• Title/Summary/Keyword: RT-PCA

Search Result 14, Processing Time 0.048 seconds

Evaluation of Combined Quantification of PCA3 and AMACR Gene Expression for Molecular Diagnosis of Prostate Cancer in Moroccan Patients by RT-qPCR

  • Maane, Imane Abdellaoui;El Hadi, Hicham;Qmichou, Zineb;Al Bouzidi, Abderrahmane;Bakri, Youssef;Sefrioui, Hassan;Dakka, Nadia;Moumen, Abdeladim
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.17 no.12
    • /
    • pp.5229-5235
    • /
    • 2016
  • Prostate cancer (PCa) remains one of the most widespread and perplexing of all human malignancies. Assessment of gene expression is thought to have an important impact on cancer diagnosis, prognosis and therapeutic decisions. In this context, we explored combined expression of PCa related target genes AMACR and PCA3 in 126 formalin fixed paraffin embedded prostate tissues (FFPE) from Moroccan patients, using quantitative real time reverse transcription-PCR (RT-qPCR). This quantification required data normalization accomplished using stably expressed reference genes (RGs). A panel of twelve RG was assessed, data being analyzed using GenEx V6 based on geNorm, NormFinder and statistical methods. Accordingly, the hnRNP A1 gene was identified and selected as the most stably expressed RG for reliable and accurate gene expression quantification in prostate tissues. The ratios of both PCA3 and AMACR gene expression relative to that of the hnRNP A1 gene were calculated and the performance of each target gene for PCa diagnosis was evaluated using receiver-operating characteristics. PCA3 and AMACR mRNA quantification based on RT-qPCR may prove useful in PCa diagnosis. Of particular interesting, combining PCA3 and AMACR quantification improved PCa prediction by increasing sensitivity with retention of good specificity.

Determination of Tyrosinase mRNA in Melanoma by Reverse Transcription-PCR and Optical Mirror Resonance Biosensor

  • Taeboo Choe;Park, Inchul;Seokil Hong
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.7 no.4
    • /
    • pp.212-215
    • /
    • 2002
  • Tyrosinase transcript In the blood Is known as the marker of malignant melanoma and it has been often determined by using reverse transcription-polymerase chain reaction (RT-PCA) . However, after the PCR process, the quantification of amplified CDMA by the gel electrophoresis is not reliable and time-consuming. for this reason, we tried to quantify the PCR product using a cuvette-type biosensor, where the oligonucleotide probe was immobilized on the cuvette surface and the single strand CDMA, the denatured PCH product, was then hybridized onto the immobilized probe to give a response signal. The response was Immediate and takes 15 min to obtain a stable signal. The biosensor was much more sensitive comparing to the gel electrophoresis method. The quantification of PCR product using a cuvette-type biosensor was feasible and rapid.

Rhus Trichocarpa Suppresses IgE-mediated Allergic Response In Vitro and In Vivo (개옻나무 추출물의 IgE 매개성 알레르기 반응 억제 효과 및 기전)

  • Lim, Hannah;Kim, Young Mi
    • Korean Journal of Pharmacognosy
    • /
    • v.44 no.2
    • /
    • pp.118-124
    • /
    • 2013
  • Mast cells is the key effector cells for IgE-mediated allergic responses. In this study, we investigated whether Rhus trichocarpa extract (RT) inhibited IgE-mediated allergic responses in mast cells and an allergic animal model. We further tried to find its mechanism of action in mast cells. We found that RT suppressed antigen-stimulated degranulation and production of TNF-${\alpha}$ and IL-4 in rat basophilic leukemia (RBL)-2H3 mast cells and bone marrow-derived mast cells (BMMC), as well as IgE-mediated passive cutaneous anaphylaxis (PCA) in mice. As the mechanism of action of RT, it inhibited the activation of spleen tyrosine kinase (Syk), a pivotal signaling molecule for activation of mast cells and that of LAT, a downstream adaptor molecule of Syk in $Fc{\varepsilon}RI$-mediated signal pathways. RT also suppressed the activation of mitogen-activated protein (MAP) kinases and Akt. The current results demonstrated for the first time that RT has the anti-allergic effect through inhibiting degranulation and secretion of cytokines by suppression of Syk in antigen-stimulated mast cells. Therefore, RT might be useful for allergic diseases.

