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1.
Chinese Journal of Experimental and Clinical Virology ; (6): 29-31, 2009.
Article in Chinese | WPRIM | ID: wpr-332439

ABSTRACT

<p><b>OBJECTIVE</b>To test the infeciton efficiency of recombinant adenoviral vector carrying HBsAg-HSP70 chimeric gene and to abserve its biological characteristics.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMC) were separated from healthy blood donor and they were infected by Ad-HSP70-HBsAg on the first day of isolation. DCs were induced in medium with cytokines IL-4, GM-CSF and TNF-alpha in vitro. The biological characteristics of DC induced were analyzed by inverted fluorecent microscope, RT-PCR, flow cytometer (FACS), and mixed lymphocyte reaction (MLR).</p><p><b>RESULTS</b>The traced gene-GFP were abserved in DCs by inverted fluorecent microscope and HSP70-HBsAg gene mRNA expression was detected by RT-PCR after the Ad-HSP70-HBsAg infection. FACS analysis shown that the expression of CD1a, CD80, CD86 and HLA-DR on surfece of two groups of DCs were similar. MLR showed that there are not a statitic difference of stimulated index (SI) between two groups.</p><p><b>CONCLUSION</b>Results indicated that Ad-HSP70-HBsAg can effectively infected DCs without affecting its biological characteristics.</p>


Subject(s)
Humans , Adenoviridae , Genetics , Physiology , Cells, Cultured , Cytokines , Genetics , Allergy and Immunology , Dendritic Cells , Allergy and Immunology , Virology , Genetic Vectors , Genetics , HSP70 Heat-Shock Proteins , Genetics , Allergy and Immunology , Hepatitis B Surface Antigens , Genetics , Allergy and Immunology , Lymphocyte Culture Test, Mixed , Recombinant Fusion Proteins , Genetics , Allergy and Immunology
2.
Chinese Journal of Medical Genetics ; (6): 42-47, 2007.
Article in Chinese | WPRIM | ID: wpr-285036

ABSTRACT

<p><b>OBJECTIVE</b>To detect the PAX3/PAX7-FKHR fusion transcripts to identify genetic alteration in embryonal rhabdomyosarcoma (ERMS) and alveolar rhabdomyosarcoma (ARMS) tissues.</p><p><b>METHODS</b>One-step reverse transcription- polymerase chain reaction (RT-PCR) were used to detect the expression of the PAX3/PAX7-FKHR fusion transcrips in 16 cases of rhabdomyosarcoma (7 cases of ARMS, 9 cases of ERMS) and 16 specimens were compared to the surrounding normal tissue. Comparative genomic hybridization (CGH) was employed to detect the genomic imbalance (DNA loss or amplification) in 16 RMS cases.</p><p><b>RESULTS</b>PAX3-FKHR fusion transcripts were positive in 3/7 and PAX 7-FKHR fusion transcripts were positive in 2/7 of ARMS patients, respectively, and were all negative in ERMS and Control tumors. There were different chromosome variations for each RMS, chromosome amplification was frequently seen in 1p36 (69%), 5q32 (56%), 8q21 (63%), 13q14 (69%), 19q (63%), 20q (56%). Chromosome loss was frequently seen in 3p21-pter (56%), 9p23-pter (50%), 10q (69%), 16/16q24 (56%).</p><p><b>CONCLUSION</b>One-step RT-PCR assay for detection specific fusion gene provides a useful tool for confirmation of the diagnosis of RMS in diagnostically difficult cases and in retrospective studies. Chimeric gene transcript resulting from specific chromosomal translocations is a reliable index for the molecular diagnosis of RMS.</p>


Subject(s)
Humans , Chromosome Aberrations , Comparative Genomic Hybridization , Forkhead Box Protein O1 , Forkhead Transcription Factors , Genetics , Gene Expression Regulation, Neoplastic , Oncogene Proteins, Fusion , Genetics , PAX3 Transcription Factor , PAX7 Transcription Factor , Genetics , Paired Box Transcription Factors , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Rhabdomyosarcoma , Genetics
3.
Chinese Journal of Oncology ; (12): 915-919, 2006.
Article in Chinese | WPRIM | ID: wpr-316267

ABSTRACT

<p><b>OBJECTIVE</b>To establish stable techniques of comparative genomic hybridization (CGH) and apply them to elucidate the genetic characteristics of hepatoblastoma (HB), and to explore the characteristics and clinical significance of loss of heterozygosity (LOH) at 1p36 in HB.</p><p><b>METHODS</b>CGH was employed to detect the genomic imbalance (DNA loss or amplification) in 20 cases of HB, and PCR-simple repeated sequence polymorphism was employed in 30 cases of HB to detect the loss of heterozygosity for 6 satellites at chromosome 1p36.</p><p><b>RESULTS</b>There were different chromosome variations for each HB. chromosome amplification was frequently seen in 1q, 2q,2p, 8q, 8p, 12q and 22q. Chromosome loss was often seen in 1p, 4q, 4p, 16q, 17p and 18q. The frequency of LOH at 6 loci on chromosome 1 was 63.3% totally (19/30), with the highest D1S199 (66.7%) and D1S450 next to it (46.7%).</p><p><b>CONCLUSION</b>There were chromosome zones with DNA amplification or loss in hepatoblastoma. There are extensive LOH at 1p36 in hepatoblastoma. The corresponding amplification of oncogene and loss of antioncogene may take part in the development of hepatoblastoma.</p>


Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Chromosome Aberrations , Chromosome Deletion , Chromosomes, Human, Pair 1 , Genetics , Hepatoblastoma , Genetics , Liver Neoplasms , Genetics , Loss of Heterozygosity , Nucleic Acid Hybridization , Methods
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