ABSTRACT
Objective To observe the effect of recombinant human tumor necrosis factor receptor-Fc fusion protein (rhTNFR-Fc, etanercept) on the expression of transforming growth factor-β1 (TGF-β1) in bleomycin induced interstitial lung disease of rats. Methods Forty-five male Sprague-Dawley (SD) rats were randomly divided into three groups (control group, model group and rhTNFR-Fc treatment group, 15 rats in each), on the 7th, 14th and 28th days, five rats of each group were killed. The lungs were incised to make pathological sections which were stained with HE and Masson, and the expression of TGF-β1 was detected by immunohistochemical technique. Results There was no collagen deposition, alveolitis and fibrosis changes in the control group. The alveolitis and fibrosis of the treatment group was less severe than that in the model group (P<0.01). The expression of TGF-β1 in the model group was significantly higher than that in the control group (P<0.01). In the 7th and 14th days, the expression of TGF-β1 in the treatment group was signific-antly higher than that in the control group (P<0.01). Although that in the 28th day was a slightly higher but no statistical significance (P>0.05) could be detected. In the treatment group, the expression of TGF-β1 was lower in the 7th day (P>0.05) and was significantly lower in the 14th and 28th days than that in the model group (P<0.01). Conclusion Recombinant human tumor necrosis factor receptor-Fc fusion protein can alleviate the severity of alveolitis and pulmonary fibrosis induced by Bleomycin-A5 in rats, which may be due to the inhibition of TGF-β1 overexpression.
ABSTRACT
Objective The present study demonstrates a novel,simple and cost-effective method for detecting known SNP genotyping by using ShineRoar probes.Methods The SNP of target genes detected by using the ShineRoar probes and melting curve analysis.Tumor necrosis factor receptor Ⅱ (TNFR Ⅱ) and apolipoprotein M (apoM) had been employed as target genes to describe the method in details.The PCR products of TNFR Ⅱ and apoM were collected and sequenced.Results The melting temperatures (TM) were significantly different between mutated genotypes and wild-type genotype.A biallelic SNP marker (T/ G) at position 196 in exon 6 of TNFR Ⅱ gene showed two melting valleys with the appropriate TMs at (52.84?0.75)℃ and (58.38?0.61)℃,respectively.For apoM T-778C,TMs of homozygous T genotype and C genotype were (42.55?0.73)℃ and (49.19?0.57)℃,respectively.Moreover,this genotyping method was validated by the DNA sequence analyses (Kappa=1,P=0.000).Conclusion It is concluded that this novel method is simple and economical and it is suitable for a large-scale genotyping screening.