Expression and Significance of Microsomal Prostaglandin Synthase-1 (mPGES-1) and Beclin-1 in the Development of Prostate Cancer

  • Xu, Lu-Wei;Qian, Ming;Jia, Rui-Peng;Xu, Zheng;Wu, Jian-Ping;Li, Wen-Cheng;Huang, Wen-Bin;Chen, Xing-Guo
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.4
    • /
    • pp.1639-1644
    • /
    • 2012
  • The aim of this study was to investigate the expression and significance of microsomal prostaglandin synthase-1 (mPGES-1) and Beclin-1 in the development of prostate cancer (PCa). Immunohistochemistry was performed on paraffin-embedded sections with rabbit polyclonal against mPGES-1 and Beclin-1 in 40 PCa, 40 benign prostatic hyperplasia (BPH) and 10 normal prostate specimens for this purpose. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied for mRNA expression of mPGES-1 and Beclin-1, while MTT assays were used to ascertain the best working concentration of the mPGES-1 inhibitor (CAY10526). The effect of CAY10526 treatment on expression of Beclin-1 in DU-145 cells was studied using Western blot analysis. Localization of Beclin-1 and mPGES-1 was in endochylema. Significant differences in expression was noted among PCa, BPH and normal issues (P<0.05). Beclin-1 expression inversely correlated with mPGES-1 expression in PCa tissue (P<0.05). CAY10526 could significantly block mPGES-1 expression and the proliferation of DU-145 cells (P<0.05), while increasing Beclin-1 levels (P<0.05). Overexpression of mPGES-1 could decrease the autophagic PCa cell death. Inhibiting the expression of mPGES-1 may lead to DU-145 cell death and up-regulation of Beclin-1. The results suggest that inhibition of mPGES-1 may have therapeutic potential for PCa in the future.

The Transcranial Doppler(TCD) Assesment of Vertebrobasal Vascular Blood Flow in Cerebral Infarction (Transcranial Doppler를 이용(利用)한 급성기뇌경색(急性期腦硬塞)이 추골기저동맥계(椎骨基底動脈係) 혈관(血管)의 혈류속도(血流速度)에 미치는 영향(影響))

  • Ahn, Teck-Won;Kim, Yoon-Sik
    • The Journal of Internal Korean Medicine
    • /
    • v.21 no.5
    • /
    • pp.805-811
    • /
    • 2000
  • Objectives : To determine the effects on blood flow of vertebrobasal vascular system in cerebral infarction, Methods : We altered 33 normal patients and 12 patients who were diagnosed cerebral infarction on Rt. middle cerebral artery(MCA) region and 8 patients who were diagnosed cerebral infarction on Lt, MCA region, and measured the mean velocity, systolic to diastolic rate(SD rate), asymmetrical index(A/I) by TCD. Results : The mean velocity of posterior cerebral artery(PCA), vertebral artery was increased in same direction as infarcted site and the mean velocity of basal artery was more increased than control, and the SD rate of PCA, vertebral artery, basal artery was larger than control. The A/l of PCA, vertebral artery was revealed that mean velocity of vertebrobasal vascular system is increase the same direction as infarcted area. Conclusion : TCD examination within 7 days(acute stage) after stroke can help to predict the infarcted direction.

  • PDF

The Functional Relevance of Prepro-melanin Concentrating Hormone (pMCH) to Skin Color Change, Blind-side Malpigmentation and Feeding of Oliver Flounder Paralichthys olivaceus

