Expression of C/EBPα and PU.1 Gene In MDS and Their Significance Analysis

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4649-4649
Author(s):  
Wencan Ji ◽  
Kaiyang Ding ◽  
Zimin Sun
Keyword(s):  
Polymerase Chain Reaction ◽  
High Risk ◽  
Real Time ◽  
Research Funding ◽  
Low Risk ◽  
Expression Level ◽  
Chain Reaction ◽  
Expression Levels ◽  
Polymerase Chain ◽  
Normal Controls

Abstract Abstract 4649 Objective This study was aimed to investigate the expression levels and significance of CCAAT/enhancer binding protein alpha gene and transcription factor PU.1 gene in myelodysplasyic syndromes (MDS). Methods A real time quantitative reverse transcriptase polymerase chain reaction (Real-time PCR) method was used to detect the expression levels of C/EBPα mRNA in bone marrow mononuclear cells (BMMNCs) of 33 patients with MDS and 14 normal controls, Reverse transcription polymerase chain reaction (RT-PCR) was established for detecting PU.1 mRNA expression levels in BMMNCs of above-mentioned samples. Results The expression of C/EBPα was significantly lower in low-risk and high-risk MDS than those of normal controls (P<0.01, P<0.001, respectively), moreover, high-risk MDS showed lower C/EBPα expression than those of low-risk MDS (P<0.05). The expression level of PU.1 significantly decreased in low-risk MDS, as compared with normal controls and high-risk MDS (P<0.001), but there was no significant difference in expression level of PU.1 between high-risk MDS and normal controls (P>0.05). Conclusion reduced C/EBPα and PU.1 expression level have closely associated with the pathogenesis of MDS, they may be important molecular biological markers of MDS; The degree of down-regulated C/EBPα has closely related to the progression of MDS, the detection of C/EBPα gene expression level may be useful for assessing disease progress and prognostic of patients with MDS. Disclosures: Ding: Anhui Provincial Natural Science Foundation (070413257X): Research Funding; Fund of Anhui Provincial “115” Industrial Innovation Program: Research Funding.

Detection of ERG and MN1 Gene Expression in Adult Patients with De Novo Acute myeloid Leukemia(AML) by Real-Time Quantitative Polymerase Chain Reaction and Its Clinical Significance.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4706-4706
Author(s):  
Bing Xu ◽  
Xutao Guo ◽  
Guoshu Chen ◽  
Xiaoyan Song ◽  
Pengcheng Shi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Clinical Significance ◽  
De Novo ◽  
Risk Group ◽  
Expression Level ◽  
Chain Reaction ◽  
Expression Levels ◽  
Polymerase Chain

Abstract Abstract 4706 Introdution Current study found that certain genetic abnormalities are related to the prognosis of AML, such as ERG and MN1 gene expression. However, little is known about the clinical significance of ERG and MN1 expression level in Chinese patients with AML and and their correlation. Patients and methods A real time quantitative reverse transcriptase polymerase chain reaction method was established for detecting ERG and MN1 gene expression levels in 87 de novo AML patients. Results The median expression levels of ERG and MN1 in AML patient were statistically higher than that in normal control group (P<0.001). ERG gene expression had no correlation with MN1 gene expression (P=0.194) in AML patients. ERG and MN1 gene expressions were equally distributed among the FAB subtypes (P=0.850 and 0.870). Spearman's rank correlation showed that leukocyte counts and lactate dehydrogenase were significantly related to high ERG expression (P=0.005 and 0.032), but not significant correlation was found between hemoglobin and platelet counts. The ERG expression level of cases with middle and low risk group was lower than that of cases with high risk group (P<0.036). There was no significant difference of CR rates in high and low ERG and MN1 expression groups after chemical theropy (P=0.968 and 0.695). During the follow-up of one year, the cumulative relapse rates of high ERG expression groups was significantly higher than that of low ERG expression groups(P=0.039), and high ERG expression cases have a significantly worse OS than low ERG expression cases(P=0.005). Conclusions There isn't a linear correlation between ERG gene expression and MN1 gene expression in AML. Overexpression of ERG gene is a independent prognositic factor for AML. Quantification of the two gene expression together is not more effective to the judgement of prognosis in AML. Disclosures: No relevant conflicts of interest to declare.

