BACKGROUND:Recent studies found that some factors play important role in the process of denervated muscle atrophy, especial y the feak-headbox transcription factor, is the key element to regulate the denervated muscle atrophy.
OBJECTIVE:To investigate the effect of RNA interference on inhibiting feak-headbox 3a gene expression in vitro.
METHODS:The myoblast cel line L6 were cultured in the 6-wel cel culture plates, then pEGFP-N1 and smal
interfering RNA recombinant plasmid with the same ratio was transfected under the Lipofectamine2000 mediation to optimize the transfection efficiency of the detection system;2μg smal interfering RNA recombinant plasmid of feak-headbox 3a gene were transfected with myoblast cel line L6 for 48 and 72 hours.
RESULTS AND CONCLUSION:At 48 hours after pEGFP-N1 and siRNA recombinant plasmid transfection, a large number of bright green fluorescent displayed under fluorescence microscope with higher transfection efficiency. Real-time quantitative PCR analysis showed that there were significant differences in the sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ, feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after trasfection (P<0.05), and the inhibition effect was more significant at 72
hours after transfection when compared with that at 48 hours after transfection. Western Blot gray analysis showed that there were significant differences in sequences of feak-headbox 3a-Ⅰ, feak-headbox 3a-Ⅱ, feak-headbox 3a-Ⅲ,
feak-headbox 3a-Ⅳ on feak-headbox 3a mRNA when compared with the control group at 48 and 72 hours after
trasfection (P<0.05), and the inhibition effect was more significant at 72 hours after transfection when compared with that at 48 hours after transfection, which was same with the effect on mRNA level. RNA interference in vitro can
significantly inhibit the fork-head transcription factor feak-headbox 3a gene expression, and the inhibition effect of feak-headbox 3a gene smal interfering RNA recombinant plasmid transfected with the sequence on the mRNA and protein level of feak-headbox 3a is not clear, which can provide new idea for the gene therapy of RNA mediated denervated skeletal muscle atrophy.

Objective To investigate the expression of slit diaphragm proteins of glomerular podocyte,such as NEPH1 and Nephrin in type Ⅴ lupus nephritis (V-LN). Methods Twenty-five patients with V-LN and 18 patients with idiopathic membranous nephritis (IMN) were enrolled into the study, and 5 normal renal samples were the normal control group. Twenty-four hours urine protein excretion, serum albumin, creatinine, triglyceride, total cholesterol, serum C3, C4, urine C3 and NAG were tested respectively.Glomerular lesions were measured by light microscopy. The expressions of NEPH1 and Nephrin were determined by indirect immunofluorescent staining. The statistical treatment was used t-test. Results Compared to the IMN group, the 24 hours urine protein excretion and the concentrations of serum albumin, creatinine, urine C3 were not significantly different while the triglyceride, total cholestorel, serum C3, C4 were significantly decrease in the V-LN group (P<0.05). Urine NAG was increased in the V-LN group (P<0.01). By indirect immuno-fluorescent histochemitry examination, the glomerular expressions of NEPH1 and Nephrin were significantly decreased in both V-LN and IMN. Compared with the IMN group, the decrease of NEPH1 and Nephrin expression was more remarkable in the V-LN group. Conclusion The expression changes of NEPH1 and Nephrin may play an important pathogenic role in proteinuria of Ⅴ lupus nephritis. Renal tubular epithelial cell damage may play a role in proteinuria of V-LN.