• Journal of Animal Science and Technology
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• v.49 no.5
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• pp.549-558
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• 2007

C4B and BAT2, assigned to the SLA class III region, were recently reported on relation with human diseases. The primers for RT-PCR and RACE-PCR for CDS analysis of these genes of pig were designed by aligning the CDSs of humans and mice from GenBank. After we amplified and sequenced with these primers and cDNAs, the full-length CDSs of pig were determined. The CDS lengths of C4B and BAT2 were shown as 5226 bp and 6501 bp. In addition, the identities of nucleotide sequences with human and mouse were 76% to 87%, and the identities of amino acids were 72% to 90%. After we carried out the alignment with determined CDSs in this study and pig genomic sequences from GenBank, the primers for cSNP detection in genome were designed in intron regions that flanked one or more exons. Then, we amplified and directly sequenced with genomic DNAs of six pig breeds. Four cSNPs from C4B and three 3 cSNPs from BAT2 were identified. In addition, amino acid substitution occurred in six cSNP positions except for C4248T of C4B. By the Multiplex-ARMS method, we genotyped seven cSNPs with DNA samples used for direct sequencing. We verified that this result was the same as that analyzed using direct sequencing. To demonstrate recrudescence, we performed both direct sequencing and Multiplex-ARMS on two randomly selected DNA samples. The genotype of each sample showed the same result from both methods. Therefore, seven cSNPs were identified from C4B and BAT2 and could be used as the basic data for haplotype analysis of SLA class III region. Moreover, the Multiplex-ARMS method should be powerful for genotyping of genes assigned to the whole SLA region for the xenograft study.

• Research in Plant Disease
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• v.15 no.1
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• pp.1-7
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• 2009

Viruses diagnosed on crops including rice plants from farmers or agricultural extension agencies cover the country were 11 species including Broad bean wilt virus 2 (BBWV2) in 2008. Tomato spotted wilt virus (TSWV) was the most important virus having the detection rate of 22.9%. Two viruses of Tomato yellow leaf curl virus (TYLCV) and Tobacco leaf curl virus (TLCV) inducing leaf yellow and curl diseases on tomatoes were occurred newly with the detection rate of 12.2% and 4.0%, respectively, in 2008. Rice stripe virus (RSV) was occurred on 869.5 ha mainly at Jindo and Haenam areas in Jeollanamdo province. At Jindo area, 12 plots were damaged severely with the infected hill rate of 83.8%. At the main production area of oriental melon at Seongju, almost all fruits from whole sale market at Seongju were infected with Cucumber green mottle mosaic virus (CGMMV) as the detection rate of 87%. The areas occurred TSWV in Korea were 25 totally from 2003 including 7 areas newly reported in 2008 including Naju in Jeoallanamdo. TSWV could be reduced as 0.1 % from 5.3% by covering insect proof net in vinyl house after chemical soil sterilization. Tomato yellow leaf curl disease was occurred on April in 2008 at Tongyoung area in Kyeongsangnamdo, and detected continuously at 13 areas, 7 in Kyeongsangnamdo, 4 in Jeollabukdo and 2 in Jejudo. Potato spindle tuber viroid (PSTVd) was occurred abruptly in a confined space of a civil breeding greenhouse and a cultivar evaluation field followed by disuse 17.4 M/T of potato tubers. No PSTVd was detected at 17 fields cultivated the related potatoes to the bred company by RT-PCR.

