• The Journal of Korean Medicine
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• v.40 no.2
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• pp.35-50
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• 2019

Objectives: Orostachys japonicas (O. japonicus) has been known for its anti-tumor effect. In the present study, it was investigated whether O. japonicus EtOH extracts could induce apoptosis and autophagy which are part of the main mechanism related to anti-tumor effect in THP-1 cells. Methods: Cells were treated with various concentrations of O. japonicus EtOH extracts ($0-300{\mu}g/ml$) for 24, 48, and 72h. Cell viability was evaluated by MTS/PMS assay and apoptosis rate was examined by flow cytometry and ELISA assay. The mRNA expression of apoptosis-related genes (Bcl-2, Mcl-1, Survivin, Bax) and autophagy-related gene (mTOR) was evaluated using real-time PCR. The protein expression of Caspase-3, Akt, LC3 II, Beclin-1, Atg5, $NF-{\kappa}B$, p38, ERK was evaluated using western blot analysis. Results: O. japonicus EtOH extracts inhibited cell proliferation and apoptosis rate was increased in both flow cytometry and ELISA assay. Bcl-2, Mcl-1, Survivin (anti-apoptosis factors) mRNA expressions were decreased and Bax (pro-apoptosis factor) mRNA level was increased. mTOR mRNA expressions was decreased and LC3 II protein expressions was increased. Activation of $NF-{\kappa}B$ was decreased and phosphorylation of p38 was increased. Conclusion: O. japonicus is regarded to inhibit cell proliferation, to induce apoptosis and to regulate autophagy-related genes in THP-1 cells via $NF-{\kappa}B$ and p38 MAPK signaling pathway. This suggests O. japonicus could be an effective herb in treating acute myeloid leukemia.

• Journal of Plant Biotechnology
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• v.6 no.1
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• pp.25-37
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• 2004

Successful genetic transformation of plants requires non-chimeric selection of transformed tissues and their subsequent regeneration. With rare exceptions, most transformation protocols still rely heavily on antibiotics for selecting transgenic cells that contain an antibiotic-degrading selectable marker gene. Here, the morphogenic capacity of in-vitro explants of chrysanthemnum and tobacco stems and leaves (control and transgenic) changed with the addition of aminoglycoside antibiotics (AAs), In a test of 6 AAs, phytotoxicity occurred at concentrations of 10 to 25 and 50 to 100$\mu\textrm{g}$ $mL^{-1}$ in chrysanthemum and tobacco explants, respectively. Light conditions as well as explant source and size also had significant effects. The use of transverse thin cell layers (tTCLs), in conjunction with high initial AA selection levels, supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow-cytometric analyses revealed no endodu-plication in chrysanthemum, even at high AA levels. However, this phenomenon was observed in tobacco calli(8C or more), even at low AA concentrations (i.e., 5 to 10 $\mu\textrm{g}$ mL$^{-1}$ ).

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.2
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• pp.178-183
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• 2007

China has numerous native domestic goat breeds, but so far there has been no extensive study on genetic diversity, population demographic history, and origin of Chinese goats. To determine the origin and genetic diversity of Chinese goats, we analyzed the complete mtDNA D-loop sequences of 183 goats from 13 breeds. The haplotype diversity value found in each breed ranged from 0.9333 to 1.0000. The nucleotide diversity value ranged from 0.006337 to 0.025194. Our results showed that there were four mtDNA lineages (A, B, C and D), in which lineage A was predominant, lineage B was moderate, and lineages C and D were at low frequencies. Lineages C and D were observed only in the Tibetan breed. The results revealed multiple maternal origins of Chinese domestic goats. There was weaker geographical structuring in the 13 Chinese goat populations, which suggested that there existed high gene flow among goat populations caused by the extensive transportation of goats in the course of history.

• Animal cells and systems
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• v.14 no.1
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• pp.31-36
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• 2010

The pinewood nematode Bursaphelenchus xylophilus causes pine wilt disease and is a serious economic concern for the forest industry of South Korea. To achieve effective control with limited resources, it is necessary to clarify the transmission routes and mechanisms of dispersal of this organism. Highly polymorphic and easy-to-use molecular markers can be used for investigating this aspect. In this study, we evaluated the usefulness of amplified fragment length polymorphisms (AFLPs) for investigating the genetic variations of B. xylophilus and related individuals from China, Japan, and South Korea. The AFLP patterns obtained in our study were similar to the microsatellite patterns reported in a previous study; our AFLP patterns indicated high genetic variability and cryptic genetic structure, but did not indicate any peculiar geographic structure. Moreover, the genetic distances between individuals suggested that the Korean population was affected to a greater extent by the Chinese population than the Japanese population. Further, the gene flow among the related species appeared to be limited; however, there may be also the possibility of genetic introgression among species. These results confirm the usefulness of AFLPs for understanding the epidemiology of pine wilt disease, thereby contributing to the effective control of this disease.

• BMB Reports
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• v.46 no.5
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• pp.270-275
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• 2013

It is essential to analyze rare mutations in many fields of biomedical research. However, the detection of rare mutations is usually failed due to the interference of predominant wild-type DNA surrounded. Herein we describe a sensitive and facile method of detecting rare point mutation on the basis of allele-specific amplification in emulsion PCR. The identification and selective amplification of rare mutation are accomplished in one-pot reaction. The allele-specific primers coupled on magnetic beads allow the exclusive amplification and enrichment of the mutant amplicons. The productive beads bearing mutant amplicons are subsequently stained with the fluorescent dyes. Thus, the rare point mutations with a percentage as low as 0.1%, can be detected by fluorescent analysis. The relative percentages of mutation among different samples can be roughly accessed by counting the fraction of fluorescent positive beads through flow cytometry.

