• Title/Summary/Keyword: Giemsa staining

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Effects of Several Medicine Herb Prescriptions on Lung Carcinoma Cells (수종의 한약처방이 폐암세포에 미치는 영향)

  • Kang, Pil-Gu;Park, Dong-Il;Choi, Won-Choul;Jeon, Jong-Choul
    • The Journal of Internal Korean Medicine
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    • v.21 no.4
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    • pp.621-631
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    • 2000
  • Objective : In this study, the medicine herbs above were directly treated to cultured normal lung cells and lung carcinoma cells, and the effects were investigated to develope new cancer treatments with increased anti-cancer efficiency as well as decreased side-effects and to suggest more useful clinical therapies. Materials and Methods : In the experiment, WI-26 VA lung normal cell line and A-427 lung carcinoma cell line were cultured. To observe the morphological change of the treated cells, were subjected to Giemsa staining and observed under Reflected Fluorescence microscope. To examine whether cell death occurred, cells observed under Reflected Fluorescence microscope, To investigate the degree of cell death in the nucleus, cells were screened by Laser cytometry ACAS 570. Results : Samgibopye-tang(Shenqibufei-tang) stimulated not only the growth of the normal cells but also that of the carcinoma cells, Wikyeung-tang(Weijing-tang) induced morphological change such as cytoplasmic constriction in the normal cells and the carcinoma cells, but it did not show any strong inhibitory effect on the cell growth. Samso-eum(Shensu-yin) caused severe cell damage in both cell lines, Eunkyo San(Yinqiao-san) significantly damaged the nuclei and caused weak cytoplasmic constriction in both cell lines, Normal cells treated with Gilgyeung-tang(Jingeng-tang) did not show any significant morphological change while some Gilgyeung-tang(Jingeng-tang) treated carcinoma cells were observed to have a normal cell-like shape, interestingly, conclusions : As the results above, Samgibopye-tang(Shenqibufei-tang), Wikyeung-tang(Weijing-tang), and Gilgyeung-tang(Jingeng-tang) helped the growth of both cell lines, and especially Samgibopye-tang(Shenqibufei-tang) showed the best effect, However, Samso-eum(Shensuyin) and Eunkyo-san(Yinqiao-san) caused lethal damage in the normal cells and also showed strong toxicity in the carcinoma cells.

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Comparative Studies to Evaluate Relative in vitro Potency of Luteolin in Inducing Cell Cycle Arrest and Apoptosis in HaCaT and A375 Cells

  • George, Vazhapilly Cijo;Kumar, Devanga Ragupathi Naveen;Suresh, Palamadai Krishnan;Kumar, Sanjay;Kumar, Rangasamy Ashok
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.631-637
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    • 2013
  • Luteolin is a naturally occurring flavonoid present in many plants with diverse applications in pharmacology. Despite several studies elucidating its significant anti-cancer activity against various cancer cells, the mechanism of action in skin cancer is not well addressed. Hence, we investigated the effects of luteolin in HaCaT (human immortalized keratinocytes) and A375 (human melanoma) cells. The radical scavenging abilities of luteolin were determined spectrophotometrically, prior to a cytotoxic study (XTT assay). Inhibitory effects were assessed by colony formation assay. Further, the capability of luteolin to induce cell cycle arrest and apoptosis were demonstrated by flow cytometry and cellular DNA fragmentation ELISA, respectively. The results revealed that luteolin possesses considerable cytotoxicity against both HaCaT and A375 cells with $IC_{50}$ values of 37.1 ${\mu}M$ and 115.1 ${\mu}M$, respectively. Luteolin also inhibited colony formation and induced apoptosis in a dose and time-dependent manner by disturbing cellular integrity as evident from morphological evaluation by Wright-Giemsa staining. Accumulation of cells in G2/M (0.83-8.14%) phase for HaCaT cells and G0/G1 (60.4-72.6%) phase for A375 cells after 24 h treatment indicated cell cycle arresting potential of this flavonoid. These data suggest that luteolin inhibits cell proliferation and promotes cell cycle arrest and apoptosis in skin cancer cells with possible involvement of programmed cell death, providing a substantial basis for it to be developed into a potent chemopreventive template for skin cancer.

