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Inhibition of Neointima Formation and Migration of Vascular Smooth Muscle Cells by Anti-vascular Endothelial Growth Factor Receptor-1 (Flt-4) Peptide in Diabetic Rats (당뇨병 쥐에서 혈관내피 성장인자 수용체-1 차단 펩타이드를 이용한 신내막 형성과 혈관평활근세포 이동의 억제)

  • Jo, Min-Seop;Yoo, Ki-Dong;Park, Chan-Beom;Cho, Deog-Gon;Cho, Kue-Do;Jin, Ung;Moon, Kun-Woong;Kim, Chul-Min;Wang, Young-Pil;Lee, Sun-Hee
    • Journal of Chest Surgery
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    • v.40 no.4 s.273
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    • pp.264-272
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    • 2007
  • Background: Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis, including stimulating the proliferation and migration of vascular smooth muscle cells (VSMCs). It has been known that diabetes is associated with accelerated cellular proliferation via VEGF, as compared to that under a normal glucose concentration. We investigated the effects of selective blockade of a VEGF receptor by using anti-Flt-1 peptide on the formation and hyperplasia of the neointima in balloon injured-carotid arteries of OLETF rats and also on the in vitro VSMCS' migration under high glucose conditions. Material and Method: The balloon-injury method was employed to induce neointima formation by VEGF. For f4 days beginning 2 days before the ballon injury, placebo or vascular endothelial growth factor receptor-1 (VEGFR-1) specific peptide (anti-Flt-1 peptide), was injected at a dose of 0.5mg/kg daily into the OLETF rats. At 14 days after balloon injury, the neointimal proliferation and vascular luminal stenosis were measured, and cellular proliferation was assessed by counting the proliferative cell nuclear antigen (PCNA) stained cells. To analyze the effect of VEGF and anti-Flt-1 peptide on the migration of VSMCs under a high glucose condition, transwell assay with a matrigel filter was performed. And finally, to determine the underlying mechanism of the effect of anti-Flt-1 peptide on the VEGF-induced VSMC migration in vitro, the expression of matrix metalloproteinase (MMP) was observed by performing reverse transcription-polymerase chain reaction (RT-PCR). Result: Both the neointimal area and luminal stenosis associated with neointimal proliferation were significantly decreased in the anti-Flt-1 peptide injected rats, ($0.15{\pm}0.04 mm^2$ and $ 36.03{\pm}3.78%$ compared to $0.24{\pm}0.03mm^2\;and\;61.85{\pm}5.11%$, respectively, in the placebo-injected rats (p<0.01, respectively). The ratio of PCNA(+) cells to the entire neointimal cells was also significantly decreased from $52.82{\pm}4.20%\;to\;38.11{\pm}6.89%$, by the injected anti-Flt-1 peptide (p<0.05). On the VSMC migration assay, anti-Flt-1 peptide significantly reduced the VEGF-induced VMSC migration by about 40% (p<0.01). Consistent with the effect of anti-Flt-1 peptide on VSMC migration, it also obviously attenuated the induction of the MMP-3 and MMP-9 mRNA expressions via VEGF in the VSMCS. Conclusion: Anti-Flt-1 peptide inhibits the formation and hyperplasia of the neointima in a balloon-injured carotid artery model of OLETF rats. Anti-Flt-1 peptide also inhibits the VSMCs' migration and the expressions of MMP-3 and MMP-9 mRNA induced by VEGF under a high glucose condition. Therefore, these results suggest that specific blockade of VEGFR-1 by anti-Flt-1 peptide may have therapeutic potential against the arterial stenosis of diabetes mellitus patients or that occurring under a high glucose condition.

