Adult ; China ; Genes, Viral ; Genotype ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Humans ; Methadone ; therapeutic use ; Middle Aged ; Opioid-Related Disorders ; virology

BACKGROUNDChemotherapy is the most frequently adopted adjuvant therapy of pancreatic ductal adenocarcinoma (PDAC), but the development of drug resistance reduces its effectiveness. Clarification of the mechanism of multidrug resistance (MDR) development in PDAC is needed to improve the therapeutic effect of chemotherapy. This study was aimed to investigate the molecular mechanism of MDR of PDAC and to identify genes associated with MDR development.

METHODSThe gene expression profiles of cell line SW1990 and three drug-selected pancreatic chemoresistant sub-lines, SW1990/5-Fu, SW1990/ADM and SW1990/GEM, were obtained using an oligonucleotide microarray (Affymetrix HG U133 2.0 plus) that contained approximately 38,000 human genes. The microarray results were validated by real-time quantitative polymerase chain reaction and Western blot analysis.

RESULTSThere were 165 genes and expressed sequence tags, some of which have never been linked to drug resistance, that were up- or down-regulated at least 2-fold in all resistant sub-lines when compared with SW1990. According to Gene Ontology annotation, differentially expressed genes related to MDR in pancreatic cancer belong to many functional families and with diverse biological processes. Genes related to antioxidant activity, apoptosis, the cell cycle, signal transduction and intracellular adhesion may undergo epigenetic changes preceding MDR development. A hierarchical clustering was conducted and several interesting clusters were discovered that may be primarily related to cell cycle and developmental regulation. A prediction rule was built from the expression profiles of 117 genes after support vector machine (SVM) analysis, and the prediction result was examined by cytotoxic testing. As a result, a differential gene expression pattern was constructed in multidrug resistant pancreatic cancer cells.

CONCLUSIONSThe findings of this study prove that construction of a chemoresistance prediction rule, based on gene expression patterns, is practical. These data provide new insights into the molecular mechanism of pancreatic cancer MDR development and may be useful for the detection and treatment of MDR in pancreatic cancer patients.

Cell Cycle Proteins ; genetics ; Cell Line, Tumor ; Computational Biology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Gene Expression Profiling ; Glutathione Peroxidase ; genetics ; Glutathione Transferase ; genetics ; Humans ; Microtubule-Associated Proteins ; genetics ; Oligonucleotide Array Sequence Analysis ; methods ; Pancreatic Neoplasms ; drug therapy ; genetics ; Tankyrases ; genetics

OBJECTIVETo study the prevalence of hepatitis C virus (HCV) infection and characteristics on molecular biology related to HCV among patients who were enrolled in a Methadone maintenance clinic in Wuhan.

METHODSSerum samples from 332 injection drug users (IDUs) were obtained and anti-HCV IgG was detected by enzyme linked immunosorbrent assay(ELISA), together with 86 anti-HCV positive specimens genotyped. A reverse transcriptase-polymerase chain reaction (RT-nPCR) assay using conserved primers deduced from the core-envelopel (C-E1) region of the HCV genome was employed to amplify a 474 bp fragment. Phylogenetic analysis of the C-E1 sequences was conducted by direct sequencing of the RT-nPCR products and alignment with determined by nucleotide sequencing followed by composition of a phylogenetic tree.

RESULTSThere were 313 cases (94.3%) appeared positive anti-HCV IgG in the 332 patients from a Methadone maintenance clinic in Wuhan. It was demonstrated that there were four different subtypes of HCV in that clinic in Wuhan, including 6a--71 cases (82.5%), 3b--7 cases (8.2%), 1a--5 cases (5.8%) and 1b--3 cases (3.5%).

CONCLUSIONInfection of 6a genotype HCV was predominant in patients from the Methadone maintenance clinic in Wuhan, followed by HCV 3b, 1a and 1b.

Adult ; Antibodies, Viral ; analysis ; China ; Enzyme-Linked Immunosorbent Assay ; Female ; Genotype ; Hepacivirus ; classification ; genetics ; Humans ; Male ; Methadone ; therapeutic use ; Middle Aged ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Substance Abuse Treatment Centers ; Substance-Related Disorders ; drug therapy ; rehabilitation ; Young Adult

Alzheimer Disease ; complications ; pathology ; therapy ; Amyloid beta-Peptides ; metabolism ; Humans ; Mitochondria ; physiology ; Mitochondrial Diseases ; etiology ; tau Proteins ; metabolism