• The Korea Journal of Herbology
• /
• v.27 no.6
• /
• pp.49-54
• /
• 2012

Objective : We recently reported that OMC-2010 has an immuno-modulatory effects via inhibiting tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-5. However, we did not find out which constituents play an important role in immuno-modulatory effect of OMC-2010. Thus, this study was performed to estimate the effects of constituents of OMC-2010 on cytokine production in mouse spleen cells, then ultimately reach to find out effective constituents regulating splenic cytokine production. Methods : Mouse spleen cells were pre-treated with water and ethanol extract of constituents of OMC-2010 such as Rehmannia glutinosa (RG), Pinellia ternata (PT), Citrus unshiu Markovich (CUM), Glycyrrhiza uralensis (GU), Platycodon grandiflorum (PG), Schisandra chinensis (SC). After 1 h, the cells were stimulated with lipopolysaccharide (LPS, 1 ${\mu}g/ml$) for 48 h. Then the cells were harvested for real-time reverse transcription polymerase chain reaction to detect cytokine productions. Results : The water extract of RG extract significantly inhibited the LPS-induced inTNF-${\alpha}$ and IL-5 mRNA expressions, but the water extract of PT, CUM, GU, PG, and SC did not. The ethanol extract of RG, PT, and SC significantly inhibited the LPS-induced TNF-${\alpha}$, and IL-5 mRNA expressions, but the ethanol extract of CUM, GU, and PG did not. Conclusions : Theses results could suggest that the water extract of RG and the ethanol extract of RG, PT, and SC inhibited the expression of TNF-${\alpha}$ and IL-5, which means that the possible candidate of OMC-2010 water extract's action might be RG, and ethanol extract's action might be RG, PR, and SC.

• The Korea Journal of Herbology
• /
• v.28 no.5
• /
• pp.87-93
• /
• 2013

Objectives : We recently have reported that constituents of OMC-2010 have an immuno-modulatory effects via inhibiting tumor necrosis factor (TNF)-alpha and interleukin (IL)-5. In this study, based on previous data, we investigated the effects of combinations with each OMC constituents on splenocyte cytotoxicity, cytokine productions, and ovalbumin (OVA) induced experimental allergic asthma. Methods : Mouse splenocytes were pre-treated with ethanol extract of constituents of Rehmannia glutinosa (RG), Pinellia ternata (PT), Schisandra chinensis (SC). We made 4 combinations using RG, PT, and SC (A;1:1:1, B;2:1:1, C;1:2:1, D;1:1:2). The cells were pretreated with A, B, C, or D for 1 h, then stimulated with lipopolysaccharide (LPS, $1{\mu}g/ml$) for 48 h. Then the cells were harvested for real-time reverse transcription polymerase chain reaction to detect cytokine productions. Then using effective combination from RG, PR and SC, we administrated the combination orally, then challenged with OVA to induce asthma. Then we analyzed the airway hyper-reactivity (AHR), lung histology and lung TNF-${\alpha}$ and IL-5 mRNA. Results : A. B. C. and D did not showed significant cytotoxicity on splenocytes. Pre-treatment of A inhibited the expression of TNF-${\alpha}$ and IL-5 significantly, but not B, C, and D. In experimental asthma, administration of A significantly inhibited the increase of AHR, lung damage, TNF-${\alpha}$ and IL-5 expression. Conclusions : Theses results could suggest that inhibitory effects of the ideal combination with RG, PT and SC (1:1:1) could be applied to treatment of asthma and study of asthma mechanisms.

• Journal of Food Hygiene and Safety
• /
• v.31 no.6
• /
• pp.393-398
• /
• 2016

The objective of this study was to isolate pathogenic Escherichia coli from fresh vegetables with selective media and Petrifilm, and identify a suspicious colony using biochemical identification. Twenty gram of lettuce, twenty gram of cabbage and ten gram of sprout were prepared, and a 5-strain mixture of pathogenic E. coli (Enterohemorrhagic E. coli NCCP11142, Enterotoxigenic E. coli NCCP14037, Enteropathogenic E. coli NCCP14038, Enteroaggregative E. coli NCCP14039, Enteropathogenic E. coli NCCP15661) was inoculated to obtain 1, 2 and 3 log CFU/g. Eighty to ninety milliliter of buffered peptone water (BPW) was placed and pummeled for 60 s. As a results, the Petrifilm method was all positive, but enrichment method of qualitative analysis was negative except for 3-log CFU/g inoculated lettuce. Regarding biochemical identification of pathogenic E. coli, the identification rates were dependent on type of methods and vegetables; lettuce: API 20E 100% (44/44), Microgen GNA 100% (44/44) and Food System 66.7% (10/15), cabbage: API 20E 64.7% (22/34), Microgen GNA 50% (16/32) and Food System 60% (9/15), sprout: API 20E 65.1% (28/43), Microgen GNA 62.3% (27/43) and Food System 53.3% (8/15). These results could be useful in determining an appropriate method to detect pathogenic E. coli in fresh vegetables.

