ABSTRACT
Hepatitis B Virus [HBV] DNA polymerase transactivated protein 1 [HBVDNAPTP1] is a novel protein transactivated by HBV DNA polymerase, screened by suppression subtractive hybridization technique [GenBank accession no: AY450389]. The biological function of HBVDNAPTP1 was investigated in this study. We constructed a vector pcDNA3.1 [-]/myc-His A-HBVDNAPTP1 and used it to transfect acute monocytic leukemia cell line THP-1. HBVDNAPTP1 expression was detected by western blot analysis in the cells. A cDNA library of genes transactivated by HBVDNAPTP1 in THP-1 cells was made in pGEM-T Easy using suppression subtractive hybridization [SSH]. The cDNAs were sequenced and analyzed with BLAST search against the sequences in GenBank. Some sequences, such as CIP4, might be involved in apoptosis development. mRNA and protein expression of CIP4 was identified by Real time RT-PCR and western blot in THP-1 cells. HBVDNAPTP1 could down-regulate the expression of CIP4 at both transcription and translation levels. HBVDNAPTP1 may be involved in the positive regulation on the initiation of monocyte apoptosis. The result contribute to reveal the HBVDNAPTP1 biological functions and provide new evidences for further exploration of the regulatory mechanism of HBVDNAPTP1
ABSTRACT
Hespintor is an unknown function protein that was got from hepatoblastoma cell lines HepG2 by suppression subtractive hybridization technique (SSH), sequence analysis showed that the protein is a new member of secretory type of Kazal type serine protease inhibitor (Serpin) family, and has high homology with esophageal cancer related gene 2 (ECRG2). The coding sequence of Hespintor's Kazal domain was subcloned into prokaryotic expression vector pET-40b(+), then transformed into Rosetta (DE3). A recombinant protein about 42 kDa in the form of inclusion body was optimization expressed by inducing with 0.25 mmol/L IPTG, 30 degrees C for 5 h. and its specificity was confirmed via Western blotting. The recombinant protein was purified by metal chelate affinity chromatography (MCAC) and anion-exchange chromatography. The preliminary experimental result showed that the recombinant protein can inhibit trysin hydrolysis activity specifically. The result clearly demonstrated that Hespintor, as a novel member of Serpin, would be valuable in developing anti-tumor agents.
Subject(s)
Humans , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Genetics , Hep G2 Cells , Proteinase Inhibitory Proteins, Secretory , Recombinant Proteins , Genetics , Serine Peptidase Inhibitors, Kazal Type , Serine Proteinase Inhibitors , Classification , GeneticsABSTRACT
Objective To observe the influence of the short effect antihypertension drugs- nifedipine and medial effect antihypertension drugs- extended release nifedipine on the blood pressure variability (BPV) in essential hypertension(EH). Methods Twenty-five EH patients were underwent 24-hour noninvasive ambulatory blood pressure monitoring (ABPM) and observed their BPV respectively before taking drugs, after taking nifedipine and extended release nifedipine. Meantime,25 normotensive controls (NC) were observed. Results (1)BPV in EH group was higher than that in controlled group and the severer the rise of blood pressure, the more obvious the increase of BPV (P 0.05). Conclusions Nifedipine could increase BPV but extended release nifedipine did not change BPV while they decreased blood pressure. Effect of extended release nifedipine was better than nifedipine in decreasing blood pressure.