Distribution of Marine Bacteria and Coliform Groups in Puksin Bay, Korea (북신만의 대장균군 및 해양세균의 분포)

  • CHOI Jong-Duck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.2
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    • pp.202-208
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    • 1995
  • Puksin Bay located in the northwestern pan of Tongyeong, Korea has been contaminated by municipal wastewaters. In Puksin Bay, red tides have occured almost every year since the early 1980's. This experiment was carried out two times in a month in the winter in 1990, the summer' 1991, and the winter in 1994 so as to clarify the distribution marine bacteria and coliform groups in Puksin Bay. The water quality of Puksin Bay was not only move polluted than that of any other costal area around Tongyeong, but also its water quality was investigated to keep going bad. Viable cell counts in Puksin Bay were $4.9\times10^3/ml$ in 1990's winter, $3.6\times10^6/ml$ in 1991's summer, and $2.1\times10^4/ml$ in 1994's winter. The variation of seasonal total and fecal coliform MPN/100ml in Puksin Bay were $6.7\times10^2\;and\;2.6\times10^2$ in 1990's winter, $1.5\times10^4$ and $5.4\times10^3$ in 1991's summer, and $1.5\times10^3$ and $5.6\times10^2$ in 1994's winter, respectively. The changes of stational total coliform MPN/100m1 from station 1 to station 8 in Puksin Bay were 95,000, 1600, 1,000, 182, 151, 94, 43 and 13 in winter, and 110,000, 29,000, 2,400, 4,100, 1,700, 1,700, 810 and 150 in 1991's summer, and 3,381, 1928, 1582, 256, 161, 59 and 23 in 1994's winter. During the study period, the number of viable cell was ranged from $10^4\;to\;10^7/ml$ and 307 bacteria strains were isolated from Puksin Bay. The dominant species were Acinetobacter spp. 86 $(28.3\%)$, Pseudomonas spp. 51 $(16.6\%)$, Flavobacterium spp. 41 $(13.4\%)$, Escherichia coli 36 $(11.7\%)$, and Vibrio spp. 27$(8.8\%)$. The results obtained in this study indicate that this bay is getting to contaminate far more with municipal wastewaters and cultivation of the shellfish and finfish in this bay is not proper. When municiple wastewaters keep flowing into this bay, any other coastal area around Tongyeong may be contaminated.

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Study on the Dissolution of Sandstones in Gyeongsang Basin and the Calculation of Their Dissolution Coefficients under CO2 Injection Condition (이산화탄소 지중 주입에 의한 경상분지 사암의 용해반응 규명 및 용해 반응상수값 계산)

  • Kang, Hyunmin;Baek, Kyoungbae;Wang, Sookyun;Park, Jinyoung;Lee, Minhee
    • Economic and Environmental Geology
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    • v.45 no.6
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    • pp.661-672
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    • 2012
  • Lab scale experiments to investigate the dissolution reaction among supercritical $CO_2$-sandstone-groundwater by using sandstones from Gyeongsang basin were performed. High pressurized cell system (100 bar and $50^{\circ}C$) was designed to create supercritical $CO_2$ in the cell, simulating the sub-surface $CO_2$ storage site. The first-order dissolution coefficient ($k_d$) of the sandstone was calculated by measuring the change of the weight of thin section or the concentration of ions dissolved in groundwater at the reaction time intervals. For 30 days of the supercritical $CO_2$-sandstone-groundwater reaction, physical properties of sandstone cores in Gyeongsang basin were measured to investigate the effect of supercritical $CO_2$ on the sandstone. The weight change of sandstone cores was also measured to calculate the dissolution coefficient and the dissolution time of 1 g per unit area (1 $cm^2$) of each sandstone was quantitatively predicted. For the experiment using thin sections, mass of $Ca^{2+}$ and $Na^+$ dissolved in groundwater increased, suggesting that plagioclase and calcite of the sandstone would be significantly dissolved when it contacts with supercritical $CO_2$ and groundwater at $CO_2$ sequestration sites. 0.66% of the original thin sec-tion mass for the sandstone were dissolved after 30 days reaction. The average porosity for C sandstones was 8.183% and it increased to 8.789% after 30 days of the reaction. The average dry density, seismic velocity, and 1-D compression strength of sandstones decreased and these results were dependent on the porosity increase by the dissolution during the reaction. By using the first-order dissolution coefficient, the average time to dissolve 1 g of B and C sandstones per unit area (1 $cm^2$) was calculated as 1,532 years and 329 years, respectively. From results, it was investigated that the physical property change of sandstones at Gyeongsang basin would rapidly occur when the supercritical $CO_2$ was injected into $CO_2$ sequestration sites.

