• Title/Summary/Keyword: IS-PCR

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No Association of $CYP2D6^*4$ and $CYP2D6^*10$ Polymorphisms with Tardive Dyskinesia in Korean Schizophrenics (한국인 정신분열병 환자의 지연성 운동장애와 $CYP2D6^*4$$CYP2D6^*10$ 다형성들의 연합에 대한 고찰)

  • Woo, Sung-Il;Kang, Dong-Woo;Seo, Han-Gil;Kim, Bong-Jo;Lee, In-Sang;Jeong, Geun-Hoa;Park, So-Young;Jung, Chi-Yeong;Lee, Hwan-Cheol;Jeong, Kyeong-Cheon;Sohn, Jin-Wook
    • Korean Journal of Biological Psychiatry
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    • v.7 no.2
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    • pp.140-146
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    • 2000
  • P450 CYP2D6 enzyme(=debrisoquine hydroxylase) is known to metabolize many neuroleptics and some genetic polymorphisms in the CYP2D6 gene were reported to be associated with tardive dyskinesia(TD). We investigeted the association of two genetic polymorphisms in the CYP2D6 gene, $CYP2D6^*4$ and $CYP2D6^*10$, with TD in Korean schizophrenic subjects. Subjects consisted of 71 Korean schizophrenics and TD was evaluated using the Abnormal Involuntary Movement Scale (AIMS). There were no statistically significant differences in the demographic variables of age, male to female percentage and the current antipsychotic(CPZ equivalent) dose between the group with TD and the group without TD. But the duration of antipsychotic drug exposure was significantly higher in the group without TD(p=0.000, by independent t-test). The mean AIMS score in the group with TD was $11.2{\pm}6.6$(S.D.). Genotypings for the presence of $CYP2D6^*4$ and $CYP2D6^*10$ were done using PCR amplifications and endonuclease digestions. There were no statistically significant genotypic and alleleic associations between TD and $CYP2D6^*4$(by chisquare tests), and between TD and $CYP2D6^*10$(by chi-square tests). These results indicate that the $CYP2D6^*4$ and $CYP2D6^*10$ polymorphisms have no significant roles in the causation of TD.

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Molecular Cloning, Characterization and Expression of a Novel Trehalose-6-phosphate Synthase Homologue from Ginkgo biloba

  • Wu, Weisheng;Pang, Yongzhen;Shen, Guo-An;Lu, Jie;Lin, Juan;Wang, Jin;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.39 no.2
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    • pp.158-166
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    • 2006
  • In many organisms, trehalose acts as protective metabolite against harsh environmental stresses, such as freezing, drought, nutrient starvation, heat and salt. Herein a cDNA (designated as GbTPS, GenBank Accession Number AY884150) encoding a trehalose-6-phosphate synthase homologue was isolated and characterized from the living fossil plant, Ginkgo biloba, which is highly tolerant to drought and cold. GbTPS encoded an 868-amino-acid polypeptide with a predicted isoelectric point of 5.83 and molecular mass of 97.9 kD. Amino acid sequence alignment revealed that GbTPS shared high identity with class II trehalose-6-phosphate synthase homologues (67% identical to AtTPS7), but had only 17% and 23% of identity with OstA from Escherichia coli and ScTPS1 from S. cerevisiae, respectively. DNA gel blot analysis indicated that GbTPS belonged to a small multi-gene family. The expression analysis by RT-PCR showed that GbTPS expressed in a tissue-specific manner in G biloba and might involve in leaf development. GbTPS was also found to be induced by a variety of stresses including cold, salt, drought and mannitol.