• Clinical and Experimental Pediatrics
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• v.45 no.2
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• pp.183-191
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• 2002

Purpose : X-linked agammaglobulinemia(XLA) is an immunodeficiency caused by abnormalities in Bruton's tyrosine kinase(Btk), and is characterized by a deficiency of peripheral blood B cells. We studied the cytoplasmic expression of Btk protein and analyzed the Btk gene in peripheral blood mononuclear cells from two siblings and one cousin with XLA, as well as additional family members. Methods : Btk protein expression was analyzed by flow cytometry. Isolation of the coding sequence of the Btk gene was performed by amplification using the reverse transcription-polymerase chain reaction(RT-PCR) technique. Sequence alterations were screened by the single-stranded conformation polymorphism(SSCP) method and characterized by standard sequencing protocols. Results : Cytoplasmic expression of Btk protein in monocytes was not detected in three patients with XLA. In addition, Btk protein analysis clearly showed cellular mosaicism in monocytes from four obligate carriers, findings further supported by SSCP. A single base pair mutation(T to C) in Btk-exon three, which encodes the PH domain, was identified in four XLA patients. A diagnostic sequencing analysis was established to detect heterozygotic pattern in 4 carrier females. Furthermore, we found significant clinical heterogeneity in individuals with the same gene mutation. Conclusion : The implicating genetic alteration provided valuable clues to the pathogenesis of XLA in Korea and the flow cytometric analysis was suggested as a useful tool for rapid detection of XLA patients and carriers. The present study has identified a genetic mutation in the Btk coding region and demonstrated heterogeneity in clinical manifestations among patients with the same mutation. A flow cytometric analysis was found to be informative in establishing a deficiency of Btk protein in both patients and carriers and is recommended as a frontline procedure in the molecular diagnosis and work-up of XLA.

• BMB Reports
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• v.40 no.5
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• pp.611-616
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• 2007

$E^{rns}$ is an envelope glycoprotein of classical swine fever virus (CSFV) and has an unusual feature of RNase activity. In the present study, we demonstrate that $E^{rns}$ counteracts Newcastle disease virus (NDV)-mediated induction of IFN-$\beta$. For this purpose, $E^{rns}$ fused to the enhanced green fluorescent protein (EGFP) was transiently expressed in porcine kidney 15 (PK15) cells. In luciferase activity assay, $E^{rns}$-EGFP was found to prevent IFN-$\beta$ promoter-driven luciferase expression and block the induction of IFN-$\beta$ promoter mediated by NDV in a dose-dependent manner. Through IFN-specific semi-quantitative RT-PCR detection, obvious decrease of IFN-$\beta$ mRNA in NDV-infected PK15 cells was observed in the presence of $E^{rns}$-EGFP. In contrast, EGFP alone showed none of this block capacity. In addition, $E^{rns}$-EGFP mutations with RNase inactivation were also found to block NDV-mediated induction of IFN-$\beta$. These evidences establish a novel function for CSFV $E^{rns}$ glycoprotein in counteraction of the IFN-$\beta$ induction pathway.

• The Journal of Internal Korean Medicine
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• v.29 no.4
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• pp.1048-1060
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• 2008

Cirsii Japonici Herba(CJ) is a wild perennial herb found in many areas of Korea as well as China and Japan, which has been used to treat bleeding and inflammation. Silibinin is the main flavonoid extracted from milk thistle (Cirsii Japonici Herba). It exhibits potent antioxidant activity and anti-inflammatory effect. In this study, the effect of CJ and silibinin extract on lipopolysaccharide-induced inflammation was investigated using MTS assay, RT-PCR, western blot, and nitric oxide detection on mouse BV2 microglial cell lines. In the present results, CJ and silibinin extract suppressed nitric oxide production by inhibiting the lipopolysaccharide-stimulated enhancement of COX-2 and iNOS gene expression in BV2 cells. Moreover, CJ and silibinin also repressed some lipopolysaccharide-induced signaling molecules. Importantly, catalase-induced COX-2 and iNOS expression needed activations of $NF-{\kappa}B$, PI3K/Akt, and MAPK, which were important for the transcriptional up-regulation of COX-2 and iNOS. CJ and silibinin interaction on BV2 cells down-regulated $NF-{\kappa}B$-dependent proinflammatory cytokine (IL-2,IL-6) expression. They are involved in the regulation of inflammatory responses. These data shows that CJ and silibinin exerts anti-inflammatory and analgesic effects, probably by suppression of COX-2 and iNOS synthase expression in BV2 microglial cells.

