• Title/Summary/Keyword: CC-PCR

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TGF-β-activated Kinase-1: A Potential Prognostic Marker for Clear Cell Renal Cell Carcinoma

  • Wei, Can;Lai, Yong-Qing;Li, Xian-Xin;Ye, Jiong-Xian
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.315-320
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    • 2013
  • Background: TGF-${\beta}$-activated kinase-1 (TAK1) has been found to be over-expressed in a variety of solid malignancies and related to tumor growth. The aim of this study was to evaluate the expression level of TAK1 in clear cell renal cell carcinoma (ccRCC) and assess its value as a novel prognostic marker. Methods: TAK1 mRNA was assessed in 51 paired ccRCC tissues and adjacent normal tissues (ADTs) by real-time PCR. Tissue TAK1 protein was also assessed in 91 ADTs and 177 samples of ccRCC immunohistochemically for evaluation of relationships with clinical characteristics. Results: RT-PCR showed that TAK1 RNA level was significantly higher in ccRCC tissues than in the paired ADTs and immunohistochemistry confirmed higher expression of TAK1 protein in ccRCC samples compared with ADTs. TAK1 protein expression in 177 ccRCC samples was significantly correlated with T stage, N classification, metastasis, recurrence and Fuhrman grade, but not age and gender. Patients with low TAK1 levels had a better survival outcome. TAK1 expression and N stage were independent prognosis factors for the overall survival of ccRCC patients. Conclusions: Overexpression of TAK1 predicts a poor prognosis in patients with ccRCC, so that TAK1 may serve as a novel prognostic marker.

Detection of Enterovituses from Surface Water by Combined Cell Culture-PCR (지표수로부터 세포배양-연계 PCR법에 의한 장바이러스의 검출)

  • 정은영;정종문;류재익;신판세;전홍기;장경립
    • Journal of Life Science
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    • v.10 no.5
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    • pp.484-489
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    • 2000
  • Enterovirues may cause gastrointestinal symptoms, cold, and fever, mainly in young children. They are also recognized as important agents in acute infections of the central nervous system such as meningitis and encephalitis, and in subacute and chronic infections of the cardiovascular system such as pericarditis, myocarditis and cardiomyopathy. They also can lead to postviral fatigue syndrome. For the detection of enteroviruses from the environmental samples, the combined cell culture-polymerase chain reaction (CC-PCR) technique was employed. In contrast to EPA standard method which mainly depends on the cell culture, it involved the use of cell culture, followed by PCR to improve the sensitivity and the accuracy of the test. According to the results of survey, from 1999 to 2000, for the presence of enteroviruses in the surface water samples from Nak-dong river, four out of twelve samples were positive for viruses. The titer of viruses in the surface water was ranged from 25 to 250 MPN. All of the viruses isolated were poliovirus type I with 98% nucleotide sequence homology. The result also clearly suggests the seasonal difference in the distribution of the waterborne enteroviruses in surface water because most of the viruses were mainly detected from the summer through the early autumn.

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The variation of insulin like growth factor 2 maker is associated with growth traits in Thai native (Kradon) pigs

  • Kessara Ampaporn;Rattikan Suwannasing;Pitchayanipa Phongphanich;Supanon Tunim;Monchai Duangjinda
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1350-1356
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    • 2023
  • Objective: This study was conducted to investigate polymorphisms of the melanocortin-4 receptor (MC4R) and insulin like growth factor 2 (IGF2) genes and to evaluate the growth traits affected by such polymorphisms in Thai native (Kradon) pigs. Methods: Blood samples and productive data from 91 Kradon pigs were collected. DNA was extracted and quantified, the IGF2 and MC4R genes were amplified, and the polymerase chain reaction (PCR) produces were digested using the PCR-restriction fragment length polymorphism (PCR-RFLP) technique. Genotyping was performed, and the association between genotypes and growth traits on the birth and weaning weights were evaluated. Results: The IGF2 intron7 g.162G>C variations in Kradon pigs were found in three genotypes: i) GG, ii) GC, and iii) CC. The GG genotype frequency was the highest followed by the GC and CC genotypes. The frequencies of the G and C alleles were 0.703 and 0.297, respectively. The MC4R genotype was found in only one genotype (GG). The IGF2 gene pattern was not associated with birth weight traits, whereas the IGF2 gene pattern was related to the weaning weight trait in Kradon pigs. Pigs with the CC and GC genotypes had higher weaning weights than ones with the GG genotype (p<0.001). Conclusion: Thai native Kradon pigs with the CC and GC genotypes of the IGF2 gene have higher weaning weights than pigs with the GG genotype.

