Erythropoietin (EPO), a glycoprotein hormone produced from primarily cells of the peritubular capillary endothelium of the kidney, is responsible for the regulation of red blood cell production. We have been investigating the roles of glycosylation site added in the biosynthesis and function of recombinant protein. In this study, we analyzed by immunohistochemical methods adaptive mechanisms to excessive erythrocytosis in transgenic (tg) mice expressing dimeric human erythropoietin (dHuEPO) gene. Splenomegaly was observed over 11~21 times in the tg mice. The 2,672 candidate spleen-derived genes were identified through the microarray analysis method, and decreased genes were higher than increased genes in the spleen. The specific proteins in the increased and decreased genes were analyzed by immunohistochemical methods. Our results demonstrate that problems of abnormal splenomegaly would solve in tg mice overexpressing dHuEPO gene.
Purpose: Vascular endothelial growth factor (VEGF) is known as a growth factor of endothelium and fibroblast. The purpose is to know the VEGF effects on fibroblast proliferation and fibroblast's notch receptor expression. Methods: CCD-986sk fibroblast was purchased from the Korean Cell Bank and was used in XTT assay for proliferation and wound healing assay for migration. Immunofluorescent (IF) staining and western blotting were used in testing notch expression of fibroblast. Semiquantitative RT-PCR was used in checking notch 1 mRNA production by fibroblast. Student-t test was used for analyzing results. Results: Cell proliferation assay using XTT showed significant higher proliferation in VEGF treated fibroblast, $2.324{\pm}0.0026$ vs. $2.463{\pm}0.017$ (p=0.002). Wound healing assay showed longer migration in VEGF treated fibroblast (p=0.062). The fluorescence was brighter in VEGF treated cells of notch 1 IF staining. Notch 1 expressions and mRNA productions increased more in VEGF treated cells. Conclusion: VEGF stimulates fibroblast to proliferate, migrate and to express Notch 1 simultaneously. Notch receptor could be related to VEGF mediated wound healing.
Irfan, Muhammad;Kwak, Yi-Seong;Han, Chang-Kyun;Hyun, Sun Hee;Rhee, Man Hee
Journal of Ginseng Research
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v.44
no.4
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pp.538-543
/
2020
Cardiovascular diseases are a rapidly growing epidemic with high morbidity and mortality. There is an urgent need to develop nutraceutical-based therapy with minimum side effects to reduce cardiovascular risk. Panax ginseng occupies a prominent status in herbal medicine for its various therapeutic effects against inflammation, allergy, diabetes, cardiovascular diseases, and even cancer, with positive, beneficial, and restorative effects. The active components found in most P. ginseng varieties are known to include ginsenosides, polysaccharides, peptides, alkaloids, polyacetylene, and phenolic compounds, which are considered to be the main pharmacologically active constituents in ginseng. P. ginseng is an adaptogen. That is, it supports living organisms to maintain optimal homeostasis by exerting effects that counteract physiological changes caused by physical, chemical, or biological stressors. P. ginseng possesses immunomodulatory (including both immunostimulatory and immunosuppressive), neuromodulatory, and cardioprotective effects; suppresses anxiety; and balances vascular tone. P. ginseng has an antihypertensive effect that has been explained by its vasorelaxant action, and paradoxically, it is also known to increase blood pressure by vasoconstriction and help maintain cardiovascular health. Here, we discuss the potential adaptogenic effects of P. ginseng on the cardiovascular system and outline a future research perspective in this area.
