• Title/Summary/Keyword: Cell volume decrease

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Condurango (Gonolobus condurango) Extract Activates Fas Receptor and Depolarizes Mitochondrial Membrane Potential to Induce ROS-dependent Apoptosis in Cancer Cells in vitro -CE-treatment on HeLa: a ROS-dependent mechanism-

  • Bishayee, Kausik;Mondal, Jesmin;Sikdar, Sourav;Khuda-Bukhsh, Anisur Rahman
    • Journal of Pharmacopuncture
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    • v.18 no.3
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    • pp.32-41
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    • 2015
  • Objectives: Condurango (Gonolobus condurango) extract is used by complementary and alternative medicine (CAM) practitioners as a traditional medicine, including homeopathy, mainly for the treatment of syphilis. Condurango bark extract is also known to reduce tumor volume, but the underlying molecular mechanisms still remain unclear. Methods: Using a cervical cancer cell line (HeLa) as our model, the molecular events behind condurango extract's (CE's) anticancer effect were investigated by using flow cytometry, immunoblotting and reverse transcriptase-polymerase chain reaction (RT-PCR). Other included cell types were prostate cancer cells (PC3), transformed liver cells (WRL-68), and peripheral blood mononuclear cells (PBMCs). Results: Condurango extract (CE) was found to be cytotoxic against target cells, and this was significantly deactivated in the presence of N-acetyl cysteine (NAC), a scavenger of reactive oxygen species (ROS), suggesting that its action could be mediated through ROS generation. CE caused an increase in the HeLa cell population containing deoxyribonucleic acid (DNA) damage at the G zero/Growth 1 (G0/G1) stage. Further, CE increased the tumor necrosis factor alpha ($TNF-{\alpha}$) and the fas receptor (FasR) levels both at the ribonucleic acid (RNA) and the protein levels, indicating that CE might have a cytotoxic mechanism of action. CE also triggered a sharp decrease in the expression of nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$) both at the RNA and the protein levels, a possible route to attenuation of B-cell lymphoma 2 (Bcl-2), and caused an opening of the mitochondrial membrane's permeability transition (MPT) pores, thus enhancing caspase activities. Conclusion: Overall, our results suggest possible pathways for CE mediated cytotoxicity in model cancer cells.

The Effect of the Storage Duration and Temperature of EDTA Specimen for CBC and WBC Differential Count in SE-9000 Automated Cell Counter (SE-9000 자동 혈구계산기에서 EDTA 검체의 보관기간 및 온도가 CBC 및 백혈구 감별계산에 미치는 영향)

  • Hong, Seung-Bok;Kim, Jong-Seok;Shin, Kyeong-Seob
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.147-151
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    • 2006
  • Although various automated CBC analyzers with different WBC analytical principles were consequently introduced to clinical laboratory, the specific information concerning the suitability or unsuitability of aging samples is scarce. For this reason, we studied the effect of storage duration and temperature on CBC parameter in SE-9000 (SYSMEX Medical Electronics Co., Ltd., Kobe, Japan), automated CBC analyzer. We tested 32 K3-EDTA specimens with SE-9000 during 72 hours. Specimens were kept at room temperature (RT) and refrigerated and were analyzed at 0 hr, 4 hr, 8 hr, 24 hr, 48 hr and 72 hr after the collection of the specimens. The percentage changes from initial value for each parameters were calculated. Among the CBC parameters, hemoglobin, red blood cell count, mean corpuscular hemoglobin and platelets were stable for the study period at both temperatures. The mean corpuscular volume (MCV), hematocrit (Hct) and red cell distribution (RDW) increased and the mean corpuscular hemoglobin concentration (MCHC) decreased over time at room temperature. These parameters were stable when refrigerated. The leukocyte count was stable during 72hr at RT and when refrigerated. At room temperature, the relative percentages of neutrophils tend to increase, whereas those of lymphocyte and monocytes tend to decrease after 48 hours. When refrigerated, those of neutrophils and monocytes tend to increase, whereas those of lymphocytes tend to decreased over time. CBC parameters of refrigerated specimen were reliable for 72 hr for the exception of differential count from 24 hr but many CBC parameters, such as MCV, Hct, MCHC, RDW and differential count of leukocyte of blood stored at room temperature for 24 hr were unreliable.