• The Journal of Internal Korean Medicine
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• v.33 no.4
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• pp.587-598
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• 2012

Objectives : This study was performed to elucidate the pro-apoptotic effect of Artemisiae capillaris herba extracted with ethanol on biliary tract cancer cells. Materials and Methods : The biliary tract cancer cell line SNU-1196 was used in this study. Cells were treated with different concentrations of Artemisiae capillaris herba for 24, 48 and 72 hours. After the treatment, cell viability, apoptosis, caspase activities and the mRNA expressions of the Bcl-2, Bax, P53, and P21 were measured by using MTT assay, cell cycle analysis, apoptosis assay, and RT-PCR. The cell cycle analysis was done by flow cytometry and apoptosis assay by cell death detection ELISA kit. Results : Artemisiae capillaris herba inhibited proliferation of SNU-1196 in long-time culture group with dose-dependent manner. All cells treated with Artemisiae capillaris herba showed increased apoptosis with dose- and time-dependent manner. Exposure of SNU-1196 to Artemisiae capillaris herba induced caspase-3 activation. However, apeoptosis was blocked when SNU-1196 was treated together with the pan-caspase inhibitor Z-VAD-FMK and the caspase-3 inhibitor Z-DEVE-FMK. After the treatment of Artemisiae capillaris herba, the mRNA expressions of caspase -3, -8, -9, p53, and p21 was increased in all cells. Artemisiae capillaris herba resulted in a significant decrease in Bcl-2 and an increase in Bax mRNA levels. Conclusions : These results suggest that Artemisiae capillaris herba would be beneficial in the treatment of biliary tract cancer.

• Journal of Physiology & Pathology in Korean Medicine
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• v.29 no.2
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• pp.168-173
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• 2015

Bangpungtongseong-san(BPTSS, Fangfengtongsheng-san in Chineses) is a traditional herbal formula comprising 18 medicinal herbs. In the present study, we performed the simultaneous analysis for four compounds of BPTSS and examined anti-allergic effects in human keratinocytes and mouse splenocytes. The column for separation of four compounds was used Luna C18 column and maintained at 40℃. The mobile phase for gradient elution consisted of two solvent systems. The analysis was carried out at a flow rate of 1.0 mL/min with PDA detection at 254 and 280 nm. To evaluate production and expression of Th2 chemokines, ELISA and RT-PCR were conducted in tumor necrosis factor (TNF)-α and interferon (IFN)-γ-stimulated HaCaT cells with or without BPTSS or silymarin, a positive control for skin inflammation. To measure Th2 cytokines, primary mouse splenocytes were treated with BPTSS and performed ELISA for interleukin (IL)-4, 5, 13. Calibration curves were acquired with r2>0.9999. The contents of geniposide, liquiritin, baicalin, and glycyrrhizin in BPTSS were 5.06 ㎎/g, 7.33 ㎎/g, 27.56 ㎎/g, and 7.81 ㎎/g, respectively. BPTSS reduced TARC and RANTES production and mRNA expression in TNF-α and IFN-γ-treated HaCaT cells. BPTSS inhibited IL-4, 5, and 13 production in mouse splenocytes. Our data will be a helpful information to upgrade quality control and anti-allergic effects of BPTSS.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.4
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• pp.510-522
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• 2008

