Objective To identify hepatitis B virus X-interactive proteins by comparative proteomics method and to understand the molecular mechanism of HBx in the development of hepatocellular carcinoma(HCC).Methods Two-dimensional gel electrophoresis(2-DE)was used to separate the total proteins of HBx-transfected human hepatoma cell lines HepG2-Px and its parental cell lines HepG2-P_0.PDQuest software was applied to analyze 2 DE images.The differentially expressed protein spots between the two cell lines were identified by matrix-assisted laser desorptionionization time of flight mass spectrometry(MALDI-TOF-MS)and electrospray ionization tandem mass spectrometry(ESI-Q-TOF).Then,the differential expression levels of some identified proteins were determined by Western blot.The data were compared using t test.Results The well-resolved,reproducible 2-DE patterns of HepG2-Px and HepG2-P_0 total proteins were established.A total of 32 differential proteins were identified in HepG2-Px cell,including 25 up-regulated proteins,such as heat shock protein(HSP)90AB1,Bcl-2 associated athanogene(BAG)-2,nucleophosmin(B23),chloride intracellular channel(CLIC)-1,matrix metalloproteinase(MMP)-3,melanoma antigen(MAGE)-12,and 7 down-regulated proteins,such as Wnt-5a.The differential expression levels of some proteins between the two cell lines were confirmed by Western blot analysis.Conclusions Most of the identified proteins are involved in many processes,such as transcription,signal transduction,cell proliferation,cell cycle regulator,apoptosis,DNA repair,metabolisms and immunity.These differential proteins may play a role in tumor genesis and HC development.The data are valuable for further study on the role of HBx in human hepatocellular carcinoma.

OBJECTIVETo analyze the functions of differentially expressed genes between abdominal aortic aneurysm and normal aortic tissue by cDNA microarray.

METHODSTotal RNAs were respectively isolated from the normal aorta and aortic aneurysm, purified into mRNAs by oligotex. Subsequently they were reverse-transcribed into cDNAs incorporated with fluorescent dUTP to make hybridization probes, which were hybridized as the cDNA microarray for scanning of fluorescent signals and differentially expressed genes between the normal aortic and aortic aneurysm by using GenePix Pro 3.0 software.

RESULTSA total of 18 differentially expressed genes were detected, accounting for 0.44% of total genes. Among these genes, 11 were related to cell cycle and the remaining 7 to cell apoptosis. The number of upregulated genes in the aortic aneurysm was 9 (mean ratio: 3.860) and that of the downregulated 9 (mean Ratio: 0.294). Bio-informative analysis showed that these 18 genes might influence the growth and apoptosis of smooth muscle cells in abdominal aortic aneurysms.

CONCLUSIONSDuring the development of abdominal aortic aneurysms, modulations of multi-gene expression would undergo various changes. Cell cycle and apoptosis-related genes were related to the growth and apoptosis of smooth muscle cells in abdominal aortic aneurysms. Further research into these genes will clarify the mechanisms of abdominal aortic aneurysms.

Aortic Aneurysm, Abdominal ; genetics ; pathology ; Apoptosis ; physiology ; Cell Cycle ; genetics ; Cells, Cultured ; Gene Expression ; Gene Expression Profiling ; Humans ; Myocytes, Smooth Muscle ; metabolism ; pathology ; Oligonucleotide Array Sequence Analysis

Objective To establish a mouse model treated with busulfan and to investigate its effects on the metabo-lism of spermatogonial stem cells(SSCs)of mouse testis.Methods C57BL/6J male mice with age of 8 weeks were injected with 10 mg/kg of busulfan intraperitoneally,then Thy1 positive cells were selected by immunomagnetic beads on day 0,day 5 and day 10 and followed by identification for purity and metabolomic analysis.Results The testis weight ratio decreased and the tissue structure of testis was damaged(P＜0.05).Based on the results of principal component analysis(PCA)and partial least squares discriminant analysis(PLS-DA),there were signifi-cant metabolic differences between the sample groups treated for 0 d,5 d and 10 d.A total of 89 differential metabolites were identified including glutathione(GSH),arginine and unsaturatedfatty acids(UFAs),and their important metabolic pathways involved glycerophospholipid metabolism,arginine and proline metabolism.Conclu-sions Affecting the specific metabolic pathway may result in obvious reproductive toxicity and lead to decrease of testicular weight as well as tissue structure damage in mice.Metabolomic analysis showed that the potential repro-ductive toxicity mechanism of SSCs may be related to the metabolic pathways such as lipid metabolism,arginine and proline metabolism.