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Composition of Free Sugars, Free Amino Acids, Non-Volatile Organic Acids and Tannins in the Extracts of L. chinensis M., A. acutiloba K., S. chinensis B. and A. sessiliflorum S. (구기자(Lycium chinensis Miller), 당귀(Angelica acutiloba Kitag), 오미자(Schizandra chinensis Bailon), 오갈피(Acanthopanax sessiliflorum Seeman) 추출물의 유리당, 유리아미노산, 유기산 및 타닌의 조성)

  • Oh, Sang-Lyong;Kim, Sung-Soo;Min, Byong-Yong;Chung, Dong-Hyo
    • Korean Journal of Food Science and Technology
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    • v.22 no.1
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    • pp.76-81
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    • 1990
  • For the determination of principal taste components in L. chinensis M., A. acutiloba K., S. chinensis B. and A. sessiliflorum S., which were extracted with water and ethanol, contents and composition of free sugars, free amino acids, non-volatile organic acids and tannins were analyzed. The extraction yield of them was high when using the water as an extraction solvent, the optimal ethanol concentration with high yield was in the range of 50-75%. The contents of free amino acids in their extract $(25^{\circ}Bx)$ was high when using the ethanol as the extraction solvent. The amino acid content was in the range of 123-159 mg%. in samples. The free sugar of extracts contained 2-12% in samples which were consisted of glucose, fructose and sucrose. The pH and acidity range of extracts were 4.4-5.1 and 1.01-2.27% respectively, and especially extracts of S. chinensis B. showed to be strong acid as pH 2.9 and addity 11.93%. The composition of non-volatile organic acid in extracts of S. chinensis B. contained citric acid and malic acid 3.90% and 3.92% respectively as major components. The tannin content of extracts was considerably high when extracting with 50% ethanol, especially A. sessiliflorm S. contained $3.35%{\sim}3.85%$.

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Analysis on the Components in Stem of the Lespedeza bicolor (싸리(Lespedeza bicolor) 줄기의 성분 분석)

  • Lee, Yang-Suk;Joo, Eun-Yong;Kim, Nam-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1246-1250
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    • 2005
  • This study was carried out to analyze the components of the stem of bush clover, Lespedeza bicolor to obtain basic data on the studies of the physiological function and a possibility of utilizing L. bicolor. General components of L. bicolor were 82.35$\%$ carbohydrate, 13.38$\%$ moisture, 2.80$\%$ crude protein, 0.86$\%$ crude ash and 0.61$\%$ crude fat. The content of reducing sugar was 147.58 mg/100 g. The total amount of free sugar was 52.4 mg/100 g that was composed of 31.0 mg/100 g fructose, 16.7 mg/100 g glucose and 4.7 mg/100 g sucrose. In the results of mineral analysis, the content of Ca was the highest (472.97 mg/100 g) and followed by 206.70mg/100 g of K. The total amount of hydrolyzed amino acid was 148.95 mg/100 g that consist of 31.74 mg/100g of essential amino acid and 117.24 mg/100 g of non-essential amino acid. Total free amino acids were contained 106.39 mg/100 g that was composed of 8.41 mg/100 g essential amino acids and 97.98 mg/100 g the non-essential, and proline (62.92 mg/100 g) was the highest that account for 59.8$\%$ of total free amino acids. Total content of amino acid derivatives was 30.01 mg/100g and that of $\gamma$ -aminoisobutyric acid was the highest as 12.57 mg/100 g among them.

Effect of Hormones and Short Chain Fatty Acids on CYP7A1 Gene Expression in HepG2 Cell (호르몬과 단쇄지방산이 HepG2 Cell 내에서 CYP7A1 발현에 미치는 효과)

  • Yang, Jeong-Lye;Lee, Hyun-Jung;Kim, Yang-Ha
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.5
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    • pp.573-580
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    • 2005
  • Cholesterol $7\alpha-hydroxylase$ (CYP7A1) is the rate-limiting enzyme in the conversion of cholesterol to bile acids and plays a central role in regulating cholesterol homeostasis. We previously showed that a fermentable $\beta-glucan$ ingestion decreased plasma cholesterol levels due to fecal bile acid excretion elevation involved inincrease of cholesterol $7\alpha-hydroxylase$ mRNA expression and activity. It is proposed that short chain fatty acids (SCFA) produced by cecal and colonic fermentation of soluble fiber are associated with cholesterol-lowering effect of fiber. In the present study, we investigated whether CYP7A1 expression is up-regulated by short chain fatty acids or by hormones in cultured human hepatoma (HepG2) cells. Confluent HepG2 cell were incubated with acetate, propionate, or butyrate at 1 mM concentration for 24 hrs. Acetate as well as propionate increased to 1.8-fold expression of CYP7A1 mRNA than the control. Butyrate also increased 1.5-fold expression of CYP7A1 mRNA. Our data show for the first time that SCFA increase expression of CYP7A1 mRNA. Adding insulin, dexamethasone and triiodothyronine $(1\;{\mu}M)$ to HepG2 cell increased the expression of CYP7A1 mRNA to $150\%,\;173\%,\;141\%$, respectively. These results suggest that SCFA produced by cecal fermentation stimulate enteric nervous system, in which secreted some neuropeptides may be responsible for change in cholesterol and bile acid metabolism. These findings suggest that SCFA are involved in lowering plasma cholesterol levels due to the up-regulation of CYP7A1 and bile acid synthesis.

