• Korean Journal of Acupuncture
• /
• v.41 no.1
• /
• pp.7-15
• /
• 2024

Objectives : This study aimed to predict the effectiveness and potential of Schizonepeta tenuifolia as an anticancer treatment for non-small cell lung cancer through network-based pharmacology and cellular experiment. Methods : To identify the major bioactive compounds in Schizonepeta tenuifolia, we used the Traditional Chinese Medicine Systems. The target genes for the cancer treatment were selected using the UniProt database and the networked using Cytoscape. We performed functional enrichment analysis based on the Gene Ontology Biological Process and Kyoto Encyclopedia of Genes and Genomes Pathways to predict the mechanisms. To investigate the effect of Schizonepeta tenuifolia on lung cancer cell growth, we treated A549 cells, a lung cancer cell line, with different concentrations of the drug and used the MTT assay for cell viability. Results : Research has shown that the most effective mechanism of active compounds from Schizonepeta tenuifolia is through the pathway of cancer. The results of the network pharmacology analysis indicate that Schizonepeta tenuifolia has potential medicinal value as an adjuvant in anticancer treatment. The concentration-dependent inhibition of cell viability was observed on A549 cells. Furthermore, synergistic anticancer activity with Doxorubicin was also observed. Conclusions : Through a network pharmacological approach, Schizonepeta tenuifolia was predicted to have potential as an anticancer agent, and its efficacy was experimentally demonstrated using A549 cells. These findings suggest that Schizonepeta tenuifolia is a promising candidate for future research.

• Journal of Animal Science and Technology
• /
• v.54 no.1
• /
• pp.1-8
• /
• 2012

This study aimed to investigate the single nucleotide polymorphisms (SNPs) of the porcine MC4R gene and validate the effect of the MC4R genotype for marker assisted selection (MAS). Six amplicons were produced to analyze the entire base sequences of the porcine MC4R gene and six SNPs were detected (c.-780C>G, c.-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, c.892A>G-Asp298Asn, and c.*430A>T). Linkage disequilibrium (LD) of the six SNPs was analyzed by performing haploid analysis. There was a perfect linkage disequilibrium in c.-780C>G, c.-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, and c.*430A>T. Only the c.892A>G (Asp298Asn) SNP showed a very low LD with an $r^2$ value of 0.028 and the D' value of 0.348. As a result, the two SNPs-c.707A>G (Arg236His) and c.892A>G (Asp298Asn)-were selected to extract the genotype frequencies from the 5 pig breeds by using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) genotype analysis method. The SNP frequency of c.707A>G (Arg236His) indicated the presence of the A (His) allele only in Yorkshire, while the G allele was fixed in the KNP, Landrace, Berkshire, and Duroc. Association analysis was carried out in 484 pigs with the c.707A>G (Arg236His) SNP and the meat quality traits of four different pig cross populations: a significant association was noted in crude fat, sirloin moisture, meat color, and the degree of red and yellow coloration. The frequency of the c.892A>G(Asp298Asn) SNP genotype varied among the breeds; while Duroc showed the highest frequency of the A (Asn) allele, KNP showed the highest frequency of the G (Asp) allele. Association analysis was carried out in 1126 pigs with the c.892A>G (Asp298Asn) SNP and the meat quality traits of four pig populations: a highly significant linkage was noted in the back-fat thickness (P<0.002). It was found that the back-fat thickness was higher in individuals with the AA genotype than in those with the AG or GG genotype. Thus, in this study, we verified that the c.892A>G (Asp298Asn) SNP in the pig MC4R gene has a sufficient effect as a gene marker for MAS in Korean pork industry.

