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Lee, Hee-Ju,Pan, Cheol-Ho,Kim, Eun-Sil,Kim, Chul-Young The Korean Society for Applied Biological Chemisty 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.2
Online high performance liquid chromatography (HPLC)-2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation ($ABTS^+$) based assay system for phenolic antioxidants in Salsola komarovii extracts was applied. HPLC elute was mixed with a stabilized solution of $ABTS^+$ reagents, and the negative peaks, indicating the antioxidant activity, were monitored by measuring the decrease in absorbance at 734 nm. HPLC with electrospray ionization mass spectrometry was used to identify the seven flavonoids and two phenolic amides in the S. komarovii extracts. The antioxidant compounds were identified as rutin, isoquercitrin, astragalin, and isorhamnetin.
Yang, Ji-Young,Jang, Soon-Young,Kim, Hyun-Ku,Park, Seon-Joo The Korean Society for Applied Biological Chemisty 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.2
Rubi Fructus (Bokbunja in Korean) is a traditional oriental medicine often prescribed as the immature fruit of Rubus coreanus in the Korean pharmacopoeia or Rubus chingii in the Chinese pharmacopoeia. However, Rubus crataegifolius is presently the most popular form in the market. Correctly identifying these fruits is difficult due to morphological similarities in their dried immature forms. Therefore, nucleotide sequences of nuclear ribosomal DNA (nrDNA), internal transcribed spacer, chloroplast DNA (cpDNA) trnL-F, and psbA-trnH intergenic spacer were analyzed to develop objective methods for discrimination among R. coreanus, R. crataegifolius, and R. chingii. Based on trnL-F intergenic sequences, three sequence characterized amplified region (SCAR) markers (CorF/CorR, HwaF2/HwaR2, SanF/SanR) were developed to distinguish the three species. Furthermore, phylogenetic analysis of the trnL-F intergenic sequences showed that R. crataegifolius is widely distributed in the Korean Rubi Fructus market as Korean product and imported goods from China. Single nucleotide polymorphism (SNP) that can be used to discriminate Korean Bokbunja medicine from those imported from China was developed, established the SNP marker, and conducted real-time polymerase chain reaction with a designed inflorescence probe. The developed SCAR marker and SNP probe were useful for distinguishing and authenticating the Korean and Chinese species that constitute Rubi Fructus medicines.
Cho, Choa-Hyoung,Jeon, Tae-Woog,Kim, Myung-Kon The Korean Society for Applied Biological Chemisty 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.1
Effects of gamma irradiation on color characteristic and biological activities of ethanol extract of Lonicera japonica irradiated at 0, 10, 20, and 30 kGy were investigated. Hunter color $L^*$- and $a^*$-value increased by irradiation in a dose-dependent manner, resulting in lighter color than the nonirradiated, whereas Hunter color $b^*$-value decreased with increasing irradiation dosage (p <0.05). The L. japonica extract showed inhibition effects against tyrosinase, xanthine oxidase, and nitrite-scavenging ability. Tyrosinase inhibition effect of L. japonica was higher in the gamma-irradiated sample than in the nonirradiated sample, and the effect was increased by increasing irradiation dosages. The L. japonica extracts had a higher inhibitory effect against xanthine oxidase, and the effect was not greatly changed by irradiation. Nitrite-scavenging activity was the highest in L. japonica extract at pH 1.2. Overall, the gamma irradiation may not greatly influence the xanthine oxidase inhibition effect and nitrite-scavenging ability of the L. japonica extract, except for the tyrosinase inhibition effect when irradiated up to 30 kGy.