Molecular Cloning and Characterization of a Novel Stem-specific Gene from Camptotheca acuminata

  • Pi, Yan;Liao, Zhihua;Chai, Yourong;Zeng, Hainian;Wang, Peng;Gong, Yifu;Pang, Yongzhen;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.39 no.1
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    • pp.68-75
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    • 2006
  • In higher plants, P450s participate in the biosynthesis of many important secondary metabolites. Here we reported for the first time the isolation of a new cytochrome P450 cDNA that expressed in a stem-specific manner from Camptotheca acuminata (designated as CaSS), a native medicinal plant species in China, using RACE-PCR. The full-length cDNA of CaSS was 1735 bp long containing a 1530 bp open reading frame (ORF) encoding a polypeptide of 509 amino acids. Bioinformatic analysis revealed that CASS contained a heme-binding domain PFGXGRRXCX and showed homology to other plant cytochrome P450 monooxygenases and hydroxylases. Southern blotting analysis revealed that there was only one copy of the CaSS present in the genome of Camptotheca acuminata. Northern blotting analysis revealed that CaSS expressed, in a tissue-specific manner, highly in stem and lowly in root, leaf and flower. Our study suggests that CaSS is likely to be involved in the phenylpropanoid pathway.

Immune-triggering effect of the foodborne parasite Kudoa septempunctata through the C-type lectin Mincle in HT29 cells

  • Shin, Ji-Hun;Yang, Jung-Pyo;Seo, Seung-Hwan;Kim, Sang-Gyun;Kim, Eun-Min;Ham, Do-Won;Shin, Eun-Hee
    • BMB Reports
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    • v.53 no.9
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    • pp.478-483
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    • 2020
  • Kudoa septempunctata is a myxozoan parasite that causes food poisoning in individuals consuming olive flounder. The present study aimed to investigate the currently insufficiently elucidated early molecular mechanisms of inflammatory responses in the intestine owing to parasite ingestion. After Kudoa spores were isolated from olive flounder, HT29 cells were exposed to spores identified to be alive using SYTO-9 and propidium iodide staining or to antigens of Kudoa spores (KsAg). IL-1β, IL-8, TNF-α and NFKB1 expression and NF-κB activation were assessed using real-time PCR, cytokine array and western blotting. The immunofluorescence of FITC-conjugated lectins, results of ligand binding assays using Mincle-Fc and IgG-Fc, CLEC4E expressions in response to KsAg stimulation, and Mincle-dependent NF-κB activation were assessed to clarify the early immune-triggering mechanism. Inflammatory cytokines (IL-1β, GM-CSF and TNF-α), chemokines (IL-8, CCL2, CCL5 and CXCL1) and NF-κB activation (pNF-κB/NF-κB) in HT29 cells increased following stimulation by KsAg. The immunofluorescence results of spores and lectins (concanavalin A and wheat germ agglutinin) suggested the importance of Mincle in molecular recognition between Kudoa spores and intestinal cells. Practically, data for Mincle-Fc and KsAg binding affinity, CLEC4E mRNA expression, Mincle immunofluorescence staining and hMincle-dependent NF-κB activation demonstrated the involvement of Mincle in the early immune-triggering mechanism. The present study newly elucidated that the molecular recognition and immune-triggering mechanism of K. septempunctata are associated with Mincle on human intestinal epithelial cells.

Breeding of Pleurotus eryngii with a high temperature tolerance trait (고온적응성 큰느타리(새송이)버섯 품종육성)

  • Im, Chak Han;Kim, Min-Keun;Kim, Kyung-Hee;Cho, Soo Jeong;Lee, Jong Jin;Joung, Wan-Kyu;Lee, Sang Dae;Choi, Young Jo;Ali, Asjad;Ryu, Jae-San
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.187-192
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    • 2014
  • In order to breed a new P. eryngii cultivar with high temperature tolerance trait to cope with climate change, strains and cultivars were characterized at $20^{\circ}C$ that is $5^{\circ}C$ higher than normal condition followed by screening for the characteristics such as required days to harvest, quality and yield. Monokaryons from the selected strains were crossed. $Da-32{\times}KNR2322-15$ derived from the crosses between KNR2322 having characteristics of short growing day and Da($Ga5Na5-4{\times}KNR2312-7$) having charateristics of high guality and yield at $20^{\circ}C$, showed 14.9 days for harvest, 120.6 g yield, and 7.0 quality in the first trial. The strains were named as Taeyangsongi and cultivated on a large scale to compare with Kenneutari No. 2 at a mushroom farm. Yield of Taeyangsongi (109 g) was significantly different(P=0.001) from Kenneutari No.2. Quality of the new (6.6) and the reference cultivar (3.5) was also statistically different (P=0.001) The brightness of pileus of Taeyangsongi (59.5) was 10 points less than the reference cultivar due to which it has an inability to bear high temperature stress. Thus, PCR reactions with URP2 discriminated between Taeyansongi and reference cultivars (Keneutari No. 2 and Aeryni).