• Tuberculosis and Respiratory Diseases
• /
• v.58 no.4
• /
• pp.367-374
• /
• 2005

Background : The fact that only 10-20% of chronic cigarette smokers develop chronic obstructive pulmonary disease (COPD) reflects the presence of genetic factors associated with the susceptibility to COPD. Recently, it was reported that the surfactant protein A increases the secretion of matrix metalloprotease 9, which degrades extracellular matrices of the lung, through a Toll-like receptor 2 (TLR2). In this context, possible role of TLR2 in the pathogenesis of COPD was postulated, and a functional dinucleotide repeat polymorphism in intron II of TLR2 was evaluated for any association with COPD. Method : Male patients with COPD and male smokers with a normal pulmonary function were enrolled in this study. The number of Guanine-Thymine repeats in intron II of the TLR2 gene were counted. Because the distributions of the repeats were trimodal, the alleles were classified into three subclasses, 12-16 repeats: short (S) alleles; 17-22 repeats: medium length (M) alleles; and 23-27 repeats: long (L) alleles. Result : 125 male patients with COPD and 144 age- and gender-matched blood donors with a normal lung function were enrolled. There were no differences in the distribution of each allele subclass (S, M and L) between the COPD and control group (p=0.75). The frequencies of the genotypes with and without each allele subclass in the COPD and control group were similar. Conclusion : A microsatellite polymorphism in intron II of TLR2 gene was not associated with the development of COPD in Koreans.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.8
• /
• pp.1271-1283
• /
• 2007

A fibrinolytic protease (PoFE) was purified from the cultured mycelia of the edible oyster mushroom Pleurotus ostreatus, using a combination of various chromatographies. The purification protocol resulted in an 876-fold purification of the enzyme, with a final yield of 6.5%. The apparent molecular mass of the purified enzyme was estimated to be 32 kDa by SDS-PAGE, fibrin-zymography, and size exclusion using FPLC. The optimal reaction pH value and temperature were pH 6.5 and $35^{\circ}C$, respectively. PoFE effectively hydrolyzed fibrinogen, preferentially digesting the $A{\alpha}$-chain and the $B{\beta}$-chain over the ${\gamma}$-chain. Enzyme activity was enhanced by the addition of $Ca^{2+},\;Zn^{2+},\;and\;Mg^{2+}$ ions. Furthermore, PoFE activity was potently inhibited by EDTA, and it was found to exhibit a higher specificity for the chromogenic substrate S-2586 for chymotrypsin, indicating that the enzyme is a chymotrypsin-like metalloprotease. The first 19 amino acid residues of the N-terminal sequence were ALRKGGAAALNIYSVGFTS, which is extremely similar to the metalloprotease purified from the fruiting body of P. ostreatus. In addition, we cloned the PoFE protein, encoding gene, and its nucleotide sequence was determined. The cDNA of cloned PoFE is 867 nucleotides long and consists of an open reading frame encoding 288 amino acid residues. Its cDNA showed a high degree of homology with PoMEP from P. ostreatus fruiting body. The mycelia of P. ostreatus may thus represent a potential source of new therapeutic agents to treat thrombosis.

• Journal of Life Science
• /
• v.26 no.4
• /
• pp.406-413
• /
• 2016

Abalones are gastropod mollusks belonging to the genus Haliotis. Pacific abalones are regarded as a very important marine gastropod mollusk in Korea, Japan, China, and also in food industries around the world. In Korea, 6 species of abalone have been reported to occur along the coasts: Haliotis discus hannai, Haliotis discus discus, Haliotis madaka, Haliotis gigantea, Haliotis diversicolor supertexta, and Haliotis diversicolor diversicolor. This study was performed to discriminate the genetic variances by the partial sequences of the mitochondrial cytochrome c oxidase subunit I (COI) genes and random amplified polymorphic DNA (RAPD) analysis against four species of Pacific abalone (H. discus hannai, H. discus, H. madaka, H. gigantea). COI gene is reasonably well conserved and has been sequenced in various invertebrate taxa. The RAPD analysis technique is a relatively simple and low cost method that allows differentiation of taxa without the need to know their genomes. In this study, we investigated the genetic diversity, phylogenetic relationships within each species. The COI and RAPD analysis were able to distinguish between H. gigantea and the other three species. However, these analysis methods were inadequate to distinguish between H. discus and H. madaka. These results are believed to be able to provide a basis data for future hybrid breeding research by defining the genetically closely related four species of abalone, which is to develop new hybrid abalone for export using hybrid breeding.

