ABSTRACT
AIM: Utilizing mRNA fluorescent differential display RT-PCR in our previous study,we found that the mRNA expression level of an intermediate protein(IF)-desmuslin was dramatically down-regulated in the abnormal veins of patients with primary vein incompetence.In this study,we testified the alteration of desmuslin expressing style at gene transcription and translation levels.METHODS: Specific PCR primers were designed according to the sequence of desmuslin mRNA.The cDNA fragments of desmuslin obtained from differential display were labeled by DIG as specific probes.Then semi-quantitative RT-PCR and Northern blotting techniques were applied to investigate the expression level of desmuslin mRNA in normal and abnormal veins.Simultaneously,the specific single-cloned antibody bestowed by Yuji Mizuno was used to evaluate the amount of DMN protein in the two groups by Western blotting and immunohistochemical techniques.RESULTS: In the abnormal veins isolated from the patients with primary vein incompetence,the expression of desmuslin mRNA was significantly down-regulated,compared with that in control group(semi-quantitative RT-PCR: 0.19?0.05 vs 0.83?0.08,P