Structure and differentiation mechanism of the seed coat of Panax ginseng are studied with light and electron microscopes to clarify the developmental processes of seed coat and the structural changes during the differentiation of the seed. The seed coat of ginseng is differentiated from the inner cell layers of ovary wall, which can be compared with the seed coat differentiated from integument(s) in other plants. The single integument is differentiated into endothelium, which is degenerated to one layer of 4${\mu}{\textrm}{m}$ in thickness, composed of remants of cell wall components in fully ripened seed. The ripened seed coat is composed of three layers; fringe layer, inner layer and palisade layer, and all of the them are crossed at right angles with one another. This may be the cause of protection of the kernel from other mechanical injuries. The thickness of fully ripened seed coat is about 300~600 ${\mu}{\textrm}{m}$, and arrangements of sclereids are irregular. However, the raphe region of seed coat is thin about 200 ${\mu}{\textrm}{m}$ in thickness and sclereids in that region are arranged regularly. This is the important cause for the cleavage of the seed coat during post-maturation process. The vascular bundles on the raphe are still remaining after sarcocarps are removed, and one of the branches of vascular bundles entered into the seed coat through the hilum and extended to chalazal region. During post-maturation process, the supply of water being necessary for growth of embryo may be accompolished by the vascular bundles entered into the seed coat through the opened hilum.
Objectives Cigarette smoking had been recorded as the main cause of impaired endothelium-dependent vasodilation in smokers by reducing nitric oxide (NO), a production of endothelial nitric oxide synthase (eNOS). However, the mechanism of NO impairment via eNOS activity is unclear until now. In this study, cell passage is suggested to be a relevant factor to eNOS expression under cigarette smoking stress. Methods Bovine aortic endothelial cells (BAECs) were chosen as the research subject with passages ranking from 6 to 9 (6P to 9P). After exposure of cigarette smoking extract (CSE) solution, MTT assay and Western blot method were performed to check the cell viability as well as eNOS protein concentration. In these experiments, four concentrations of CSE at 0.5, 1, 2, and 4% were selected for treatment. Results Our results showed that cells almost died at 4% of CSE. Besides, eNOS protein mass had a linear decrease under the increase of CSE concentration. In addition, the effect of CSE on eNOS expression was dissimilar between different passages. Conclusions This study indicated that CSE had effect on both cell viability and eNOS expression. Besides, a reduction in protein mass was matched with the decrease of cell viability due to CSE tress. Last but not least, the response of eNOS protein to different concentration of CSE at different passages was disparate, making the hypothesis about cell passage related inhibition of eNOS caused by CSE solution.
Erythropoietin (EPO), a glycoprotein hormone produced from primarily cells of the peritubular capillary endothelium of the kidney, is responsible for the regulation of red blood cell production. We have been investigating the roles of glycosylation site added in the biosynthesis and function of recombinant protein. We constructed three EPO mutants ($\Delta$69, $\Delta$105 and $\Delta$69,105), containing an additional oligosaccharide chains. EPOWT and EPO$\Delta$69 were effectively expressed in transient and stably transfected CHO-K1 cell lines. But, it wasn't detected any protein in the culture medium of EPO$\Delta$105 and EPO$\Delta$69,105 mutants. The growth and differentiation of EPO-dependent human leukemic cell line (F36E) were used to measure the cytokine dependency and in vitro bioactivity of rec-hEPO. MTT assay values were increased by survival of F36E cells at 24h. To analysis biological activity in vivo, two groups of ICR-mice (7 weeks old) were injected subcutaneously with 10 IU per mice of rec-hEPO molecules on days 0 and 2. Red blood cell and hematocrit values were measured on 6 days after the first injection. The hematocrit values were remarkably increased in all treatment groups. The pharmacokinetic analysis was also affected in the mice injected with rec-hEPO molecules 2.5 IU by tail intravenous. Protein samples were detected by Western blotting. An EPO$\Delta$69 protein migrated as a broad band with an average apparent molecular and detected slightly high band. Enzymatic N-deglycosylation resulted in narrow band and was the same molecular size. The biological activity of EPO$\Delta$69 was enhanced to compare with wt-hEPO. The half-life was longer than wt-hEPO. The results suggest that hyperglycosyalted recombinant human erythropoietin (EPO$\Delta$69) may have important biological and therapeutic good points.