• The Korean Journal of Mycology
• /
• v.43 no.3
• /
• pp.200-205
• /
• 2015

Perilla frutescens var. japonica Hara was collected from farmland in Seongju-gun. Fifteen endophytic fungal strains with different colony morphologies were isolated from roots of P. frutescens. Waito-c rice seedlings were treated with the concentrated culture filtrates (CF) of endophytic fungi for observation of their plant growth-promoting activities. In the results, the CF of Y2H001 fungal strain promoted the growth of the waito-c rice seedlings. The phylogenetic tree of Y2H001 strain was analyzed by the combined sequences of the partial internal transcribed spacer region (ITS) and partial betatubulin gene. Molecular and morphological studies identified the Y2H001 strain as belonging to Aspergillus flavus. In gas chromatography mass spectrometry (GC/MS) analysis of the CF of Y2H001 strain, gibberellic acid (GA) was detected and quantified. Therefore, we describe Y2H001 strain as a new $GA_3$-producing A. flavus based on morphological, molecular characteristics and analysis of secondary metabolite.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.65 no.4
• /
• pp.274-283
• /
• 2020

Seed dormancy is an adaptive trait in which seeds do not germinate under unfavorable environmental conditions. Low dormancy seeds are easily germinated under optimal environmental conditions, and these characteristics greatly reduce the yield and quality of crops. In the present study, we compared the pre-harvest sprouting (PHS) rate of two cultivars, Joun and Jopyeong, using the Winkler scale after heading day and temperature of the test. The PHS rate increased as the Winkler scale after heading day increased from 700℃ to 1100℃ and the temperature of the test increased. In all conditions, the PHS rate of Jopyeong was higher than that of Joun. RNA-sequencing was used to analyze the cause of the high PHS rate. We analyzed the biological metabolic processes related to the abscisic acid (ABA) metabolite pathway using the KEGG mapper with selected differentially expressed genes in PHS seeds. We found that the expression of ABA biosynthesis genes (OsNCEDs) was down-regulated and that ABA catabolic genes (OsCYP707As) was up-regulated in PHS seeds. However, the quantitative real-time PCR results showed that Joun had a higher expression of OsNCEDs than that of Jopyeong, but OsCYP707As did not yield a significant result. Joun displayed higher ABA content than that of Jopyeong not only during ripeness time but also during PHS treatment. Taken together, we provided evidence that the ABA content remaining in the seed is important to the PHS rate, which is determined by the expression level of the ABA biosynthesis gene OsNCEDs.

• BMB Reports
• /
• v.37 no.3
• /
• pp.376-382
• /
• 2004

The Epstein-Barr-transformed B lymphoblastoid cell lines, LCL, which express antigens, are potential antigen-presenting cells (APCs) for the induction of cytotoxic T lymphocytes in vitro. However, transfecting LCL with subsequent selection by antibiotics is notoriously difficult because the plating efficiencies of LCL are reported to be 1% or less. Therefore, this study investigated the optimal conditions for increasing the transduction efficiency of a recombinant adenovirus to LCL for use as a source of APCs. The transduction efficiencies were < 13% (SD $\pm$ 2.13) at a multiplicity of infection (MOI) of 100, while it was increased to 28% (SD $\pm$ 9.43) at an MOI of 1000. Moreover, its efficiencies to LCL that expressed the coxsackie adenovirus receptor were increased to 60% (SD $\pm$ 6.35) at an MOI of 1000, and were further increased to 70% (SD $\pm$ 4.56) when combined with the centrifugal method. The cationic liposome or anionic polymer had no effect on the transduction efficiency when compared to that of the centrifugal method. These results may be used as a convenient source of target cells for a CTL assay and/or autologous APCs for the induction of the in vitro CTL responses that are specific to viral and tumor antigens.