The Effects of Microcurrent Stimulation on the Astrocytes Proliferation at Injured Brain of Rabbit (극저전류자극이 손상된 토끼 뇌의 별아교세포 증식에 미치는 효과)

  • Kim, Ji-Sung;Min, Kyoung-Ok
    • Journal of Korean Physical Therapy Science
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    • v.9 no.3
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    • pp.107-119
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    • 2002
  • Astrocyte, which shares the greatest part of the brain (about 25%), is a land of glial cell that composes the central nervous system along with microglia, ependymal cell and oligodendroglia. It has 7-9nm of fibers in its cytoplasma, which are composed of glial fibrillary acidic protein (GFAP) and vimentin. As for the functions of the astrocyte, it has, so far, been supposed that the astrocyte will play a cytoskeletal role in maintaining the structure of the cerebrum, play a role as a blood-brain barrier so that it can induce migration of the neuron in its development and substances in the blood cannot go into the nervous tissue, and a role of immunology and phagocytosis. However, it was revealed today that it will be a role in preventing expansion of injury by attaching itself to the connective tissue such as the vessel and the pia mater when the nervous tissue or the arachnoid is injured. Microcurrent stimulation can control current, on the basis of A unit. That is, with such devices using it, it is possible to sense, from the outside, the injured current(wound current) of the lesion and to change it into the normal current, thereby promoting the restoration of the cells. In order to examine the effects of microcurrent stimulation on the injured astrocytes in the rabbits, this study was conducted with 24 New Zealand White Rabbit as its subjects, which were divided into 8 animals of the experiment group and 16 animals of the control group. After the animals in the experiment group were fixed to the stereotaxic apparatus, their hair was removed and their premotor area(association area) perforated by the micro-drill for skull-perforation with the depth of 8mm from the scalp. In one week after the injury, 4 animals in the control group and 8 animals in the experiment group were sacrificed and examined with immunohistochemical method. And in three weeks, the remaining 4 animals in the control group and 8 animals in the experiment group were also sacrificed and examined with the same way. The conclusion has been drawn as follows : In the control group sacrificed in one week after the injury, the astrocytes somewhat increased, compared with the normal animals, and in the group sacrificed in three weeks after the injury, they increased more (p < 0.05). The experiment group A in one week showed a little increase, but there was no significant differences, but the experiment group in three weeks showed more increase, compared with the experiment group in one week (p < 0.05). The experiment group B in one week showed more increase than the control group or the experiment group A, and the experiment group in three weeks showed more increase than the experiment group in one week (p < 0.05). Among the astrocytes, fibrous astrocytes were mostly observed, increasing as they are close to the lesion, and decreasing as they are remote from it. The findings show that microcurrent can cause the astrocytes to proliferate and that it will be more effective to stimulate the cervical part somewhat remote from the lesion rather than to directly stimulate the part of the lesion. Thus, microcurrent stimulation can be one of the methods that can activate the reaction of astrocytes, which is one of the mechanism for treating cerebral injury with hemorrhage. Therefore, this study will be used as basic research data for promoting restoration of functions in the patient with injury in the central nervous system.