• Research in Plant Disease
• /
• v.15 no.2
• /
• pp.83-87
• /
• 2009

Direct Polymerase Chain Reaction (PCR) method that combines biological and enzymatic amplification of PCR targets was developed for the detection of Xanthomonas axonopodis pv. glycines on soybeen seeds without DNA isolation. Primers Xag F1 and Xag R1 were designed to specifically amplify a 401 bp fragment of the glycinecin A gene of X axonopodis pv. glycines. Xag F1 and Xag R1 were used to carry out the PCR analysis with genomic DNA from 45 different bacterial strains including phylogenetically related bacteria with X axonopodis pv. glycines, and other bacterial strains of different genus and species. The PCR assay using this set of primers were able to detect X axonopodis pv. glycines with DNA concentration as low as 200 fg and $1.8{\times}10^3$ cfu/ml. The Xag was detected from the seed samples incubated for 2 hrs with shaking and the intensity of the band was increase with the incubation time of seeds. The Direct PCR assay method without DNA isolation makes detection of X. axonopodis pv. glycines on soybean seeds easier and more sensitive than other conventional methods. The developed seed assay using direct PCR method will be useful for the specific detection of X. axonopodis pv. glycines in soybean seed samples.

• Journal of Life Science
• /
• v.22 no.11
• /
• pp.1456-1462
• /
• 2012

Sorafenib is a multikinase inhibitor and an oral anticancer drug approved for the treatment of patients with advanced renal cell carcinoma and those with unresectable hepatocellular carcinoma. The purpose of this study was to develop an efficient method of the determination of sorafenib in human plasma using tandem mass spectrometry coupled with liquid chromatography (LC/MS/MS) and validate the method by the guidelines of the Korean Food and Drug Administration (KFDA). Plasma samples ($100{\mu}l$) were added with chlorantraniliprole as an internal standard and then mixed with the 0.1% formic acid-containing extraction solution composed of isopropyl alcohol and ethyl acetate (1:4, v/v). After centrifugation, the supernatant was concentrated at $45^{\circ}C$ under negative pressure and centrifugal force. The residue was reconstituted with a mobile phase and injected into the HPLC instrument using a reverse phase Waters XTerra$^{TM}$ C18 column (particle size $3.5{\mu}m$). Liquid chromatography was carried out within the run time of 5 min using a mobile phase composed of buffer (0.1% formic acid and 10 mM ammonium formate), methanol, and acetonitrile (1:6:3, v/v/v). The analytes were monitored by tandem mass spectrometry in the multiple reaction monitoring method programmed to detect sorafenib at 'm/z 465.2 ${\rightarrow}$ 252.5' and chlorantraniliprole at 'm/z 484.4 ${\rightarrow}$ 286.2' with positive electrospray ionization mode ($ES^+$). The result showed the proper linearity ($r^2$ > 0.99) over the range of 2,000-5,000 ng/ml with good accuracy (90.7-103.9%) and precision (less than 10%). The newly developed method using LC/MS/MS was validated by the guideline of KFDA and identified as more sensitive compared to the previous methods.

• Korean Journal of Clinical Laboratory Science
• /
• v.51 no.3
• /
• pp.309-315
• /
• 2019

Cervical cancer is the fourth most common malignant neoplasm in women worldwide. Most cases of cervical cancer are caused by an infection by the human papillomavirus. Molecular diagnostic methods have emerged to detect the HPV for sensitivity, specificity, and objectivity. In particular, real-time PCR has been introduced to acquire a more sensitive target DNA or RNA. RNA extraction and complementary DNA synthesis are proceeded before performing real-time PCR targeting RNA. To identify an adequate and sensitive cDNA synthesis kit, this study evaluated the two commonly used kits for cDNA synthesis. The results show that the $R^2$ and efficiency (%) of the two cDNA synthesis kits were similar in the cervical cancer cell lines. On the other hand, the Takara kit compared to Invitrogen kit showed P<0.001 in the $10^2$ and $10^3$ SiHa cell count. The Takara kit compared to the Invitrogen kit showed P<0.001 in the $10^1$ and $10^2$ HeLa cell count. Furthermore, 8, 4, 2, 1, and 0.5 ml of forty exfoliated cell samples were used to compare the cDNA synthesis kits. The Takara kit compared to the Invitrogen kit showed P<0.01 in 8, 4, and 1 ml and P<0.05 in 0.5 mL. The study was performed to identify the most appropriate cDNA synthesis kit and suggests that a cDNA synthesis kit could affect the real-time PCR results.