Inhibitory Effect of Mix proportion of Root of Scutellaria baicalensis and Coptis chinensis on LPS-induced type-I interferon Production in RAW264.7 Cells (LPS로 자극한 RAW267.4 세포에서 황금(黃芩), 황련(黃連) 배합 비율에 따른 TYPE-1 interferon 억제효과)

  • Kook, Yoon-Bum
    • Herbal Formula Science
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    • v.16 no.2
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    • pp.155-162
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    • 2008
  • Objectives : The present study was designed to investigate corelation between mix proportion of Scutellaria baicalensis (SB) and Coptis chinensis (CC) on lipopolysaccharide (LPS)-induced TYPE-1 interferon production. Methods : I examined TYPE-1 interferon, interferon regulating factor (IRF)-1,7 and interleukin(IL)-10 production on LPS-induced RAW264.7 cells to evaluate inhibitory effect of mix proportion of SB and CC using real time PCR. Results : Mixture of SB and CC regulated TYPE-1 interferon and IRF-1,7 mRNA expression with SB dose dependent manner, while maintained IL-10 mRNA expression on LPS-induced RAW264.7 cells. Conclusion : In mixture of SB and CC, SB plays a key role in reducing TYPE-1 interferon through inactivation IRF-1,7. Furthermore mixture of SB and CC maintained IL-10 mRNA level. Collectively, this results suggest that SB confer beneficial effects in autoimmune diseases clinically.

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Occurrence of Plant-parasitic Nematodes in Major Potato Production Areas and PCR Identification of Root-knot Nematodes (감자 주산지 식물기생선충 분포와 뿌리혹선층의 PCR 동정)

  • Cho Myoung-Rae;Lee Young-Gyu;Kim Jum-Soon;Yoo Dong-Lim
    • Korean journal of applied entomology
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    • v.45 no.1 s.142
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    • pp.79-85
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    • 2006
  • This study was conducted to get basic information on the occurrence of plant-parasitic nematodes for the establishment of nematode management strategy in major potato production areas in Korea. Nationwide soil collection was done in 11 areas of Cheju, Yesan, Gimchun, Goryoung, Hong chun, Pyungchang, Gimjae, Milyang, Namwon, Gangnung, and Inje in 2004-2005. Root-hot nematode juveniles(J2) were detected in 30 samples among the 50 samples. The average density was 12-69 J2/100cc soil. Pratylenchus sp., Helicotylenchus sp., Ditylenchus sp., Tylenchus sp., and Tylenchorhynchus sp. were also detected in various locations, however, their densities were very low. Root-knot nematode females were collected from tomato roots inoculated with the potato field soils for PCR-RFLP identification. The females from Cheju, Milyang, and Goryung showed PCR products of 500 bp. And the Dra I restriction enzyme digestions showing 290 bp and 230 bp fragments confirmed their identity as Meloidogyne hapla.

Biosynthesis of (R)-phenyl-1,2-ethanediol by using Single Recombinant Epoxide Hydrolase from Caulobacter Crescentus (재조합 epoxide hydrolase를 단일 생촉매로 사용한 광학수렴 가수분해반응을 통한 광학활성 (R)-phenyl-1,2-ethanediol 생합성)

  • Lee, Ok Kyung;Lee, Eun Yeol
    • Applied Chemistry for Engineering
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    • v.18 no.3
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    • pp.279-283
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    • 2007
  • Epoxide hydrolase (EH) gene of Caulobacter crescentus was cloned by PCR and expressed in Escherichia coli. The C. crescentus EH (CcEH) primarily attacked at the benzylic carbon of (S)-styrene oxide, while the CcEH preferentially attacked at the terminal carbon of (R)-styrene oxide, thus leading to the formation of (R)-phenyl-1,2-ethanediol as the main product. (R)-phenyl-1,2-ethanediol was obtained with 85% enantiomeric excess and yield of 69% from racemic styrene oxide via enantioconvergent hydrolysis by using recombinant CcEH as the single biocatalyst.