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CHANGES IN THE SHAPE AND ULTRASTRUCTURE OF THE ARTICULAR DISK OF RAT FOLLOWING POSTURAL HYPERPROPULSOR (백서 하악골의 기능적 전방위가 악관절 원판에 미치는 영향)

  • Jang, Byung-Chun;Kyung, Hee-Moon;Sung, Jae-Hyun;Bae, Yong-Chul
    • The korean journal of orthodontics
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    • v.24 no.4 s.47
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    • pp.917-932
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    • 1994
  • This study was conducted to examine the changes in the shape of the Sprague-Dawley rats' articular disk following postural hyperpropulsion by observing their articular specimens through light and electronic microscopes after following 2-week and 4-week postural hyperpropulsion from their four weeks of age. The findings of this study are summarized as follows. It was shown that as compared with the control group, the experimental group indicated a significant increase in thickness of the 2-week groups' anterior and postreior portion of the articular disc. The experimental group showed statistically more significant increase in thickness of the 4-week groups' anterior portion of the articular disc than the control group. Light micrograph showed that the experimental group had more fibroblast in the anterior portion of the 2-week and 4-week groups than the comparing group. The 2-week groups showed in the findings through the electronic microscope that there were found the well developed and dilated RER which seems to actively synthesize the extracellular matrix including collagen, the cells with the well developed RER without distention which seems to actively synthesize the intracellular microfilaments due to the well developed free ribosome, and the typical chondroid cells. In addition, there was more fibroblast cell with the distended and well developed RER in the anterior area of the experimental group than that of the control group. The 4 week experimental group's anterior area of the disk had more cells than that of the control group while fibroblast with the well developed RER and free ribosome was quite abundat. Based on the above result of this study, it was shown that the functional hyperpropulsion of the mandible causes the changes in the nature of the mechanical load to the certain portion of the articular disk. As a result, it seems that there may be occurred some changes in morphology of the disc by adaptation or confrontation with these changes at the cellular level.

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Cryosurgery of Lung with 2.4 mm Cryoprobe: An Experimental in vivo Study of the Cryosurgery in Canine Model (냉동침을 이용한 폐 냉동수술의 동물실험: 냉동수술 방법의 비교 실험)

  • Kim Kwang-Taik;Chung Bong-Kyu;Lee Sung-Ho;Cho Jong-Ho;Son Ho-Sung;Fang Young-Ho;Sun Kyung;Park Sung-Min
    • Journal of Chest Surgery
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    • v.39 no.7 s.264
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    • pp.520-526
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    • 2006
  • Background: The clinical application of cryosurgery the management of lung cancer is limited because the response of lung at low temperature is not well understood. The purpose of this study is to investigate the response of the pulmonary tissue at extreme low temperature. Material and Method: After general anesthesia the lungs of twelve Mongrel dogs were exposed through the fifth intercostal space. Cryosurgical probe (Galil Medical, Israel) with diameter 2.4 mm were placed into the lung 20 mm deep and four thermosensors (T1-4) were inserted at 5 mm intervals from the cryoprobe. The animals were divided into group A (n=8) and group B (n=4). In group A the temperature of the cryoprobe was decreased to $-120^{\circ}C$ and maintained for 20 minutes. After 5 minutes of thawing this freezing cycle was repeated. In group B same freezing temperature was maintained for 40 minutes continuously without thawing. The lungs were removed for microscopic examination on f day after the cryosurgery. In four dogs of the group A the lung was removed 7 days after the cryosurgery to examine the delayed changes of the cryoinjured tissue, Result: In group A the temperatures of T1 and T2 were decreased to the $4.1{\pm}11^{\circ}C\;and\;31{\pm}5^{\circ}C$ respectively in first freezing cycle. During the second freezing period the temperatures of the thermosensors were decreased lower than the temperature during the first freezing time: $T1\;-56.4{\pm}9.7^{\circ}C,\;T2\;-18.4{\pm}14.2^{\circ}C,\;T3\;18.5{\pm}9.4^{\circ}C\;and\;T4\;35.9{\pm}2.9^{\circ}C$. Comparing the temperature-distance graph of the first cycle to that of the second cycle revealed the changes of temperature-distance relationship from curve to linear. In group B the temperatures of thermosensors were decreased and maintained throughout the 40 minutes of freezing. On light microscopy, hemorrhagic infarctions of diameter $18.6{\pm}6.4mm$ were found in group A. The infarction size was $14{\pm}3mm$ in group B. No viable cell was found within the infarction area. Conclusion: The conductivity of the lung is changed during the thawing period resulting further decrease in temperature of the lung tissue during the second freezing cycle and expanding the area of cell destruction.

