotein and osteonectin was found in dentin while no or weaker staining was observed loose connective tissue of the pulp. 4. These results suggest that develpment and repair is achieved through a series of cell differentiation and attachment by the specific ECM components.

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Advanced tube formation assay using human endothelial colony forming cells for in vitro evaluation of angiogenesis

  • Lee, Hyunsook;Kang, Kyu-Tae
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.6
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    • pp.705-712
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    • 2018
  • The tube formation assay is a widely used in vitro experiment model to evaluate angiogenic properties by measuring the formation of tubular structures from vascular endothelial cells (ECs). In vitro experimental results are crucial when considered the advisability of moving forward to in vivo studies. Thus, the additional attentions to the in vitro assay is necessary to improve the quality of the pre-clinical data, leading to better decision-making for successful drug discovery. In this study, we improved the tube formation assay system in three aspects. First, we used human endothelial colony forming cells (ECFCs), which are endothelial precursors that have a robust proliferative capacity and more defined angiogenic characteristics compared to mature ECs. Second, we utilized a real-time cell recorder to track the progression of tube formation for 48 hours. Third, to minimize analysis error due to the limited observation area, we used image-stitching software to increase the microscope field of view to a $2{\times}2$ stitched area from the $4{\times}$ object lens. Our advanced tube formation assay system successfully demonstrated the time-dependent dynamic progression of tube formation in the presence and absence of VEGF and FGF-2. Vatalanib, VEGF inhibitor, was tested by our assay system. Of note, $IC_{50}$ values of vatalanib was different at each observation time point. Collectively, these results indicate that our advanced tube formation assay system replicates the dynamic progression of tube formation in response to angiogenic modulators. Therefore, this new system provides a sensitive and versatile assay model for evaluating pro- or anti-angiogenic drugs.

Morphological Observation of Alexandrium tanarense (Lebour) Balech, A. catenella (Whedon et Kofoid) Balech and One Related Morphotype (Dinophyceae) in Korea

  • Kim, Keun-Yong;Matoko Yoshida;Yasuwo Fukuyo;Kim, Chang-Hoon
    • ALGAE
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    • v.17 no.1
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    • pp.11-19
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    • 2002
  • Twenty-nine culture strains belonging to the genus Alexandrium Halim (Dinophyceae) were established from water column or sediments in Korea. Seventeen isolates were identified as A. tamarense (Lebour) Balech, eight isolates as A. sp. cf. catenella and one as A. catenella (Whedon et Kofoid) Balech according to the presence or absence of a ventral pore, the shape of the posterior sulcal plate and the sulcal width. Three isolates were unable to be identified due to considerable distortion of thecal plates and lack of enough materials, but typical of A. tamarense and/or A. catenella. The overall cell shape of A. tamarense was usually longer than wide. The posterior sulcal plate was definitely longer than wide dorsoventrally, and sulcus extended posteriorly without apparent widening. They were distributed in three major coasts of Korea. In contrast, the cell shape of A. sp.cf. catenella was generally anterior-posteriorly flattened. The transversal axis of the posterior sulcal plate was always longer than the longitudinal, or both axes were nearly equal in length. Its sulcus was broader than that of A. tamarense and widened in the direction of antapex about 1.5 times. This morphotype existed in nearshore and offshore waters of the southern Korea sea. One of A. catenella isolates from Jinhae Bay showed no conspicuous differences with A. sp. cf. catenella except for the consistent absence of a ventral pore.

Effect of Bunsimgieumgagambang on the Stress Due to the Maternal Separation in Rats (분심기음가감방(分心氣飮加減方)이 모성분리(母性分離) stress 백서(白鼠)에 미치는 영향)

