By using AdEasy system, which is based on the homologous recombination in bacteria, an EGFP labeled recombinant adenovirus vector containing hVEGF165 was constructed quickly and efficiently expressed in mouse haematopoietic cells, First, hVEGF165 coding sequence was subcloned into shuttle plasmid pAdTrack-CMV, then cotransformed with adenoviral backbone vector pAdEasy-1 into E. coli strain BJ5183. The recombinant adenoviral plasmid was transfected into HEK293 cells to assembly replication-defective adenovirus Ad-EGFP/hVEGF165. The expression of EGFP could be easily detected. The rate of EGFP positive mouse bone marrow mononuclear cells by flow cytometric analysis was 27.3% (MOI=100), and the expression of hVEGF165 protein in the supernatant was (1385±332) pg/10⁶ cells. These results suggest that the construction of adenovirus vector by homologous recombination in bacteria features high efficiency and simplicity. The prepared high titer Ad-EGFP/hVEGF165 can be used an efficient helpful vector to infect hematopoietic cells.

To observe the inhibitory effects of an antisense u-PAR vector on invasion of highly invasive PC-3M cell subclones, the effects of the antisense u-PAR on activity of MMP-9 in those highly invasive cell subclones were detected by a quantitative RT-PCR and zymography. The monolayer invasion assay and colony formation assay in soft agar were used. And tumorigenesis rate and invasions by the cell subclones with or without the antisense u-PAR were observed in nude mice. It was found thatin vitro growth of highly invasive PC-3M cell subclones transfected with the antisense u-PAR was declined, and the ability of anchorage-independent growth of those cell subclones was found decreased sharply, with the inhibiting rate becoming 79% and 60%, respectively. Although the antisense u-PAR didn’t change MMP-9 gene transcription, they could inhibit the activation of MMP-9 of highly invasive PC-3M cell subclones. Moreover, the tumorigenesis rate of the cell subclones with the antisense u-PAR decreased and the growth of a neoplasm also slowed down. Thet tests showed the difference between experimental and control groups was statistically significant (P<0.01). The antisense u-PAR vector could not only inhibit the invasion ability of highly invasive PC-3M cell subclonesin vitro but also restrain the growth of those cell subclonesin vivo.

To investigate whether apoptosis is associated with cell adhesion in bronchial epithelium, and whether it contributes to the kinetics of injury and repair of surface epithelia, this study was performed for E-cadherin expression by using immunohistochemistry technique and for apoptosis by TUNEL method. An animal model of smoking was used for this study. The results showed that epithelial cells with membrane anchored E-cadherin decreased remarkably at several time points during 6 months of exposure to smoke (P<0.01) and then restored to normal level. This fluctuation was associated exclusively with the alteration in number of apoptotic cells (P<0.01). There was no significant difference in activation of nuclear transcription factor NF-kappa B among groups (P>0.05). All these suggested that apoptosis is associated with E-cadherin expression in bronchial epithelium of smoking mouse.

In situ hybridization and immunocytochemical techniques were employed to examine the expression of matrix metalloproteinases-1 (MMP-1) and to identify the pattern of its distribution in rat pancreas. The results indicated that the signal of MMP-1 mRNA and MMP-1 positive immunoreaction were detected in some fiberoblasts around interlobular ducts and exocrine cell in margin acinus of some lobules, but the signal of MMP-1 mRNA and MMP-1 positive immunoreaction could not be detected in most of other acinus and islets of pancreas. It is concluded that the expression of MMP-1 in above cells of rat might play an important role in acinar proliferation and differentiation of rat pancreatic tissues.