Effects of BCG on Gastric Chief Cells of the Mouse Implanted with Ehrlich Carcinoma Cells (BCG가 Ehrlich 암세포를 이식한 생쥐 위점막 으뜸세포의 미세구조에 미치는 영향)

  • Ryoo, In-Sang;Ahn, E-Tay;Park, Kyung-Ho;Park, Dae-Kyoon;Kim, Myeong-Soo;Ko, Jeong-Sik
    • Applied Microscopy
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    • v.35 no.3
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    • pp.153-163
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    • 2005
  • This experiment was performed to evaluate the morphological responses of the gastric chief cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG (Bacillus Calmette-Guerin). Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (experimental control group and BCG treated group). In the experimental groups, each mouse was inoculated with $1x10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the last injection, each mouse was sacrificed. Pieces of the tissue were taken from the stomach, prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. The ultrathin sections were stained with uranyl acetate and lead citrate. The size of zymogen granule and the size of the mitochondrion of the gastric chief cells were observed and calculated. In the BCG treated group, most chief cells did not show any difference in ultrastructure, except that myelin figures were more frequently observed, in comparison with that of nornmal control group. The size of zymogen granule in the gastric chief cells of normal control, experimental control and BCG-treated groups were $0.98({\pm}0.108){\mu}m,\;1.05({\pm}0.092){\mu}m\;and\;0.93({\pm}0.053){\mu}m$, respectively. And the mitochondrial size of the gastric chief cells of normal control, experimental control and BCG-treated groups were $0.80({\pm}0.130){\mu}m,\;0.83({\pm}0.143){\mu}m\;and\;0.72({\pm}0.078){\mu}m$, respectively. From the above results, it was concluded that BCG may slightly suppress function of the gastric chief cells.

Effects of BCG on the Absorptive Cells in the Appendix of the Mouse Implanted with Ehrlich Carcinoma Cells (BCG가 Ehrlich 암세포를 이식한 생쥐의 막창자꼬리점막 흡수세포의 미세구조에 미치는 영향)

  • Lee, Woon-Woo;Park, Kyung-Ho;Kim, Myeong-Soo;Park, Dae-Kyoon;Ko, Jeong-Sik
    • Applied Microscopy
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    • v.37 no.3
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    • pp.157-166
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    • 2007
  • This experiment was performed to evaluate the ultrastructural responses of the absorptive cells in the appendix of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG (Bacillus Calmette-Guerin). Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (experimental control group and BCG treated group). In the experimental groups, each mouse was inoculated with $1{\time}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.5mL of saline or BCG (0.5 mL/25gm B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8CFU$) were injected subcutaneously to the animals every other day. The day following the last injection, each mouse was sacrificed. Pieces of the tissue were taken from the appendix, prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. The ultrathin sections were stained with uranyl acetate and lead citrate. In the normal control, experimental control and BCG treated mice, general morphology of the absorptive cells of appendix were similar. But myelin figures and intramitochondrial dense granules were more frequently observed in the absorptive cells of BCG treated mice than normal control ones. Above results show that BCG did show slight ultrastructural alterations in the absorptive cell of the appendix. These results that BCG may slightly suppress function of the absorptive cells of the appendix.

