• Title/Summary/Keyword: PCR (Polymerase Chain Reaction)

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Overexpression of Cyclooxygenase-1 Correlates with Poor Prognosis in Renal Cell Carcinoma

  • Yu, Zu-Hu;Zhang, Qiang;Wang, Ya-Dong;Chen, Jing;Jiang, Zhi-Mao;Shi, Min;Guo, Xin;Qin, Jie;Cui, Guang-Hui;Cai, Zhi-Ming;Gui, Yao-Ting;Lai, Yong-Qing
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3729-3734
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    • 2013
  • The aim of this study was to evaluate expression of COX-1 in renal cell carcinoma (RCC) and its prognostic value. mRNA of COX-1 was detected in 42 paired RCC and adjacent normal tissues with quantitative realtime polymerase chain reaction (qRT-PCR). Expression of COX-1 was also evaluated in 196 RCC sections and 91 adjacent normal tissues with immunohistochemistry. Statistical analysis was performed to assess COX-1 expression in RCC and its prognostic significance. The results of qRT-PCR showed mRNA levels of COX-1 in RCC tissues to be significantly higher than that in adjacent normal tissues (p < 0.001). Immunohistochemical assays also revealed COX-1 to be overexpressed in RCC tissues (p < 0.001). Statistical analysis demonstrated high expression of COX-1 was correlated with tumour size (p = 0.002), pathological stage (p = 0.003), TNM stage (p = 0.003, 0.007, 0.027, respectively), and tumour recurrence (p < 0.001). Survival analysis indicated patients with high expression of COX-1 had shorter survival time (p < 0.001), and COX-1 was an independent predictor. This is the first study to reveal overexpression of COX-1 in RRC and point to use as a prognostic marker in affected patients.

Effects of Ipriflavone on bone remodeling in the rat calvarial cell (백서 두개관세포에서 Ipriflavone이 골조직 개조에 미치는 영향)

  • Lee, Yong-Seung;Kim, Young-Jun;Lee, Ki-Heon;Hwang, Hyeon-Shik
    • The korean journal of orthodontics
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    • v.35 no.4 s.111
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    • pp.275-285
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    • 2005
  • Ipriflavone (isoprofoxyisoflavone), a synthetic derivative from soy isoflavone diazein, has been shown to inhibit bone resorption and perhaps stimulate bone formation This study was performed to examine the effects of ipriflavone on the proliferation and bone remodeling in rat calvarial cells in vitro The rat calvarial cells were isolated from fetus aged 20 to 21 days and cultured In BGJb media The graded concentration of ipriflavone $(10^{-9}\;10^{-5}M)$ was administered into cultured cells. When the cell proliferation was estimated through the measurement of MTT assay, there was no increase in cellular proliferation of the rat calvarial cell at any ipriflavone concentration. The cellular activity was evaluated through the formation of mineralized nodules stained by alizarin red. The formation of mineralized nodules significantly increased at concentrations of $10^{-8}M,\;10^{-7}M\;and\;10^{-6}M$ ipriflavone. Reverse transcription-polymerase chain reaction analyses (RT-PCR) were done at 7 and 14 days after culture to detect the expression of Bone Sialoprotein (BSP), Type I Collagen (COL I) and Osteocalcin(OCN) As a result, the expressions of BSP and COL I increased on the 7th day of culture and the expression of OCN increased on the 14th day of culture. These results indicate that ipriflavone facilitates the bone remodeling process bvy promoting rat calvarial cell differentiation aid stimulating mineralization through increased expression of extracellular matrix genes. such as BSP. COL I and OCN.

Association of the Interleukin-1$\beta$ and Interleukin-1 Receptor Antagonist Genetic Polymorphism and Korean Gastric Cancer (Interleukin-1$\beta$ 및 Interleukin-1 Receptor Antagonist의 유전적 다형성과 한국인 위암과의 연관 관계)

