• Bulletin of the Korean Chemical Society
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• v.24 no.5
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• pp.559-565
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• 2003

An analytical method the determination of benzo(a)pyrene (BaP) and its hydroxylated metabolites, 1-hydroxybenzo(a)pyrene (1-OHBaP), 3-hydroxybenzo(a)pyrene (3-OHBaP), benzo(a)pyrene-4,5-dihydrodiol (4,5-diolBaP) and benzo(a)pyrene-7,8-dihydrodiol (7,8-diolBaP), in rat urine and plasma has been developed by HPLC/FLD and GC/MS. The derivatization with alkyl iodide was employed to improve the resolution and the detection of two mono hydroxylated metabolites, 1-OHBaP and 3-OHBaP, in LC and GC. BaP and its four metabolites in spiked urine were successfully separated by gradient elution on reverse phase ODS $C_{18}$ column (4.6 mm I.D., 100 mm length, particle size 5 ㎛) using a binary mixture of MeOH/H₂O (85/15, v/v) as mobile phase after ethylation at 90 ℃ for 10 min. The extraction recoveries of BaP and its metabolites in spiked samples with liquid-liquid extraction, which was better than solid phase extraction, were in the range of 90.3- 101.6% in n-hexane for urine and 95.7-106.3% in acetone for plasma, respectively. The calibration curves has shown good linearity with the correlation coefficients (R²) varying from 0.992 to 1.000 for urine and from 0.996 to 1.000 for plasma, respectively. The detection limits of all analytes were obtained in the range of 0.01-0.1 ng/mL for urine and 0.1-0.4 ng/mL for plasma, respectively. The metabolites of BaP were excreted as mono hydroxy and dihydrodiol forms after intraperitoneal injection of 20 mg/kg of BaP to rats. The total amounts of BaP and four metabolites excreted in dosed rat urine were 3.79 ng over the 0-96 hr period from adminstration and the excretional recovery was less than 0.065% of the injection amounts of BaP. The proposed method was successfully applied to the determination of BaP and its hydroxylated metabolites in rat urine and plasma for the pharmacokinetic studies.

• The Korean Journal of Mycology
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• v.19 no.1
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• pp.79-84
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• 1991

Fruit bodies of Ganoderma lucidum have been used for most pharmacological studies, but pharmacological effects are likely variable because the habitats and strains of Ganoderma lucidum are different. Therefore, their fermentation is required to produce constant and reliable pharmacolo­gical constituents from Ganoderma lucidum. During the studies of medium for industrial application. it was found that ginseng root residues, remaining after being extracted with ethanol, were a good carbon source for a fermentation of Genoderma lucidum and a corn steep liquor was also economical for the nitrogen source. Yield of the mycelial cultured in ginseng root residues and corn steep liquor was 2.5 times higher than that in glucose and peptone, known as a conventional medium of Ganoderma lucidum. The polysaccharide content of the extracts from the cultured mycelia was higher than that from fruit bodies, but protein content was vice versa. Extracts of the cultured mycelia were more effective and lasting than extracts of the fruit bodies in decreased hypertention of spontaneously hypertensive rats (SHR).

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.22 no.1
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• pp.76-88
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• 2009

Back ground and Objective : There is a need for objectification and scientific verification of Pattern identification in Oriental medicine. The purpose of this study was to investigate the skin characteristics of Qi deficiency and Blood deficiency animal models. Material and Methods : Sprague-Dawley rats were divided into three groups: normal group, Qi deficiency group and Blood deficiency group. The Qi deficiency animal model was induced through restriction of food (75g/kg/day) for 20 days. Blood deficiency animal model was induced by bleeding from tail vein(0.3 ml/time) 8 times. The normal animal model was kept without any intervention. The general condition was observed by measuring body weight, body temperature, blood pressure, pulse rate, and hematological and biochemical parameters. The skin characteristics were observed by measuring the erythema index(EI), melanin index(Ml), transepidermal water loss(TEWL) and dermal microcirculation. Results : 1. In the Qi deficiency group, body weight was lower than the other groups. In the Qi deficiency group, blood pressure was lower than in the Normal group. There was no difference in body temperature and pulse rate between the three groups. 2. In the Qi deficiency group, blood sugar was lower than in the Blood deficiency group. There was no difference in triacylglycerol between the three groups. In the Qi deficiency group, the WBC count was lower than in the Blood deficiency group. RBC count was highest in the Qi deficiency group, Normal group and Blood deficiency group respectively. In the Qi deficiency group, Hb and Hct were higher than the other groups. 3. EI and MI were decreased in the Qi deficiency group, and EI showed a significant decrease. 4. EI and MI were increased in the Blood deficiency group, and MI showed a significant increase. 5. TEWL was significantly increased in the Qi deficiency group, while it was decreased in the Blood deficiency group, TEWL was highest in the Qi deficiency group, Normal group and Blood deficiency group respectively and all three groups showed significant difference. 6. In the Qi deficiency group, dermal microcirculation was lower than the other groups. Conclusion : The above results show that the erythema index decreases in the Qi deficiency model, and the melanin index increases in the Blood deficiency model. The Qi deficiency animal model shows an increase in transepidermal water loss, while the Blood deficiency animal model shows a decrease. Further studies should develop new models of Pattern Identification that are more specific.