Immunohistochemical Study on the Activation of Cell mediated immunity in Murine Lymph node on Allergic Contact Dermatitis by DNCB -Based on the change of T lymphocytes and Il-2 receptors- (알러지성 접촉피부염 유발 피부 주변 림프절에서의 세포성 면역 활성에 관한 면역조직화학적 연구 - T 림프구와 IL-2 수용기의 분포 변화를 중심으로 -)

  • kim, Jin-Taek;Ahn, Sang-Hyun;Park, In-Sick;Chung, Jae-Man;Kim, Ho-Hyun
    • The Journal of Dong Guk Oriental Medicine
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    • v.7 no.1
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    • pp.33-41
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    • 1998
  • Lymph node tissues of BALB/C mouse treated with DNCB were immunohistochemically observed to investigate the activation of cell mediated immunity in lymph node of murine with allergic contact dermatitis. The inguinal region of BALB/C mice were sensitized by one application of $25{\mu}l$ of 5% 2,4-dinitrochlorobenzene(DNCB) onto an abdominal skin and 2 weeks later, the mice were challenged with $4{\mu}l$ of 2.5% DNCB. The inguinal lymph node were obtained at hour 24, 48, and 72 after 2nd DNCB treatment and embedded with paraffin, and then stained by following ABC method that used monoclonal antibody including L3T4(CD4), Ly2(CD8), IL-2R(CD25). The distribution of helper T lymphocytes, cytotoxic T lymphocytes and IL-2 receptors began to increase at hour 24 after after 2nd DNCB treatment and these increase appeared in paracortical area and medullary sinius. These increase were greatest at hour 48. These results indicated that the IL-2 secretion began to increase by activation of helper T lymphocytes in lymph node of DNCB re-exposure area and subsequently to activate suppress T lymphocytes.

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Diagnostic Significance of TNF-$\alpha$ in Tuberculous and Non-Tuberculous Pleural Effusion (결핵성 및 비결핵성 흉막삼출액에서 TNF-$\alpha$ 농도의 진단적 의의)

  • Na, Hyun-Joo;Park, Seog-Chea;Kang, Kwang-Won;Park, Hyeong-Kwan;Kim, Young-Chul;Choi, In-Seon;Park, Kyung-Ok
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.611-620
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    • 1997
  • Objectives : The differentiation of tuberculous effusion from the other causes of exudative pleural effusion remained difficult even with aids of biochemical analyses and pleural biopsy. As the pathophysiology of tuberculous pleural effusion is an enhanced cell mediated immunity, Adenosine deaminase(ADA) and various eytokines including Inteferon-$\gamma$, tumor necrosis factor alpha(TNF-$\alpha$) are considered as useful diagnostic tools in differentiating exudative pleural effusion. The author would like to demonstrate the diagnostic usefulness of TNF-$\alpha$ in the differentiation of exudative pleural effusion, and compared the discriminating ability of TNF-$\alpha$ with ADA. Methods : Pleural fluids obtained from 80 patients (tuberculous : 39, malignant : 31, parapneumonic : 10) with exudate pleural effusions were processed for cell counts and biochemical analysis including ADA and TNF-$\alpha$. Results : Tuberculous pleural fluid showed higher levels of ADA and TNF-$\alpha$, $48.7{\pm}32.7U/L$ and $184.1{\pm}214.2pg/mL$ than that of non-tuberculous effusion $26.0{\pm}41.3U/L$ and $44.1{\pm}114.2pg/mL$, respectively (ADA, TNF-$\alpha$, p < 0.05, p < 0.01). Receiver operating characteristics(ROC) curves were generated for ADA and TNF-$\alpha$ and the best cut-off value for adenosine deaminase and TNF-$\alpha$were considered as 30U/L and 15pg/ml, respectively. Comparing the area under the ROC curves, there was no significant difference between ADA and TNF-$\alpha$. Conclusion : For the differential diagnosis of tuberculous pleural effusion from the other causes of exudative pleural effusions, TNF-$\alpha$ as well as ADA was considered as useful diagnostic method. However adding TNF-$\alpha$ to ADA has no further diagnotic benefit than ADA alone.

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