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Study on the Reproductive Function in Transgenic Pig Harboring Human Erythropoietin (hEPO) Gene

  • Lee, Hyun-Gi;Lee, Hwi-Cheul;Chung, Hak-Jae;Hwang, In-Sul;Choi, Myoung-Seob;Byun, Sung-June;Lee, Seung-Hoon;Kim, Min-Ji;Woo, Jae-Seok;Chang, Won-Kyong;Lee, Poong-Yeon;Lee, Hoon-Taek;Park, Jin-Ki
    • Reproductive and Developmental Biology
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    • v.32 no.2
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    • pp.117-121
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    • 2008
  • Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma $17{\beta}$-estradiol ($E_2$) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and $E_2$ level were measured by Hemavet 950 and $E_2$ ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma $E_2$ level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma $E_2$ level, thereby causing reproductive disorders.

Differences in Clinical Laboratory Data between the Elderly and the Young Adults (노인군과 청장년군 간의 정상 검사치의 비교 분석)

  • Lee, Kun-A;Lee, Keun-Mi;Jung, Seung-Pil;Bae, Seong-Wook
    • Journal of Yeungnam Medical Science
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    • v.14 no.2
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    • pp.430-442
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    • 1997
  • Due to the lowering of biological functions resulted from old age, the elderly is known to have many different clinical laboratory data compared with the young adults. But, in korea, such study is lacking. This research is to find the differences between the elderly and the young adults, and also to know the sexual differences, by comparing the outcomes of the clinical. laboratory data. Along with that, it is to help clinical usage of the data in the future. The age of the elderly was between 60 and 83(average age 63.8), and that of the young. controls was between 20 and 35. In both sexes, MCV, MCH, ESR, CRP, AST, ALT, ${\gamma}$GTP, ALP, BUN, total cholesterol were significantly higher in the elderly than in the controls. And lymphocyte count(%), total bilirubin, direct bilirubin, total protein, albumin, $T_3$ were significantly lower in the elderly than in the controls(P<0.05). Hemoglobin, Hct, platelet count, $T_4$ were significantly lower only in the male elderly, and eosinophil count(%), creatinine were significantly higher only in the female elderly(P<0.05). HDL cholesterol was significantly higher only in the male elderly(P<0.01). There were no significant difference between two groups regarding WBC count, segment neutrophil count(%), monocyte count(%), TSH. Many clinical laboratory data are different between the elderly and the young adults, and some clinical laboratory data also have sexual differences.

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A Study on the Hazardousness and the TLV in Working Environments of Benzine (벤진의 유해 위험성과 작업환경 노출기준 연구)

  • Kim, Hyeon-Yeong;Lee, Sung-Bae;Han, Jung-Hee;Shin, Jea-Hoon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.16 no.3
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    • pp.233-244
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    • 2006
  • Of many volatile organic detergents for metals, benzine(CAS No. 8030-30-6), of which the toxicity has not yet been proven, has been used as an alternative of the halide compounds in the consideration of toxic effects, global warming and the destruction of ozone layer. In order to evaluate the effects of the benzine on human body by investigating the subchronic inhalation toxicity, to obtain the basic data for establishing the criteria of exposure in working environments and to classify the hazardousness in compliance with the Industrial Safety and Health Act by evaluating the hazardousness, repeated inhalation exposure test was carried with SD rats. The rats were grouped by 10 females and males each. The repetitive inhalation exposures were carried out at 4 levels of concentration of 0 ppm, 60 ppm, 300 ppm, and 1,500 ppm, for 6 hours a day, 5 days a week, for 13 weeks. The results are described hereunder. 1. No death of the animals of the exposed and controlled groups in the test period. Not any specific clinical symptoms, change in feed intake quantity, abnormality in eye test, or change in activity were observed. 2. In the 300 ppm and 1,500 ppm groups, weight reduction in the female groups and weight increase of liver and kidney in the male groups compared with control group were observed with statistical significance(p<0.05). 3. In the blood test, the HCT increased in the male 300 ppm group and the number of hematocyte increased, MCV and MCH decreased in the male 1,500 ppm group. In the female 1,500 ppm group, the HB decreased and the distribution width of the hematocyte particle size increased. In the blood biochemistry test, the TP in the male 1,500 ppm group and the LDH in the female 1,500 ppm group were increased with statistical significance(p<0.05). 4. Under the test conditions of the present study with SD rats, the NOEL was evaluated to be from 60 ppm to 300 ppm for both male and female groups. By extrapolation, the NOEL for human who work 8 hours a day was evaluated to be from 128 ppm to 640 ppm 5. Since the NOEL evaluated in this study do not exceed 60ppm(0.184 mg/L) the test material does not belong to the classification of the hazardous substance "NOEL${\leq}$0.5mg/L/6hr/90day(rat), for continuous inhalation of 6hours a day for 90 days" nor to the basic hazardous chemical substance class 1(0.2 mg/L/6hr/90day(rat) defined by the GHS which is a criteria of classification and identification of chemical compounds. However, considering the boiling point($30-204^{\circ}C$), flashing point($-40^{\circ}C$), vapor pressure(40 mmHg), and the inflammable range(1.0 - 6.0 %), sufficient care should be taken for handling in the safety aspects including fire or explosion.