  • Park Jik Young;Cho Young Gu;Kim Chang Jae;Park Yong Kyu;Kim Young Sil;Park Cho Hyun;Lee Sug Hyung;Yoo Nam Jin;Lee Jung Young;Park Won Sang
    • Journal of Gastric Cancer
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    • v.2 no.3
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    • pp.163-167
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    • 2002
  • Purpose: Interleukin 1$\beta$ (IL-1$\beta$) polymorphisms are associated with hypochlorhydria, atrophic gastritis, and increased risk of gastric cancer in Caucasians. We tried to determine whether the IL-1.. and IL-1 receptor antagonist (IL-1 RN) genetic polymorphisms contribute to the development of gastric cancer and the specific type of gastritis in Korean. Materials and Methods: The study population was comprised of 128 gastric cancer patients with histologically proven carcinoma and 63 normal healthy individuals. Sixty-eight carcinomas were of intestinal-type and sixty tumors were of diffuse-type. No patient had a familial gastric cancer history. The 511 bp and 31 bp polymorphisms in the IL-1.. were genotyped by polymerase chain reaction (PCR)-restriction fragment length polymorphism. The polymorphism of the IL-1 RN was analyzed with variable number tandem repeat after PCR. Results: The genotype of 511C/-31T of IL-1$\beta$ and allele 1 of IL-1 RN was dominant in the present subjects. The allelic frequencies of the C allele IL-1$\beta$, which is a high risk genotype for gastric cancer, were 0.551 and 0.429 in gastric cancer and normal controls, respectively. Statistically, significant difference in allelic frequencies of three polymorphic sites between gastric cancer patients and normal controls, and between intestinal-type and diffuse-type was not observed. Conclusions: These results suggest that the polymorphisms of IL-1$\beta$ and IL-1 RN may not contribute to the development of Korean gastric caner and that other endogenous or exogenous factors will be important for gastric carcinogenesis.

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Analysis of Genetic Diversity in Soybean Varieties Using RAPD Markers (사료작물로 이용이 가능한 한국 재배콩의 RAPD 표지인자에 의한 유전적 다양성 분석)

  • Lee, Sung-Kyu;Kim, Bum-Jun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.4
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    • pp.277-284
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    • 1998
  • Random amplified polymorphic DNA (RAPD) analysis was used to detect the genetic diversity of soybean (Glycine max (L.) Merr.) varieties and field bean (Glycine soza Sieb. and Zucc.) Five soybean varieties and one field bean were analysed with random primers using the polymerase chain reaction (PCR). Nine primers of a total twenty random primer were selected to amplify DNA segments. A total of 74 PCR products were amplified and 67.6% of which were polymorphic. The size of DNA molecule is ranged 0.13~2.0Kb and typically generated four to eight major bands. Specific genetic marker were revealed in primer sequence 5'-CAG GCC CIT C-3', 5'-TGC TCT GCC C-3' and 5'-GTC CAC ACG G-3', respectively. Genetic similarity between each of the varieties were calculated from the pair-wise comparisons of amplification products and a dendrogram was constructed by an unweighted pair-group method with arithmethical means (UPGMA). The results indicate that intervarietal relationships of soybean have a narrow genetic base and between the varieties, Hwanggum-kong and Seckryang-bootkong is more closely related than the rest of varieties, and field bean is related quite distant.

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Metabolic Gene Expression in Lipid Metabolism during Cotyledon Development in Cucumbers and the Possibility of a Secondary Transport Route of Acetyl Units (오이 떡잎의 발달에서 지방 대사관련 유전자의 발현과 아세틸 단위체의 2차 경로 가능성)

  • Cha, Hyeon Jeong;Kim, Dae-Jae
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1055-1062
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    • 2014
  • We investigated the expression of cucumber genes involved in lipid mobilization and metabolism during cotyledon development to compare gene activity and to study the direction of carbon (acetyl unit) transport between glyoxysomes and mitochondria. The core metabolic pathway involving 10 genes was examined in four intracellular compartments: glyoxysomes (peroxisomes), mitochondria, chloroplasts, and cytosol. Additionally, we tested the early germination response of dark-grown seedlings and the immediate light response for a further 3 days. According to the reverse transcription polymerase chain reaction (RT-PCR), 3-L-ketoacyl-CoA thiolase 2 (Thio2), isocitrate lyase (ICL), and malate synthase (MS), the genes involved in storage lipid mobilization showed a similar and consistent pattern of gene expression in seedling development. Furthermore, coordinate expression of the A BOUT DE SOUFFLE (BOU) gene with ICL and MS during seedling emergence pointed to a possible secondary route of acetyl unit (acetyl-CoA) transport between peroxisomes and mitochondria in cucumber. The expression of the BOU gene was light dependent, as shown by BOU activity in Arabidopsis, suggesting that the dark condition also results in weak membrane biogenesis. In addition, several genes were active throughout the development of the green cotyledon, even during senescence. In conclusion, this study summarizes oil-seed germination and gene expression during cucumber cotyledon development and proposes an additional route for acetyl unit transport.

