OBJECTIVEGene therapy has been becoming one of the most attractive medical areas. But the using of gene therapy in plastic surgery is relatively scarce. Our purpose was to investigate the effect of naked plasmid encoding Vascular Endothelial Growth Factor on the viability of the random skin flap by directly injected subcutaneously.

METHODS30 female Sprague-Dawley rats randomly divided into three groups. A random dorsal skin flap of 3 cm x 9 cm was elevated in each of the rats. And 1 ml double-distilled water solution was injected subcutaneously, which was only water in group 1 during the operation, 200 micrograms VEGF cDNA plasmid in group 2 during the operation, 200 micrograms pcDNA3.1/zeo(+)--VEGF in group 3, 24 hours before the operation, respectively. 7 days after the operation, all the animals were sacrificed by overdose anesthetic. The survival tissue was measured with planimetry. Two samples were harvested from each group for pathological check and immunohistochemical test.

RESULTSImmunohistochemical staining demonstrated that there was human VEGF deposited around the capillary in the flaps treated with VEGF gene. The flaps treated with VEGF gene had a larger percentage of survival skin (group 1 = 47% +/- 5.4%, group 2 = 65.4% +/- 6.3%, group 3 = 72.3% +/- 8.5%, P < 0.05).

CONCLUSIONVEGF gene directly injected into subcutaneous can express VEGF. It makes the gene therapy simple and practical and will be promising future in the tissue transplantation.

Animals ; Endothelial Growth Factors ; genetics ; Female ; Genetic Therapy ; Injections, Subcutaneous ; Lymphokines ; genetics ; Rats ; Rats, Sprague-Dawley ; Surgical Flaps ; physiology ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

IFN-λ1 is a member of a new family of interferons called type Ⅲ IFNs with similar functions to type ⅠIFNs. Previously we obtained recombinant soluble human rhIFN-λ1 from Pichia pastoris. However, the hyper-glycosylation from P. pastoris brings immunogenicity and low purification recovery rate. To overcome this disadvantage, in this study, we constructed an rhIFN-λ1 mutant (rhIFN-λ1-Nm) devoid of the potential N-glycosylation sites by site-directed mutagenesis. rhIFN-λ1-Nm was successfully expressed and secreted extracellularly in P. pastoris (GS115) using methanol inducible AOX1 promoter with α-mating factor signal sequence. rhIFN-λ1-Nm was purified and characterized. There was no significant difference between rhIFN-λ1-Nm and rhIFN-λ1 in structure and bioactivity. The molecular weight was low after N-glycosylation mutation whereas the glycosylation was much lower. The mutational rhIFN-λ1 (rhIFN-λ1-Nm) could legitimately be developed as substitutes for rhIFN-λ1, and thus it may be developed into a more promising therapeutic reagent in the future.

To explore the clinical significance of the altered expression of polycomb group (PcG)-associated protein RYBP in hepatocellular carcinoma (HCC) specimens.
 Methods: The expression levels of RYBP in tumor tissues and adjacent normal tissues in 77 HCC cases were detected by immunohistochemistry (IHC), and the relationships between RYBP expression levels and HCC clinicopathological characteristics, five-year survival rates or prognosis of HCC patients were analyzed.
 Results: RYBP expression level was significantly decreased in HCC tumor tissues than that in the adjacent normal tissues (P<0.05). The expression levels of RYBP in HCC specimens were highly correlated with HBsAg, ALT, GGT, Type III procollagen, tumor size, distant metastasis, and tumor differentiation (P<0.05). The RFS and OS for patients with RYBP-low expression were markedly lower than those with RYPB-high expression (P<0.05). Both age and RYBP expression level were protective factors for RFS, while GGT, lymph node metastasis, TNM stage, tumor differentiation and tumor size were risk factors for RFS (P<0.05). As to OS, RYBP expression level was a protective factor, while tumor number, ALT, GGT, AFP, pCEA, lymph node metastasis, TNM stage, tumor differentiation and tumor size were risk factors (P<0.05). The age, GGT, lymph node metastasis and TNM stage were independent prognostic factors for RFS (P<0.05), and both lymph node metastasis and TNM stage were independent risk factors for OS (P<0.05). Comparing to serum alpha fetoprotein (AFP) level, RYBP expression level in tumor tissues was applied to predict the prognosis of HCC patients more accurately.
 Conclusion: PcG associated protein RYBP displays a reduced expression in HCC tissues, which is related to poor prognosis of HCC patients. It might be a promising therapeutic target for HCC treatment.
Biomarkers, Tumor ; Carcinoma, Hepatocellular ; Humans ; Immunohistochemistry ; Intracellular Signaling Peptides and Proteins ; metabolism ; Liver Neoplasms ; Lymphatic Metastasis ; Prognosis