  • Kim Ki-Bong;Kim Jang-Hyun;Chang Gyu-Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.5
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    • pp.1303-1310
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    • 2005
  • This study was performed to investigate the effect of Bunsimgieumgagam on the stress due to the maternal separation in rat. In this study, we researched in 'the behavioral observation', 'the changes of body weight', 'quantitative analysis of the number of BrdU-positive cells per section in dentate gyrus of hippocampus', 'free radical scavenging assay' and 'MTT-based cytotoxicity assay of SK-N-SH cell line', in order to figure out the effect on which Bunsimgieumgagam has the increase of neuron in dentate gyrus of hippocampus damaged by the stress due to the maternal separation. In the behavioral Observation, Bunsimgieumgagam was also efficacious against the decline of one's behavior and anorexia derived from the stress by the maternal separation. In the change of body weight, it showed that the Bunsimgieumgagam is effective in the recovery of weight loss caused by heavy stress(p<0.05). Also, Bunsimgieumgagam had an increasing effect, which is similar to a normal state, on DG's neuron in hippocampus (P<0.001). In free radical scavenging assay, Bunsimgieumgagam had a superior free radical scavenging effect. And it showed a significant result with the high cell proliferation effect in MTT-based cytotoxicity assay(P<0.01, p<0.001) This result suggest that Bunsimgieumgagam has an anti-stress effect and a proliferation effect of neuron in dentate gyrus of hippocampus, and it shows the potential of Bunsimgieumgagam in the treatment for the various disorders derived from children's stress.

Morphological Review of Red Blood Cells After X-ray Irradiation (방사선 조사 후 적혈구의 형태적 고찰)

  • Tae-Jeong Ji
    • Journal of the Korean Society of Radiology
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    • v.17 no.2
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    • pp.223-230
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    • 2023
  • This study attempted to concider the morphological change of red blood cells after whole body irradiation. Blood samples used red blood cells of white mice and mouse after irradiation. Transmission electron microscope observation results, Anisocytosis was observed in red blood cells 20 days after 5 Gy irradiation. Triangle and tetrapod were observed for small red blood cell types. Poikilocytosis, sickle-shaped Drepanocyte, and Acantocyte were observed in general-sized red blood cells. Schizocyte was observed in red blood cells 20 days after 7 Gy irradiation. Scanning electron microscope observation results, Dacryocyte was observed with microcytes. It was also confirmed that red blood cells were get tangled with each other. In addition, polygonal shapes and half-moon shapes were also observed. In conclusion, it is judged that the modified form of pathological study is more important than the numerical change in the study of red blood cells by radiation exposure. In conclusion, it was confirmed that modified morphological studies are more important than numerical changes in the study of red blood cells by radiation exposure.

Suppressive Effect of GJB-7 on Collagen-Induced Arthritis in Mice (관절7호방이 Type II collagen으로 유발된 흰쥐의 관절염에 미치는 영향)

  • Min, Boo-Ki;Oh, Min-Seok
    • The Journal of Korean Medicine
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    • v.31 no.4
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    • pp.63-78
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    • 2010
  • Objectives: This study was carried out to know the effects of Gwan-Jul-Bang-7 (hereafter referred to GJB-7) on the inhibition of arthritis induced by collagen on the mouse. Methods: To assess the effects of GJB-7 on mouse with arthritis induced by collagen II, we conducted several experiments such as analysis of cytotoxicity, hepatotoxicity, arthritis index, total cell number of draining lymph nodes and paw joints, value of immunocyte in PBMC (peripheral blood mononuclear cell), DLN (draining lymph node) and paw joint, measurement of cytokine and anti-collagen II, observation of the histological changes of joint. Results: 1. Cytotoxicity against HFC (human fibroblast cells) was not observed in any concentration and hepatotoxicity was not observed in the GJB-7 treated group. 2. The incidence of arthritis significantly decreased. 3. Total cell number of draining lymph nodes significantly increased and total cell number of paw joints significantly decreased. 4. The percentage of $CD8^+$ cells in PBMC (peripheral blood mononuclear cell) significantly increased. The percentage of $CD3^+/CD69^+$, and $CD3^+/CD49b^+$ cells in PBMC significantly decreased. 5. The percentage of $CD19^+,\;CD3^+$, and $CD4^+/CD25^+$ cells in DLN (draining lymph nodes) significantly increased. The percentage of $B220^+/CD23^+$ cells in DLN significantly decreased. 6. The percentage of $CD3^+,\;CD4^+$, and $CD11b^+/Gr-1^+$ cells in paw joints significantly decreased. 7. The production of TNF-$\alpha$, IL-6, and MCP-1 significantly decreased. 8. Anti-collagen II in serum significantly decreased. 9. With the hematoxylin and eosin stain, inflammation of joint decreased. Under Masson's trichrome stain, the cartilage destruction and synovial cell proliferation and the expression of collagen fibers decreased. Conclusions: Comparison of the results for this study showed that GJB-7 had immunomodulatory effects. So we expect that GJB-7 could be used as an effective drug for not only rheumatoid arthritis but also another auto-immune diseases.

