• Title/Summary/Keyword: digital PCR

Search Result 57, Processing Time 0.087 seconds

Expression of angiogenin, TGF-${\beta}$, VEGF, APEX and TNF-${\alpha}$ in oral squamous cell carcinoma

  • Lee, Ho-Sun;Kim, Kyoung-Won;Kim, Wun-Jae
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
    • /
    • v.32 no.1
    • /
    • pp.8-18
    • /
    • 2006
  • Purpose: The purpose of this study was to verify that the expressions of angiogenin, transforming growth factor-beta(TGF-${\beta}$), vascular endothelial growth factor(VEGF), human apurinic/apyrimidinic endonuclease(APEX) and tumor necrosis factor-alpha(TNF-${\alpha}$) were associated with the tumorigenesis of the oral squamous cell carcinoma(OSCC). Materials and Methods: Fifty-one samples of OSCC and fifteen normal oral mucosae were obtained to analyze the expression levels of above five factors. mRNA expressions were quantified by the quantitative competitive PCR(QC-PCR) method. After 2% agarose gel electrophoresis stained with ethidium bromide, the concentration of mRNA was calculated by a digital image analysis system. The expression levels of angiogenin, TGF-${\beta}$, VEGF, APEX and TNF-${\alpha}$ were compared by unpaired Student's t-tests between cancer and normal tissues. We analyzed statistically to find the cut-off values that would be useful as diagnostic markers, and the linear regression analysis between every two factors of these five factors by SAS system. Results: All of these five factors (angiogenin: P<0.0037, TGF-${\beta}$: P<0.0001, VEGF: P<0.0102, APEX: P<0.0023, TNF-${\alpha}$: P<0.0074) were significantly correlated with OSCC. In the analysis to find the cut-off values for the diagnosis, we could not find any value that had a reasonable sensitivity and specificity. In the linear regression analysis, there were correlations between angiogenin and TNF-${\alpha}$, TGF-${\beta}$ and VEGF, TGF-${\beta}$ and APEX, TGF-${\beta}$ and TNF-${\alpha}$, VEGF and APEX, VEGF and TNF-${\alpha}$, APEX and TNF-${\alpha}$. Conclusion: Our results suggest that not only angiogenin, TGF-${\beta}$, VEGF, APEX and TNF-${\alpha}$ are significantly associated with the tumorigenesis, but also the close relationship between these factors might enhance the tumorigenesis of OSCC. We can not find clinical availability for diagnosis.

Selection of Reference Genes for Real-time Quantitative PCR Normalization in the Process of Gaeumannomyces graminis var. tritici Infecting Wheat

  • Xie, Li-hua;Quan, Xin;Zhang, Jie;Yang, Yan-yan;Sun, Run-hong;Xia, Ming-cong;Xue, Bao-guo;Wu, Chao;Han, Xiao-yun;Xue, Ya-nan;Yang, Li-rong
    • The Plant Pathology Journal
    • /
    • v.35 no.1
    • /
    • pp.11-18
    • /
    • 2019
  • Gaeumannomyces graminis var. tritici is a soil borne pathogenic fungus associated with wheat roots. The accurate quantification of gene expression during the process of infection might be helpful to understand the pathogenic molecular mechanism. However, this method requires suitable reference genes for transcript normalization. In this study, nine candidate reference genes were chosen, and the specificity of the primers were investigated by melting curves of PCR products. The expression stability of these nine candidates was determined with three programs-geNorm, Norm Finder, and Best Keeper. $TUB{\beta}$ was identified as the most stable reference gene. Furthermore, the exopolygalacturonase gene (ExoPG) was selected to verify the reliability of $TUB{\beta}$ expression. The expression profile of ExoPG assessed using $TUB{\beta}$ agreed with the results of digital gene expression analysis by RNA-Seq. This study is the first systematic exploration of the optimal reference genes in the infection process of Gaeumannomyces graminis var. tritici.

