• Title, Summary, Keyword: Cell counting method

Search Result 87, Processing Time 0.052 seconds

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
    • The Korean Journal of Thoracic and Cardiovascular Surgery
    • /
    • v.40 no.4
    • /
    • pp.264-272
    • /
    • 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.

Evaluation on the Usefulness of Filter in Sentinel Lymphoscintigraphy Using $^{99m}Tc$-Phytate (Phytate를 이용한 감시림프절 검사 시 Filter의 유용성 평가)

  • Jeong, Yeong-Hwan;Seo, Han-Kyung;Shim, Cheol-Min;Lim, Seong-Dong;Han, Dong-Hyeon;Park, Yung-Sun;Kim, Dong-Yun
    • The Korean Journal of Nuclear Medicine Technology
    • /
    • v.14 no.1
    • /
    • pp.35-39
    • /
    • 2010
  • Purpose: The aim of this study was to investigate distribution of particle size in phytate kit and compare filtered method with non-filtered method using 200 nm filter for sentinel lymphoscintigraphy (SLS). Materials and Methods: Five phytate kit of having the same available period was measured by particle size analyzer. For in-vivo experiment, $^{99m}Tc$-phytate was injected intradermally at both foot to perform lymphoscintigraphy. Imaging was acquired at 1hour after injection. Region of interest (ROI) was drawn in inguinal and background area for analysis. RAW 264.7 cells (Murine macrophage cell) were prepared for measurement of celluar uptake as a representative of macrophages. Paired t-test was performed using SPSS (SPSS Inc, USA) for statistical analysis. Results: The size of most particle in Techne phytate kit was distributed in 130~650 nm(90.5 %). In-vivo study, the ROI analysis showed similar result between filtered and non-filtered sample, and the numerical value of count/pixel were $58.3{\pm}5.97$ and $60.2{\pm}4.88$. In-vitro study, cellular uptake study also showed no difference between filtered and non-filtered sample by gamma counting. Conclusion: The present study demonstrates that there was no meaning of 200 nm filtered method for SLS using $^{99m}Tc$-phytate.

  • PDF

The Anti-bacterial Effects and Epidermal Permeability Barrier Function of Red Onion Juice Produced in Jeon-Nam province in Korea (전남 함평산 자색양파 추출물의 항균효과와 피부장벽 보호효과)

  • Youn, Dae-Hwan;Shin, Heon-Tae
    • Journal of Society of Preventive Korean Medicine
    • /
    • v.16 no.1
    • /
    • pp.43-56
    • /
    • 2012
  • Objective : In this study, we assessed the anti-bacterial effects and epidermal permeability barrier function of red onion juice comparing to yellow onion juice and $Houttuynia$ $cordata$ extract $in$ $vitro$. Methods : 3types of red and yellow onion juice were prepared as antibacterial agent candidates with Houttuynia cordata hot water extract using 4 different bacterial strains ($Escherichia$ $coil$, $Salmonella$ $enterica$ $subsp.$ $enterica$, $Staphylococcus$ $epidermidis$, $Staphylococcus$ $aureus$ $subsp$) by colony counting method. The expression of filaggrin, a marker of keratinocyte differentiation, and serine palmitoyl transferase (SPT), a marker of the formation of the stratum corneum lipid barrier, in human HaCat keratinocytes were analyzed using HaCaT cell line. The expression of COX-2 and AP-1 which is a factor of COX-2 transcription were also analyzed by western blotting method. Results : There was detectable anti-bacterial effects on $Staphylococcus$ $epidermidis$, $Staphylococcus$ $aureus$ $subsp$ among 1%, 5%, 10% extracts of yellow and red onion.(81%-100%) The bacteriocidal effects were not shown on $Escherichia$ $coil$, $Salmonella$ $enterica$ $subsp.$ $enterica$ among $Houttuynia$ $cordata$, yellow onion and red onion extracts. The in vitro results showed the concentration-dependent effects on the expression of both filaggrin and SPT in HaCat cells among 0.01%, 0.05%, 0.1%, 0.5% extracts in Houttuynia cordata and red onion, reflecting the notion that $Houttuynia$ $cordata$ and red onion can induce epidermal keratinocyte differentiation and improve the recovery of skin barrier functions. The concentration-dependent effects also have been shown on the expression of both COX-2 and AP-1 among 0.01%, 0.05%, 0.1%, 0.5% extracts in $Houttuynia$ $cordata$ and red onion, while slight effect in yellow onion. Conclusion : Red onion juice could be a potential candidate enhanser for the skin care and cosmetology.