Analysis of Manifestation of CC and CXC Chemokine Genes in Olive Flounders (Paralichthys olivaceus) Artificially Infected with VHSV during the Early Developmental Stage

  • Kim, Kyung-Hee;Kim, Woo-Jin;Park, Choul-Ji;Park, Jong-Won;Noh, Gyeong Eon;Lee, Seunghyung;Lee, Young Mee;Kim, Hyun Chul
    • Development and Reproduction
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    • v.22 no.4
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    • pp.341-350
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    • 2018
  • Chemokines is a small protein that plays a major role in inflammatory reactions and viral infections as a chemotactic factor of cytokines involved in innate immunity. Most of the chemokines belong to the chemokine groups CC and CXC. To investigate the immune system of the olive flounder (Paralichthys olivaceus), an expression pattern specifically induced in the early developmental stages of analysis is examined using qRT-PCR. We also examined tissue-specific expression of both CC and CXC chemokine in healthy olive flounder samples. CC and CXC chemokine shows increased expression after immune-related organs are formed compared to expression during early development. CC chemokine was more highly expressed in the fin, but CXC chemokine showed higher expression in the gills, spleen, intestines, and stomach. Spatial and temporal expression analysis of CC and CXC chemokine were performed following viral hemorrhagic septicemia virus (VHSV) infection. CC chemokine showed high expression in the gills, which are respiratory organs, whereas CXC chemokine was more highly expressed in the kidneys, an immune-related organ. These results suggest that CC and CXC chemokine play an important role in the immune response of the olive flounder, and may be used as basic data for the immunological activity and gene analysis of it as well as other fish.

Diagnosis and Monitoring of Chronic Myeloid Leukemia: Chiang Mai University Experience

  • Tantiworawit, Adisak;Kongjarern, Supanat;Rattarittamrong, Ekarat;Lekawanvijit, Suree;Bumroongkit, Kanokkan;Boonma, Nonglak;Rattanathammethee, Thanawat;Hantrakool, Sasinee;Chai-Adisaksopha, Chatree;Norasetthada, Lalita
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.2159-2164
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    • 2016
  • Background: A diagnosis of chronic myeloid leukemia (CML) is made on discovery of the presence of a Philadelphia (Ph) chromosome. The success of the treatment of this form of leukemia with tyrosine kinase inhibitor (TKI) is monitored by reduction of the Ph chromosome. Objective: To compare the role of conventional cytogenetic (CC) methods with a real time quantitative polymerase chain reaction (RQ-PCR) and fluorescence in situ hybridization (FISH) for diagnosis and treatment monitoring of CML patients. The secondary outcome was to analyze the treatment responses to TKI in CML patients. Materials and Methods: This was a retrospective study of CML patients who attended the Hematology clinic at Chiang Mai University Hospital from 2005-2010. Medical records were reviewed for demographic data, risk score, treatment response and the results of CC methods, FISH and RQ-PCR. Results: One hundred and twenty three cases were included in the study, 57.7% of whom were male with a mean age of 46.9 years. Most of the patients registered as intermediate to high risk on the Sokal score. At diagnosis, 121 patients were tested using the CC method and 118 (95.9%) were identified as positive. Five patients failed to be diagnosed by CC methods but were positive for BCR-ABL1 using the FISH method. Imatinib was the first-line treatment used in 120 patients (97.6%). In most patients (108 out of 122, 88.5%), a complete cytogenetic response (CCyR) was achieved after TKI therapy and in 86 patients (70.5%) CCyR was achieved long term by the CC method. Five out of the 35 analyzed patients in which CCyR was achieved by the CC method had a positive FISH result. Out of the 76 patients in which CCyR was achieved, RQ-PCR classified patients to only CCyR in 17 patients (22.4%) with a deeper major molecular response (MMR) in 4 patients (5.3%) and complete molecular response (CMR) in 55 patients (72.4%). In the case of initial therapy, CCyR was achieved in 95 patients (79.1%) who received imatinib and in both patients who received dasatinib (100%). For the second line treatment, nilotinib were used in 30 patients and in 19 of them (63.3%) CCyR was achieved. In half of the 6 patients (50%) who received dasatinib as second line or third line treatment CCyR was also achieved. Conclusions: CML patients had a good response to TKI treatment. FISH could be useful for diagnosis in cases where CC analysis failed to detect the Ph chromosome. RQ-PCR was helpful in detecting any residual disease and determining the depth of the treatment response at levels greater than the CC methods.