Anti-inflammatory Metabolites of Agrimonia pilosa Ledeb. and Their Mechanism

  • Park, Mi Jin;Ryu, Da Hye;Cho, Jwa Yeoung;Kang, Young-Hwa
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.13-13
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    • 2018
  • The anti-inflammatory (INF) compounds (1-15) were isolated from Agrimonia pilosa Ledeb. (APL) by activity-guided isolation technique. The isolated compounds (1-15) were identified as quercetin-7-O-rhanmoside (1), apigenin-7-O-glycoside (2), kaempferol-7-O-glycoside (3), apigenin-7-O-[6"-(butyl)-glycoside] (4), querceitn (5), kaempferol (6), apigenin (7), apigenin-7-O-[6"-(pentyl)-glycoside] (8), agrimonolide (9), agrimonolide-6-O-glucoside (10), desmethylagrimonolide (11), desmethylagrimonolide-6-O-glucoside (12), luteolin (13), vitexin (14) and isovitexin (15). Flavonoids, compound 2, 3, 11, and 14-15 have been found in APL for the first time. Furthermore, two novel flavone derivatives, compound 4 and 8, have been isolated inceptively in plant. In the no cytotoxicity concentration ranges of $0-20{\mu}M$, nitric oxide (NO) production level of 1-15 was estimated in LPS-treated Raw 264.7 macrophage cells. The flavone aglycones, 7 (apigenin, $IC_{50}=3.69{\pm}0.34{\mu}M$), 13 (luteolin, $IC_{50}=4.62{\pm}0.43{\mu}M$), 6 (kaempferol, $IC_{50}=14.43{\pm}0.23{\mu}M$) and 5 (quercetin, $IC_{50}=19.50{\pm}1.71{\mu}M$), exhibited excellent NO inhibitory (NOI) activity in dose-dependent manner. In the structure activity relationship (SAR) study of apigenin-derivatives (APD), apigenin; Api, apigenin-7-O-glucoside; Api-G, apignenin-7-O-[6"-(butyl)-glycoside]; Api-BG and apignenin-7-O-[6"-(pentyl)-glycoside]; Api-P, from APL on INF activity was investigated. The INF mediators level such as NO, INF-cytokines, NF-KB proteins, iNOS and COX-2 were sharply increased in Raw 264.7 cells by LPS. When pretreatment with APD in INF induced macrophages, NOI activity of Api was most effective than other APD with $IC_{50}$ values of $3.69{\pm}0.77{\mu}M$. And the NOI activity was declined in the following order: Api-BG ($IC_{50}=8.91{\pm}1.18{\mu}M$), Api-PG ($IC_{50}=13.52{\pm}0.85{\mu}M$) and API-G ($IC_{50}=17.30{\pm}0.66{\mu}M$). The NOI activity of two novel compounds, Api-PG and Api-BG were lower than their aglycone; Api, but more effective than Api-G (NOI: Api-PG and Api-BG). And their suppression ability on INF cytokines such as $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 mRNA showed the similar tendency. Therefore, the anti-INF mechanism study of Api-PG and Api-BG on nuclear factor-kappa B ($NF-{\kappa}B$) pathway, representative INF mechanism, was investigated and Api was used as positive control. Api-BF was more effectively prevent the than phosphorylation of $pI{\kappa}B$ kinase (p-IKK) and p65 than Api-PG in Raw 264.7 cells. In contrast, Api-PG and Api-BG were not reduced the phosphorylation of inhibitor of kappa B alpha ($I{\kappa}B{\alpha}$). Moreover, pretreatment with Api-PG and Api-BG, dose-dependently inhibited LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNAs and proteins in macrophage cells, and their expression were correlated with their NOI activity. Therefore, APL can be utilized to health promote agent associated with their AIN metabolites.