  • Kang, Duk-Young;Kim, Hyo-Chan;Kang, Han-Seung
    • Fisheries and Aquatic Sciences
    • /
    • v.17 no.3
    • /
    • pp.325-337
    • /
    • 2014
  • To assess the functional structure of prepro-melanin-concentrating hormone (pMCH), we isolated and cloned pMCH (of-pMCH) mRNA from the brain of the olive flounder, Paralichthys olivaceus, and compared its amino acid sequence with those from other animals. In addition, to examine whether activation of the brain of-pMCH gene is influenced by background color, density, and feeding, we compared pMCH mRNA activities against different background colors (bright and dark) and at different densities (100% PCA and 200% PCA). To examine whether the pMCH gene is related with malpigmentation of blind-side skin and appetite, we compared pMCH gene expression between ordinary and hypermelanic flounders, and between feeding and fasting flounders. The of-pMCH cDNA was 405 bp in the open reading frame [ORF] and encoded a protein of 135 amino acids; MCH was 51 bp in length and encoded a protein of 17 amino acids. An obvious single band of the expected size was obtained from the brain and pituitary by RT-PCR. In addition, of-pMCH gene activity was significantly higher in the bright background only at low density (< 100% PCA) making the ocular skin of fish whitening, and in ordinary fish. However, the gene activity was significantly decreased in dark background, at high density (>200% PCA), and in hypermelano fish. These results suggest that skin whitening camouflage of the flounder is induced by high MCH gene activity, and the density disturbs the function of background color in the physiological color change. Moreover, our data suggest that a low level of MCH gene activity may be related to malpigmentation of the blind-side skin. In feeding, although pMCH gene activity was significantly increased by feeding in the white background, the pMCH gene activity in the dark background was not influenced by feeding, indicating that the MCH gene activity increased by feeding can be offset by dark background color, or is unaffected by appetite. In conclusion, this study showed that MCH gene expression is related to ocular-skin whitening camouflage and blind-skin hypermelanosis, and is influenced by background color and density.

The Effect of Moutan Cortex on Pro-inflammatory Cytokines through NF-κB & MAPKs pathway in HMC-l (목단피의 NF-κB와 MAPKs 억제를 통한 Pro-inflammatory Cytokines 활성에 미치는 효과)

  • Park, Joo-Ho;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
    • /
    • v.22 no.2
    • /
    • pp.1-18
    • /
    • 2009
  • Objective : Moutan Cortex (the root bark of Paeonia suffruticosa Andr.) is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how MC(Moutan Cortex) affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted MC on the mast cell-mediated inflammatory responses. Method : We observed the effect of MC on compound 48/80-induced histamine release of rat peritoneal mast cells and the effect of administering MC on PCA in rat. We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of MC. The TNF-$\alpha$ protein levels were analysised by Western blot. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. NF-$\kappa$B, phospho-I$\kappa$B and MAPKs were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by luciferase assay. Result : 1. Enzyme immunoassay indicated that MC suppressed histamine secretion of rat peritoneal mast cells. 2. In PCA dependent on IgE, MC had anti-allergic effect of the internal surface of rat skin. 3. Western blot indicated that MC decreased TNF-$\alpha$ protein levels. 4. ELISA indicated that MC decreased TNF-$\alpha$, IL-6 but MC had no significant effect on IL-8 in HMC-1 cells. 5. RT-PCR indicated that MC decreased TNF-$\alpha$, IL-8 but MC had no significant effect on IL-6 in HMC-l cells. 6. Western blot indicated that MC suppressed the induction of MAPKs, NF-$\kappa$B & phospho-I$\kappa$B activity in HMC-1 cells. 7. Luciferase assay indicated that MC suppressed the PMA plus A23187-induced NF-$\kappa$B promoting activityin HMC-1 cells. Conclusion : In this study, we have found that MC is an inhibitor of NF-$\kappa$B, MAPKs & cytokines on the mast cell-mediated inflammatory responses.

  • PDF

A study on stroke patient's characteristics and damage (뇌혈관 손상환자의 특성 및 장애에 대한 연구)

  • Choi, Young-Deog
    • Journal of Korean Physical Therapy Science
    • /
    • v.5 no.4
    • /
    • pp.785-794
    • /
    • 1998
  • We have made a survey of 40 patients in the university hospitals and oriental medical centers in Seoul from Sep. 1, 1997 to Mar. 1, 1998. We sampled 25 of them and the result shows that there were 12 MCA damaged patients(48%), 5 SAH(20%), 5 ACA(20%), 2 PCA (8%), 1 PCOA(4%). The number of MCA patients were the most. 1. As the cause of each disease, 4 of the 12 MCA damaged patients(33.35%) have infarction and cerebral hemorrhage, 2 of 5 SAH patients(40%) have cerebral hemorrhage and head injury, 3 ACA damaged patients have cerebral hemorrhage. 11 of 25 brain bloodvessel damaged patients(44%) were hemorrhage patients. 2. Rt. hemiparesis was the main symptom of 6 of 12 MCA damaged patients(50%) and 3 of 5 SAH patients(60%), and the main symptom of 3 of 5 ACA patients(60%) was Lt. hemiparesis. The main symptom of 13 of 25 brain bloodvessel damaged patients(52%) was Lt. hemiparesis 11 of them(44%) Rt. hemiparesis, and 1 of them(8.3%) Quadriplegia. 3. Language was the most well preserved function. 12 MCA damaged patients could understand language. 4. Retraction of shoulder girdle, among VIE flexor synergy, was the most frequent element because 9 of 12 MCA damaged patients had it. Among VIE flexor synergy, 5 SAH patient's most frequent synergy was Elbow flexion because all of them had it. All of 5 ACA damaged patients have shoulder girdle elevation, shoulder joint, hyperextension, abduction, and external rotation among VIE flexor synergy. 5. 7 of 12 MCA damaged patients(58.3%) were stereognosis handicapped patients, 3 of 5 SAH patients(60%) have handicap of position sense, light touch, and temperature, 3 of 5 ACA patients(60%) have position handicap. 13 of brain bloodvessel damaged patients(52%) have light touch handicap. 6. 8 of MCA damaged patients(66.7%) have facial palsy, 4 of SAH damaged patients(80%) have memory and action decline, and 3 of ACA damaged patients(60%) have action decline and facial palsy. The problem of Hemiplegia is very extensive from muscle weakness, atrophy, or deformation to psychical problems. Therefore physical therapists should have sufficient interest in psychological handicap as well as physical handicap as they deal with adult hemiplegia.