Physiological Activity and Antiproliferation Effects of Citron Seed Extracts on Cancer Cells (유자씨 추출물의 생리활성과 암세포 성장 억제효과)

  • Lee, Yoon-Jeong;Hwang, In-Guk;Joung, Eun-Mi;Kim, Hyun-Young;Park, Eui-Seok;Woo, Koan-Sik;Jeong, Heon-Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.12
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    • pp.1672-1678
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    • 2009
  • This study was carried out to investigate the total polyphenol, total flavonoid content, antioxidant activity, angiotensin I-converting enzyme (ACE) inhibitory activity, $\alpha$-glucosidase inhibitory activity, and antiproliferation activity of the citron seed. The citron seed were separated to hull and embryo, and extracted with n-hexane and 70% ethanol. Antioxidant activity of ethanol extract was higher than that of n-hexane extract. IC50 value for DPPH radical scavenging activity of ethanol extract of hull (CSE1) and embryo (CSE2) were 3.18 and 8.43 mg/mL, and those of total antioxidant activity were 19.96 and 11.28 mg AA eq/g, respectively. ACE inhibitory activity and $\alpha$-glucosidase inhibitory activity on CSE1 showed the highest values of 31.61 and 45.17%, respectively. Antiproliferation effects on the MCF7, HepG2, H460, HCT-116, and PC3 cell line showed the highest values of 14.09, 19.12, 12.29, 9.78, and 9.12% in extract concentration of 5 mg/mL, respectively. These results suggested that citron seed can be used for development of functional food material which have biological activities.

Effects of Ethyl Acetate Extracts of Folium Mori on Cadmium Inhalation Toxicity (상엽 Ethyl Acetate 추출물이 카드뮴의 흡입독성에 미치는 영향)

  • Jeung Jaeyeal;Song Young Sun;Lee Ki Nam
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.3
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    • pp.700-710
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    • 2003
  • For the experiment of the effects between cadmium aerosol inhalation toxicity and ethyl acetate extracts of Folium Mori, 4 inhalation exposure groups of rat were exposed to cadmium aerosol in air by whole-body inhalation exposure for 6 hours/day, 5 days/week, and 4 weeks. Cadmium concentration in the air was 0.96㎎/㎥ and mass median diameter (MMD) was 2.48㎛ with 1.85 of geometric standard deviation(GSD). Intraperitoneal injections of ethyl acetate extracts of Folium Mori to inhalation exposure groups were performed for 4 weeks and the results were as follows: The highest body weight gain for 4 weeks and food intake per day were 159.29/4 weeks in treated group III and 18.45g/day in treated group I, respectively. The highest lung and liver weights were 1.31 g in treated group I and 9.42g in treated group III, respectively. The highest kidney weight was 2.21g from treated group I. The lowest cadmium content in lung was 86.39㎍/g from treated group III and the lowest cadmium concentration in blood was 2.72㎍/㎗ from treated group II. Cadmium concentrations of 22.09㎍/g in liver and 24.82㎍/g in kidney were the lowest from inhalation exposure group I and III, respectively. For weekly cadmium concentration in urine, the value of the fourth week from treated group III was the highest, 1.35㎍/㎖. For weekly cadmium concentration in feces, the values of the second and fourth week from treated group I were the highest, 1.11㎍/g. The highest metallothionein concentration in lung was 31.85㎍/g from treated group III and the highest metallothionein concentration in liver was 205.77㎍/g from treated group III. The highest metallothionein concentration in kidney was 206.55㎍/g from treated group III. The highest Hct and Hb values were 38.26% and 11.63g/㎗ from treated group III, respectively. The highest RBC and WBC values were 7.68×106/㎣ and 9.85×10³/㎣ from treated group I, respectively.

Acute Pulmonary Toxicity and Body Distribution of Inhaled Metallic Silver Nanoparticles

  • Kwon, Jung-Taek;Minai-Tehrani, Arash;Hwang, Soon-Kyung;Kim, Ji-Eun;Shin, Ji-Young;Yu, Kyeong-Nam;Chang, Seung-Hee;Kim, Dae-Seong;Kwon, Yong-Taek;Choi, In-Ja;Cheong, Yun-Hee;Kim, Jun-Sung;Cho, Myung-Haing
    • Toxicological Research
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    • v.28 no.1
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    • pp.25-31
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    • 2012
  • The purpose of this study was to determine the acute pulmonary toxicity of metallic silver nanoparticles (MSNPs, 20.30 nm in diameter). Acute pulmonary toxicity and body distribution of inhaled MSNPs in mice were evaluated using a nose-only exposure chamber (NOEC) system. Bronchoalveolar lavage (BAL) fluid analysis, Western blotting, histopathological changes, and silver burdens in various organs were determined in mice. Mice were exposed to MSNPs for 6 hrs. The mean concentration, total surface area, volume and mass concentrations in the NOEC were maintained at $1.93{\times}10^7$ particles/$cm^3$, $1.09{\times}10^{10}\;nm^2/cm^3$, $2.72{\times}10^{11}\;nm^3/cm^3$, and 2854.62 ${\mu}g/m^3$, respectively. Inhalation of MSPNs caused mild pulmonary toxicity with distribution of silver in various organs but the silver burdens decreased rapidly at 24-hrs post-exposure in the lung. Furthermore, inhaled MSNPs induced activation of mitogen-activated protein kinase (MAPK) signaling in the lung. In summary, single inhaled MSNPs caused mild pulmonary toxicity, which was associated with activated MAPK signaling. Taken together, our results suggest that the inhalation toxicity of MSNPs should be carefully considered at the molecular level.