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Effects of Herbs and Green Tea on the Sensory and the Antioxidative Qualities of Beef-Yukwonjeon (서양 향신료 및 녹차를 첨가한 쇠고기 육원전의 관능적 특성과 항산화 효과)

  • Lee, Joo-Hee;An, Lee-Hwa
    • Journal of the East Asian Society of Dietary Life
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    • v.17 no.6
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    • pp.808-815
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    • 2007
  • Four different herbs, including rosemary, oregano, basil and sage, along with green tea were individually added to ground beef at different percentages: 0.5, 1.0, and 1.5, respectively. The sensory qualities of the five natural herbs and their antioxidant effects were evaluated using Beef-Yukwonjeon. For the sensory evaluation results, the 0.5% herb addition and the control group generally showed higher acceptability scores than the 1.0% or 1.5% herb addition groups. In particular, the Yukwonjeon made with 0.5% green tea showed higher preferences in the acceptance tests and ranking tests. However, the sensory differences among samples were reduced after 3 days of storage at $6^{\circ}C$. For the results of lipid oxidation without storage, the TBARS values of the herb addition groups were slightly lower than the value of the control group, but not significantly(p>0.05). However, the antioxidant effects of each herb showed higher values over a longer storage period as well as at the higher herb concentrations. The antioxidant effects of the herbs during 7 days' of storage at $6^{\circ}C$ were evaluated as $10.0{\sim}38.5%$ reductions in TBARS values as compared to the group without herbs. Furthermore, the antioxidant effects of the herbs during 30 days' of storage at $-20^{\circ}C$ were shown as $16.8{\sim}27.8%$ reductions in the TBARS value. The green tea showed the highest antioxidant activity against lipid oxidation, because it was acceptable at higher concentrations in the sensory evaluation. These results indicate the potential to produce better quality beef-Yukwonjeon, with herbs, especially green tea.

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The Experimental Study on the Immuno-regulatory effect of ASARI HERBA CUM RADICE Herbal- acupuncture at Pyesu(BL13) on OVA-induced asthma in mice (폐유(肺兪) 세신약침(細辛藥鍼)이 OVA-induced Asthma Mouse Model의 면역조절작용(免疫調節作用)에 미치는 영향)

  • Ryu, Eun-sang;Lee, Hyun;Lee, Byung-ryul
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.185-200
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    • 2005
  • Objective : The aim of this study was to investigate the asthma-suppressive and immune-regulatory effect of AHCR-HA(ASARI HERBA CUM RADICE Herbal-acupuncture) at Pyesu(BLl3) on OVA(ovalbumin)-induced asthma in mice. Methods : C57BL/6 mice out of all the experimental groups, except the Normal group and the AHCR-HA group, were sensitized and challenged with OVA The mice in the AHCR-HA group and the OVA-AHCR-HA group were treated with AHCR-HA(1%) at Pyesu(BL13). The mice in the OVA-Saline group were injected with saline at Pyesu(BL13). The mice in the OVA-Needle-Prick group were treated with a single prick with an injection needle at Pyesu(BL13). AHCR-HA saline injection and needle prick were administered for 8 weeks, three times a week. Result : 1. The populations of granulocytes, CD3e-/CCR3+ cells, CD69+/CD3e+ cells, CD4+ cells and CD23+/B220+ cells in the OVA-induced asthmatic mouse lungs decreased significantly by AHCR-HA. 2. The lung weight, total cells in lung, total leukocytes in BALF, eosinophils in BALF, collagen accumulation in the lung sections of the OVA-AHCR-HA group decreased significantly. 3.The concentrations of IL-4, IL-5, IL-13, IgE in BALF and serum of the OVA-AHCR-HA group decreased significantly. 4. The numbers of Gr-1+/CD11b+, CCR3+, CD3e+, CD19+, CD3e+/CD69+cells in the OVA-AHCR-HA group decreased significantly. 5. The mRAN expressions of $TNF-{\alpha}$, IL-5, IL-4 and IL-13 in lung of the OVA-AHCR-HA group decreased significantly. 6. The AHCR-HA group didn't show any considerable difference from the Normal group. The OVA-saline group and the OVA-Needle prick group showed suppressive effects on OVA-induced asthma however they were not statistically significant. Conclusion : These results suggest that AHCR-HA at Pyesu(BL13) is considered to be effective in treating asthma and to be put to practical use in the future asthma clinic.