• Proceedings of the Ginseng society Conference
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• 1988.08a
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• pp.28-32
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• 1988

Recently, we isolated a toxic substance named 'toxohormone-L' from ascites fluid of patients with various malignant tumors. The toxohormone-L stimulated lipolysis in rat adipocytes and induced anorexia in rats. Both the lipolytic and the anorexigenic actions of toxohormone-L were found to be inhibited by ginsenoside $Rb_2$ in Korean red ginseng. Isolated rat adipocytes are well known to possess opposite pathways of lipid metabolism: lipolysis and lipogenesis. Both of the metabolism respond to various biologically active substances such as epinephrine, ACTH and insulin. Epinephrine and ACTH stimulate lipolysis and insulin accelerates lipogenesis. Recently. Korean red ginseng powder was found to contain adenosine and an acidic substance which inhibited epinephrine-induced lipolysis and stimulated insulin-mediated lipogenesis from added glucose. The chemical structure of this acidic substance is determined to be pyro-glutamic acid (Pyro-Glu), Pyro-Glu exhibits selective modulations toward the opposite metabolic pathways in rat adipocyte; It inhibits the lipolysis but stimulates the lipogenesis. We call these substances (adenosine, Pyro-Glu) 'selective modulators' or 'insulin-like substances'. Based on these results, physiological significances of these substances in Korean red ginseng were discussed.

• Proceedings of the Ginseng society Conference
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• 1988.08a
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• pp.81-86
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• 1988

The effect of three polyacetylene compounds. panaxydol. panaxynol and panaxytriol isolated from Korean ginseng on $CCI_4-induced$ lipid peroxidation in vitro and in vivo hepatic microsomal lipid peroxidation were investigated. Lipid peroxide levels both in serum and liver and serum enzyme (GOT. GPT. LDH) activities of normal or $CCI_4-treated$ mice and rats were also determined after administration of polyacetylenes. Hepatic microsomal cytochrome P-450 content and activities of aniline hydroxylase and aminopyrine demethylase were measured after treatment of polyacetylenes with or without carbon tetrachloride. As results. treatment with polyacetylenes to control mice did not influence the levels of lipid peroxides and serum enzyme activities while panaxynol did. Panaxynol itself inhibited liver lipid peroxidation in normal mice. Polyacetylene compounds protected hepatic lipid peroxidation and lowered serum lipid peroxide levels induced by $CCI_4$ Polyacetylenes prevented leakage of LDH to serum but elevated GOT and GPT levels caused by $CCI_4$ were not changed by polyacetylene pretreatment. $CCI_4$ caused losses in the content of cytochrome P-450 and activities of aniline hydroxylase and aminopyrine demethylase. When polyacetylenes were treated without $CCI_4$ panaxydol and panaxynol induced aniline hydroxylase and all three polyacetylenes induced aminopyrine demethylase. Cytochrome P-450 contents were not affected by polyacetylenes. In vitro hepatic microsomal lipid peroxidation was inhibited by polyacetylenes and $DL-{\alpha}-tocopherol$ in a concentration-dependent manner.

• Journal of Ginseng Research
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• v.17 no.1
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• pp.52-60
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• 1993