Analysis of p53-Dependency of Differentially Expressed Genes by Capsaicin in Human Colorectal Cancer Cell (인간 대장암 세포주에서 capsaicin 처리에 의한 차별적인 유전자 발현의 p53 의존성 분석)

  • Kim, Hyo-Eun;Jang, Min-Jeong;Lim, Seung-Hyun;Kim, Hyo-Rim;Kim, Soon-Young;Lee, Gun-Joo;Kim, Jong-Sik
    • Journal of Life Science
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    • v.20 no.2
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    • pp.213-218
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    • 2010
  • In the present study, we investigated anti-proliferative activities of capsaicin and gene expression changes in response to capsaicin treatment in human colorectal HCT116 cells. The results showed that capsaicin decreased cell viabilities in a dose dependent manner and induced global gene expression changes. We found that 103 genes were up-regulated more than twofold, whereas 153 genes were down-regulated more than twofold by $100\;{\mu}M$ capsaicin treatment. Among the up-regulated genes, we selected 4 genes (NAG-1, DDIT3, GADD45A and PCK2) and performed RT-PCR to confirm the microarray data. We found that $100\;{\mu}M$ of capsaicin increased tumor suppressor p53 gene expression. In addition, the results showed that NAG-1, DDIT3 and GADD45A expressions were not dependent on p53 presence, whereas PCK2 expression. The results of this study may help to increase our understandings of the molecular mechanism of anti-proliferative activity mediated by capsaicin in human colorectal cancer cells.

Berchemia floribunda-mediated Proteasomal Degradation of CyclinD1 via GKS3β-dependent Threonine-286 Phosphorylation in Human Colorectal Cancer Cells (인간 대장암 세포에 대한 먹넌출 추출물의 GSK3β 의존성 threonine-286 인산화를 통한 Cyclin D1 분해)

  • Kang, Yeongyeong;Eo, Hyun Ji;Kim, Da Som;Park, Youngki;Song, Jeong Ho;Park, Gwang Hun
    • Korean Journal of Plant Resources
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    • v.33 no.4
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    • pp.271-278
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    • 2020
  • In this study, we evaluated the anti-cancer activity and potential molecular mechanism of 70% ethanol extracts of the Berchemia floribunda (BF) which belongs to Rhamnaceae against human colorectal cancer cells. The treatment of BF decreased the cell proliferation in HCT116 cell and suppressed cellular accumulation of Cyclin D1 protein. Inhibition of proteasomal activity by MG132 attenuated BF-mediated Cyclin D1 downregulation and Cyclin D1 was decreased in the cell treated with BF. These findings indicates that BF-mediated Cyclin D1 downregulation may be result from Cyclin D1 proteasomal degradation. Additionally, BF-mediated Cyclin D1 degradation was blocked in the presence of LiCl, a GSK3β inhibitor, but not PD98059, SP600125, SB203580, Bay11-7082, LY294002 an ERK1/2 inhibitor, JNK inhibitor, p38 inhibitor, IκK inhibitor and PI3K inhibitor. Furthermore, BF phosphorylated Cyclin D1 at threonine-286 (Thr286), and LiCl-induced GSK3β inhibition reduced the BF mediated phosphorylation of Cyclin D1 at Thr286. These results suggested that BF may downregulate Cyclin D1 expression as a potential anti-cancer target through GSK3β dependent Cyclin D1 degradation. Therefore, this study provides that the extract of BF has anticancer activity against human colorectal cancer cells.

The overexpression of Arachis hypogaea resveratrol synthase 3 (AhRS3) modified the expression pattern of phenylpropanoid pathway genes in developing rice seeds

  • Lee, Choonseok;Jeong, Namhee;Kim, Dool-Yi;Ok, Hyun-Choong;Choi, Man-Soo;Park, Ki-Do;Kim, Jaehyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.167-167
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    • 2017
  • Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_ Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

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The overexpression of Arachis hypogaea resveratrol synthase 3 (AhRS3) modified the expression pattern of phenylpropanoid pathway genes in developing rice seeds

  • Lee, Choonseok;Jeong, Namhee;Kim, Dool-Yi;Ok, Hyun-Choong;Choi, Man-Soo;Park, Ki-Do;Kim, Jaehyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.105-105
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    • 2017
  • Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

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