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Effects of antler, red ginseng, safflower, ipriflavone and estrogen on hormones, Ca, P and ALP levels in ovariectomized rats (난소적출 rat에 녹용, 홍삼, 홍화, ipriflavone 및 estrogen을 투여 했을 때 호르몬, Ca, P 및 ALP수준에 미치는 영향에 관한 연구)

  • 유상식;김민수;박상훈;김상근
    • Korean Journal of Veterinary Service
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    • v.23 no.3
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    • pp.247-254
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    • 2000
  • This study was performed to elucidate the effects of antler, red ginseng, safflower seed, ipriflavone and estrogen on ovariectomized rats. The rats were fed with Ca and P deficient diet for five weeks to induce osteoporosis. After this period, these animals were fed with normal feed and treated every other day with antler(600 mg/kg, p.o), red ginseng(200 mg/kg, p.o), safflower(200 mg/kg, p.o), ipriflavone(80 mg/kg, p.o) and estrogene(400$\mu\textrm{g}$/kg, i.m) for 5 weeks. During the treatment, the rats were examined for serum concentrations of estradiol, calcitonin, Ca, p and alkaline phosphatase(ALP) activities. The results are summarized as follows 1. The levels of serum 17 $\beta$-estradiol after five weeks of treatment were showed 39.6$\pm$3.0 pg/ml by antler, 33.2$\pm$2.5pg/ml by red ginseng, 34.9$\pm$2.4pg/ml by safflower, 28.1$\pm$3.1 pg/ml by ipriflavone and 40.6$\pm$3.0pg/ml by estrogen-treated group. They were lower than 50.8$\pm$3.1pg/ml of normal control group which had not received ovariectomy. They, however, were significantly higher than 26.8$\pm$1.8pg/ml of ovariectomized non-treatment group(p<0.05). 2. The levels of serum calcitonin after five weeks of treatment were showed 0.60$\pm$0.02 ng/ml by antler, 0.55$\pm$0.04ng/ml by red ginseng, 0.59$\pm$0.02ng/ml by safflower, 0.56$\pm$0.04ng/ml by ipriflavone and 0.62$\pm$0.02ng/ml by estrogen-treated group. They were lower than 0.67$\pm$0.03pg/ml of normal control group. However, they were significantly higher than 0.45$\pm$0.05ng/ml of ovariectomized non-treatment group(p<0.05). 3. The levels of serum Ca of the rats after five weeks of treatment with antler, red ginseng, safflower, ipriflavone and estrogen were 23.51$\pm$2.19$\mu\textrm{g}$/$m\ell$, 25.22$\pm$3.44$\mu\textrm{g}$/$m\ell$, 23.20$\pm$0.02$\mu\textrm{g}$/$m\ell$, 24.76$\pm$3.57$\mu\textrm{g}$/$m\ell$, 23.07$\pm$3.66$\mu\textrm{g}$/$m\ell$, respectively. They were a bit higher than 21.43$\pm$2.22$\mu\textrm{g}$/$m\ell$ of normal control group. And non-treatment group showed 26.12$\pm$0.29$\mu\textrm{g}$/$m\ell$ which was significantly higher than that of control group(P<0.05). 4. The serum P concentraions after five weeks of treatment were showed 12.11$\pm$2.14$\mu\textrm{g}$/$m\ell$ by antler, 13.18$\pm$1.64u91m4 by red ginseng, 12.67 $\pm$2.31$\mu\textrm{g}$/$m\ell$ by safflower, 12.38$\pm$2.07$\mu\textrm{g}$/$m\ell$ by ipriflavone, 11.86$\pm$1.93$\mu\textrm{g}$/$m\ell$ by estrogen-treated group. They were a bit higher than 11.29$\pm$1.23$\mu\textrm{g}$/$m\ell$ of normal control group. And non-treatment group showed 13.42$\pm$1.87$\mu\textrm{g}$/$m\ell$ which was higher than that of control group but not significant. 5. The levels of serum ALP after five weeks of treatment were showed 164.8$\pm$3.8IU/ml by antler, 277.7$\pm$4.8IU/ml by red ginseng, 288.5$\pm$4.5IU/ml by safflower, 214.7$\pm$5.7IU/ml by ipriflavone and 159.4$\pm$5.4IU/ml by estrogen-treated group. They were significantly higher than 144.1$\pm$3.5IU/ml of normal control group(p<0.05). However, they were significantly lower than 336.9 $\pm$12.7IU/ml of ovariectomized non-treatment group(p<0.05). Antler and safflower elevated serum estradiol and calcitonin, and decreased serum ALP significantly. Therefore they were thought to have therapeutic effect on osteoposis by making inhibitory effect on osteoclasts rather than activating osteoblasts.