Macerating Properties of Fruits and Vegetables for Suspensions Containing Single Cells (단세포 함유 반응물 제조를 위한 과일과 채소류의 가공 특성)

  • Park, Yong-Kon;Kang, Yoon-Han
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.64-68
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    • 2004
  • Cell-separating enzyme (Sumyzyme MC) was used to investigate enzymatic maceration of strawberry, sweet persimmon, kiwi, onion, garlic, and cucumber, Maceration rate, volume, brix, color, particle size distribution, and viscosity were determined, and microscopic observation made on suspensions containing single cells. Sweet persimmon and strawberry showed over 90% meceration rates, and kiwi showed 80%. Color, storage test, and sensory evaluations of single-cell suspensions and their filtrates were performed before and after sterilization. Total dietary fiber contents of raw material and single-cell suspension of garlic were 30.77 and 18.55%, respectively, Results indicate fruit and vegetable suspensions produced through enzymatic disintegration using cell-separating enzyme can be utilized as basic materials in the manufacture of single-cell foods.

Anti-Cancer Effects and Apoptosis by Korean Medicinal Herbs

  • Ko Seong Gyu;Jun Chan Yong;Park Chong Hyeong;Bae Hyun Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.3
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    • pp.819-825
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    • 2003
  • pharbitis nil and Taraxacum mongolicum are representative herbs that have been used for cancer treatment in Korean traditional medicine. To understand the molecular basis of the antitumor function, we analyzed the effect of these herbs on proliferation and apoptosis of tumor cells using a gastric cancer cell line AGS. Cell counting assay showed that pharbitis nil strongly inhibit cell proliferation Of AGS whereas Taraxacum mongolicum exhibit no detectable effect on cellular growth. [³H]thymidine uptake analysis also demonstrated that DNA replication of AGS is suppressed in a dose-dependent manner by treatment with pharbitis nil. Additionally, tryphan blue exclusion assay showed that Pharbitis nil induce apoptotic cell death of AGS in a dose-dependent. To explore whether anti antiproliferative and/or proapototic property of Pharbitis nil is associated with their effect on gene expression, we performed RT-PCR analysis of cell cycle- and apoptosis-related genes. Interestingly, mRNA expression levels of c-Jun, c-Fos, c-Myc, and Cyclin D1 were markedly reduced by Pharbitis nil. Taraxacum mongolicum also showed inhibitory action on expression of these growth-promoting protooncogene but there effects are less significant, as compared to Pharbitis nil. Furthermore, it was also found that Pharbitis nil activates expression of the p53 tumor suppressor and its downstream effector p21Waf1, which induce G1 cell cycle arrest and apoptosis. Collectively, our data demonstrate that Pharbitis nil induce growth inhibition and apoptosis of human gastric cancer cells and these effects are accompanied with down-and up-regulation of growth-regulating protooncogenes and tumor suppressor genes, respectively. This observation thus suggests that the anticancer effect of Pharbitis nil might be associated with its regulatory capability of tumor-related gene expression.