Molecular and cytogenetic findings in 46,XX males

  • Choi, Soo-Kyung;Kim, Young-Mi;Seo, Ju-Tae;Kim, Jin-Woo;Park, So-Yeon;Moon, In-Gul;Ryu, Hyun-Mee;Kang, Inn-Soo;Lee, You-Sik
    • Journal of Genetic Medicine
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    • v.2 no.1
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    • pp.11-15
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    • 1998
  • This paper reports 3 cases with 46,XX sex reversed male. Three 46,XX hypogonadal subjects showed complete sex reversal and had normal phallus and azoospermia. We studied them under clinical, cytogenetic and molecular aspects to find out the origin of the sex reversal. Patients had markedly elevated serum follicle-stimulating hormone (FSH) and lutenizing hormone (LH) and decreased or normal range of serum testosterone. The testicular volumes were small (3-8ml). Testicular biopsy showed Leydig cell hyperplasia and atrophy of seminiferous tubules. We obtained the results of normal 46,XX, and the presence of Y chromosome mosaicism was ruled out through XY dual fluorescent in situ hybridization (FISH). By using polymerase chain reaction (PCR), we amplified short arm (SRY, PABY, ZFY and DYS14), centromere (DYZ3), and heterochromatin (DYZ1) region of the Y chromosome. PCR amplification of DNA from these patients showed the presence of the sex-determining region of the Y chromosome (SRY) but didn't show the centromere and heterochromatin region sequence. The SRY gene was detected in all the three patients. Amplification patterns of the other regions were different in these patients; one had four amplified loci (PABY+, SRY+, ZFY+, DYS14+), another had two loci (SRY+, ZFY+) and the other had two loci (PABY+, SRY+). We have found that each patient's translocation elements had different breakpoints at upstream and downstream of the SRY gene region. We conclude that the testicular development in 46,XX male patients were due to insertion or translocation of SRY gene into X chromosome or autosomes.

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A study on association of progesterone receptor gene polymorphism (PROGINS) with Endometriosis (자궁내막증과 progesterone receptor gene polymorphism (PROGINS)과의 연관성에 관한 연구)

  • Kim, Yun-Jin;Noh, Ji-Hyun;Koh, Jae-Whoan;Kim, Yong-Bong
    • Journal of Genetic Medicine
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    • v.4 no.2
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    • pp.128-132
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    • 2007
  • Purpose : Endometriosis is a steroid dependent disease with a particular genetic background but the location of possible genomic aberrations are still poorly clarified. This study was designed to investigate the associations between the polymorphism of the progesterone receptor gene (PROGINS) and endometriosis. Methods : 100 women with surgically diagnosed and histologically confirmed endometriosis were enrolled as a patient population and a total of 110 female control subjects undergoing health examination were enrolled as control population. DNA extraction and polymerase chain reaction (PCR) were used to genotype women for the presence of the PROGINS polymorphism in peripheral blood samples. The x2-test was used to compare genotype distributions between endometriosis and controls. Results : T1/T2 heterozygote was found to be one patient in each group, and the rest of the subjects were all T1/T1 homozygotes. There was no difference in the genotype distribution between the endometriosis group and the control group. Conclusion : These results suggest that the progesterone receptor gene PROGINS is not associated with the risk for endometriosis.

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Factors Influencing Agrobacterium-Mediated Transformation Efficiency in Perennial Ryegrass (Agrobacterium 매개에 의한 페레니얼 라이그라스의 형질전환에 영향을 미치는 요인)

  • Lee, Ki-Won;Kim, Ki-Yong;Lee, Joung-Kyong;Park, Hyung-Soo;Kim, Kyung-Hee;Lee, Byung-Hyun;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.3
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    • pp.165-170
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    • 2009
  • A system for the production of transgenic plants has been developed for perennial ryegrass (Lolium perenne L.) via Agrobacterium-mediated transformation. Included in this study were two factors which may affect the gene transfer efficiency: concentrations of acetosyringone (AS, 0 to 300 ${\mu}M$), and co-culture period (1 to 7 days). Both factors were very important to achieve high efficiency gene transformation in the perennial ryegrass. The highest transformation efficiency was obtained when embryogenic calli were inoculated with Agrobacterium in the presence of 100 ${\mu}M$ AS with the culture medium for 5 days. Phosphinothricin resistant calli were developed with into complete plants. GUS histochemical assay, polymerase chain reaction (PCR) and Northern blot analysis of transgenic plants demonstrated that transgenes were integrated into the genome of perennial ryegrass. Using this protocol, it was possible to obtain transformants efficiently for further study.