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SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT (인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Hwang, Soon-Jeong;Choung, Pill-Hoon;Kim, Myung-Jin;Kim, Nam-Yeol;Yoo, Sang-Bae;Jahng, Jeong-Won;Kim, Hyun-Man;Kim, Joong-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.3
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

The Protective Effect of Inhaled Heparin, Cromolyn, Budesonide, and Furosemide on Exercise-induced Asthma (운동유발성 천식의 기관지 수축에 대한 Heparin, Cromolyn, Budesonide, Furosemide 흡입 치료의 효과)

  • Lee, Sin-Hyung;Shim, Jae-Jeong;Lee, Sang-Youb;Cho, Jae-Youn;In, Kwang-Ho;Yoo, Se-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1188-1198
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    • 1998
  • Background : The purpose of the present study was to determine the protective effect of antiasthmatic activity of inhaled heparin, cromolyn sodium, budesonide, furosemide in exercise-induced asthma(EIA). The other important considerable point of this study was the mechanism of bronchoconstriction on EIA. Methods : Eight subjects with a history of EIA were studied on 5 different experiment days. After obtaining baseline $FEV_1$ and FVC, subjects performed a standardized exercise challenge. EIA was assessed by measurement of $FEV_1$ before and after exercise. On experiment day 4, the exercise challenge was performed after the subjects inhaled either heparin (1,000 units/kg/day for 5 days), furosemide (1mg/kg for 5 days), cromolyn (4mg/day for 5 days), or budesonide ($400{\mu}g/day$ for 5 days). On experiment day 5, the methacholine bronchial provocation test was performed. On experiment day 3, activated partial thromboplastine time(aPTI) was checked. Results : Maximum decrements of $FEV_1$ (mean${\pm}$SE) among 0 to 120 minutes after exercise were as follows : heparin was $83.1{\pm}4.81%$ (p=0.010), furosemide was $80.5{\pm}6.87%$ (p=0.071), cromolyn was $86.8{\pm}6.53%$ (p=0.340), and budesonide was $79.4{\pm}7.31%$ (p=0.095). Above medications were compared to the control value ($72.5{\pm}18.2%$) by paired t-test. No medications had effect on $PD_{20}$ of methacholine bronchial provocation test The results were control $1.58{\pm}0.49{\mu}mol$), heparin ($4.17{\pm}1.96{\mu}mol$), furosemide ($1.85{\pm}0.86{\mu}mol$), cromolyn ($2.19{\pm}0.89{\mu}mol$), and budesonide ($3.38{\pm}1.77{\mu}mol$), respectively(p>0.05). The inhaled heparin had no effect of anticoagulation. Conclusion : These data demonstrate that inhaled heparin has a protective effect on EIA. The effect of inhaled cromolyn was statistically absent with manufacture's recommended dosage on EIA. So, the dosage of cromolyn should be carefully evaluated in future. Although inhalation of budesonide and furosemide have no statistical significance compared to control, these drugs also have some protective effects on EIA.

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Detection of rpoB Gene Mutation in Rifampin-Resistant M. Tuberculosis by Oligonucleotide Chip (Oligonucleotide chip을 이용한 Rifampin 내성 결핵균의 rpoB 유전자 돌연변이 검출)

  • Park, Soon-Kew;Lee, Min-Ki;Chung, Byung-Seon;Kim, Cheol-Min;Chang, Chul-Hun L.;Park, Hee-Kyung;Jang, Hyun-Jung;Park, Seung-Kyu;Song, Sun-Dae
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.5
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    • pp.546-557
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    • 2000
  • Background : Oligonucleotide chip technology has proven to be a very useful tool in the rapid diagnosis of infectious disease. Rifampin resistance is considered as a useful marker of multidrug-resistance in tuberculosis. Mutations in the rpoB gene coding $\beta$ subunit of RNA polymerase represent the main mechanism of rifampin resistance. The purpose of this study was to develop a diagnosis kit using oligonucleotide chip for the rapid and accurate detection of rifampin-resistance in Mycobacterium tuberculosis. Method : The sequence specific probes for mutations in the rpoB gene were designed and spotted onto the glass slide, oligonucleotide chip. 38 clinical isolates of Mycobacterium were tested. A part of rpoB was amplified, labelled, and hybridized on the oligonucleotide chip with probes. Results were analyzed with a laser scanner. Direct sequencing was done to verify the results. Result : The low-density oligonucleotide chip design어 to determine the specific mutations in the rpoB gene of M. tuberculosis accurately detected rifampin resistance associated with mutations in 28 clinical isolates. Mutations at codons 531, 526, and 513 were confirmed by direct sequencing analysis. Conclusion : Mutant detection using oligonucleotide chip technology is a reliable and useful diagnostic tool for the detection of multidrug-resistance in M. tuberculosis.

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