Multisystemic Aspergillosis with Granulomas in Layer Chickens (산란계에서의 전신 다발성 육아종성 Aspergillosis 발생)

  • Kim, Ji-Ye;Kim, Jong-Man;Mo, In-Pil
    • Korean Journal of Poultry Science
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    • v.38 no.1
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    • pp.45-50
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    • 2011
  • A case of aspergillosis in 39-day-old layer chickens having a history of gradual emaciation and subsequently death with nervous signs such as torticollis and lack of equilibrium was documented. Based on the results from serology and polymerase chain reaction (PCR) test, this flock was not affected with known viral or bacterial diseases. On postmortem examination of the affected birds, multiple white to yellow nodules measuring 1~5 mm in diameter were observed in the lungs, cerebrum, liver and kidney. Microscopically, these nodules were identified as granulomatous lesions characterized by mixed population of multinucleated giant cells and lymphocytes. By periodic acid-schiff staining and nucleotide sequencing analysis, Aspergillus flavus with characteristic septate and branched hyphae were identified in the granuloma of lung and cerebrum. This case was a chronic and multisystemic aspergillosis specialized to central nervous system caused by Aspergillus flavus infection in the layer flocks.

A case of idiopathic thrombocytopenia in a Yorkshire terrier dog (개 특발성 혈소판감소증 1례)

  • Kim, So-La;Cho, Chang-Seok;Kim, Min-Jae;Kang, Tae-Young;Lee, Joo-Myoung;Park, Hyun Jung;Cheong, Jongtae;Lee, Kyoung-kap;Yun, Young-min
    • Korean Journal of Veterinary Research
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    • v.47 no.4
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    • pp.475-478
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    • 2007
  • A 4-year-old spayed female, Yorkshire terrier dog with a history of petechial and ecchymotic hemorrhages on the face, trunk and hind limb was referred to Veterinary Medical Teaching Hospital, Cheju National University. The complete blood count revealed a marked thrombocytopenia ($96{\times}10^3/{\mu}l$). The biochemical profile showed only slightly increased glucose. The coagulation profile such as prothrombin time and activated partial thromboplastin time, was within the reference range. In Giemsa staining, there were no endoparasites like as Babesia spp. on the RBC. This case was diagnosed as a idiopathic thrombocytopenia in a Yorkshire terrier dog. She was treated with prednisone (1 mg/kg BW, IM q 12 h) and cephalosporin (10 mg/kg BW, IM, q 12 h). When rechecked on day 6, the platelet was within reference range ($507{\times}10^3/{\mu}l$) and also petechial and ecchymotic hemorrhages on the body were gradually improved without any complications. The dose of prednisone was decreased to 0.5 mg/kg BW, q 24 h. On day 17, we finished treatment because all the clinical signs, blood and serum chemistry were reference range and platelet count was dramatically increased.

Study on natural killer cell activity and its characteristics during hepatocarcinogenesis in rats (랫드의 간암 발생과정에서 분리한 자연살해세포의 활성측정 및 특성연구)

  • Jeong, Ja-young;Lee, Kuk-kyung;Kil, Jwang-sup;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.169-176
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    • 1999
  • The purposes of this study were to set up the method of the natural killer(NK) cell activity assay using the flow cytometer and to examine the characteristics and distribution of the NK cell during rat hepatocarcinogenesis. Forty five male 6 week-old specific pathogen free(SPF) Sprague-Dawley rats were randomly divided into three groups. Group I was the non-treated control and given normal diet and water. Group II was treated with diethylnitrosamine(DEN, 200mg/kg, i.p.) and partial hepatectomy. Group III was treated with DEN, partial hepatectomy and 0.05% phenobarbital sodium in water from 3 to 16 weeks. All animals were examined the morphology of the large granular lymphocyte(LGL), the LGL percent of the total lymphocytes and the LGL conjugation rate with YAC-1 cell in peripheral blood, spleen and liver. Moreover, activity of the LGL isolated from peripheral blood lymphocytes was determined using the flow cytometer. As results, LGL were observed in the peripheral blood, spleen and liver. LGL were observed the relatively faintly staining basophilic cytoplasm with granules, and eccentric, often kidney-shaped nuclei in Giemsa stain. Its size was $11{\sim}13{\mu}m$. LGL percentage of the isolated lymphocytes in peripheral blood, spleen and liver were 1.8~2.3%, 1.3~1.4% and 0.87~0.99%, respectively. LGL conjugation rate with YAC-1 cell was shown to be peripheral blood(9.3~10.3 %) > spleen(7.7~8.7%) > liver(5.6~7.0%). The activity of the LGL isolated from peripheral blood lymphocytes in Group I, II and III was 33.7%, 30.5% and 35.4%, respectively. However, all values were not significantly between groups.