Panax ginseng has been extensively used in the traditional oriental medicine as a restorative, tonic and Prophylactic agent. Recently, several reports regarding to anticancer effects of Panax ginseng has accumulated. These studies emphasized the fact that the anticancer activities might be due to a glycoside group called ginsenoside or pan.u saponin which has a water soluble characteristic. However, the authors and collaborates demonstrated that a highly lipid soluble component in extract of Panax ginseng roots contains a considerable cytotoxic activities against marine leukemic cells (L1210, P388) and human censer cells (HRT-18, HT-29, HCT48). This study was devised to observe the cytotoxic activities of Petroleum-ether extract of Panax giuseng roots (crude GBD and its Partially Purified fraction from silicic acid column chromatography (7 : 3 GX) against sarcoma-180 (5-180) and Walker carcinosar- coma 256 (Walker 256) in vivo, and murine leukemic Lymphocytes (L1210) and human rectal cancer cells (HRT-18) and human colon cancer cells (HT-29 and HCT48) in vitro. Each cell-line was cultured in medium containing serial concentration of the crude GX or 7 : 3 GX in vitro. A highly lipid soluble compound in the extract of Panax ginseng root was cytocidal to murine leukemic cells and human colon and rectal cancer cells in vitro. In the meantime, ginseng saponin derivatives did not have cytotoxic effects at its corresponding concentration. The growth rates of the cancer cells in medium containing ginseng extracts were inhibited gradually to a significant degree roughly in proportion to the increase of the extract concentration. The cytotoxic activity of 7 : 3 GX was about 3 times more potent than that of crude GX, one unit of cytotoxic activity against L1210 cells being equivalent to 2.54 Ug and 058 Ug for the crude GX and 7 : 3 GX, respectively. The Ri value of the active compound on silica- gel thin layer chromatography with petroleum-ether/ethyl ether/acetic acid mixture (90 : 10 : 1, v/v/v) as a developing so lvent was 053. While, the Panaxydol and Panaxynol as active compounds were purified from Petroleum-ether extract of Panax ginseng root by Drs. Ahn and Kim, and author found out that the one unit of cytotoxic activity of the Panaxydol and Panaxynol against L1210 cells being equivalent to 056 Ug and 0.3918 respectively. The survival times of mice inoculated with S-180 cells were extended about 1.5 to 2 times by the 7 : 3 GX treatment compared with their control group. The significantly decreased hemoglobin values of rats after inoculation with Walker 256 were recovered to normal range by oral administration of the crude Gt The synthetic levels of protein, DNA and RNA in human colon and rectal cancer cells were significantly diminished by treatment with the crude GX, which can explain a part of the origin of its anticancer activity.

• Journal of Ginseng Research
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• v.35 no.2
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• pp.176-190
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• 2011

There seems to be some controversy about the effect of total ginseng saponin (TGS) on the secretion of catecholamines (CA) from the adrenal gland. Therefore, the present study aimed to determine whether TGS can affect the CA release in the perfused model of the adrenal medulla isolated from spontaneously hypertensive rats (SHRs). TGS (15-150 ${\mu}g/mL$), perfused into an adrenal vein for 90 min, inhibited the CA secretory responses evoked by acetylcholine (ACh, 5.32 mM) and high $K^+$ (56 mM, a direct membrane depolarizer) in a dose- and time-dependent fashion. TGS (50 ${\mu}g/mL$) also time-dependently inhibited the CA secretion evoked by 1.1-dimethyl-4 -phenyl piperazinium iodide (DMPP; 100 ${\mu}M$, a selective neuronal nicotinic receptor agonist) and McN-A-343 (100 ${\mu}M$, a selective muscarinic M1 receptor agonist). TGS itself did not affect basal CA secretion (data not shown). Also, in the presence of TGS (50 ${\mu}g/mL$), the secretory responses of CA evoked by veratridine (a selective $Na^+$ channel activator (50 ${\mu}M$), Bay-K-8644 (an L-type dihydropyridine $Ca^{2+}$ channel activator, 10 ${\mu}M$), and cyclopiazonic acid (a cytoplasmic $Ca^{2+}$-ATPase inhibitor, 10 ${\mu}M$) were significantly reduced, respectively. Interestingly, in the simultaneous presence of TGS (50 ${\mu}g/mL$) and N${\omega}$-nitro-L-arginine methyl ester hydrochloride [an inhibitor of nitric oxide (NO) synthase, 30 ${\mu}M$], the inhibitory responses of TGS on the CA secretion evoked by ACh, high $K^+$, DMPP, McN-A-343, Bay-K-8644, cyclopiazonic acid, and veratridine were considerably recovered to the extent of the corresponding control secretion compared with the inhibitory effect of TGS-treatment alone. Practically, the level of NO released from adrenal medulla after the treatment of TGS (150 ${\mu}g/mL$) was greatly elevated compared to the corresponding basal released level. Taken together, these results demonstrate that TGS inhibits the CA secretory responses evoked by stimulation of cholinergic (both muscarinic and nicotinic) receptors as well as by direct membrane-depolarization from the isolated perfused adrenal medulla of the SHRs. It seems that this inhibitory effect of TGS is mediated by inhibiting both the influx of $Ca^{2+}$ and Na+ into the adrenomedullary chromaffin cells and also by suppressing the release of $Ca^{2+}$ from the cytoplasmic calcium store, at least partly through the increased NO production due to the activation of nitric oxide synthase, which is relevant to neuronal nicotinic receptor blockade, without the enhancement effect on the CA release. Based on these effects, it is also thought that there are some species differences in the adrenomedullary CA secretion between the rabbit and SHR.

