Rongrong Jing

Cell-Free circulating DNA: a new biomarker for the acute coronary syndrome.

Background: In recent studies, concentrations of cell-free circulating DNA (cf-DNA) have been correlated with clinical characteristics and prognosis in several diseases. The relationship between cf-DNA concentrations and the acute coronary syndrome (ACS) remains unknown. Moreover, no data are available for the detection cf-DNA in ACS by a branched DNA (bDNA)-based Alu assay. The aim of the present study was to investigate cf-DNA concentrations in ACS and their relationship with clinical features. Methods: Plasma cf-DNA concentrations of 137 ACS patients at diagnosis, of 60 healthy individuals and of 13 patients with stable angina (SA) were determined using a bDNA-based Alu assay. Results: ACS patients (median 2,285.0, interquartile range 916.4–4,857.3 ng/ml), especially in ST-segment elevation myocardial infarction patients (median 5,745.4, interquartile range 4,013.5–8,643.9 ng/ml), showed a significant increase in plasma cf-DNA concentrations compared with controls (healthy controls: median 118.3, interquartile range 81.1–221.1 ng/ml; SA patients: median 202.3, interquartile range 112.7–256.1 ng/ml) using a bDNA-based Alu assay. Moreover, we found positive correlations between cf-DNA and Gensini scoring and GRACE (Global Registry of Acute Coronary Events) scoring in ACS. Conclusion: cf-DNA may be a valuable marker for diagnosing and predicting the severity of coronary artery lesions and risk stratification in ACS.

A sensitive method to quantify human cell-free circulating DNA in blood: relevance to myocardial infarction screening.

OBJECTIVES Human cell-free circulating DNA (cf-DNA) derived mainly from cell apoptosis and necrosis can be measured by a variety of laboratory techniques, but almost all of these methods require sample preparation. We have developed a branched DNA (bDNA)-based Alu assay for quantifying cf-DNA in myocardial infarction (MI) patients. DESIGN AND METHODS A total of 82 individuals were included in the study; 22 MI and 60 normal controls. cf-DNA was quantified using a bDNA-based Alu assay. RESULTS cf-DNA was higher in serum compared to plasma and there was a difference between genders. cf-DNA was significantly higher in MI patients compared to the controls. There was no correlation between cf-DNA and creatine kinase-MB (CK-MB), troponin I (cTnI) or myoglobin (MYO). In serial specimens, cf-DNA was sensitive and peaked earlier than cTnI. CONCLUSIONS The bDNA-based Alu assay is a novel method for quantifying human cf-DNA. Increased cf-DNA in MI patients might complement cTnI, CK-MB and MYO in a multiple marker format.

APRIL Induces Tumorigenesis and Metastasis of Colorectal Cancer Cells via Activation of the PI3K/Akt Pathway

A proliferation-inducing ligand (APRIL) is highly expressed in colorectal cancer (CRC) tissues and cell lines. However, the biological functions and precise signals elicited by APRIL in CRC have not been fully understood. Here, we used small interfering RNA to selectively deplete APRIL and to determine its tumorigenic effects in a CRC cell line SW480 both in vitro and in vivo. Knockdown of APRIL in SW480 cells was associated with modulation of cell proliferation as well as reduction of cell migration and invasion in vitro. Moreover APRIL-knockdown SW480 cells displayed markedly inhibited tumor growth and decreased metastasis to the liver in immunodeficient mice upon subcutaneous injection. Importantly, we observed that downregulation of APRIL in SW480 cells resulted in greatly decreased activity of phosphoinositide 3-kinase (PI3K)/Akt pathway. In addition, we observed that recombinant human APRIL mediated activation of the PI3K/Akt pathway in CRC cells resulting in induced expression of important cell cycle proteins and matrix metalloproteinases in a PI3K/Akt dependent manner. This was concurrent with marked cell growth viability as well as increased cell migration and invasion. Together, these compelling data suggest that APRIL-induced tumorigenesis and metastasis of CRC cells may be accomplished through activation of the PI3K/Akt pathway. These findings may lead to a better understanding of the biological effects of APRIL and may provide clues for identifying novel therapeutic and preventive molecular markers for CRC.