• Tuberculosis and Respiratory Diseases
• /
• v.45 no.4
• /
• pp.861-869
• /
• 1998

Backgrounds: The injury of a tissue results in the infalmmation, and the imflammed tissue is replaced by the normal parenchymal cells during the process of repair. But, constitutional or repetitive damage of a tissue causes the deposition of collagen resulting in the loss of its function. These lesions are found in the lung of patients with idiopathic pulmonary fibrosis, complicated fibrosis after diffuse alveolar damage (DAD) and inorganic dust-induced lung fibrosis. The tissue from lungs of patients undergoing episodes of active and/or end-stage pulmonary fibrosis shows the accumulation of inflammatory cells, such as mononuclear cells, neutrophils, mast cells and eosinophils, and fibroblast hyperplasia. In this regard, it appears that the inflammation triggers fibroblast activation and proliferation with enhanced matrix synthesis, stimulated by inflammatory mediators such as interleukin-1 (IL-1) and/or tumor necrosis factor (TNF). It has been well known that TGF-$\beta$ enhance the proliferation of fibroblasts and the production of collagen and fibronectin, and inhibit the degradation of collagen. In this regard, It is likely that TGF-$\beta$ undergoes important roles in the pathogenesis of pulmonary fibrosis. Nevertheless, this single cytokine is not the sole regulator of the pulmonary fibrotic response. It is likely that the balance of many cytokines including TGF-$\beta$, IL-1, IL-6 and TNF-$\alpha$ regulates the pathogenesis of pulmonary fibrosis. In this study, we investigate the interaction of TGF-$\beta$, IL-1$\beta$, IL-6 and TNF-$\alpha$ and their effect on the proliferation of fibroblasts. Methods: We used a human fibroblast cell line, MRC-5 (ATCC). The culture of MRC-5 was confirmed by immunofluorecent staining. First, we determined the concentration of serum in cuture medium, in which the proliferation of MRC-5 is supressed but the survival of MRC-5 is retained. Second, we measured optical density after staining the cytokine-stimulated cells with 0.5% naphthol blue black in order to detect the effect of cytokines on the proliferation of MRC-5. Result: In the medium containing 0.5% fetal calf serum, the proliferation of MRC-5 increased by 50%, and it was maintained for 6 days. IL-1$\beta$, TNF-$\alpha$ and IL-6 induced the proliferation of MRC-5 by 45%, 160% and 120%, respectively. IL-1$\beta$ and TNF-$\alpha$ enhanced TGF-$\beta$-induced proliferation of MRC-5 by 64% and 159%, but IL-6 did not affect the TGF-$\beta$-induced proliferation. And lNF-$\alpha$-induced proliferation of MRC-5 was reduced by IL-1$\beta$ in 50%. TGF-$\beta$, TNF-$\alpha$ and both induced the proliferation of MRC-5 to 89%, 135% and 222%, respectively. Conclusions: TNF-$\alpha$, TGF-$\beta$ and IL-1$\beta$, in the order of the effectiveness, showed the induction of MRC-5 proliferation of MRC-5. TNF-$\alpha$ and IL-1$\beta$ enhance the TGF-$\beta$-induced proliferation of MRC-5, but IL-6 did not have any effect TNF-$\alpha$-induced proliferation of MRC-5 is diminished by IL-1, and TNF-$\alpha$ and TGF-$\beta$ showed a additive effect.

• Korean Journal of Soil Science and Fertilizer
• /
• v.43 no.3
• /
• pp.390-399
• /
• 2010