Tuberculous Empyema Necessitatis with Osteomyelitis, a Rare Case in the 21st Century (늑골 골수염에 동반된 흉벽 천공성 농흉 1례)

  • Kim, Han Wool;Lim, Goh-Woon;Cho, Hye Kyung;Lee, Hyunju;Won, Tae Hee;Park, Kyoung Un;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.18 no.1
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    • pp.80-84
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    • 2011
  • Empyema necessitatis refers to empyema that extends into the extrapleural space through a defect in the pleural surface. Tuberculous empyema necessitatis is a rare complication of tuberculosis. We experienced a 21-month-old boy with tuberculous empyema necessitatis with osteomyelitis in the right $7^{th}$ rib. He presented with a mass on the right lateral chest wall, which was soft and nontender, enlarging for one month. He also had mild fever. The plain radiograph of his chest revealed soft tissue swelling and calcified lymph node on the left axilla, and his PPD skin test was positive. CT scan of the chest showed empyema necessitatis at the right lower chest and upper abdominal walls with osteomyelitis of the right $7^{th}$ rib. He did not have concurrent pulmonary tuberculosis. Surgery was performed for diagnosis and treatment. In histopathologic findings, chronic granulomatous inflammation with caseation necrosis was shown and was positive for acid fast bacilli stain. In addition, M. tuberculosis complex was found as etiology by polymerase chain reaction. The patient has been treated with anti-tuberculous medication without any specific complication.

Epidemiologic Characteristics of Human Bocavirus-Associated Respiratory Infection in Children (소아 보카바이러스 호흡기 감염증의 역학적 특징)

  • Choi, Jae Hong;Paik, Ji Yeun;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.18 no.1
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    • pp.61-67
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    • 2011
  • Purpose : This study was performed to investigate the epidemiologic characteristics of human bocavirus (HBoV)-associated lower respiratory tract infections (LRTIs) in children. Methods : Nasopharyngeal aspirate samples were obtained from 658 children who had been hospitalized for LRTIs in Seoul National University (SNU) Children's Hospital and SNU Bundang Hospital from March 2000 to September 2005. Multiplex RT-PCR was performed to detect 11 respiratory viruses including respiratory syncytial virus, adenovirus, rhinovirus, parainfluenza viruses 1 and 3, influenza viruses A and B, human metapneumovirus, HBoV, human coronavirus (HCoV) OC43/ 229E, and HCoV-NL63. Clinical data were reviewed retrospectively. Results : Overall, respiratory viruses were detected in 325 (49.4%) among 658 patients. HBoV was detected in 62 cases (9.4%) and was responsible for 19.1% of virus-positive cases. HBoV was prevalent among infants and young children aged from 3 months to 5 years with the mean age of 25.3 months. Co-detection of HBoV and other respiratory viruses was observed in 37.1% which is significantly higher than average co-detection rate (12.3%) among overall virus-positive cases (P=0.000). HBoV was identified mainly in late spring and early summer from May to July. Conclusion : This study describes epidemiologic features of HBoV in Korean children compared with those associated with other respiratory viruses. HBoV was prevalent among LRTIs in childhood, especially in late spring and early summer season in Korea.