• Korean Journal of Bronchoesophagology
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• v.6 no.1
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• pp.29-37
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• 2000

Background and Objectives : The concentration of sulfur dioxide($SO_2$) gas in the ambient air appears increasing in the industry and urban area day by day. It was known that $SO_2$ is noxious gas. $SO_2$ can be irritating to the eyes, nose, throat, upper respiratory tract and skin. It produces sulfurous acid on contact with water and is extremely irritating to the nasopharynx and respiratory tract. Laminin is a family of extracellular matrix glycoproteins localized in the basement membrane that separates epithelial cells from the underlying stroma. The biological activities of laminin are to promote cell migration, wound healing, growth and differentiation. Meterials and Methods : The histologic changes and the expression of laminin in tracheal mucosa sacrificed at every weeks (to 7 weeks) after continued $SO_2$ exposure of 250ppm for 30 minutes a day were studied in rats. Results : Pathologic tissue was formed at the tracheal mucosa and the underlying tissue by the infiltration of monocytes and epithelium was transformed to the single cell layered epithelium above 5 weeks after exposure. At the 6 weeks after exposure, epithelial cells were partially lost and epithelial cell layer was transformed to be leaf-shaped. Submucosal tissue was transformed to be lymphatic tissue. An intense positive staining for laminin was found in apical cytoplasm and lateral surface of the normal epithelial cells and basement membrane but at the 5 and 6 weeks after exposure, laminin activity was decreased to the moderate activity. At the 7 weeks after exposure, laminin activity was decreased to the weak activity. Conclusion : Our finding suggests that $SO_2$ makes histologic damage on the tracheal mucosa and decreases immunoreactivity for laminin. Longer duration of the exposure of $SO_2$ makes more histologic damage on the tracheal mucosa and decreases immunoreactivity for laminin.

• The Korean Journal of Zoology
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• v.33 no.2
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• pp.183-190
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• 1990

The present study exarnines the physiological alteradons in prolactin (PRL) messenger ribonucleic acid (mRNA) and serum PRL levels during the rat estrous cycle and the effed of naloxone, an endogenous oploid peptide receptor antagonist, on PRL gene expression during the rat estrous cycle. Adult female rats exhibiting at least two consecutive 4-day estrous cycles were used in this study. A single injection of naloxone (2mg/kg b.w.) or saline was given sc 30 mm prior to decapitation. Animals were sacrificed at 10:00 h of each stage of the estrous cycle, and at 2-h intervals from 10:00 h to 20:00 h during the proestrus. PRL mRNA and serum PRL levels were determined by a RNA-blot hybridization with the rat PRL cDNA probe and by a PRL radjoimmunoassay, respectively. PRL mRNA and serum PRL levels were not dramatically altered in the morning of each stage of diestrus I, II and proestrus, and naloxone failed to modify the two parameters. During estrus naloxone clearly suppressed serum PRL levels, but it was unable to modify PRL mRNA levels. A more detailed examination of the proestrus stage revealed that PRL mRNA and serum PRL levels were fluctuated as a function of time: PRL mRNA levels reached a maximum level at 12:00 h and gradually decreased until 18:00 h. PRL mRNA levels then rose at 20:00 h. No difference of PRL mRNA levels between the control and naloxone-treated groups was observed. Changes in serum PRL levek during proestrus were conversely related to changes in PRL mRNA: serum PRL levels were low from 10:00 h to 14:00 h, then increased and reached a maximum level at 16:00-18:00 h. Following then, serum PRL levels were decreased. Naloxone was effective in suppressing the charaderistic afternoon surge of PRL from 16:00 h to 20:00 h. These data clearly showed that alterations in PRL mRNA levels were conversely correlated with changes mn serum PRL levels on proestrus, indicating a differential regulation of PRL gene expression and secretion.

• The Korean Journal of Zoology
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• v.14 no.4
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• pp.199-204
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• 1971

The effect of 400 R total-body X-irradiation on the rate of deoxycytidine-2-$^14 C$(CdR-2-$^14 C$) into DNA and on the degradation of DNA has been studied in the liver, spleen and thymus of the rat. The postirradiation period can be divided into a radiation reaction period followed by a regeneration period. During the period of radiation reaction, which consists of days 1-2, markdely decreased CdR-2-$^14 C$ incorporation into DNA of each organ is observed. Rate of incorporation of labeled precursor in the thymus shows the most profound decrease, whereas those in the liver and spleen show similar decrease when expressed as percent of normal. The change in the amount of DNA as percent of normal exhibits a similar pattern in all organs, but the rate of decrease is larger in the spleen and thymus compared to that in the liver. The period of regeneration as judged by the incorporation experiment appears day 4 to 5, which consists of the second phase of the regeneration period. The second phase is highlighted by a markedly increased rate of CdR-2-$^14 C$ incorporation and by a slow and continued increase in the amount of DNA in all organs. The regeneration occurs faster in the liver and spleen than in the thymus which is the most radiosensitive of the all. The findings of the present experiments are strongly suggestive of the fact that the radiation-induced loss of spleen and thymus DNA as well as the radiation-caused inhibition in the CdR incorporation into DNA of the thymus are the important factors in the elevated levels of CdR in the urine and plasma.