• Asian-Australasian Journal of Animal Sciences
• /
• v.33 no.3
• /
• pp.424-435
• /
• 2020

Objective: The study was conducted to investigate variations in the immunophysiological responses to exercise-induced stress in Jeju and Thoroughbred horses. Methods: Blood samples were collected from the jugular veins of adult Jeju (n = 5) and Thoroughbred (n = 5) horses before and after 30 min of exercise. The hematological, biochemical, and immunological profiles of the blood samples were analyzed. Blood smears were stained and observed under a microscope. The concentration of cell-free (cf) DNA in the plasma was determined using real time polymerase chain reaction (PCR). Peripheral blood mononuclear cells (PBMCs) and polymorphonuclear cells were separated using Polymorphprep, and the expression of various stress-related and chemokine receptor genes was measured using reverse transcriptase (RT) and real-time PCR. Results: After exercise, Jeju and Thoroughbred horses displayed stress responses with significantly increased rectal temperatures, cortisol levels, and muscle catabolism-associated metabolites. Red blood cell indices were significantly higher in Thoroughbred horses than in Jeju horses after exercise. In addition, exercise-induced stress triggered the formation of neutrophil extracellular traps (NETs) and reduced platelet counts in Jeju horses but not in Thoroughbred horses. Heat shock protein 72 and heat shock protein family A (Hsp70) member 6 expression is rapidly modulated in response to exercise-induced stress in the PBMCs of Jeju horses. The expression of CXC chemokine receptor 4 in PBMCs was higher in Thoroughbred horses than in Jeju horses after exercise. Conclusion: In summary, the different immunophysiological responses of Jeju and Thoroughbred horses explain the differences in the physiological and anatomical properties of the two breeds. The physiology of Thoroughbred horses makes them suitable for racing as they are less sensitive to exercise-induced stress compared to that of Jeju horses. This study provides a basis for investigating the link between exercise-induced stresses and the physiological alteration of horses. Hence, our findings show that some of assessed parameters could be used to determine the endurance performance of horses.

• Journal of Microbiology and Biotechnology
• /
• v.28 no.4
• /
• pp.606-612
• /
• 2018

The enzyme xylose isomerase (E.C. 5.3.1.5, XI) is responsible for the conversion of an aldose to ketose, especially xylose to xylulose. Owing to the ability of XI to isomerize glucose to fructose, this enzyme is used in the food industry to prepare high-fructose corn syrup. Therefore, we studied the characteristics of XI from Anoxybacillus kamchatkensis G10, a thermophilic bacterium. First, the gene coding for XI (xylA) was inserted into the pET-21a(+) expression vector and the construct was transformed into the Escherichia coli competent cell BL21 (DE3). The expression of recombinant XI was induced in the absence of isopropyl-thio-${\beta}$-galactopyranoside and purified using Ni-NTA affinity chromatography. The optimum temperature of recombinant XI was $80^{\circ}C$ and measurement of the heat stability indicated that 55% of residual activity was maintained after 2 h incubation at $60^{\circ}C$. The optimum pH was found to be 7.5 in sodium phosphate buffer. Magnesium, manganese, and cobalt ions were found to increase the enzyme activity; manganese was the most effective. Additionally, recombinant XI was resistant to the presence of $Ca^{2+}$ and $Zn^{2+}$ ions. The kinetic properties, $K_m$ and $V_{max}$, were calculated as 81.44 mM and $2.237{\mu}mol/min/mg$, respectively. Through redundancy analysis, XI of A. kamchatkensis G10 was classified into a family containing type II XIs produced by the genera Geobacillus, Bacillus, and Thermotoga. These results suggested that the thermostable nature of XI of A. kamchatkensis G10 may be advantageous in industrial applications and food processing.