Cytokinetic Study of MCF-7 Cells Treated with Commercial and Recombinant Bromelain

  • Fouz, Nour;Amid, Azura;Hashim, Yumi Zuhanis Has-Yun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6709-6714
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    • 2013
  • Background: Breast cancer is a leading cause of death in women. The available chemotherapy drugs have been associated with many side effects. Bromelain has novel medicinal qualities including anti-inflammatory, anti-thrombotic, fibrinolytic and anti-cancer functions. Commercially available bromelain is obtained through tedious methods; therefore, recombinant bromelain may provide a cheaper and simpler choice with similar quality. Materials and Methods: This study aimed to assess the effects of commercial and recombinant bromelain on the cytokinetic behavior of MCF-7 breast cancer cells and their potential as therapeutic alternatives in cancer treatment. Cytotoxic activities of commercial and recombinant bromelain were determined using (sulforhodamine) SRB assay. Next, cell viability assays were conducted to determine effects of commercial and recombinant bromelain on MCF-7 cell cytokinetic behavior. Finally, the established growth kinetic data were used to modify a model that predicts the effects of commercial and recombinant bromelain on MCF-7 cells. Results: Commercial and recombinant bromelain exerted strong effects towards decreasing the cell viability of MCF-7 cells with $IC_{50}$ values of 5.13 ${\mu}g/mL$ and 6.25 ${\mu}g/mL$, respectively, compared to taxol with an $IC_{50}$ value of 0.063 ${\mu}g/mL$. The present results indicate that commercial and recombinant bromelain both have anti-proliferative activity, reduced the number of cell generations from 3.92 to 2.81 for commercial bromelain and to 2.86 for recombinant bromelain, while with taxol reduction was to 3.12. Microscopic observation of bromelain-treated MCF-7 cells demonstrated detachment. Inhibition activity was verified with growth rates decreased dynamically from 0.009 $h^{-1}$ to 0.0059 $h^{-1}$ for commercial bromelain and to 0.0063 $h^{-1}$ for recombinant bromelain. Conclusions: Commercial and recombinant bromelain both affect cytokinetics of MCF-7 cells by decreasing cell viability, demonstrating similar strength to taxol.

Effects of Rhei Rhizoma on Gastric Ulcer in Sprague-Dawley Rats (대황(大黃)이 흰쥐의 위점막 손상에 미치는 영향)

  • Kim, Bum-Hoi
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.1
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    • pp.71-77
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    • 2011
  • Gastric ulcer has multifactorial etiology, and the development of ulcer is known to be caused by gastric acidity, pepsin secretion, gastric motility and gastric mucosal blood flow. The ulcer results from the tissue necrosis and apoptotic cell death triggered by mucosal ischemia, free radical formation and cessation of nutrient delivery. The gastric mucosa is usually exposed to a wide range of aggressive insults, and has developed efficient mechanisms to repair tissue injury. The apoptotic process of gastric mucosa is triggered by the induction of such proapoptotic gene expression, such as BAX. The Bcl-2 family of proteins plays a pivotal role in the regulation of apoptosis. The maintenance of gastric mucosa integrity depends upon the ratio between cell proliferation and cell death. Stress-inducing factors may affect Bcl-2/BAX ratio and thus the rate of apoptosis through modulation of the expression of both proteins depends upon the experimental model. In addition to the regulation of apoptosis, new vessels have to be generated in order to ensure an adequate supply of oxygen and nutrients to the healing gastric mucosa. This events are regulated by several factors. Among them, such polypeptide growth factors, such as vascular endothelial growth factor (VEGF) regulates essential cell functions involved in tissue healing including cell proliferation and differentiation. The purpose of this study was carried to investigate whether Rhei Rhizoma administration might protect apoptotic cell death and promote angiogenesis in gastric mucosa. Sprague-Dawley rats were randomly divided into 4 groups; normal, saline, cimetidine and Rhei Rhizoma-treated group. The saline, cimetidine and Rhei Rhizoma extracts were orally administrated to each group and gastric ulcer was induced by HCl-EtOH solution. After 1 hour, the stomachs were collected for histological observation and immunohistochemistry. In results, Rhei Rhizoma proves to promote to heal wound in gastric ulcer in conclusion and the significant changes of BAX, Bcl-2 and VEGF quantity in gastric mucosa were observed. These results suggest that Rhei Rhizoma extract may promote incision wound healing and has protective effects on gastric ulcer in rats.