The Telescopic Findings and Clinical Manifestations of Laryngeal Tuberculosis (후두 결핵의 후두 내시경 소견과 임상 양상 분석)

  • Lee, Sang-Hyuk;Lee, Seung-Suk;Lee, No-Hee;Ban, Jae-Ho;Lee, Kyung-Chul;Jin, Sung-Min
    • Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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    • v.17 no.1
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    • pp.38-42
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    • 2006
  • Background and Objectives: There has been a gradual decline in the incidence of laryngeal tuberculosis due to effective an-tituberculous chemotherapy and improvement in the standard of living. During the last decade, however, the disease has changed its clinical pattern. We aimed to study a clinical and telescopic evaluation of laryngeal tuberculosis. Materials and Method: A retrospective clinical analysis was done for 16 patients who diagnosed laryngeal tuberculosis from January 2000 through December 2004. All patients had a complete clinical and laboratory work-up including telescopy, chest X-rays, sputum cultures, Tbc PCR(polymerase chain reaction). Laryngeal biopsies were performed in some cases. All the patients received proper antituberculous chemotherapy. Results: The patients age ranged from 21 to 59 with a mean age 43. The male to female ratio was 10:6. The prominent presenting symptom was hoarseness. The various telescopic findings were categorized: 7 patients showed ulcerative, 4 patients granulomatous, 3 patients ulcerofungative, 2 patients had nonspecific inflammatory lesions. Involving sites were true vocal cord most commonly in 12 patients. Three of 16 patients had normal chest X-ray fmding. Among 16 patients, 4 patients showed positive response for AFB stain, 13 patients showed positive for Tbc PCR. Biopsy was done for 3 patients who were not confirmed in initial tuberculous test. All patients responded satisfactorily to antituberculous medication. Conclusion: The telescopic findings and clinical manifestations of laryngeal tuberculosis have changed and seemed to be different from those of classic reports. Thus, the clinicians who deal with the various symptoms and diseases should be aware of the existence of laryngeal tuberculosis and the changing patterns of the disease.

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Refining prognostic stratification of human papillomavirus-related oropharyngeal squamous cell carcinoma: different prognosis between T1 and T2

  • Lee, Sumin;Lee, Sang-wook;Park, Sunmin;Yoon, Sang Min;Park, Jin-hong;Song, Si Yeol;Ahn, Seung Do;Kim, Jong Hoon;Choi, Eun Kyung;Kim, Su Ssan;Jung, Jinhong;Kim, Young Seok
    • Radiation Oncology Journal
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    • v.35 no.3
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    • pp.233-240
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    • 2017
  • Purpose: To validate the 8th edition of the American Joint Committee on Cancer/Union for International Cancer Control (AJCC/UICC) TNM staging system for human papillomavirus (HPV)-related oropharyngeal squamous cell carcinoma (OPSCC) and investigate whether a modified classification better reflects the prognosis. Materials and Methods: Medical records of patients diagnosed with non-metastatic HPV-related OPSCC between 2010 and 2016 at a single institution were retrospectively reviewed. HPV status was determined by immunohistochemical analysis of p16 and/or HPV DNA polymerase chain reaction (PCR). We reclassified TNM stage T0-1 and N0-1 as group A, T2-3 or N2 as B, and T4 or N3 as C. Survival analysis according to 8th AJCC/UICC TNM staging and the modified classification was performed. Results: Of 383 OPSCC patients, 211 were positive for HPV DNA PCR or p16. After exclusion, 184 patients were included in this analysis. Median age was 56 years (range, 31 to 81 years). Most primary tumors were in the palatine tonsil (148 tumors, 80%). The eighth AJCC/UICC TNM classification could not differentiate between stage I and II (p = 0.470) or II and III (p = 0.209). Applying modified grouping, the 3-year overall survival rate of group A was significantly higher than that of group B and C (98% vs. 91%, p = 0.039 and 98% vs. 78%, p < 0.001, respectively). Differentiation between group B and C was marginally significant (p = 0.053). Conclusion: The 8th AJCC/UICC TNM staging system did not clearly distinguish the prognosis of stage II from that of other stages. Including the T2N0-1 group in stage II may improve prognostic stratification.