Kim, Min-Jung;Kim, Taegyun;Suh, Gil Joon;Kwon, Woon Yong;Kim, Kyung Su;Jung, Yoon Sun;Ko, Jung-In;Shin, So Mi;Lee, A Reum
Clinical and Experimental Emergency Medicine
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v.5
no.4
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pp.211-218
/
2018
Objective This study aimed to determine whether simultaneous decreases in the serum levels of cell adhesion molecules (intracellular cell adhesion molecule-1 [ICAM-1], vascular cell adhesion molecule-1 [VCAM-1], and E-selectin) and S100 proteins within the first 24 hours after the return of spontaneous circulation were associated with good neurological outcomes in cardiac arrest survivors. Methods This retrospective observational study was based on prospectively collected data from a single emergency intensive care unit (ICU). Twenty-nine out-of-hospital cardiac arrest survivors who were admitted to the ICU for post-resuscitation care were enrolled. Blood samples were collected at 0 and 24 hours after ICU admission. According to the 6-month cerebral performance category (CPC) scale, the patients were divided into good (CPC 1 and 2, n=12) and poor (CPC 3 to 5, n=17) outcome groups. Results No difference was observed between the two groups in terms of the serum levels of ICAM-1, VCAM-1, E-selectin, and S100 at 0 and 24 hours. A simultaneous decrease in the serum levels of VCAM-1 and S100 as well as E-selectin and S100 was associated with good neurological outcomes. When other variables were adjusted, a simultaneous decrease in the serum levels of VCAM-1 and S100 was independently associated with good neurological outcomes (odds ratio, 9.285; 95% confidence interval, 1.073 to 80.318; P=0.043). Conclusion A simultaneous decrease in the serum levels of soluble VCAM-1 and S100 within the first 24 hours after the return of spontaneous circulation was associated with a good neurological outcome in out-of-hospital cardiac arrest survivors.
Park, Sun Young;Lee, Pyeng Jae;Shin, Seon Mi;Kim, Ho Hyun
Journal of Physiology & Pathology in Korean Medicine
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v.27
no.4
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pp.400-408
/
2013
This study aimed to investigate the relaxation effects and its underlying mechanisms of Rubus coreanus(RC) extract in contracted rabbit corpus cavernous tissues by phenylephrine(PE) $1{\mu}M$. In order to define the relaxation effects of RC, rabbit corpus cavernous tissues were prepared in $2{\times}2{\times}6mm$ sized strip. The dose-dependent relaxation responses of RC at 0.01-3.0 $mg/m{\ell}$ in contracted strips induced by PE were measured and also observed after endothelial denudation. To analyze the underlying mechanisms of RC-induced relaxation, indomethacin(IM), tetraethylammonium chloride(TEA), $N{\omega}$-nitro-L-arginine (L-NNA), methylene blue(MB) were treated before RC extract infused into precontracted strips induced by PE. To study the effect of RC extract on influx of extracellular $Ca^{2+}$ in corpus cavernous strips, calcium chloride(Ca) 1 mM infused into precontracted strips induced by PE after pretreatment of RC extract in $Ca^{2+}$-free krebs-ringer solution. To investigate cytotoxic activity and nitric oxide(NO) concentration of RC extract on human umbilical vein endothelial cell(HUVEC), cell viability on HUVEC was measured by MTT assay, and NO concentration was measured by Griess reagent system. The cavernous strips were significantly relaxed by RC extract at 1.0 $mg/m{\ell}$, 3.0 $mg/m{\ell}$ and the relaxation responses to RC were inhibited significantly by endothelial disruption. The pretreatment of IM, TEA didn't affect RC extract-induced endothelium-dependent relaxation, but the pretreatment of L-NNA, MB reduced RC extract-induced endothelium-dependent relaxation. When $Ca^{2+}$ was supplied the cavernous strips which were precontracted by PE in a $Ca^{2+}$-free krebs-ringer solution, contraction of strips was increased, but pretreatment of RC inhibited contractile response to $Ca^{2+}$. When RC extract was applicated on HUVEC, NO concentration was increased. Our findings show that RC extract exerts a relaxing effect on corpus cavernosum in part by suppressing influx of extracellular $Ca^{2+}$ through activating the NO-cGMP system.