• IMMUNE NETWORK
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• v.1 no.1
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• pp.70-76
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• 2001

Background: Rheumatoid arthritis is an autoimmune disease characterized by a chronic inflammatory process, primarily involving the synovial membrane of peripheral j oints, where T cell activation is found. To address the superantigen stimulation in rheumatoid arthritis, T cell clonality and the expression of activation markers were analyzed. Methods: To detect TCRB V usage, inverse PCR and sequencing were done. Monoclonal antibodies were used for flow cytometric analysis of TCRBV8 or TCRBV5. As results, a restricted usage of TCRBV3 gene was detected in synovial lymphocytes from one rheumatoid arthritis patient. However, preferential usage for TCRB V8, which may be one indicator for stimulation by staphylococcal superantigen, was not obvious although general activation of T cells was found as high DR+ percentage in synovial T cells. These data show specific antigen rather than superantigen might involve the pathogenesis of rheumatoid arthritis.

• Bulletin of the Korean Chemical Society
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• v.29 no.6
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• pp.1179-1184
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• 2008

Commercial nanometer sized silver is widely used for its antibacterial effect; however, nanoparticles may also have ecotoxicological effects after being discharged into water. Nanometer sized silver can flow into aquatic environments, where it can exert a variety of physiologically effects in living organisms, including fish. The present study aimed to investigate the effect of nanometer sized silver on the development of zebrafish embryos, analyze the properties of commercial nanometer sized silver and define the toxicity relationship between embryogenesis and hatched flies. The commercial nanometer sized silver was analyzed in the $Ag^+$ ion form. The hatch rate decreased in the nano-silver exposed groups (10 and 20 ppt); furthermore, the hatched flies had an abnormal notochord, weak heart beat, damaged eyes and curved tail. The expression of the Sel N1 gene decreased in the nano-silver exposed groups, and the catalase activities of the exposed groups increased relative to those in the control group. Therefore, the $Ag^+$ ions in commercial nanometer sized silver could accumulate in aquatic environments and seriously damage the development of zebrafish embryos.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.5
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• pp.2087-2092
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• 2015

Nowadays herbal-derived medicines are attracting attention as new sources of drugs with few side effects. Silibinin is a flavonoid compound with chemotheraputic effects on different cancers such as examples in the prostate, lung, colon and breast. In the present study, the cytotoxic effects of silibinin on MCF7 breast cancer cells were investigated. Apoptosis was determined by flow cytometry and the impact of silibinin on the expression of pivotal genes including Bak, P53, P21, BRCA1, BCL-X1 and ATM was analyzed. Treatment for 24h had a significant dose-dependent inhibitory effect on cell growth (p<0.05) with dose- and time- dependent induction of apoptosis (p<0.05). In addition, there were significant increases in BRCA1, ATM, Bak and Bcl-XL gene expression at the mRNA level with different concentrations of silibinin for 24 or 48 h (p<0.05). Taken together, the results suggest that silibinin inhibits the proliferation and induces apoptosis of MCF-7 cells by down-regulating Bak, P53, P21, BRCA1, BCL-Xl and thus may be considered as an effective adjuvant drug to produce a better chemopreventive response for the cancer therapy.

• Journal of Life Science
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• v.18 no.2
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• pp.175-179
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• 2008

Cornus controversa is a long-lived woody species mostly distributed in East Asia. Random amplified polymorphic DNA (RAPD) markers were used to investigate the genetic diversity and population structure of Korean populations of this species. A high level of genetic variation was found in seven populations of C. controversa. The mean genetic diversity (H) was 0.222 across populations, varying from 0.200 to 0.238. Eighty of the 93 loci (86.0%) showed detectable polymorphism in at least one population. Total genetic diversity values ($H_T$) varied between 0.192 and 0.231, giving an average overall polymorphic loci of 0.212. The interlocus variation of genetic diversity within populations ($H_S$) was high (0.167). Mean of genetic diversity in C. controversa was higher than average values for species with similar life history traits. The sexual reproduction, perennial habitat, and longevity are proposed as possible factors contributing to high genetic diversity. On a per locus basis, the proportion of total genetic variation due to differences among populations ($G_{ST}$) ranged from 0.169 to 0.278 with a mean of 0.216, indicating that about 21.6% of the total genetic variation was among populations. An indirect estimate of the number of migrants per generation (Nm=1.893) indicated that gene flow was extensive among Korean populations of C. controversa.

• Journal of Life Science
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• v.14 no.6 s.67
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• pp.963-966
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• 2004

The sequence of ITS (partial 16S ribosomal DNA, complete ITS1, 5.8S ribosomal DNA and ITS2, and partial 28S ribosomal DNA) was analysed by PCR and autosequencing in Sarcodon aspratus. The ITS lenght of S. aspratus was 716 base pair. As this sequence compared with other reports on S. aspratus (ace No AF335110), the sequence variation based on nucleotide deletion and substitution was $1.8\%$. This nucleotide variation rate in same species was very higher than in other species. Also, the sequence varitation rates between this S. aspratus and S. imbricatus, and S. squamus were $8\%\;and\;10\%$, respectively. This results suggested that the high sequence variation of S. aspratus might be caused specific host and inhabitat environment which limited gene flow.