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Hyperglycemia aggravates decrease in alpha-synuclein expression in a middle cerebral artery occlusion model

  • Kang, Ju-Bin;Kim, Dong-Kyun;Park, Dong-Ju;Shah, Murad-Ali;Kim, Myeong-Ok;Jung, Eun-Jung;Lee, Han-Shin;Koh, Phil-Ok
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.195-202
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    • 2018
  • Hyperglycemia is one of the major risk factors for stroke. Hyperglycemia can lead to a more extensive infarct volume, aggravate neuronal damage after cerebral ischemia. ${\alpha}$-Synuclein is especially abundant in neuronal tissue, where it underlies the etiopathology of several neurodegenerative diseases. This study investigated whether hyperglycemic conditions regulate the expression of ${\alpha}$-synuclein in middle cerebral artery occlusion (MCAO)-induced cerebral ischemic injury. Male Sprague-Dawley rats were treated with streptozotocin (40 mg/kg) via intraperitoneal injection to induce hyperglycemic conditions. MCAO were performed four weeks after streptozotocin injection to induce focal cerebral ischemia, and cerebral cortex tissues were obtained 24 hours after MCAO. We confirmed that MCAO induced neurological functional deficits and cerebral infarction, and these changes were more extensive in diabetic animals compared to non-diabetic animals. Moreover, we identified a decrease in ${\alpha}$-synuclein after MCAO injury. Diabetic animals showed a more serious decrease in ${\alpha}$-synuclein than non-diabetic animals. Western blot and reverse-transcription PCR analyses confirmed more extensive decreases in ${\alpha}$-synuclein expression in MCAO-injured animals with diabetic condition than these of non-diabetic animals. It is accepted that ${\alpha}$-synuclein modulates neuronal cell death and exerts a neuroprotective effect. Thus, the results of this study suggest that hyperglycemic conditions cause more serious brain damage in ischemic brain injuries by decreasing ${\alpha}$-synuclein expression.

THE ANTICANCER EFFECT OF PACLITAXEL($Taxol^{(R)}$) IN ORAL SQUAMOUS CELL CARCINOMA XENOGRAFT (이종 이식된 구강편평세포 암종에서 Paclitaxel ($Taxol^{(R)}$)의 항암 효과)

  • Kim, Ki-Hwan;Kim, Chul-Hwan;Han, Se-Jin;Lee, Jae-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.2
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    • pp.95-110
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    • 2006
  • The treatment for oral and maxillofacial carcinoma with chemotherapeutic agents is evaluated by many effective methods to reduce the tumor mass and cancer cell proliferation. However these chemotherapy have many serious side effects, such as bone marrow suppression, renal toxicity, G-I troubles. Therefore a possible approach to develop a clinically applicable chemotherapeutic agent is to screen anticancer activity of Taxol which is known to have very little side effect and have been used to breast cancer and ovarian carcinoma. Taxol is a new anti-microtubular anti-cancer agent extracted from the bark of the Pacific yew, Taxus brevifolia. Paclitaxel(Taxol) acts by promoting tubulin polymerization and over stabilizing microtubules agianst depolymerization. Despite the constant improvements of methods of the cancer treatment especially chemotherapy, the rate of cancer metastasis and recurrent are not decreased. Thus the investigation of new drug which have very little side effect and a possible clinically application continues to be a high priority. Considering that the Taxol have shown very effective chemotherapeutic agent with relatively low toxicity in many solid tumors, it deserves to evaluate its efficacy in oral squamous cell carcinoma. In this study, to investigate the in-vivo and in-vitro anti-cancer efficacy of Taxol in oral squamous cell carcinoma and lastly, the potency of Paclitaxel in the clinical application for oral cancer was evaluated. In vivo study, after HN22 cell line were xenografted in nude mice, the growth of tumor mass was observed, 3 mg/Kg taxol was injected intraperitoneally into nude mice containing tumor mass. The methods of these study were measurement of total volume of tumor mass, histopathologic study, immunohistochemical study, drug resistance assay, growth curve, MTT assay, flow cytometry, cDNA microarray in vivo and in vitro. The results were obtained as following. 1. The visual inspection of the experimental group showed that the volume of the tumor mass was slightly decreased but no significant difference with control group. 2. Ki-67 index was decreased at weeks 4 in experimental group. 3. Microscopic view of the xenografted tumor mass showed well differentiated squamous cell carcinoma and after Taxol injection, some necrotic tissue was seen weeks 4. 4. The growth curve of the tumor cells were decreased after 1day Taxol treatment. 5. According to the MTT assay, HN22 cell line showed relative drug resistancy above $5\;{\mu}g/ml$ concentrations of Taxol. 6. In drug resistance assay, the decrease of cell counts was seen relatively according to concentration. 7. In Flow cytometry, G2M phase cell arrests were seen in low concentration of the Taxol, while S phase cell arrests were seen in high concentration of the Taxol. 8. Using cDNA microarray technique, variable gene expression of ANGPTL4, TXNRD1, FAS, RRAGA, CTGF, CYCLINEA, P19, DUSP5, CEBPG, BTG1 were detacted in the oral squamous cell carcinoma cell after taxol treatment. In this study paclitaxel is effective against oral squamous cell carcinoma cell lines in vitro, but week effect was observed in vivo. So we need continuous study about anticancer effect of taxol in vivo in oral squamous cell carcinoma.