Kim, In-Suk;Ryu, Gun-Chang;Chae, Soo-Chul;Jeon, Chang-Jin
Applied Microscopy
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v.36
no.3
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pp.227-234
/
2006
To investigate the comparative effect of spherical and aspherical RGP lens were worn during 3 weeks on rabbit's cornea. Four white rabbits were worn right eyes with spherical lens and 4 white rabbits were worn right eyes with aspherical RGP lens. Left eyes were served as control. The rabbits were sacrificed at 3 weeks after fitting and observed morphological changes by scanning electron microscopy and also investigate proliferation rate of the corneal epithelium with RGP wearing. After spherical RGP lens wearing, the epithet layer damaged compared to aspherical lens. The superficial cell layer strip off seriously, cell size significantly changed abnormal. Both spherical and aspherical RGP lens fitting group showed so many bacteria and back surface of lens was found like a fern shape. The aspherical RGP lens original material type was some formal than spherical lens. We thought that these pattern was significantly altered with spherical lens by prohibited transmitter oxygen from atmosphere therefore the epithelium shape was changed. This suggested wearing the aspherical lens might be less physiologic than shperical lens fitting.
Histochemical experiment was carry out respectively to confirm the properties of the salis (Achatina fulica and Incilaria fruhstorferi). SDS-PAGE was carried out to compare and invertigate the distribution aspects of protein patterns between the two species. Five types(A, B, F, H and I)of gland cells with four neutral mucopolysaccharide cells and one acid mucopolysaccharide cells and one acid mucopolysaccharide cell were observed in acinous of Achatina fulica, while six types were observed in acinous of Incilaria fruhstorferi: ond acid mucopolysaccharide cell(type-A) and four neutral mucopolysaccharide cells(type-B, C, D and F) and one cell that acid mucopolysaccharide is only mimbrane that surrounded granule(type-E). The results are follows:The thpe-A fland cell is commonly observed between the two species. The type-A gland cell in Achatina fulica possesses a nucleus with a developed heterdchromatin, and the cytoplasm was filled with round granules. The granules were surrounded with an uncertain boundary mimbrane and confirmed with neutral mucopolysaccharides, but is confirmed acid mucopolysaccharide in Incilaria fruhstorferi.The type-B gland cell is obwerved in the two species, too. The type-B gland cell in Achatina fulica was round shaped, and included an evenly alrge nucleus. The uncleoplasm included granules that were confirmed in the neutral mucopolysaccharides of the two species. The type-C and D gland cells exist only in Incilaria fruhstorferi, nucleoplasm was well developed heterochromatins. The type-E gland cell appears in the acinous surrounded the salivary gland of Incilaria fruhstorferi. Thdse granules appear irregular irregular shape and size and the cytoplasm is formed in alveolar. The type-F gland cells are commonly observed in the salivary glands of the two species. They are similar with the type-B gland cell, but the granular shape is comparatively small and irregular, and possess the neutral mucos granules. The type-H gland cells are mainly seen in only Achatina, and in nucleus is a well developed heterochromatin. The cytoplasm is filled with round small granules with acid mucopolysaccharide for alcianophilia observed. The type-I cell was small cell with an irregular shape and only observed in the gland cells of Achatina fulica. The heterochromatins were developed in the nucleus and the granules are not observed in cytoplasm.Secretory ducts of saliva are composed of the interlobular duct and interlobar secretory duct. In Achatina fulica the interlobular duct consists of a simple cuboidal epithelium, while the endothelium of intralobar secretory duct of Incilaria fruhstorferi consists of a simple squamous epithelium and in the cytoplasm is filled with granules(type-G secretory cell). A SDS-PAGE was carried out to confirm that the protein band pattern consist of salivary gland. In conclusions, five more bands in Achatina fulica and three bands in Incilaria fruhstorferi were confirmed in MW<29 kDa. one main band coincides comparatively with both and is between 29-45 kDa. There are four main bands in Achatina fulica and two main bands in Incilaria fruhstorferi between 45-66.5 kDa respectively. The bands in Achatina fulica seem more complex than in incilaria fruhstorferi.
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