Two cases of Hemolytic Uremic Syndrome Associated with Esecherichia coli O114 (Escherichia coli O114에 의한 용혈성 요독 증후군 2례)

  • Ryu Su-Jeung;Hahn Hyewon;Yoo Soo Jin;Lee Byung-Sun;Lee Jae Kyoo;Kim Mi-Na;Kim Eui Chong;Park Young Seo
    • Childhood Kidney Diseases
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    • v.6 no.1
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    • pp.102-108
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    • 2002
  • We report two cases of hemolytic uremic syndrome (HUS) associated with Escherichia coli O114. Two cases were similar and showed the same clinical courses. After prodrome of diarrhea and vomiting lasting 1-2 days, azotemia persisted for about 10 days, and during that period, the patients were on peritoneal dialysis. They recovered without any sequelae after about 15 days. Direct multiplex PCR of stool culture revealed eae and stx2 gene and the result of ELISA done on the colony positive of one gene confirmed Escherichia coli O114. This is the first report of HUS associated with Escherichia coli O114. We recommend, Shiga toxin producing bacterial Infection must be considered and efforts should be made to scrutinize the organism in all diarrhea-prodrome HUS patients.(J Korean Soc Pediatr Nephrol 2002 ;6 : 102-8)

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Experimental Research on the Effect of Haeyeol-tang (해열탕(解熱湯)의 항염증(抗炎症), 항산화(抗酸化), 항노화(抗老化) 작용(作用)에 대한 실험적(實驗的) 연구(硏究))

  • Chu, Cheng-Kang;Kim, Jin-Ju;Roh, Young-Lae;Lee, Sung-Hun;Jeong, Seung-Yeon;Jung, Hee-Jae;Jung, Sung-Ki
    • The Journal of Internal Korean Medicine
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    • v.28 no.4
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    • pp.671-680
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    • 2007
  • Objectives : The purpose of this study was to examine the effect of Haeyeol-tang on infectious disease of respiratory system. Methods : Haeyeol-tang was used by cold storage after dilution with 1, 10 and $100{\mu}g/ml$ of PBS, from dried material as extracted material with extraction with 50% ethanol. Statistical significance was examined through measuring TNF-$\alpha$, RT-PCR, DPPH, nitric oxide(NO), and MTT of Raw264.7 cells activated with LPS. Results : For the toxicity experiment of Haeyeol-tang, almost no toxicity was shown on cells in the concentration ranges of 1, 10 and $100{\mu}g/ml$. For the effect of NO production, Haeyeol-tang showed significant (p<0.001) decrease of NO production, according to its concentration. For the effect on the eliminating activity of DPPH radical, Haeyeol-tang showed increasing tendency of eliminating activity of DPPH radical, according to its concentration. For the effect on revelation of NOS2 enzyme, Haeyeol-tang showed inhibiting tendency against the revelation of NOS2, according to its concentration. For the effect on the TNF-$\alpha$ production, Haeyeol-tang showed significant (p<0.001) decrease of TNF-$\alpha$ production. Conclusions : This result means that Haeyeol-tang is effective for anti-inflammation, as well as excellent effect for anti-oxidation and anti-aging.

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Different Potential of Hematopoietic Differentiation in Two Distinct Mouse Embryonic Stem Cells (두 개의 다른 마우스 배아줄기세포의 차별적인 조혈세포 분화능)

  • Kim, Jin-Sook;Kang, Ho-Bum;Song, Jee-Yeon;Oh, Goo-Taeg;Nam, Ki-Hoan;Lee, Young-Hee
    • Development and Reproduction
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    • v.9 no.2
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    • pp.105-114
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    • 2005
  • Embryonic stem(ES) cells have tremendous potential as a cell source for cell-based therapies. Realization of that potential will depend on our ability to understand and manipulate the factors that influence cell fate decision and to develop methods for getting enough cell numbers for clinical applications. Hematopoiesis has been widely studied, and hematopoietic differentiation from ES cells is a good model to study lineage commitment. In this study, we investigated stemness and compared the efficiency of hematopoietic differentiation using two different mouse embryonic stem cell lines TC-1 and B6-1. Although the two cell lines showed known stem cell properties with minor differences, the embryoid body formation efficiency in methylcellulose was much higher in TC-1 than B6-1. When measured potentials of hematopoietic differentiation using functional(colony-forming cell) and phenotypic(specific marker expression) assays, we found that TC-1 can differentiate into hematopoietic cells in methylcellulose culture but B6-1 cannot. These results imply that we can improve the efficiency of hematopoietic cell differentiation by selection of proper cell lines and this may be also applied in the differentiation of human embryonic stem cells.

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