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Anticancer Effects of Vitamin D3 Analog on Human Leukemic Cell Line(U937) and Role of Vitamin $D_3$ Analog on Immune Function of Human Peripheral Blood Lymphocytes (Human 백혈병 세포에 대한 비타민 $D_3$ 유도체의 항암효과 및 Human 임파구의 면역기능에 대한 비타민 $D_3$ 유도체의 역할)

  • 정수자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.1
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    • pp.141-148
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    • 1998
  • This study describes the effects of 1,25-dihydroxyvitamin D3[1,25(OH)2D3, calcitriol] analog, 1,25(OH)2-16ene-23yne-D3 on proliferatin and differentiatin of the human histiocytic lymphoma cell line U937. This paper also describes the effects of 1,25(OH)2-16ene-23yne-D3 on ${\gamma}$-interferon(IFN-${\gamma}$) synthesis by phytohemagglutinin-activated peripheral blood lymphocytes(PBLs). In the present investigation, 1,25(OH2)-16ene-23yne-D3 was compared to the natural metablite of vitamin D3, 1,25(OH)2D3. 1,25(OH)2-16ene-23yne-D3 was more potent than 1,25(OH)2D3 for inhibition of proliferation and induction of differentiation of U937 cells, Its effects on inhibition of proliferation was about 30-fold more potent than 1,25(OH)2D3. On induction of differentiation as measured by nonspecific esterase (NSE) activity and morphologic change, this analog morphologically and functionally differentiated U937 cells to monocyte-macrophage phenotype showing a decrease of N/C ration in Giemsa staining and the increase of adherence ability of surface. After 3 days in culture, a more significant supression of IFN-${\gamma}$ synthesis analog on supression of IFN-${\gamma}$ synthesis was a dose-dependent manner, with peak activity at 10-7M. The strong direct effects of 1,25(OH)2-16ene-23yne-D3 on cell proliferation and cell differentiation, make this compound an interesting candidate for clinical studies for several types of malignancies, and the effects on supression of IFN-${\gamma}$ synthesis provide the further evidence for a role of 1,25(OH)2-16ene-23yne-D3 in immunoregulation.

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Methamphetamine and MDMA (3,4-methylenedioxymethamphetamine) Induce Apoptosis in Both Human Serotonergic and Dopaminergic Cell Lines

  • Kim, Kyu Bong;Suh, Soo Kyung;Lee, Bo Kyung;Kim, Byung Kyu;Kim, Jae Hee;Han, Eui Sik;Park, Chang Won;Kim, Jong Won;Kim, Kwang Jin;Lee, Sun Hee
    • Biomolecules & Therapeutics
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    • v.11 no.4
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    • pp.214-223
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    • 2003
  • Methamphetamine (METH) and 3,4-methylenedioxymethamphetamine (MDMA) have become popular recreational drugs of abuse in many countries. Although the neurotoxic damage caused by METH and MDMA is characterized by degeneration of the dopaminergic and serotonergic systems in brain, the molecular and cellular mechanisms remain to be clarified. Therefore, the purposes of this study were to confirm the capability of METH and MDMA to induce apoptosis and to clarify the action of its molecular mechanism by using serotonergic JAR cells and dopaminergic SK-N-SH cells. METH and MDMA were dose-dependently cytotoxic to human serotonergic JAR cells and dopaminergic SK-N-SH cells. The morphological change of apoptosis was found in Giemsa staining and TUNEL and further verified in DNA fragmentation analysis. Immunoblotting analysis revealed proteolytic cleavage of caspase-3 and -9 and change of bcl-2 and bax proteins. These results suggest that METH and MDMA may induce caspase-dependent apoptosis via the mitochondrial cell death pathway and METH and MDMA-induced neurotoxicity may happen to broadly and independently of both dopaminergic and serotonergic systems.

The Histopathological Examination for Diagnosis of MALT Lymphoma in the Stomach

  • Lee, Tae Hee;Hyun, Sung Hee;Kim, In Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.46 no.3
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    • pp.91-98
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    • 2014
  • Primary gastric lymphoma (PGL) is derived from mucosa-associated lymphoid tissue (MALT) and it differs from nodal lymphoma in histologic features and biologic behavior. Recent studies have showed that Helicobacter pylori (H.pylori ) infection is closely related to the development of low grade gastric lymphoma, and eradication of the infection induces regression of the tumor. H. pylori infection is known to be important to the development of gastric MALT lymphoma. The aim of this study was to elucidate the histopathological behavior of PGL according to the concept of MALT and to compare the predictive value of tests frequently used for diagnosis of H. pylori. The histological features of gastric lymphoma arising from MALT are the replacement of glands by uniform dense infiltration of centrocyte-like cells in the lamina propria and lymphoidepithelial lesion. H. pylori-associated histologic changes of neutrophilic infiltration, lymphoid follicle or aggregates formation and intestinal metaplasia, and H. pylori immunoreactivity were analyzed. Detection of H. pylori in chronic active gastritis and peptic ulcer suggests a possible role of H. pylori in the pathogenesis. Giemsa, Toluidine blue and Long H&E stains were used in H. pylori detection. Histopathological examination of gastric biopsy specimens revealed lymphoepithelial lesions pathognomonic of MALT lymphoma, and immunohistochemical staining for CD20 was diffusely positive. CD3 was positive in reactive T cells. PAX-5 was negative except the follicle. Bcl-2, cytokeratin, Ki-67, and c-myc were positive. The findings may indicate a predictable transition of low grade to high grade, and c-myc may be used as a valuable marker before molecular pathology diagnosis.

