Cell Line ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Interleukin-1 ; administration & dosage ; pharmacology ; P-Selectin ; metabolism

ObjectiveTo investigate the loss of heterozygosity (LOH) on 6q in bladder tumor.MethodsD6S404 and D6434 microsatellite markers near 6q21 were tested by PCR-SSLP-stain method on tumor DNA from 31 cases of bladder tumor.ResultsAmong these 31 cases of bladder tumor,LOH was detected in tumor tissues on site for D6S404 (35.5%) and D6S434(22.6%).ConclusionOne or more tumor suppressor gene near 6q21 maybe relevant for the development of bladder tumor.

Lack of proper study models has brought difficulties in the study of the mechanism of viral infection, life cycle and pathogenic mechanism of hepatitis C virus (HCV) and also become the major obstacles in development of efficient vaccine and new drugs for hepatitis C. In recent years, the establishment of robust HCV cell culture infection system and HCV transgenic animal provide powerful tools for the analysis of host virus interactions, which facilitate the discovery of antiviral drugs and vaccines for this important human pathogen.
Animals ; Animals, Genetically Modified ; virology ; Disease Models, Animal ; Genotype ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Mice ; Mice, Transgenic ; virology

OBJECTIVETo investigate the effects of hepatitis C virus (HCV) on transcription regulation genes of host cells by gene chip assays in cultured cells with intact HCV genome.

METHODSHuh-7 hepatoma cells were cultured and infected with in vitro constructed HCV. The total RNAs, proteins and cell culture supernatants of HCV infected cells and control cells were isolated. Proteins and cell culture supernatants were used to detect the HCV replication and protein expression in cell culture system. The HCV protein expression was detected with Western blotting. Released HCV from infected cells was analyzed by real-time fluorescence quantitative PCR. Total RNA was qualified using 10 g/L agarose gel electrophoresis. cRNA was synthesized, fluorescence labeled and purified, then hybridized with Agilent oligo microarray (20173 probes). Differential expression of genes related to transcription in cell culture system was analyzed.

RESULTHCV was positive in cell culture supernatants and HCV protein expression was also positive according to Western blotting results. Eleven up-regulated and 11 down-regulated genes related to transcription were found after Agilent gene chip screening.

CONCLUSIONIntact hepatitis C virus cell culture system provides an useful tool for study on the affects of HCV infection on transcription regulation genes in host cells.

Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genome, Viral ; Hepacivirus ; genetics ; growth & development ; Hepatocytes ; virology ; Humans ; Liver Neoplasms ; genetics ; pathology ; virology ; Transcription, Genetic

OBJECTIVETo investigate the expression of keratin 8 (K8) in carbon tetrachloride (CCl(4))-induced liver injury of mice.

METHODSForty ICR mice were divided into four groups. CCl(4) 300 microl/kg body weight in olive oil was injected intraperitoneally for 0, 2, 4, 6 weeks in group A, B, C and D, respectively. Mice were sacrificed 3 d after the last injection and then the vital organs were collected and weighed. RT-PCR and immunohistochemistry methods were used to analyze the expression of keratin 8 in the liver.

RESULTSThe ratios of liver and body weight were increased significantly after administration of CCl(4), which were 5.60 %, 6.87%, 7.83 % and 7.76% at 0, 2, 4 and 6 weeks after injection, respectively. The expression of K8 was increased at the 2 w, 4 w and 6 w after CL(4) administration.

CONCLUSIONThe expression of K8 is positively correlated with the liver injury induced by CCl(4). The accumulation of K8 may be involved in the mechanism of liver injury.

Animals ; Carbon Tetrachloride ; Chemical and Drug Induced Liver Injury ; metabolism ; Female ; Keratin-8 ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; RNA, Messenger ; genetics ; metabolism

OBJECTIVETo perform a Meta-analysis on peginterferon with interferon in treatment of HIV patients coinfected with refractory genotype HCV.