Laboratory-Developed L1 Sequencing and Type-Specific, Real-Time Polymerase Chain Reaction for the Detection and Typing of Human Papillomaviruses in Formalin-Fixed, Paraffin-Embedded Tissues

2013 ◽  
Vol 137 (1) ◽  
pp. 50-54 ◽  
Author(s):  
Anne Mills ◽  
Rajeev Balasubramaniam ◽  
Teri A. Longacre ◽  
Christina S. Kong ◽  
Benjamin A. Pinsky
Keyword(s):  
Polymerase Chain Reaction ◽  
High Risk ◽  
Real Time ◽  
Clinical Laboratory ◽  
Low Risk ◽  
Chain Reaction ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain ◽  
Formalin Fixed

Context.—The detection and typing of high-risk and low-risk human papillomavirus (HPV) in archival formalin-fixed, paraffin-embedded tissues by nucleic acid amplification testing is an important adjunct to immunohistochemical staining in evaluation of squamous cell proliferations of the oropharynx, larynx, and anal canal. Objective.—To evaluate semiautomated, xylene-free extraction from formalin-fixed, paraffin-embedded tissues combined with laboratory-developed HPV L1 sequencing and type-specific HPV 6, 11, 16, and 18 real-time polymerase chain reaction for identification and typing of HPV in the clinical laboratory. Design.—We evaluated the adequacy of extraction using β-globin amplification and compared L1 sequencing and real-time polymerase chain reaction methods for typing accuracy using 68 formalin-fixed, paraffin-embedded tissues, including 56 anorectal biopsy or surgical resection specimens and 12 laryngeal papilloma specimens from patients with recurrent respiratory papillomatosis. Results.—Adequate DNA was obtained from 68 of 68 specimens analyzed and all were HPV positive. In 47 cases where L1 sequencing demonstrated that the predominant HPV type was 6, 11, 16, or 18, type-specific, real-time polymerase chain reaction provided concordant results. Sequencing revealed additional low-risk (HPV 40) and high-risk HPV types (HPV 31, 33, 56, and 58) in anorectal specimens, whereas HPV 6 or 11 were the types found in laryngeal papillomas. Conclusion.—Both L1 sequencing and type-specific, real-time polymerase chain reaction are suitable methods for routine HPV testing of formalin-fixed, paraffin-embedded tissues in a clinical laboratory setting.

Human Papillomavirus Infection of the Epididymis and Ductus Deferens: An Evaluation by Nested Polymerase Chain Reaction

2003 ◽  
Vol 127 (11) ◽  
pp. 1471-1474 ◽  
Author(s):  
Alexandr Švec ◽  
Iva Mikyšková ◽  
Ondřej Hes ◽  
Ruth Tachezy
Keyword(s):  
Polymerase Chain Reaction ◽  
High Risk ◽  
Hpv Infection ◽  
Low Risk ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Ductus Deferens ◽  
Hpv Types ◽  
Polymerase Chain ◽  
High Risk Hpv

Abstract Context.—Human papillomaviruses (HPVs) play an important role in the etiology of squamous cell carcinoma of the uterine cervix. The possible role of the male urogenital tract as a reservoir of HPV infection is not fully understood. We inferred from our previous observation of HPV-31 in epididymal tissue in a case of chronic epididymitis that HPV might be commonly present in cases of epididymitis caused by sexually transmitted pathogens. Objective.—To assess the presence of HPV in the epididymis and ductus deferens in nontuberculous epididymitis. Design.—Epididymal samples obtained from 17 patients and epididymal and ductus deferens samples from 5 patients surgically treated for nontuberculous epididymitis were analyzed by nested polymerase chain reaction for the presence of HPV DNA. In positive samples, the HPV type was determined by DNA sequencing. Setting.—Tertiary-care academic hospital and national reference laboratory for papillomaviruses. Results.—Low-risk HPV type 6 and high-risk HPV types 16, 33, 35, 55, and 73 were detected in 7 patients (31%). Neither koilocytes nor dysplastic changes were found in the epididymis and ductus deferens. Conclusion.—Low-risk and high-risk HPV types were detected in the epididymis and ductus deferens of patients with nontuberculous epididymitis. The infection was not accompanied by koilocytic atypia or dysplasia. Our findings support the hypothesis that the male urogenital tract serves as a reservoir of HPV infection.