Our earlier studies showed that estrogen was involved in the regulation of fluid reabsorption in adult mouse efferent ductules (ED), through estrogen receptor (ER) ${\alpha}$ and $ER{\beta}$ by modulating gene expression of epithelial genes involved in ion homeostasis. However, little is known about the importance of $ER{\alpha}$ in the ED during postnatal development. Based on previous findings, we hypothesized that there should be a difference in the expression of epithelial ion transporters and anion producers in the ED of postnatal wild type (WT) and estrogen receptor ${\alpha}$ knockout (${\alpha}ERKO$) mice. Using absolute, comparative and semi-quantitative RT-PCR along with immunohistochemistry, we looked at expression levels of several genes in the ED across postnatal development. The presence of estrogen in the testicular fluid was indirectly ascertained by immunohistochemical detection of the P450 aromatase in the testis. There was no immunohistochemically detectable difference in the expression of P450 aromatase in the testes and ER${\beta}$ in the ED of WT and ${\alpha}$ERKO mice. ER${\alpha}$ was only detected in the ED of WT mice. The absence of ER${\alpha}$ in the ED of postnatally developing mice resulted in differential expression of mRNAs and/or proteins for carbonic anhydrase II, $Na^+/H^+$ exchanger 3, down-regulated in adenoma, cystic fibrosis transmembrane regulator, and $Na^+/K^+$ ATPase ${\alpha}$. Our data indicate that the absence of ER${\alpha}$ resulted in altered expression of an epithelial ion producer and transporters during postnatal development of mice. We conclude that the presence of ER${\alpha}$is important for regulation of the ED function during the prepubertal developmental and postpubertal period.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.5
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• pp.1851-1855
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• 2015

Background: There are increasing data about microRNAs (miRNA) in the literature, providing abundant evidence that they play important roles in pathogenesis and development of colorectal cancer. In this study, we aimed to investigate the miRNA expression profiles in surgically resected specimens of patients with recurrent and non-recurrent colorectal cancer. Materials and Methods: The study population included 40 patients with stage II colorectal cancer (20 patients with recurrent tumors, and 20 sex and age matched patients without recurrence), who underwent curative colectomy between 2004 and 2011 without adjuvant therapy. Expression of 16 miRNAs (miRNA-9, 21, 30d, 31, 106a, 127, 133a, 133b, 135b, 143, 145, 155, 182, 200a, 200c, 362) was verified by quantitative real-time polymerase chain reaction (qRT-PCR) in all resected colon cancer tissue samples and in corresponding normal colonic tissues. Data analyses were carried out using SPSS 15 software. Values were statistically significantly changed in 40 cancer tissues when compared to the corresponding 40 normal colonic tissues (p<0.001). MiR-30d, miR-133a, miR-143, miR-145 and miR-362 expression was statistically significantly downregulated in 40 resected colorectal cancer tissue samples (p<0.001). When we compared subgroups, miRNA expression profiles of 20 recurrent cancer tissues were similar to all 40 cancer tissues. However in 20 non-recurrent cancer tissues, miR-133a expression was not significantly downregulated, moreover miR-133b expression was significantly upregulated (p<0.05). Conclusions: Our study revealed dysregulation of expression of ten miRNAs in Turkish colon cancer patients. These miRNAs may be used as potential biomarkers for early detection, screening and surveillance of colorectal cancer, with functional effects on tumor cell behavior.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.77-84
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• 2003

Objective : The aim of this study was to evaluate the influence of three different media on preimplatation embryo development and the expression of Bcl-2, Mcl-1, Bax, and Bok in mouse. Materials and Methods: Two-cell embryos were retrieved from ICR female mice (4 weeks old) at 48 hr after hCG injection and cultured in Ham's F-10, HTF, and G1.2 media. The developmental rate of 2-cell embryos was evaluated from 24 hr to 72 hr after culture. RT-PCR was performed for the detection of Bcl-2, Mcl-1, Bax, and Bok gene expression. Results: The rates of morula and blastocyst in HTF and G1.2 media (88%, 98.1%) were significantly higher than those in Ham's F-10 media (39.6%) at 48 hr. Likewise, the rates of hatching and hatched blastocyst in HTF and G1.2 media (21.9%, 52.9%) were higher than those in Ham's F-10 media (3.5%) at 72 hr. Bcl-2 and Bax mRNAs were highly detected in embryos cultured in Ham's F-10 when compared in embryos cultured in HTF and G1.2. In contrast, the expression of Mcl-1 and Bok was not significantly different. Conclusion: These results show that HTF and G1.2 culture media increase the rate of blastocyst formation and stimulate Bcl-2 and Bax gene expression in mouse preimplantation embryos.