Changes in the Blood Components Caused by Water Intake (물 섭취에 의한 혈액 성분 변화에 관한 연구)

  • Kim, Hyun-Kyung;Kim, Soo-Hwan;Ryu, Jae-Ki
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.3
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    • pp.227-232
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    • 2017
  • Although water is an essential component of the human body and is involved in many physiological processes, the effect of a steady and sufficient water intake on blood components has not well elucidated. Therefore, we investigated the changes in hematological parameters, high-sensitivity C-reactive protein (hs-CRP), and immunoglobulin G (IgG) after water intake in 13 healthy adults. They were divided into two groups: The control group (N=4), which consumed water ad libitum, and the experimental group (N=9), which consumed 2 L of water per day. Two weeks later, blood cell counts, hematocrit, and hemoglobin content had increased in the experimental group, although not significantly (p>0.05); however, there was a significant increase in the mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) (p<0.05; and p<0.01, respectively), and a significant decrease in the mean platelet volume (MPV) (p<0.05). Of the immunologic parameters, a non-significant decrease in the concentration of hs-CRP, an indicator of cardiovascular disease risk, was observed (p>0.05). However, there was a dramatic and significant increase in the concentration of IgG (p<0.05). In conclusion, a steady and sufficient water intake may contribute to alleviate anemia by increasing hemoglobin. Additionally, it may decrease the risk of cardiovascular disease by decreasing platelet activation and concentration of hs-CRP. Furthermore, a steady intake of water may improve immune function by increasing the concentration of the components of humoral immunity.

Isolation of Antifungal Lactic Acid Bacteria (LAB) from "Kunu" against Toxigenic Aspergillus flavus