  • PDF

Radical Scavenging Activities of Phenolic Compounds Isolated from Mulberry (Morus spp.) Cake

  • Shin, Young-Woong;Lee, Seong-Kwon;Kwon, Yun-Ju;Rhee, Soon-Jae;Choi, Sang-Won
    • Preventive Nutrition and Food Science
    • /
    • v.10 no.4
    • /
    • pp.326-332
    • /
    • 2005
  • A methanol extract of mulberry cake prepared from mulberry fruits (Morus spp.) was shown to have strong scavenging activities against DPPH, superoxide and hydroxyl radicals. Eleven phenolic compounds were isolated from the mulberry cake by a combination of Diaion HP-20, silica gel (or polyamide), Sephadex LH-20 column chromatographies, preparative HPLC and TLC. Their chemical structures were characterized as procatechuic acid (PCA), caffeic acid (CA), cyanidin 3-O-$\beta$-D-glucopyranoside (CyG) and cyanidin $3-O-\beta­D-rutinoside$ (CyR), rutin (RT), isoquercitrin (IQT), astragalin (AG), quercetin (QT), morin (MR), di-hydroquercetin (DHQ), and 4-prenylmoracin (PM) by spectral analysis and the published data. Most of the phenolic constituents were effective scavengers of DPPH, superoxide and hydroxyl radicals, and especially caffeic acid and 4-prenylmoracin showed potent superoxide and hydroxyl radical scavenging activity, in which their activities were higher than that of the well-known antioxidant, BHT (p< 0.05). Dehydroquercetin and quercetin also exhibited strong superoxide and hydroxyl radical scavenging activities. These results suggest that mulberry cake containing antioxidant phenolic compounds may be useful as natural antioxidants in functional foods and cosmetics.

Ethyl Acetate Extract of Korean Rice Wine Lees Inhibits IgE-Mediated Degranulation in Rat Basophilic Leukemia RBL-2H3 Cells and Passive Cutaneous Anaphylaxis in Mice (주박 에틸아세테이트 추출물의 항알러지 효과)

  • Kang, Yeo-Jin;Park, Sae-Jin;Bae, Ki-Ho;Yoo, Jung-Min;Pyo, Hyeong-Bae;Choi, Ji-Ho;Kim, Tack-Joong
    • Journal of Life Science
    • /
    • v.21 no.10
    • /
    • pp.1364-1369
    • /
    • 2011
  • Mast cells, the central effector cells involved in the allergic response, release histamine, arachidonic acid, and proinflammatory cytokines. We investigated the effect of the ethyl acetate fraction (EA), derived from Korean rice wine lees, on RBL-2H3 cell degranulation and passive cutaneous anaphylaxis in an animal model. The EA fraction suppressed the release of beta-hexosaminidase, a marker of degranulation, and the mRNA expression of interleukin-3 (IL-3) and IL-13. EA also successfully suppressed the passive cutaneous anaphylaxis (PCA) reaction in mice in a dose-dependent manner. These results suggest that EA can inhibit mast cell degranulation through the inhibition of IL-3 and IL-13 mRNA expression, and that EA may potentially serve as an anti-allergic agent.