[Effects of antisense RNA of connective tissue growth factor expressing plasmid on rat liver fibrosis].

OBJECTIVE: To observe the effects of antisense RNA of connective tissue growth factor (CTGF) on rat liver fibrosis. METHODS: Gene recombinant techniques were used to construct a rat antisense RNA of CTGF recombinant plasmid which could be expressed in eukaryotic cells. The recombinant plasmids were encapsulated with lipofectamine and then transducted into a carbon tetrachloride (CCl4) induced rat liver fibrosis model. Expression of CTGF was assessed by RT-PCR, Western blot and immunohistochemistry. Immunohistochemistry was used to identify type I and III collagens. HE stained liver slides were used for pathological study. RESULTS: The mRNA and protein expression of CTGF in the fibrotic liver transfected with antisense-CTGF were significantly decreased compared with those of the controls (P<0.01). The depositions of type I and type III collagens were also decreased (P<0.05). Antisense-CTGF also minimized the pathological fibrosis in the rat livers (P<0.01). CONCLUSION: The results demonstrate that the antisense RNA of CTGF recombinant plasmid has certain effects in preventing liver fibrosis and makes it a possible candidate for use in future gene therapy.

[Clinical significance of serum GPDA-F determined by immunoelectrophoresis in diagnosis of hepatocellular carcinoma].

BACKGROUND & OBJECTIVE: Serum fast band of glycylproline dipeptidyl aminopeptidase isoenzyme (GPDA-F) is useful to the diagnosis of hepatocellular carcinoma, especially for the cases without expression of alpha-fetoprotein (AFP). Polyacrylamide electrophoresis for detection of GPDA-F is relatively complicated and has limitation in its clinical use. This study was to establish a simple and easy method of immunoelectrophoresis to detect serum GPDA-F, and evaluate clinical value of GPDA-F in the diagnosis of hepatocelluar carcinoma. METHODS: Serum GPDA-F was purified to raise polyclonal GPDA-F antibody, and immunoelectrophoresis was established for the detection of serum GPDA-F. Serum GPDA-F in 99 specimens of hepatocellular carcinoma and 115 specimens of benign liver diseases (36 cases of liver cirrhosis, 23 cases of acute hepatitis, 38 cases of chronic hepatitis, and 18 cases of benign liver space-occupying lesions) was simultaneously detected by both polyacrylamide electrophoresis and immunoelectrophoresis. The clinical value of serum GPDA-F detected by immunoelectrophoresis was compared with that by polyacrylamide electrophoresis for the diagnosis of hepatocellular carcinoma. RESULTS: If the cut-off was set at 71 u/L, the diagnostic sensitivity, specificity, and accuracy of immunoelectrophoresis in detecting GPDA-F in hepatocellular carcinoma were 83.8%, 85.2%, and 84.6%, respectively; while those of polyacrylamide electrophoresis were 81.8%, 77.3%, and 79.4%, respectively. CONCLUSIONS: Serum GPDA-F detected by immunoelectrophoresis is useful to the diagnosis of hepatocellular carcinoma. Compared with polyacrylamide electrophoresis, this method is cheap, time-saving, and easy.