Jun, Hee-Jin,Wen, Qingcheng,Lee, Ji-Hae,Jeun, Jung-Ae,Lee, Hwa-Jung,Lee, Kwang-Geun,Oh, Sea-Kwan,Lee, Sung-Joon The Korean Society for Applied Biological Chemisty 2012 Journal of Applied Biological Chemistry (J. Appl. Vol.55 No.2
Toxicity, antioxidant activity, and biological functionality of Korean black raspberry wine (KBRW) extracts on human ARPE-19 and HepG2 cells were determined using MTT and lipid/malonaldehyde (MA) assays and reverse transcription polymerase chain reaction (RT-PCR) as well as anti-obesity effect of KBRW extracts on various cancer cells. All samples inhibited MA formation by 38.0-88.0%. Bokbunja and Sanmaesu exerted greater inhibitory effect than other wines. Both cell lines were each treated with KBRW for 24 h, and viability was measured by MTT assay. No toxicity was found, even at 500 ${\mu}g/mL$. Changes in gene expression for CYP7A1, low-density-lipoprotein (LDL) receptor, 3-hydrozy-3-methylglutaryl-CoA (HMG-CoA) reductase, and vescular endothelial growth factor (VEGF) were assessed by semi-quantitative RT-PCR. CYP7A1 and LDL receptor expressions in ARPE-19 cells were elevated, whereas HMG CoA reductase expression was decreased by Bokbunja and Sanmaesu. Expression of VEGF was decreased by Bokbunja but elevated by Sanmaesu. In HepG2 cells, gene expression was similar after Bokbunja and Sanmaesu treatments. CYP7A1 and HMG CoA expressions were elevated. LDL receptor expression was increased by Bokbunja but decreased by Sanmaesu. Bokbunja and Sanmaesu inhibited VEGF expression but elevated those of CYP7A1 and HMG CoA reductase. Bokbunja and Sanmaesu displayed anti-cholesterol effects that could be attributed to increased gene expression in CYP7A1 and HMG CoA reductase.
Lee, Hye-Ri,Kim, Eun-Hye,Moon, Joon-Kwan,Zhu, Yong-Zhe,Do, Jung-Ah,Oh, Jae-Ho,Kwon, Ki-Sung,Lee, Young-Deuk,Kim, Jeong-Han The Korean Society for Applied Biological Chemisty 2012 Journal of Applied Biological Chemistry (J. Appl. Vol.55 No.2
A precise single residue analytical method was developed for fungicide cyazofamid in various crops. Apple, mandarin, Korean cabbage, green pepper, potato, and soybean were selected as representative crops, and clean-up system, partition solvent and extraction solvent were optimized. Limit of quantitation (LOQ) of cyazofamid was 2 ng (S/N>10) and good reproducibility was observed with small coefficient of variation (<4%). Excellent linearity was achieved between 0.05 and 20 mg/kg of cyazofamid standard solutions, with coefficients of determination of 1.000. Method limit of quantitation (MLOQ) was 0.02 mg/kg. For recoveries tests, crop samples were macerated and fortified with cyazofamid standard solution at three fortification levels (MLOQ, 10 MLOQ, and 100 MLOQ). And then those were extracted with acetone, concentrated and partitioned with dichloromethane. Then the extracts were concentrated again and cleaned-up through $Florisil^{(R)}$ column with ethyl acetate : n-hexane (30:70, v/v) before concentration and analysis with HPLC. Good recoveries from 75.3 to 98.5% with coefficients of variation of less than 10% were obtained, regardless of sample type, which satisfies the criteria of KFDA. Those results were reconfirmed with LC-MS/MS. The method established in this study could be applied to most of crops as an official and general method for residue analysis of cyazofamid.
Kim, Young-Seon,Ryuk, Jin-Ah,Ko, Byoung-Seob The Korean Society for Applied Biological Chemisty 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.1
Rehmanniae Radix, from the roots of Rehmannia glutinosa Libosch has been used in traditional herb medicine for the treatment of fever and strengthening liver function, among others. Information on the phylogenetic relationship is very limited in the region of its cultivation. It is very important to know the information of the close relatives of R. glutinosa Libosch and R. glutinosa Libosch. f. hueichingensis Hsiao, R. glutinosa produced in Wen County, Meng County, Bo'ai County, Qinyang County in Henan province, China. In this study, we examined the polymorphism analysis of Rehmanniae Radix originated from both Korea and China to compare the difference at the genomic DNA level. Results revealed that ITS and rps16 region sequences of R. glutinosa in Korea and R. glutinosa in China were correspond, while randomly amplified polymorphic DNA analysis showed a difference in UBC 301 primer. The specific primer designed was amplified at 334 bp for R. glutinosa originated from China. This primer (HRgF and HRgR) would be used efficiently to distinguish R. glutinosa from different sources.
Purification and Characterization of a 34-kDa Chitobiosidase from Aeromonas sp. GJ-18
Jeong, Hyun-Cheol,Ju, Wan-Taek,Jo, Kyung-Hyun,Park, Ro-Dong The Korean Society for Applied Biological Chemisty 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.1
Chitobiosidase was purified and characterized from Aeromonas sp. GJ-18 by ammonium sulfate precipitation, anion-exchange chromatography, and gel filtration chromatography. The purified enzyme has a molecular weight of 34 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme showed an optimum pH and temperature of 6.0 and $30-50^{\circ}C$, respectively. The enzyme was stable at pH 5-8 and $50^{\circ}C$ and was completely inhibited in the presence of 10 mM $Zn^{2+}$ ions. The enzyme could efficiently hydrolyze colloidal chitin into N,N'-diacetylchitobiose as the major product, indicating that the purified enzyme is a chitobiosidase. When colloidal chitin was used as the substrate, the $K_m$ and $V_{max}$ of this enzyme were established as 3.45 mg/mL and 2.91 ${\mu}mol/min$, respectively.