METHODSA literature search of Medline was conducted to identify eligible randomized controlled trials. Meta analysis was conducted to evaluate peginterferon and interferon in treatment of coinfected HCV genotype 1 or 4 in HIV patients.

RESULTSix trials of 88 matched the selection criteria. Total 1,131 patients with coinfection of HCV genotype 1 or 4 and HIV were included. Sustain viral response was higher in patients treated with peginterferon plus ribavirin compared with that of interferon plus ribavirin (26 % compared with 8 %) or peginterferon alone (26 % compared with 13 %). Severe adverse effects and withdrawal rates were similar for patients treated with peginterferon and patients treated with interferon.

CONCLUSIONPeginterferon plus ribavirin in treatment of patients with coinfection of genotype 1 or 4 HCV and HIV can achieve higher sustain viral response and the likelihoods of serious adverse effects and withdrawal rates are similar to other therapies.

Adult ; Antiviral Agents ; administration & dosage ; Drug Therapy, Combination ; Female ; Genotype ; HIV Infections ; complications ; drug therapy ; immunology ; Hepacivirus ; classification ; genetics ; Hepatitis C, Chronic ; complications ; drug therapy ; virology ; Humans ; Interferon-alpha ; administration & dosage ; Male ; Polyethylene Glycols ; administration & dosage ; Randomized Controlled Trials as Topic ; Recombinant Proteins ; Ribavirin ; administration & dosage

OBJECTIVETo study the inhibitory effect of T-2 toxin on the expression of aggrecan and collagen II in chondrocytes and the protection of selenium against this effect.

METHODSHuman chondrocytes cultured in vitro were treated with T-2 toxin at different concentrations for varied time periods (1-5 days), and the cell viability was measured by MTT assay. Aggrecan expression was detected by toluidine blue staining and collagen II expression by immunostaining using monoclonal antibody of collagen. Aggrecan and collagen II mRNA expressions were measured by semiquantitative RT-PCR.

RESULTST-2 toxin dose- and time-dependently affected chondrocyte viability within the concentration range of 0.001-2 mg/L, the prolonged treatment time further enhanced the dose dependence of the inhibitory effect. T-2 toxin lowered aggrecan and collagen II synthesis in the chondrocytes and reduced their mRNA expressions. Selenium could partly attenuate the inhibitory effects of T-2 toxin on aggrecan mRNA expression, but showed no such effect against T-2-induced collagen II expression.

CONCLUSIONT-2 toxin can obviously inhibit aggrecan and collagen II synthesis in human chondrocytes, and selenium can partly antagonize the inhibitory effects of T-2 toxin on aggrecan.

Aggrecans ; biosynthesis ; genetics ; Cells, Cultured ; Chondrocytes ; cytology ; metabolism ; Collagen Type II ; biosynthesis ; genetics ; Dose-Response Relationship, Drug ; Fetus ; Humans ; Protective Agents ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Selenium ; pharmacology ; T-2 Toxin ; toxicity

OBJECTIVETo evaluate the relationship of expressions of nucleoside diphosphate kinase (nm23) and proliferating cell nuclear antigen (PCNA), as well as apoptosis, with the prognosis of HCC patients by analyzing their pathological and clinical data.

METHODSThe expressions of nm23 and PCNA were analyzed by immunohistochemistry and the apoptotic phenomena were detected by TUNEL technique in the liver samples from 43 HCC tissues, 39 para-neoplastic tissues, and 10 normal tissues. The mean apoptosis index (AI) and proliferative index (PI) in individual sample were calculated.

RESULTSAs shown by the detection, 32.6% of carcinomas had negative nm23 signal in tumor tissues, whereas all para-neoplastic and normal tissues had positive nm23. The AI in nm23 positive HCC was significantly higher than that in nm23 negative one, with statistical difference (P<0.05). Furthermore, the expressions of nm23, and the values of AI and PI were contrastively analyzed with some main pathological and clinical data of HCC. It revealed that HCC with extrahepatic metastasis showed remarkable correlation with the negative nm23 (P=0.013) and higher PI values of HCC (P=0.015). The disease-free survival in HCC patients with negative nm23 expression was significantly poorer than that in patients with positive nm23 expression.