Enhancement of Anti-inflammatory Activity of Lactobacillus plantarum Fermented by Achyranthes japonica on Extraction Solvents (추출 용매에 따른 Lactobacillus plantarum 발효 우슬의 항염증 효과 증진)

  • Jo, Eun Sol;Woo, Young Min;Kim, Ok Ju;Jo, Min Young;Ahn, Mee Young;Lee, Jae-Hwa;Ha, Jong-Myung;Kim, Andre
    • Applied Chemistry for Engineering
    • /
    • v.30 no.2
    • /
    • pp.145-150
    • /
    • 2019
  • In this study, we used extracts obtained from five different solvents (water, ethanol, hexane, ethyl acetate, butanol) of Achyranthes japonica (AJ) and also AJ fermented with Lactobacillus plantarum (LP) to confirm effects on the anti-inflammatory activity in RAW264.7 cells. Experiments of measuring nitric oxide (NO) and cytokine production were performed in lipopolysaccharide (LPS)-induced RAW264.7 cells, and the expression of both cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was observed by a western blot method. The cytotoxicity of RAW264.7 was confirmed by the cell counting kit (CCK) assay at a concentration of $100{\mu}g/mL$, which has no toxicity. As a result of the inhibition of NO production, the inhibition rate of AJ-LP extracted with ethanol samples was about 74% higher than that of using the control group. Interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and Interleukin-$1{\beta}$ (IL-$1{\beta}$), which are inflammatory cytokines, also showed an excellent efficacy with inhibition rates of about 57, 70, and 74%, respectively. Comparing to the results of COX-2 and iNOS expression in the AJ group, the inhibition rate of 20-hydroxyecdysone was the highest than others. On the other hand, the COX-2 expression level of AJ-LP group decreased about 16% compared to that of the control group, and the iNOS expression level was also decreased about 7%. These results suggest that the extract of AJ fermented from L. plantarum can be used as an anti-inflammatory natural material.

Epidemiological Study of Air Pollution and Its Effects on Health of Urban Population (서울시(市) 대기오염(大氣汚染)이 시민보건(市民保健)에 미치는 영향(影響)에 관(關)한 조사연구(調査硏究))

  • Chung, Kyou-Chull
    • Journal of Preventive Medicine and Public Health
    • /
    • v.2 no.1
    • /
    • pp.5-22
    • /
    • 1969
  • The urban population of our country is rapidly increasing due to many factors of social structure, and sociologists are predicting that the increase rate of the urban population will be over 50% in 1980's. Above all, the population of the capital city of Seoul, is at present more than four millions. Such centring of people in cities, together with the improvement of the standards of living, caused rapid increase in the amount of fuel consumption, and this consumption of fuels became one of the primary sources of the air pollution in cities. Moreover, the heavy traffic, construction of many tall buildings, and the increasing number of new factories due to the industrial development-all these are contributing to make the matter of air pollution worse and worse in the Metropolitan, whose geographical location is quite unfavorable considered from the view point of air pollution. Most homes in Seoul use briquet as fuel, while oils are used in tall buildings. The CO, $SO_2$, and smoke that come from burning of these fuels are a great threat to the health of the urban population. With the purpose of examining the influence of air pollution upon the public health, written inquiries were made upon respiratory diseases, and the carboxyhemoglobin saturation in the blood was measured to determine whether the air pollution may affect the health of the urban population. Method of Health Examination (1) Investigation of Respiratory Diseases Patients' records were examined to figure out the monthly ambulance rate of respiratory patients to the total number of patients treated. On the other hand, by using the questionnaire form approved by the Medical Research Council's Committee on Research into Chronic Bronchitis, investigators interviewed the examinees and inquired into the respiratory symptoms. (2) Measuring of Carboxyhemoglobin Saturation From the ear lobe of the examinees, with the use of the melangeur for the white blood cell counting, blood was taken, and after diluting it ten times with 0.1% $Na{2}CO_{3}$, again diulting it 20 times with 0.5% $Na_{2}\;CO_{3}$, its absorbancy was measured. The following results are obtained from the investigation. (1) It was found out that 7.7% of the total patients under treatment were suffering from upper respiratory infection, acute or chronic bronchitis, bronchial asthma, pulmonary emphysema and bronchiectasis. Of them all, patients with upper respiratory infection numbered the greatest with 4.8% and patients with acute or chronic bronchitis the next with 2.1%, and their monthly ambulance rate was high from December to February during the winter, and from April to May and from September to October during the changeable seasons. (2) The daily ambulance rate of respiratory patients, it was revealed, had a close connection with the concentration of $SO_2$ and CO in the air. (3) It was found out that men were more subject to respiratory disease than women, and both men and women were more liable to the diseases with the advancing of age. (4) People living at Choong-ku with the heavy traffic and in the industrial zones of Yeungdungpo had high frequency of respiratory symptoms. (5) Considered from the view point of occupations, high frequency was found among those without job, with jobs unknown, merchants and intdustrial workers, whose social status was rather low and traffic policemen who were always exposed to the exhaust gas of cars. As for women, the frequency was detected in the order of those from high to low, housewives who were exposed to briquet gas, women with jobs unknown, women without jobs, whose social status was low. (6) Ex-smokers rather than smokers, of both sexes, had higher frequency. As for men, heavy smokers had high frequency, while in women light smokers had rather high frequency which was presumed to be due to their average old age. (7) Men's average of carboxyhemoglobin saturation was 9.48%, while women's was 11.3%, higher than men's. (p<0.05). Age meant no difference in the case of men, but as for women, the saturation was remarkably high between the ages from 20 to 60. (8) No regional difference was detected in the carboxyhemoglobin saturation. (9) The carboxyhemoglobin saturation was found, in the case of men, in the order of office workers, traffic policemen, students, the unemployed, merchants and industrial workers, drivers; and as for women, the order was housewives, office workers, merchants and industrial workers. (10) No significant correlation was found between the carboxyhemoglobin saturation and the concentration of CO detected in kitchens, or between the carhoxyhemoglobin saturation and the passing of time after exposure to briquet gas. No difference of carboxyhemoglobin saturation was detected between smokers and non-smokers, and the degrees of smoking; only, significant negative correlation was found between the passing of time after smoking and the carboxyhemoglobin saturation. It is ture that air pollution causes or aggravates the respiratory diseases, increases the carboxyhemoglobin saturation in the blood, but what seems to be more hazardous to the health is the air polluted by the briquet gas in the kitchens and on-dol rooms rather than the atmospheric air pollution.