Yang, Chang-Hee,Song, Byung-Cheol,Cho, Moon-Jae The Korean Society for Applied Biological Chemisty 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.2
Hepatocellular carcinoma (HCC), one of the most prevalent cancers, and HCC-associated diseases are caused by the hepatitis B virus (HBV). Viral sequences often contain mutations in the basal core promoter (BCP), which overlaps with the open reading frame encoding HBV X protein (HBx). HBx protein plays a very important role in the development of HCC. Influence of naturally-occurring mutation of HBx (BCP1) on cell cycle progression was investigated in the Huh7 HCC line. BCP1 mutation was cloned from clinical samples into recombinant green fluorescent protein-fusion expression plasmid constructs that were then transfected into Huh7 cells for either transient expression studies or isolation of stable transfectants, along with wild-type HBx and empty vector controls. The mutant protein was more effective than wild-type in reducing expression of cyclin-dependent kinase (CDK) inhibitor p27kip1, promoting cell cycle progression, inhibiting production of cleaved poly ADP ribose polymerase (PARP) (a marker for apoptosis), and down-regulating the multifunctional cytokine transforming growth factor-${\beta}$ (TGF-${\beta}$). Since TGF-controls the cell cycle and apoptosis, the effect of BCP1 mutation on the enhanced activity of HBx to promote cell cycle progression and to inhibit apoptosis in Huh7 cells may result from down-regulation of TGF-${\beta}$.
Kim, Bong-Gyu,Lee, Eung-Ryoung,Ahn, Joong-Hoon The Korean Society for Applied Biological Chemisty 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.1
Gene encoding the key flavonoid biosynthetic enzyme, chalcone synthase, in Populus euramericana Guinier (PeCHS) was cloned and characterized. PeCHS preferentially uses feruloyl-CoA as a substrate. Expression of the flavonoid biosynthetic genes, phenylalanine ammonimum lyase from P. euramericana (PePAL), PeCHS, chalcone isomerase from P. euramericana (PeCHI), and flavonol synthase from P. euramericana (PeFLS) from P. euramericana in response to abiotic stresses such as wounding and UV-irradiation were analyzed. These genes were induced under the stress conditions. Most genes were expressed at an early stage of the stress response. In addition, total flavonoid content increased after UV-irradiation.
Shin, In-Sik,Lee, Mee-Young,Lim, Hye-Sun,Seo, Chang-Seob,Ha, Hye-Kyung,Shin, Hyeun-Kyoo The Korean Society for Applied Biological Chemisty 2012 Applied Biological Chemistry (Appl Biol Chem) Vol.55 No.2
Benign prostate hyperplasia (BPH) is the most common proliferative disorder affecting older men and results in prostate enlargement and lowered urinary tract symptoms. Jaeumganghwa-tang (JGT), an oriental traditional herbal formula, has been used in China (Zi-yin-jiang-huo-tang in Chinese), Japan (Jiin-koka-to in Japanese), and Korea for many years. Effects of JGT on prostate dihydrotestosterone (DHT) level and prostatic hyperplasia were investigated using a rat model, in which BPH was induced using testosterone propionate (TP). Rats were divided into five groups. One group was used as a normal, and four groups received subcutaneous injections of TP for 4 weeks to induce BPH. JGT (200 or 400 mg/kg) was administered daily for 4 weeks by oral gavage concurrently with TP injections, and rats were sacrificed at scheduled times. Prostates were weighed, and histopathologic examination was conducted. DHT levels in serum and the prostate were measured, and the expression of proliferating cell nuclear antigen (PCNA) protein was investigated using Western blotting. BPH animals showed increases in absolute and relative weights of the prostate, levels of DHT in serum and the prostate, and expression of PCNA in the prostate, whereas JGT-treated animals showed significant reductions in these indices compared with the BPH animals. Administration of JGT attenuated TP-induced epithelial hyperplasia. These findings indicate that JGT inhibits the development of BPH, an effect closely associated with a reduction in DHT level.