Aim To investigate the effects of cyclosporin A (CsA) on growth and collagen synthesis of cardiac fibroblasts (CFs) induced by arginine vasopressin (AVP). Methods CFs of neonatal Sprague-Dawley rats were isolated by trypsinization and cultured; growth-arrested CFs were stimulated with 1×10-7 mol·L-1 AVP in the presence or absence of CsA (0.05, 0.5 and 5 μmol·L-1). MTT and flow cytometry techniques were adopted to measure cell number and analyze cell cycle respectively. Collagen synthesis was determined by measurement of hydroxyproline content in culture supernatant with colorimetry. Calcineurin activity was estimated by chemiluminescence. Trypan blue staining to test the viability of CFs. Results 0.05, 0.5 and 5 μmol·L-1 CsA inhibited the increase of CFs number induced by 1×10-7 mol·L-1 AVP in a dose-dependent manner, with the inhibitory rates by 12%, 24% and 29%, respectively (P＜0.05). Furthermore, cell cycle analysis showed 0.5 μmol·L-1 CsA decreased the S stage percentage and proliferation index of CFs stimulated by AVP (P＜0.05). In culture medium, the hydroxyproline content induced by AVP decreased by 0.5 and 5 μmol·L-1 CsA (P＜0.05), with the inhibitory rates of 29% and 33%, respectively. CsA completely inhibited the increment of calcineurin activity induced by AVP (P＜0.01), but CsA itself had no effect on the baseline of calcineurin activity and CFs viability. Conclusion CsA inhibits proliferation and collagen synthesis of CFs by virtue of blocking calcineurin signaling pathway and might provide a novel target for prevention and treatment to cardiac fibrosis.

Background Cardiac mast cell-derived chymase is involved in myocardial fibrosis,but the underlying mechanisms remain unclear. Objective To investigate the effect of chymase on the collagen synthesis and its relationship with transforming growth factor-?1 (TGF-?1)/Smad pathway in rat cardiac fibroblasts (CFs). Methods CFs,from neonatal SD rats,were isolated by trypsinization. The collagen synthesis of CFs was determined by 3H-proline incorporation. The protein expressions of TGF-?1,phosphorylated Smad2/3 (P-Smad2/3) and total Smad2/3 were determined by immunoblotting in CFs. Results Chymase (15,30 and 60 ?g/L) increased the 3H-proline incorporation in a concentration-dependent manner. 30 ?g/L chymase stimulation increased the protein expressions of TGF-?1 and P-Smad2/3 in a time-dependently,while little effect on Smad2/3 protein expression was found. The stimulatory effect of chymase on 3H-proline incorporation elicited by 30 ?g/L chymase was blocked in the presence of TGF-?1 antibody or staurosporine,a P-Smad2/3 inhibitor. Conclusion Chymase promotes collagen synthesis of rat CFs,TGF-?1/Smad might be involved into the signal pathway.

OBJECTIVE To prove the diagnosis value for Legionnaires pneumophial pneumonia using polymerase chain reaction. METHODS L. pneumophial-DNA (LPN-DNA) from 47 spuum and 6 bronchoalveolar lavage fluid samples collected from 53 patients with atypical pneumonia was detected by PCR. RESULTS The positive rate of LPN-DNA in 53 patients with atypical pneumonia was 9.4%, while the positive rate of sputum and bronchoalveolar lavage fluid samples was 6.4%and 33.3%, respectively. CONCLUSIONS LPN-DNA detected by PCR for early diagnosis of atypical pneumonia has favorable clinical application.