Prognostic Value of Free DNA Quantification in Serum and Cerebrospinal Fluid in Glioma Patients

Unlike uniformly truncated DNA released from apoptotic nondiseased cells, free DNA released from dead tumor cells varies in size. Free DNA has been considered as a candidate biomarker for malignant tumors. We obtained serum samples from 70 patients with glioma and 22 healthy volunteers as control and cerebrospinal fluid (CSF) samples from 20 patients with glioma and eight nonneoplastic controls with hydrocephalus or arachnoid cyst and performed preoperative analysis of free DNA concentration and integrity by a quantitative polymerase chain reaction. With two primers sets amplifying short and long free DNA fragments (ALU115 and ALU247), free DNA integrity was determined by ratio of the concentration of ALU247 over ALU115 (ALU247/115). Our results indicate that free DNA integrity and the ratio of long fragments to short fragments may be a useful diagnostic assay for glioma. In summary, the CSF-free DNA concentration and integrity may serve as a new marker for the diagnosis of glioma.

Alu-based cell-free DNA: a potential complementary biomarker for diagnosis of colorectal cancer.

OBJECTIVES Many patients with colorectal cancer (CRC) present with regional or widespread metastasis, partially reflecting limitations of the current screening programs. This study was aimed to find a complementary marker that can improve the diagnostic accuracy. DESIGN AND METHODS Concentrations of cell-free DNA based on Alu (Alu-based CFD) in 31 unselected CRC patients, 30 intestinal polyp patients and 92 healthy individuals were detected by branch DNA (bDNA). Concentrations of carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) were detected by ARCHITECT assay. RESULTS There was significant difference in concentrations of CFD between CRC and intestinal polyp patients or healthy individuals (P<0.0001). There was no statistically significant difference in CFD in different subgroups of CRC patients with respect to gender, age, tumor site and pathologic stage, suggesting that CFD might be an independent marker relative to CEA and CA19-9. There was a significant correlation between pathologic stage and CEA or CA19-9. Although no significant correlation was observed between pathologic stage and CFD, CFD (the area under the receiver operating characteristic curve (AUC)=0.904) seemed to be a better indicator to distinguish CRC patients from intestinal polyp patients as compared with CEA (AUC=0.681) or CA19-9 (AUC=0.651). CFD was more accurate than CEA or CA19-9 in diagnosing CRC. CONCLUSIONS Combination of CFD, CEA and CA19-9 may be a better option for the diagnosis of CRC than any of them used alone. Discrimination CRC from intestinal polyp patients with CFD and staging with CEA and CA19-9 may substantially improve the accuracy CRC diagnosis.

Drug resistance-related microRNAs in hematological malignancies: Translating basic evidence into therapeutic strategies

Systemic chemotherapy has been used as the first-line therapy for patients with hematological malignancies. Despite the enormous progress in anti-tumor efficacy achieved during the last decades, the development of multidrug resistance (MDR) remains a major challenge in the successful treatment of hematological malignancies. Extensive investigations have discovered diverse mechanisms underlying MDR. More recently, increasing evidence demonstrates that miRNAs play a key regulatory role in MDR in hematological malignancies through modulating drug transporter-related proteins, cell cycle-related proteins, drug targets, autophagy, tumor microenvironment, cell survival signaling and apoptosis pathways. Pre-clinical evidence suggests that miRNAs may prove to be an ideal biomarker for predicting drug response or clinical outcomes, thus holding much promise for the development of targeted therapies and personalized medicines for the treatment of hematological malignancies. This review focuses on molecular mechanisms underlying miRNA-mediated chemoresistance in hematological malignancies and discusses evidence of transitional research aiming to bring drug resistance-associated miRNAs into clinical settings.