CONCLUSIONSThese data suggest that expressions of nm23 protein in tumor tissues are correlated with occurrences of metastasis and length of survival of the HCC patients, which may be an indicator for their prognosis.

Adult ; Aged ; Apoptosis ; Biomarkers, Tumor ; biosynthesis ; Carcinoma, Hepatocellular ; enzymology ; mortality ; pathology ; Case-Control Studies ; Cell Proliferation ; Disease Progression ; Disease-Free Survival ; Female ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Kaplan-Meier Estimate ; Liver ; enzymology ; pathology ; Liver Neoplasms ; enzymology ; mortality ; pathology ; Male ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; biosynthesis ; Neoplasm Metastasis ; Proliferating Cell Nuclear Antigen ; biosynthesis

OBJECTIVETo investigate the effects of T-2 toxin on expressions of Fas, p53, Bcl-xL, Bcl-2, Bax and caspase-3 on human chondrocytes.

METHODSHuman chondrocytes were treated with T-2 toxin (1-20 ng/ml) for 5 d. Fas, p53 and other apoptosis-related proteins such as Bax, Bcl-2, Bcl-xL, caspase-3 were determined by Western blot analysis and their mRNA expressions were determined by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSIncreases in Fas, p53 and the pro-apoptotic factor Bax protein and mRNA expressions and a decrease of the anti-apoptotic factor Bcl-xL were observed in a dose-dependent manner after exposures to 1-20 ng/ml T-2 toxin, while the expression of the anti-apoptotic factor Bcl-2 was unchanged. Meanwhile, T-2 toxin could also up-regulate the expressions of both pro-caspase-3 and caspase-3 in a dose-dependent manner.

CONCLUSIONThese data suggest a possible underlying molecular mechanism for T-2 toxin to induce the apoptosis signaling pathway in human chondrocytes by regulation of apoptosis-related proteins.

Apoptosis ; drug effects ; Base Sequence ; Blotting, Western ; Caspase 3 ; genetics ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; metabolism ; DNA Primers ; genetics ; Gene Expression ; drug effects ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects ; T-2 Toxin ; toxicity ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism ; bcl-X Protein ; genetics ; metabolism ; fas Receptor ; genetics ; metabolism

BACKGROUNDTo study the full length L and M sequence of Hantavirus Q32 strain gene and explore its molecular characters.

METHODSThe L and M segment cDNA of Hantavirus Q32 strain was amplified by RT-PCR. The purified PCR products were sequenced directly or cloned into pGEM-T Vector and then sequenced.

RESULTSThe L genome segment of Q32 virus was found to be 6533 nucleotides in length. One large open reading frame was found located at bases 38 to 6493. This was predicted to encode an L protein 2151 amino acids in length with a molecular mass of 2.46 x 10(5). The M genome segment was 3616 nucleotides in length. One open reading frame was located at bases 41 to 3488. This was predicted to encode an M protein 1135 amino acids with a molecular mass of 1.26 x 10(5).

CONCLUSIONThe nucleotides sequence of M and L segments of strain Q32 was similar to that of other Hantavirus M and L segments. Deduced amino acid sequences of glycoprotein and RNA polymerase revealed high homologue to other Hantavirus.

Amino Acid Sequence ; Animals ; Cercopithecus aethiops ; DNA, Complementary ; chemistry ; genetics ; Hantavirus ; genetics ; Molecular Sequence Data ; Murinae ; Phylogeny ; RNA, Viral ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Vero Cells ; Viral Matrix Proteins ; classification ; genetics ; Viral Proteins ; genetics