ABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of focal adhesion kinase (FAK) in adhesion and migration of hepatic stellate cells (HSC).</p><p><b>METHODS</b>Two recombinant plasmids expressing short hairpin RNAs (shRNAs) targeting FAK were constructed and one plasmid substantially suppressing FAK expression in HSC was selected. Real-time PCR and Western blot were used to detect the knockdown effects of FAK gene. After 48-hour treatment with FAK shRNA, toluidine blue colorimetric assay was used to detect the cell adhesion. Wound-healing assay and improved Boyden double-chamber were used to detect the cell migration induced by FN.</p><p><b>RESULTS</b>The recombinant plasmid expressing FAK shRNA was successfully constructed and transfected into HSC. Compared with the controls, the expression of FAK mRNA and protein in HSC treated with FAK shRNA was markedly down-regulated by 76.82% and 72.53%, respectively. The expression of p-FAK (Tyr397) protein was also decreased by 62.71% 48 h posttransfection. The adhesion of HSC was inhibited by 58.69% at 48 h after shRNA transfection. FAK gene silencing could also dramatically inhibit FN-stimulated HSC migration, and the cell migration distance and the cell number of crossing membrane were decreased by 58.27% and 83.70%, respectively.</p><p><b>CONCLUSIONS</b>FAK gene silencing suppresses adhesion and migration of HSC, and FAK may be a potential target for novel anti-fibrosis therapies.</p>
Subject(s)
Animals , Rats , Blotting, Western , Cell Adhesion , Cell Line , Cell Movement , Down-Regulation , Fibronectins , Focal Adhesion Kinase 1 , Genetics , Metabolism , Genetic Vectors , Hepatic Stellate Cells , Cell Biology , Liver Cirrhosis , Pathology , Plasmids , Genetics , Polymerase Chain Reaction , RNA Interference , RNA, Messenger , Genetics , Metabolism , TransfectionABSTRACT
Objective To study the onset features and axiom of children disease in Duanguan region.Methods A statistic study was carried on 1 782 cases of children disease which confirmed by pathological examinations.The sex,age and pathology(congenital malformation,inflammatory disease,tumor-like lesions,benign tumors,malignant tumors)were registered.Results The inflammatory disease occupied the first place in incidence rate(44.61%),followed by tumor-like lesions(25.03%),benign tumors(23.34%),congenital malformation(5.22%)and malignant tumors(1.80%).Among inflammatory disease,appendicitis was the most common,the next was chronic tonsillitis and tuberculosis.Among tumor-like lesions,cystic disease was the most common;the next was polyps.Among benign tumors,vasculoma was the most common,the next was pilomatricoma,osteoid and chonodroid tumors.Among congenital malformation,ear fistula was the most common,the next was diverticulosis and intestineal duplicateion.Among the malignant tumors,the sarcoma of soft tissue was the most common,the next was the malignant peripheral nerve tumors and lymphoma.Conclusion Children's disease have its own characteristics which varied in age,sex and pathological types,and differ from those of adults.