scholarly journals Comparison of real-time quantitative Polymerase Chain Reaction (PCR) and digital droplet PCR for quantification of hormone membrane receptor FSHR, GPER and LHCGR transcripts in human primary granulosa lutein cells

2020 ◽  
Author(s):  
Samantha Sperduti ◽  
Claudia Anzivino ◽  
Maria Teresa Villani ◽  
Gaetano De Feo ◽  
Manuela Simoni ◽  
...  
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Membrane Receptor ◽  
Reproductive Endocrinology ◽  
Chain Reaction ◽  
Expression Levels ◽  
Polymerase Chain

AbstractBackgroundQuantitative real time polymerase chain reaction (qPCR) and droplet digital PCR (ddPCR) are methods used for gene expression analysis in several contexts, including reproductive endocrinology.ObjectivesHerein, we compared qPCR and ddPCR technologies for gene expression analysis of hormone membrane receptor-encoding genes, such as follicle-stimulating hormone (FSHR), G protein-coupled estrogen (GPER) and choriogonadotropin receptors (LHCGR), as well as the commonly used RPS7 housekeeping gene, in order to identify the most reliable method to be applied for gene expression analysis in the context of human reproduction.MethodsTotal RNA was extracted from human primary granulosa cells of donor patients undergoing assisted reproduction and used for gene expression analysis by qPCR and ddPCR, after finding the optimal annealing temperature.ResultsBoth techniques provided results reflecting the low number of FSHR and GPER transcripts, although ddPCR detected also unspecific transcripts in using RPS7 primers and quantified the low-expressed genes with major accuracy, thanks to its higher reaction efficiency. The absolute FSHR and GPER transcript number was also determined by ddPCR, resulting in 50- to 170-fold lower amount than LHCGR transcripts.ConclusionThese results suggest that ddPCR is the candidate technology for analysis of genes with relatively low expression levels and provides useful insights for characterizing hormone receptor expression levels in the context of reproductive endocrinology.

Effect of TRIM28 on proliferation, apoptosis and histone H3K9 trimethylation in bovine fibroblasts

2019 ◽  
Author(s):  
Panpan Ma ◽  
Xiaxia Man ◽  
Shubao Yang ◽  
Xiaoqing Dong ◽  
Liyan Su ◽  
...  
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Cell Proliferation ◽  
Real Time ◽  
Chain Reaction ◽  
Expression Levels ◽  
H3k9 Trimethylation ◽  
Polymerase Chain ◽  
Apoptosis Gene ◽  
Hp1 Proteins

TRIM28 is a co-repressor, which interacts with HP1 proteins and chromatin repressive complexes leading to gene expression silencing. In this study, after we used Lipofectamine3000-mediating siRNA and restructured p EGFP-IRES2-TRIM28 to transfect bovine fibroblasts, the expression levels of TRIM28, HP1BP3, DNMT, SETDB1, TP53, BAX, Bcl and SOD were detected by real-time polymerase chain reaction (PCR). In addition, the influence of p EGFP-IRES2-TRIM28 on histone H3K9me3 was also observed. In the siRNA transfected group, TRIM28 was down-regulated (P less than 0.01), which inhibited cell proliferation and reduced the level of H3K9me3 expression. In the p EGFP-IRES2-TRIM28 transfected group, TRIM28 was over-expressed (P less than 0.01), and the pro-apoptosis gene BAX was significantly decreased (P ess tahn 0.01), but there was no significant change in other genes. Our results demonstrate that TRIM28 plays an important role in bovine fibroblasts and might be a valuable biomolecule for proliferation, apoptosis, and histone H3K9 trimethylation.

Detection of human papillomavirus types by the polymerase chain reaction and the differentiation between high-risk and low-risk cervical lesions

1992 ◽  
Vol 62 (1) ◽  
pp. 167-171 ◽  
Author(s):  
Marion T. E. Cornelissen ◽  
Tom Bots ◽  
Maarten A. Briët ◽  
Maarten F. Jebbink ◽  
Arie P. H. B. Struyk ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Papillomavirus ◽  
High Risk ◽  
Low Risk ◽  
Cervical Lesions ◽  
Chain Reaction ◽  
Polymerase Chain
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