  • Olonisakin, Oluwafunmilayo Oluwakemi;Jeff-Agboola, Yemisi Adefunke;Ogidi, Clement Olusola;Akinyele, Bamidele Juliet
    • Preventive Nutrition and Food Science
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    • v.22 no.2
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    • pp.138-143
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    • 2017
  • The antifungal activity of isolated lactic acid bacteria (LAB) from a locally fermented cereal, "Kunu", was tested against toxigenic Aspergillus flavus. The liquid refreshment, "Kunu", was prepared under hygienic condition using millet, sorghum, and the combination of the two grains. The antifungal potential of isolated LAB against toxigenic A. flavus was carried out using both in vitro and in vivo antifungal assays. The LAB count from prepared "Kunu" ranged from $2.80{\times}10^4CFU/mL$ to $4.10{\times}10^4CFU/mL$ and Lactobacillus plantarum, Lactobacillus delbrueckii, Lactobacillus fermentum, Pediococcus acidilactici, and Leuconostoc mesenteroides were the isolated bacteria. Inhibitory zones exhibited by LAB against toxigenic A. flavus ranged from 5.0 mm to 20.0 mm. The albino mice infected with toxigenic A. flavus showed sluggishness, decrease in body weight, distortion of hair, and presence of blood in their stool, while those treated with LAB after infection were recovered and active like those in control groups. Except for the white blood cell that was increased in the infected mice as $6.73mm^3$, the packed cell volume, hemoglobin, and red blood cell in infected animals were significantly reduced (P<0.05) to 29.28%, 10.06%, and 4.28%, respectively, when compared to the treated mice with LAB and control groups. The antifungal activity of LAB against toxigenic A. flavus can be attributed to the antimicrobial metabolites. These metabolites can be extracted and used as biopreservatives in food products to substitute the use of chemical preservatives that is not appealing to consumers due to several side effects.

The effect of Westergren tube angle, temperature and osmobility with the different percent of NaCl solution on erythrocyte sedimentation rate (적혈구 침강속도에 대한 Westergren tube 각도, 온도 및 NaCl 농도가 미치는 영향)

  • Kim, Do-kyun;Kim, Young-hong;Yu, Chang-jun
    • Korean Journal of Veterinary Research
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    • v.36 no.1
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    • pp.75-82
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    • 1996
  • Erythrocyte sedimentation rate is influenced by plasma protein, red cell itself and physiological conditions and it is clear that the measurement of erythrocyte sedimentation rate varies with technique and various environmental factors. The effect of temperature, angle of test tube, and osmobility with different percent of NaCl solution on erythrocyte sedimentation rate on sheep was determined by modified Westergren method. In sheep, as the angle of Westergren tube was decreased from $90^{\circ}$ to $45^{\circ}$, erythrocyte sedimentation rate was increased in both diluted plasma and NaCl solutions. As temperature was increased from $4^{\circ}C$ to $20^{\circ}C$ at $45^{\circ}$ angle. erythrocyte sedimentation rate was increased. In ruminants, this modified Westergren methods, erythrocyte 1 : plasma 9 instead of erythrocyte 4 : plasma 6(whole blood), enable them to have meanings as dog therefor, using this method, clinical can determine the erythrocyte sedimentation rate of ruminants for diagnosis. Because erythrocyte sedimentation rate was changed according to the angle of Westergren tube, temperature as erythrocyte diluted with NaCl, this study detected that the change of temperature, the angle of Westergren tube could change erythrocyte sedimentation rate by effecting red cell itself. The increase of osmobility owing to change of NaCl percent resulted in the decrease of rapid erythrocyte sedimentation rate. So this fact indicate that deformibility and the change of red cell volume have meaning in the change of erythrocyte sedimentation rate.

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Effects of Daejo-hwan(Tatsao-wan) on L-NAME Induced Learning and Memory Impairment and on Cerebral Ischemic Damage of the Rats (L-NAME으로 유발된 학습.기억장애와 뇌허혈 손상에 관한 대조환의 효과)

  • 김근우;구병수
    • The Journal of Korean Medicine
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    • v.21 no.2
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    • pp.25-36
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    • 2000
  • Objectives : This study demonstrates the effects of Daejo-hwan on learning and memory impairment induced by L-NAME (75 mg/kg) treatment and on cerebral ischemic damage induced by middle cerebral artery (MCA) occlusion in rats. Methods : Daejo-hwan emulsion (73.3 mg/100 g/l ml) was administered to rats along a timed study schedule. The Moms water maze was used for learning and memory test of the rats. The MCA was occluded by using the intraluminal thread method. The brain slices were stained by 2 % triphenyl tetrazolium chloride (TTC) and 1 % cresyl violet solution. Infarct size, neuron cell number and size in penumbra was measured by using computer image analysis system. Results : 1. The escape latency of the Daejo-hwan treated group decreased significantly with respect to the control group. 2.The memory score of the Daejo-hwan treated group showed increase tendency, And the swimming distance was not different between the normal, the control, and the Daejo-hwan treated group. 3. The infarct size of the Daejo-hwan treated group decreased significantly with respect to the control group. 4. The total infarct volume of the Daejo-hwan treated group showed decrease tendency. And the brain edema index of the Daejo-hwan treated group decreased significantly with respect to the control group. 5. The neuron cell number and cell size in penumbra of the Daejo-hwan treated group increased significantly with respect to the control group. Conclusions : According to the above results, it is supposed that Daejo-hwan is clinically applicable to the vascular dementia.