  • PDF

The Influence of Water Temperature and Salinity on Filtration Rates of the Hard Clam, Gomphina veneriformis (Bivalvia) (수온과 염분의 변화에 따른 연령별 대복 (Gomphina veneriformis: Bivalvia) 의 여과율 변동)

  • Shin, Hyun-Chool;Lee, Jung-Ho;Jeong, Hyo-Jin;Lee, Jung-Sick;Park, Jung-Jun;Kim, Bae-Hoon
    • The Korean Journal of Malacology
    • /
    • v.25 no.2
    • /
    • pp.161-171
    • /
    • 2009
  • The present study was performed to describe the influence of water temperature and salinity on filtration rates of the venus clam, Gomphina veneriformis, a suspension-feeding (filter-feeding) bivalve species. The calmswere collected from the eastern coastal area of Sokcho, Gangneung and Jumunjin at Kangwon-do, Korea, during December 2006 and May 2007. Isochrysis galbana (KMCC H-002) cells as food organisms were indoor-cultured by f/2 medium, and were used to measure the filtration rate of clam. Filtration rates of clam were measured by indirect method. Cell concentration of food organisms were determined by direct counting cells used the hemacytometer under the light microscope. The filtration rates of clams by water temperature sharply increased with temperatures up to $15^{\circ}C$ as optimum temperature and above this temperature, the filtration rates decreased exponentially. Venus clams showed very low filtration rates at low salinity (10-15 psu) and maximum values at high salinity (30-35 psu). Regardless of water temperature and salt change, 2-year class clams showed high filtration rates, but low in 4-year-class. Polynomial regression curves with water temperature were shifted to the left in low temperature region. Thermal coefficient $Q_{10}$ values showed much higher values at low temperature range than at high temperature range, too. These results indicate that the venus clam is more sensitive in cold water. Polynomial regression curves with salinity were shifted to the right in high saline region. According to this study, the venus clam Gomphina veneriformis, subtidal filter-feeding bivalve, was the stenothermal organism, inhabited mainly in low temperature and the stenohaline, in high saline waters.

  • PDF

Rabbit's Cervical Tracheal Replacement with Cryopreserved Homograft - Effects on the Viability and Rejection - (토끼 경부기관의 초냉동보관 동종이식편 기관 이식술 - 생육성 및 거부반응에 미치는 영향 -)

  • 원태희;서정욱;성숙환
    • The Korean Journal of Thoracic and Cardiovascular Surgery
    • /
    • v.31 no.12
    • /
    • pp.1127-1133
    • /
    • 1998
  • Background: There are no ideal substitutes for tracheal replacement. Therefore we investigated the possibility of clinical use of cryopreserved tracheal homograft with special interest in the viability and rejection of the epithelial cell and cartilage. Material and Method: Rabbit's trachea was sected and stored in liquid nitrogen tank for 1 month. Tracheal replacement was done in 45 rabbits with autograft(n=15, Group 1), fresh allograft(n=15, Group 2) and cryopreserved homograft(n=15, Group 3). After 7, 14, and 30 days, 5 rabbits in each group were sacrificed and the regeneration of epithelium and cartilage and the degree of rejection were assessed by counting the monocellular infiltration. Result: Investigation at day 7, showed no difference in epithelial regeneration, however, at days 14 and 30, Group 1 showed better regeneration of epithelium than groups 2 and 3. There was no difference of epithelial regeneration between group 2 and 3. There was little rejection at day 7, but at days 14 and 30, there was significant rejection in group 2 and group 3.(P<0.05). Group 3 showed lesser rejection than group 2 at days 14 and 30, but it was not statistically significant. Cartilage showed no rejection and maintained its viability in groups 2 and 3. Conclusion: Cryopreserved tracheal homograft can maintain its viability, therefore it may represent a possibility of clinical application for tracheal replacement. However, cryopreservation can not eliminate the antigenicity of the trachea completely. Furthere studies for lowering the antigenicity and rejection should be performed for an ideal substitute for tracheal replacement.

  • PDF