Non-photochemical quenching (NPQ) values for utilizing them to detect osmotic tolerance in plants were examined with two different soybean cultivars, an osmotic tolerant soybean (Shinpaldalkong 2) and a control soybean (Taekwangkong). Two different stresses were applied to the cultivars as the restricted irrigations of 200 and 50 ml water $pot^{-1}\;d^{-1}$ for 5 days for a control and a drought stress, respectively, and a sodium chloride solution of 200 mmol for 6 days for a salt stress. The intact leaves of the two cultivars after treatment were used to measure chlorophyll fluorescence parameters, maximum efficiencies of photosystem II photochemistry (Fv/Fm), efficiencies of photosystem II photochemistry (${\Phi}_{PSII}$), $CO_2$ assimilation rate ($P_N$), and NPQ. Leaf water potentials of the two cultivars decreased from - 0.2 to - 0.8MPa by a drought treatment and from - 0.7 to - 1.7MPa by a salt treatment. Leaf water content of Shinpaldalkong 2 after a salt treatment was less decreased than that of Taekwangkong. $F_v/F_m$ values of both cultivars were not changed, while ${\Phi}_{PSII}$ and $P_N$ were decreased proportionally to leaf water potential decrease. The response of NPQ was occurred in Shinpaldalkong 2 under the drought and salt stresses. With Taekwangkong cultivar, only drought stress referred NPQ response. The cultivar differences on chlorophyll fluorescence parameters were found in the relationships between ${\Phi}_{PSII}$ and $P_N$, and between NPQ and ${\Phi}_{PSII}$. Although the positive relationships between ${\Phi}_{PSII}$ and $P_N$ were established on all treatments of both cultivars, the decreasing rate of ${\Phi}_{PSII}$ to $P_N$ was smaller in Shinpaldalkong 2 than Taekwangkong. The NPQ was increased according to the decrease of ${\Phi}_{PSII}$ by osmotic treatments in Shinpaldalkong 2. The complementary relationships between NPQ and ${\Phi}_{PSII}$ were well maintained at all treatments in Shinpaldalkong 2, while these relationships were lost at a salt treatment in Taekwangkong. Taken together, the results suggest that analysis of complementary relationships between ${\Phi}_{PSII}$ and NPQ could be more valuable and applicable for determining osmotic tolerance than single analysis of each parameter such as $F_v/F_m$, ${\Phi}_{PSII}$ and NPQ.

• Journal of Sericultural and Entomological Science
• /
• v.16 no.1
• /
• pp.85-92
• /
• 1974

Many of studies on the transovarial transmission of occult virus and their activation due to various stresses such as cold or heat treatment, chemical feeding, and nutritional deficiency, etc., in the silkworm, Bombyx mori L. have been made, but any attempts have been not made to control virus diseases by detection of the occult virus-carried moths in the production of silkworm egg of hybrids, because of difficulty to detect occult virus in any stage. Therefore, it may be worth while to disclose whether a sublethal infection of the moths from which active virus are detectable, has the same level of induction rate as that of occult virus activation, thus to apply its results for the reduction of the occurence of virus diseases in silkworm rearing. For these purposes, the following experiment was conducted as one of preliminary steps. In this study, investigations on the generation-to-generation transmission of occult virus and a sublethal infection, and the role of chromosomal gene of the host, Jam 103 and Jam 104 in the Previous generation, and Jam 103 x Jam 103 and Jam 104 f Jam 104 in the next generation were made for the induction of virus diseases due to the transmitted virus. The frequency of cytoplasmic polyhedrosis due to the induction in the F$_1$ generation was markedly higher in the cross-batches, male$\times$female and male$\times$female in which inoculated individuals were used as fem ale parents than in the cross-batches, male$\times$female and male$\times$female in which virus has been not inoculated or inoculated only to male in the previous generation. The tendency of increasing rate was observed in any treatments; such as the inoculations of cytoplasmic polyhedrosis virus (10$\^$5/, 10$\^$6/ 10$\^$7, and 10$\^$8//ml ill different concentration of inocula) , cold-treatment (5$^{\circ}C$, 12hrs or 24hrs), and formalin-feeding treatment (2％ or 3％). The shape of polyhedra (tetragonal in outline) examined in the F, larvae was identified as that of the inoculated polyhedra with partial application of immunofluorescent techniques. These results suggests that the cytoplasmic polyhedrosis virus in B. meri L. are transmitted to the next generation through the egg, apparently in the occult state. And the experimental results of various cross-batches revealed the egg cytoplasm plays an important part i the transmission of the occult virus of the cytoplasmic polyhedrosis virus,

• The Korean Journal of Pharmacology
• /
• v.24 no.2
• /
• pp.233-240
• /
• 1988

A simple and selective isocratic HPLC method for the analysis of tissue polyamine contents is described and applied on the study of the changes of the hepatic polyamine contents after partial hepatectomy in male rats. The hepatic polyamines are extracted with 0.4 M perchloric acid containing 2 mM disodium EDTA, and then the extract is redissolved in 100 ul of 1 M sodium carbonate and incubated with 300 ul of FNBT-dimethylsulfoxide (1: 100) mixture. The N-2'-nitro-4'-trifluoromethylphenyl drivatives of polyamines are separated through a ERC-ODS column in an isocratic mode with an acetonitrile-water (80:20) mobile phase within 20 min. per a sample, while monitoring the effluent at 242 nm. This improved method which could detect subnanogram of each polyamines is highly specific and reproducible as evidenced by the application of it on the study of the changes of polyamine contents in the regenerating rat liver after partial hepatectomy.