Inhibitory effect of Cinnamomi Cortex extract on motility of prostate cancer cells through reducing YAP activity (육계의 전립선암세포에서 YAP 활성 억제를 통한 전이 저해 효능 연구)

  • Jung, Hyo Won;Kim, Ok-Hyeon;Wang, Tsu Yu;Kim, Seong Eun;Park, Yong-Ki;Lee, Hyun Jung
    • The Korea Journal of Herbology
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    • v.34 no.3
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    • pp.55-61
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    • 2019
  • Objectives : Recently, natural bioactive components catch a major attention for their potent anticarcinogenic activity. In this study, the inhibitory effect of Cinnamomi Cortex (CC) was examined in PC3 prostate cancer cells. Methods : The toxicity of CC extract was evaluated with cell viability and cell morphology. The activity of Yes associated protein (YAP) was tested with qRT-PCR for the target gene expression such as CTGF and AMOTL2. Western blotting was performed for the evaluation of phospho-YAP level. For cell motility analysis, cellular motility was imaged by live imaging system for 6 hr. Successive images were used for the generation of movie file. Using this movie file, cellular migration was manually tracked and analyzed using time-lapse microscope and Fiji software. Results : Cytotoxicity of CC extract was not detected at $500{\mu}g/m{\ell}$ or below concentration. Although $500{\mu}g/m{\ell}$ of CC extract reduced CTGF and AMOTL2 gene expression as YAP target genes, it was not statistically significant (CTGF expression P=0.0605, AMOTL2 expression P=0.4478). However, phosphorylated YAP was highly enhanced by CC extract treatment, when normalized with total YAP protein expression, suggesting YAP activation was inhibited. Finally prostate cancer cell motility was markedly reduced by $500{\mu}g/m{\ell}$ of CC extract. Conclusions : CC extract suppresses cancer cell motility and migration ability through inhibiting YAP activation without prostate cancer cell death, suggesting that this herb might be effective therapeutic drug for prostate cancer metastasis.

Genetic Variation in MicroRNAs and Risk of Oral Squamous Cell Carcinoma in South Indian Population

  • Sushma, PS;Jamil, Kaiser;Kumar, P Uday;Satyanarayana, U;Ramakrishna, M;Triveni, B
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7589-7594
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    • 2015
  • Background: MicroRNAs (miRNAs) are small non-coding RNA molecules, implicated in several activities like initiation, progression and prognosis of various cancers. Single nucleotide polymorphisms (SNPs) in miRNA genes can lead to alteration in mRNA expression, resulting in diverse functional consequences. The aim of our study was to investigate the association of miR-149C>T and miR-196a2C>T SNPs with susceptibility to development of oral squamous cell carcinoma (OSCC) in South Indian subjects. Materials and Methods: 100 OSCC patients and 102 healthy controls from the general population were recruited for the study. Genetic analysis was performed by polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) as per a standard protocol. Results: The genotype frequencies in miR-196a2 polymorphism, of TT, CT and CC in the OSCC patients were 69%,10% and 22% respectively while for control group it was 80%, 15% and 5% respectively. The CC genotype of miR196a2 polymorphism was significantly associated with oral squamous cell carcinoma. The genotype frequencies in miR-149 polymorphisms of CC, CT and TT in the oral squamous cell carcinoma (OSCC) patients were 72%, 22% and 6% respectively and for control group 88%, 12% and 0% respectively. CT and TT genotypes of miR149 polymorphism were found to be significantly associated with OSCC (p = 0.05 and 0.07). Conclusions: Our study suggests that miR-196a2C>T and miR-149C>T polymorphisms may play crucial roles in the development of OSCC in South Indian subjects.