1, 25(OH)$_2$-23ene-$D_3$ : Effects on Proliferation and Differentiation of U937 Cells in vitro and on Clcium Metabolism of Rat in vivo (1, 25(OH)$_2$-23ene-$D_3$ : in vitro에서 U937 세포의 증식과 분화 및 in vivo에서 쥐의 칼슘대사에 미치는 영향)

  • 정수자;서명자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.1
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    • pp.1-9
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    • 1995
  • 1, 25(OH)2-23ene-D3 is a novel vitamine D3 analog which has a double bond between C-23 and C-24. We describe the effects of this analog on cell differentiation and cell proliferation in vitro using the human histiocytic lymphoma cell line U937, and on calcium metabolism in rats in vivo. In the present investigation 1, 25(OH)2-23ene-D3 was compared to the natural metabolite of vitamin D3, 1$\alpha$, 25-dihydroxycholecalciferol[1, 25(OH)2-23ene-D3 was more potent than 1, 25(OH)2-23ene-D3 for inhibition of proliferation and induction of differentiation of U937 cells. Especially, its effect on induction of differentiation, as measured by superoxide production and nonspecific esterase(NSE) activity, was about 20-fold more potent that 1, 25(OH)2-23ene-D3. This analog morphologically and functionally differentiated U937 cells to monocyte-macrophage phenotype showing a decrease of N/C ratio in Giemsa staining and the increase of adherence ability to surface. Intraperitoneal administration of 1, 25(OH)2-23ene-D3 to rats showed that the compound had at least 50 times less activity than 1, 25(OH)2-23ene-D3 in causing hypercalcemia and hypercalciuria. The strong direct effects of 1, 25(OH)2-23ene-D3 on cell proliferation and cell differentiation, coupled with its decreased activity of calcium metabolism make this compound an interesting candidate for clinical studies including patients with leukemia, as well as several skin disorders, such as psoriasis.

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Effect of Deer Antler Extract on Muscle Differentiation and 5-Aminoimidazole-4-Carboxamide Ribonucleoside (AICAR)-Induced Muscle Atrophy in C2C12 Cells

  • Jo, Kyungae;Jang, Woo Young;Yun, Beom Sik;Kim, Jin Soo;Lee, Hyun-Sun;Chang, Yeok Boo;Suh, Hyung Joo
    • Food Science of Animal Resources
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    • v.41 no.4
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    • pp.623-635
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    • 2021
  • The effect of deer antler extract on muscle differentiation and muscle atrophy were evaluated to minimize muscle loss following aging. Various deer antler extracts (HWE, hot water extract of deer antler; FE, HWE of fermented deer antler; ET, enzyme-assisted extract of deer antler; UE, extract prepared by ultrasonication of deer antler) were evaluated for their effect on muscle differentiation and inhibition of 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR)-induced muscle atrophy in C2C12 cells. Morphological changes according to the effect of antler extracts on muscle differentiation were confirmed by Jenner-Giemsa staining. In addition, the expression levels of genes related to muscle differentiation and atrophy were confirmed through qRT-PCR. In the presence of antler extracts, the length and thickness of myotubes and myogenin differentiation 1 (MyoD1) and myogenic factor 5 (Myf5) gene expression were increased compared to those in the control group (CON). Gene expression of AMP-activated protein kinase (AMPK), MyoD1, and myogenin, along with the muscle atrophy factors muscle RING finger-1 (MuRF-1) and forkhead box O3a (FoxO3a) upon addition of deer antler extracts to muscle-atrophied C2C12 cells was determined by qRT-PCR after treatment with AICAR. The expression of MuRF-1 and FoxO3a decreased in the groups treated with antler extracts compared to that in the group treated with AICAR alone. In addition, gene expression of MyoD1 and myogenin in the muscle atrophy cell model was significantly increased compared that into the CON. Therefore, our findings indicate that antler extract can increase the expression of MyoD1, Myf5 and myogenin, inhibit muscle atrophy, and promote muscle differentiation.