Cell-free DNA: Characteristics, Detection and its Applications in Myocardial Infarction

Much attention has been focused on cell-free DNA (cf-DNA) and its levels in the plasma, serum and body fluids of pregnant women and patients with cancer or autoimmune disease, and those with severe trauma; however, knowledge of its biology and relationship with myocardial infarction (MI) is still at an early stage. This paper introduces general aspects of the nomenclature, structure, function, release mechanisms and methodology of cf-DNA and summarizes the current literature on concentration variation and regulatory mechanisms in MI.

Tissue-Specific Expression Profiling of Receptor for Advanced Glycation End Products and Its Soluble Forms in Esophageal and Lung Cancer

The receptor for advanced glycation end products (RAGE) interacts with several ligands and is involved in various human diseases. Several splicing forms of the RAGE gene have been characterized, and two general mechanisms are usually responsible for the generation of soluble receptors. However, variants distribution and respective roles in different tumors are not clear. We analyzed RAGE and hRAGEsec mRNA expression in esophageal and lung cancer by RT-polymerase chain reaction. The Agilent clipper 1000 Bioanalyzer using lab-on-a-chip technology was applied to size and quantify the polymerase chain reaction products. Western blotting was performed to measure total soluble RAGE protein levels. The results showed that RAGE and its splice variants increased in esophageal cancers and decreased in lung cancers. We conclude that RAGE presents as a major isoform; soluble RAGE may also play certain roles in esophageal cancer and lung cancer.

Changes of Serum Lactate Dehydrogenase and Potassium Levels Produced by a Pneumatic Tube System

Objective: Errors in laboratory measurements could be derived from many pre-analytical factors. The aim of this study was to assess the influence of the hospital’s pneumatic tube system (PTS) on serum lactate dehydrogenase (LDH) and potassium. Methods: Forty-five healthy blood donors were involved. We studied LDH and potassium delivered to the laboratory by a PTS with different carrier inserts and transport times. In addition, influences of the PTS sending different types of specimens on LDH and potassium were determined. Results: Blood specimens sent via PTS several times or without carrier inserts had statistical changes in LDH; the potassium had a slightly rising trend. Of the under-filled blood draw tubes or lithium heparin tube specimens, changes were caused by the PTS, but there were no effects on pure serum specimens. Conclusions: Many minor shakings derived from the transportation of the PTS inevitably influenced LDH and potassium.

Rheumatoid Factor, Anti-Cyclic Citrullinated Peptide Antibody, C-Reactive Protein, and Erythrocyte Sedimentation Rate for the Clinical Diagnosis of Rheumatoid Arthritis

OBJECTIVE To explore the value of rheumatoid factor (RF), anti-cyclic citrullinated peptide (CCP) antibody, C-reactive protein (CRP), and the erythrocyte sedimentation rate (ESR) in the diagnosis of rheumatoid arthritis (RA). METHODS Using rate nephelometry, chemiluminescence microparticle immunoassay (CMIA), and Westergren sedimentation rate testing, we detected RF, anti-CCP antibody, CRP, and ESR in 134 patients with RA and 50 healthy control individuals. RESULTS We observed significant differences in RF, anti-CCP antibody, CRP, and ESR concentrations between the RA and control groups (P <.01). The sensitivity, specificity and accuracy in the diagnosis of RA were 91.0%, 74.4%, and 87.0%, respectively, for RF; 88.0%, 90.4%, and 88.1%, respectively, for anti-CCP antibody; and 90.2%, 83.3%, and 89.5%, respectively, for the detection of RA via the combination of RF and anti-CCP antibody. CONCLUSION Anti-CCP is more specific than the other parameters we reviewed for the diagnosis of RA. Combined detection of the 4 parameters is beneficial when confirming a diagnosis of RA.