OBJECTIVE: To observe the means of fibronectin(FN) alternative splicing and the expression of EIIIA-FN variant in rat skin after bruise, for the sake of providing some help for forensic estimation of wound interval. METHODS: Total RNA was isolated from wounded skin, and reverse transcription polymerase chain reaction was conducted to amplify target segments. RESULTS: Detectable EIIIA+(526 bp) segments, lacked in normal organize, was amplified at 1 h after experimental wound, and the levels were increased within 24 h. CONCLUSION The alternative splicing EIIIA-fibronectin variant would be a satisfied criterion for research of skin injury.
Alternative Splicing ; Animals ; Epithelium/metabolism* ; Fibronectins/genetics* ; Forensic Medicine ; Integrin alpha4beta1/biosynthesis* ; RNA, Messenger/metabolism* ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Skin/metabolism* ; Time Factors

To investigate the effect of vascular endothelial growth factor (VEGF) on beta1 integrin (VLA-4 and VLA-5) activation ability in K562 cells transfected with antisense VEGF121 cDNA, K562 cells were transfected with antisense (As), sense (S) and vector (V, pcDNA(3)). Flow cytometry was used to evaluate the expression rate of VLA-4 (CD49d/CD29) and VLA-5 (CD49e/CD29) and beta1 integrin activation ability in the transfected K562 cells. The results showed that the expression rates of VLA-4 and VLA-5 were more than 92% in the transfected K562 cells and there was no difference among the K562/V, K562/S and K562/As cells. However, beta1 integrin activated 9EG7 expression rate in K562/As cells was higher than that in K562/V cells [(75.6 +/- 10.5)% vs (41.2 +/- 2.1)%, P < 0.01)] after activation with beta1 integrin activator 8A2. It is concluded that function of beta1 integrin to be activated is restored in K562 cells transfected with antisense VEGF121 cDNA.
DNA ; genetics ; DNA, Antisense ; genetics ; Endothelial Growth Factors ; genetics ; metabolism ; Flow Cytometry ; Humans ; Integrin alpha4beta1 ; biosynthesis ; Integrin alpha5beta1 ; biosynthesis ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; K562 Cells ; Lymphokines ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

To explore the effect of ligustrazine on the expression of adherent molecule VCAM-1/VLA-4 of bone marrow cells in syngenic bone marrow transplantation (BMT) mice, the mice were divided into 3 groups: normal group (which received no treatment), BMT control group and ligustrazine-treated groups. BMT mouse models were established. The BMT control group and the ligustrazine-treated group were orally administered 0.2 ml saline per mouse and 2 mg ligustrazine per mouse, respectively, twice a day. On the day 7, 14, 21, 28 after BMT, mice were respectively killed. Bone marrow nucleated cells were detected, and then the expression of VCAM-1/VLA-4 was assayed by immunohistochemistry, RT-PCR and flow cytometry analysis, respectively. The results showed that in ligustrazine-treated group, the accounts of bone marrow nucleated cells on the day 7, 14, 21, 28 after BMT were all higher than that in BMT control group. The expression level in the ligustrazine-treated group was significantly higher than that in the BMT control group (P < 0.05 or P < 0.01). It is concluded that ligustrazine can enhance VCAM-1/VLA-4 expression in bone marrow after syngenic bone marrow transplantation in mice, which may be related to the mechanisms underlying the ligustrazine accelerating hematopoietic reconstitution in allogenic bone marrow transplantation.
Animals ; Bone Marrow Cells ; drug effects ; metabolism ; Bone Marrow Transplantation ; methods ; Flow Cytometry ; Immunohistochemistry ; Integrin alpha4beta1 ; biosynthesis ; genetics ; Male ; Mice ; Mice, Inbred BALB C ; Pyrazines ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Isogeneic ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics