Objective To screen the possible regulatory proteins showing the ability for interaction with serum response factor ( SRF) in the progress of myofibroblast activation, and to see if the proteinprotein interaction is contributing to induce the expression of smooth muscle αactin ( α-SMA) . Methods Phage display cDNA libraries were constructed from the transdifferentiated airway epithelial cells and parental cells. Phage clones were then selectively amplified during the biopanning procedure by using SRF as a bait protein for the two cDNA libraries. Following four rounds of biopanning, recovered cDNAs were sequenced and the obtained sequences were aligned by BLAST tool to select the candidate gene. PAI-RBP1 of the candidate gene was cloned and sub-cloned into pcDNA3. 0 plasmid. Transient transfection and RT-PCR analysis were performed for investigation of the expression of α-SMA. Results Three candidate proteinbinding partners, PAI-RBP1, Nucleolin, and HF1OO, were identified. Among them, PAI-RBP1 pcDNA3. 0 plasmid was subjected to transient co-transfection with SRF, showing up-regulation of α-SMA expression. Conclusions Combined with phage display technique, through protein-protein interaction between core transcription factor and unknown proteins to find a newtranscriptional regulator may serve as an effective strategy. Three novel SRF binding proteins were found from transdifferentiated cells. This study indicates that PAI-RBP1 involves in the activation of myofibroblast by induction of α-SMA expression.
ObjectiveTo evaluate the clinical values of phage amplified biologically assay (PhaB) for diagnosis of tuberculosis by comparatively analyzing the diagnostic performances of PhaB, acid fast stain and culture. MethodsThe samples of random sputum and morning sputum from 157 tuberculosis patients diagnosed between January and December 2014 were detected by mycobacteria culture, PhaB, acid fast stain and culture method. The differences of diagnostic performances were analyzed by chi-square test. ResultsThe diagnostic sensitivity was 89.8% (mycobacteria culture), 68.2% (PhaB) and 52.2% (acid fast stain); according to the gold standard of culture method, the positive coincident rate was 74.5% and 57.4%, respectively in PhaB and acid fast stain (P<0.05), and the general coincident rate was 75.8% and 60.5% (P<0.05); of those patients with two negative sputum smears, the positive rate was 33.3% (25/75) in PhaB; the detection time was 1 hour (acid fast stain), 46 hours (PhaB) and 9.5 days (mycobacteria culture), respectively. ConclusionBecause of its high sensitivity, high specificity and short turn around time, simple operation, distinguishing dead isolates and live isolates and drug resistance detection, PhaB is a new method for screening test of tuberculosis or as an effective complementary testing for traditional assays.