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Attenuation of Anemia by Relmα in LPS-Induced Inflammatory Response

  • Lee, Mi-Ran
    • Journal of the Korea Society of Computer and Information
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    • v.23 no.10
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    • pp.135-141
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    • 2018
  • In this paper, we propose to evaluate the effect of resistin-like molecule alpha ($Relm{\alpha}$) on the progression of anemia of inflammation. Anemia of inflammation is a common feature of inflammatory disorders, including chronic kidney disease, infections, and rheumatoid arthritis. $Relm{\alpha}$ is highly up-regulated in various inflammatory states, especially those involving asthma, intestinal inflammation, and parasitic diseases, and regulates the pathogenesis of those diseases. However, the role of $Relm{\alpha}$ in anemia of inflammation is unknown. To explore the roles of $Relm{\alpha}$ in anemia of inflammation in vivo, we generated mouse model of the disease by injecting 0.25 mg/kg lipopolysaccharides (LPS) intraperitoneally into $Relm{\alpha}-deficient$ and wild-type (WT) mice daily for 10 days. Research data was expressed as differences between LPS-treated $Relm{\alpha}-deficient$ and WT mice by a two-tailed non-parametric Mann-Whitney U-test using GraphPad Instat program. The results of the study are as follows: LPS-treated $Relm{\alpha}-deficient$ mice had significantly (p<0.05) lower hemoglobin contents, hematocrit levels and red blood cell indices including mean corpuscular volume, mean corpuscular hemoglobin than WT controls. This decrease was accompanied by significant (p<0.05) increase in total white blood cell and monocyte counts in the blood. However, there was no significant difference in mRNA levels of hepatic hepcidin and renal erythropoietin between the two animal groups. Taken together, these results indicates that $Relm{\alpha}$ deficiency exacerbates the anemia by increasing inflammation, suggesting therapeutic value of $Relm{\alpha}$ in the treatment of anemia of inflammation.

Analysis on the Decrease of Planning Target Volume in the Case of Lung Radiation Surgery with the Application of Respiratory Gated Radiotherpy Method (폐암 환자의 방사선수술 시 호흡연동 방사선치료 방법의 적용을 통한 계획용표적체적 감소 효과 분석)

  • Song, Ju-Young;Nah, Byung-Sik;Chung, Woong-Ki;Ahn, Sung-Ja;Nam, Taek-Keun;Yoon, Mee-Sun;Jung, Jae-Uk
    • Progress in Medical Physics
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    • v.19 no.4
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    • pp.263-268
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    • 2008
  • The application of a respiratory gated radiotherpy method to the lung radiation surgery was evaluated compared with the conventional method in which the whole tumor motion range is considered in the delineation of PTV (Planning target volume). The four dimensional CT simulation images were acquired for the five NSCLC (Non-small cell lung cancer) patients for radiation surgery. The respiratory gated plan was prepared with the 50% phase CT images and the conventional method was planned based on the ITV (Internal target volume) which include all the target volumes created in each phase CT images within a whole respiratory period. The DVH (Dose volume histogram) of OAR (Organ at risk) which calculated in each method was compared for the evaluation of the plan properness. The relative decrease of OARs' DVH were verified in the application of respiratory gated method. The average decrease rate were $16.88{\pm}9.97%$ in the bronchus, $34.13{\pm}19.15%$ in the spinal cord, $28.42{\pm}18.49%$ in the chest wall and $32.48{\pm}16.66%$ in the lung. Based on these results, we can verified the applicability and the effectiveness of the respiratory gated method in the lung radiation surgery.

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