Convergence Studies of NO Homeostasis in Cellular Signalling (세포의 신호전달 과정에서 NO 항상성에 관한 융복합 연구)

  • Oh, Hee-Kyun;Do, Eun-Young;Park, Hae-Ryoung
    • Journal of Digital Convergence
    • /
    • v.13 no.12
    • /
    • pp.461-467
    • /
    • 2015
  • Saussurea lappa is known for a variety of physiological activities as a component but has not known to show the effect of the cellular signaling pathway. We investigated the anti-inflammatory effects by Saussurea lappa ethanol extract on the LPS(lipopolysaccharide) induced nitric oxide (NO) production by RAW 264.7 cell line. It shows the expressions of iNOS and COX-2 at the transcriptional level (RT-PCR). The Saussurea lappa ethanol extract showed transcriptional expression levels of pro-inflammatory cytokine TNF- and IL-$1{\beta}$ induced by LPS(lipolysaccharide) in RAW264.7 cell line. Saussurea lappa ethanol extract reduced the LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the RNA levels in a concentration-dependent manner. The finding that ethanol extract of Saussurea lappa has an influence on NO (nitric oxide) homeostasis in the study of the action mechanism on the macrophage-mediated inflammatory reaction was considered in terms of convergence. And it is to provide an important basis for the prevention and treatment of inflammatory diseases in the future.

Effect of Astragali Radix Pharmacocupuncture for Wound Healing of Full-Thickness Skin Wound in Mouse (황기약침이 전층피부손상 마우스의 치유에 미치는 영향)

  • Hong, Chang-Ho;Kang, Kyung-Hwa;Yoon, Hwa-Jung;Yoon, Hyoun-Min;Song, Choon-Ho;Kim, Cheol-Hong
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
    • /
    • v.31 no.1
    • /
    • pp.22-31
    • /
    • 2018
  • Objectives : In this study, we investigated whether Astragali Radix Pharmacocupuncture (ARP) has an effective on the full thickness defect wound healing process of mouse. Methods : A total of 50 mice (ICR mouse, 7 week-old male) were divided into control group and ARP group. A single full thickness skin defect was made on the dorsal side of the each mouse using an 8mm diameter biopsy punch. Control group were treated with 0.2㎖ saline and ARP group were treat with 0.2ml ARP at 8 points around the wound every three days total 4 times during the experimental period. The change in wound size, contraction rate, healing rate, and epithelization rate was measured by digital images taken on days 3, 6, 9, and 13, and evaluated using a digital image analysis program. Tissues were collected for histological analysis, RT-PCR, and Western blot on days 3, 6, 9, and 15. Results : The results are as follows. ARP group accelerated the rate of wound contraction, wound healing and epithelization compared to the control group. ARP group showed the decrease of inflammatory cells in early inflammatory phase compared to the control group. ARP group upregulated PECAM-1 mRNA and protein expression compared to the control group. ARP group inhibited the scar width and area compared to the control group. Conclusions: ARP showed positive effects on wound healing through the inhibition of inflammatory reaction and increase of PECAM-1 expression related to the wound healing process.

The Effect of Oral Health Care Program Based on Motivational Interviewing (동기면담을 적용한 구강 관리 프로그램의 효과)

  • Han, Ye-Seul;Bae, Hyun-Sook;Cho, Young-Sik
    • Journal of dental hygiene science
    • /
    • v.14 no.3
    • /
    • pp.287-294
    • /
    • 2014
  • The purpose of this study is to use basic data of dental hygiene curriculum with a new technique called motivational interviewing of communication skill to demonstrate the effectiveness of the method of oral health care. In this study, we performed oral health care program that has been made in dental hygiene department to university students. It was assigned to the control group and 66 and 32 experimental group based on the date of the first visit time. It conducted motivational interviewing of a total of three times in the experimental group. The analytical results of the measurements obtained in the oral examination and questionnaires. The results were as follows: The experimental group measured value was reduced after the intervention compared to before the PSR to evaluate the state of periodontal, gingival index, calculus index, plaque control record (PCR; O'Leary plaque index), simple plaque scor of Quantitative Light Induced Fluorescnece Digital measurement value (p<0.05). Experimental group decreased more and more the result of changes in the reduction of the average of the PCR. But control group was reduced to 3 weeks and increased back to the middle 16 weeks. There was also support interaction between the measurement point and the groups (p<0.05). Re-visit adherence of fit, 12.1% in the control group, the experimental group was 43.7% in the period of participation in the oral health care program. Thus, visit adherence of the experimental group was higher. In this study, a group that has motivational interviewing, It was able to confirm the improvement of oral health state. Discussion of the motivational interviewing can be applied to oral health care program.

Identification and characterization of the phytocystatin family from Brassica rapa

  • Hong, Joon-Ki;Hwang, Jung-Eun;Park, Tae-Ho;Zang, Yun-Xiang;Lee, Sang-Choon;Kwon, Soo-Jin;Mun, Jeong-Hwan;Kim, Hyun-Uk;Kim, Jin-A;Jin, Mi-Na;Kim, Jung-Sun;Lee, Soo-In;Lim, Myung-Ho
    • Journal of Plant Biotechnology
    • /
    • v.35 no.4
    • /
    • pp.317-327
    • /
    • 2008
  • Phytocystatins, which are inhibitors of plant cysteine peptidases, are involved in the regulation of protein turnover and in the defense against insect pests and pathogens. Extensive searches in the Brassica rapa genome allowed the prediction of at least eight different phytocystatin genes on seven chromosomes in the B. rapa genome. Structure comparisons based on alignments of the all BrCYS ($\underline{B}$. $\underline{r}apa$ $phyto{\underline{cys}}tatin$) proteins using the CLUSTALW program revealed conservation of the three consensus motifs known to interact with the active site of cysteine peptidases. According to the phylogenetic analysis based on the deduced amino acid sequences, the eight BrCYS proteins were divided into several clusters related to the orthologous phytocystatin. The predicted three-dimensional structure models of the eight BrCYS proteins demonstrate that all of these proteins are similar to the reported crystal structure of oryzacystatin-I (OC-I). Digital northern and RT-PCR analyses indicated that the eight BrCYS genes exhibit different expression patterns in B. rapa tissues and respond differently to abiotic stimuli. The differences in gene structure and expression between the eight BrCYS genes suggest that these proteins may play diverse physiological roles in B. rapa and may interact with cysteine peptidases through different mechanisms.

Analysis of the Gene Expression by Laser Capture Microdissection (III) -Microarray Analysis of the Gene Expression at the Mouse Uterine Luminal Epithelium of the Implantation Sites during Apposition Period- (Laser Capture Microdissection을 이용한 유전자 발현 연구 (III) -생쥐 착상 부위 자궁 내강상피 조직에서 배아 병치 기간 동안 일어나는 유전자 발현에 관한 Microarray 분석-)

  • Yoon, Se-Jin;Jeon, Eun-Hyun;Park, Chang-Eun;Ko, Jung-Jae;Choi, Dong-Hee;Cha, Kwang-Yul;Kim, Se-Nyun;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.29 no.4
    • /
    • pp.323-335
    • /
    • 2002
  • Object: The present study was accomplished to obtain a gene expression profile of the luminal epithelium during embryo apposition in comparison of implantation (1M) and interimplantation (INTER) sites. Material and Method: The mouse uterine luminal epithelium from IM and INTER sites were sampled on day 4.5 (Day of vaginal plug = day 0.5) by Laser Captured Microdissection (LCM). RNA was extracted from LCM captured epithelium, amplified, labeled and hybridized to microarrays. Results from microarray hybridization were analyzed by Significance Analysis of Microarrays (SAM) method. Differential expression of some genes was confirmed by LCM followed by RT-PCR. Results: Comparison of IM and INTER sites by SAM identified 73 genes most highly ranked at IM, while 13 genes at the INTER sites, within the estimated false discovery rate (FDR) of 0.163. Among 73 genes at IM, 20 were EST/unknown function, and the remain 53 were categorized to the structural, cell cycle, gene/protein expression, immune reaction, invasion, metabolism, oxidative stress, and signal transduction. Of the 24 structural genes, 14 were related especially to extracellular matrix and tissue remodeling. Meanwhile, among 13 genes up-regulated at INTER, 8 genes were EST/unknown function, and the rest 5 were related to metabolism, signal transduction, and gene/protein expression. Among these 58 (53+5) genes with known functions, 13 genes (22.4%) were related with $Ca^{2+}$ for their function. Conclusions: Results of the present study suggest that 1) active tissue remodeling is occurring at the IM sites during embryo apposition, 2) the INTER sites are relatively quiescent than IM sites, and 3) the $Ca^{2+}$ may be a crucial for apposition. Search for human homologue of those genes expressed in the mouse luminal epithelium during apposition will help to understand the implantation process and/or implantation failure in humans.