Purpose: Pulmonary fibrosis (PF) is a severe lung disease causing significant morbidity and mortality. PF pathogenesis is attributed to the fibroblast-to-myofibroblast transition (FMT) driven by the most potent pro-fibrogenic factor TGF-beta 1 activating the Smad3-dependent TGF-beta 1 canonical pathway. Iguratimod (IGU) is a novel anti-rheumatic drug that suppresses the secretion of inflammatory factors, but is also able to modulate the differentiation of multiple cells. Therefore, the aim of this work was to investigate the effect of IGU on FMT. Materials/methods: PF mouse model was induced in C57BL/6 male mice by bleomycin. The effect of IGU was assessed through the evaluation of lung morphology by H&E and through the collagen accumulation in the lung by Masson staining. Primary human lung fibroblasts (pHLFs) were also used to evaluate the effect of IGU in vitro on TGF-beta 1-stimulated cells, and proliferation, migration and invasion were measured, together with genes and proteins involved in FMT. Results: IGU attenuated bleomycin-induced PF in mice and improved the pathological changes in their lungs. In addition, IGU significantly inhibited proliferation, migration and invasion in TGF-beta 1-stimulated pHLFs without causing apoptosis. Moreover, IGU significantly reduced TGF-beta 1-induced increase of collagen I and III mRNA expression, thus reducing lung function impairment, and alpha-SMA, Smad2 and Smad3 phosphorylation, fibronectin expression and F-actin microfilament formation, thus attenuating FMT through the inhibition of the Smad3 pathway. Conclusions: Our results collectively revealed the beneficial effect of IGU on the inhibition of FMT, thus suggesting that it might act as an effective anti-fibrotic agent in preventing the progression of PF.

Objective: Pirfenidone (PFD) is an oral anti-fibrotic drug used for idiopathic pulmonary fibrosis (IPF) therapy. We determined the role of activating transcription factor 3 (ATF3) and the effect of PFD on fibroblast to myofibroblast transition (FMT) in rheumatoid arthritis-associated interstitial lung disease (RA-ILD). Methods: RA-ILD lung specimens were obtained by CT-guided percutaneous transthoracic biopsy. A pulmonary fibrosis mouse model was established by the intratracheal injection of bleomycin. Pathological variation and the expression of alpha-SMA and ATF3 were observed by H&E, Masson and immunofluorescence staining. Primary human lung fibroblasts (pHLFs) were isolated from lung tissues that were pathologically confirmed to be normal by pneumonectomy. Cell viability was detected using an MTT assay. Cell migration and invasion were detected using a Transwell chamber. The protein levels of alpha-SMA, ATF3, Smad3 and p-Smad3 were measured by Western blot. HLFs were infected with lentiviruses expressing ATF3 or scrambled shRNA. Results: ATF3 was dramatically upregulated in lung tissues from both bleomycin-induced mice and patients with RA-ILD compared with controls. The upregulation of ATF3 and the accumulation of collagen in the lung tissues of mice with pulmonary fibrosis were reduced by PFD. PFD significantly inhibited increases in the proliferation, invasion and migration of pHLFs stimulated by TGF-beta 1. Moreover, we observed the inhibitory effect of PFD on FMT via the downregulation of ATF3, which was further confirmed in ATF3 knockdown (KD) pHLFs. Conclusions: This work shows the inhibitory effect of PFD on FMT in pHLFs, which is mediated by the downregulation of ATF3. Our findings suggest that PFD might have therapeutic potential for the treatment of RA-ILD.

Objective: By analysis the correlation between images of high-resolution computed tomography (HRCT) and histopathology patterns of different connective tissue diseases associated interstitial lung diseases (CTD-ILDs), we aimed to investigate the feasibility of deduce the histopathology pattern from HRCT imaging and thereby avoid lung biopsy in such patients. Methods: Patients with a diagnosis of CTD in Guangdong General Hospital from November 2008 to June 2014 were screened and those with a diagnosis of ILD were enrolled in this study. The clinical data and findings on HRCT of those patients were reviewed; histopathology patterns of lung biopsy samples from 43 patients obtained by CT guided percutaneously transthoracic approaches were analyzed and compared with their HRCT images. Results: 2320 patients were screened and 325 of those were enrolled. Three major imaging patterns were identified: usual interstitial pneumonia (UIP)(76 patients), non-specific interstitial pneumonia (NSIP) (121 patients) and indeterminate (128 patients). UIP and NSIP were more common in SSc-ILD patients, and rarely in SLE/PM-ILD patients. Histopathology patterns from 43 lung biopsies could also be divided to three types: UIP (20 samples), NSIP (20 samples), undefined (3 samples). Statistical analysis showed positive correlation between histopathology patterns and imaging features on HRCT in CTD-ILD patients (P<0.05). Conclusion: Fibrosis is the main finding of HRCT in patients with SSc/RA-ILD and which is rarely seen in SLE/PM-ILD. Since there was a very good correlation between HRCT imaging findings and histopathology results, HRCT features of those patients with CTD may be a good indication of ILD and may avoid lung biopsy in those patients. Correlation between Imaging Features of High-resolution Computed Tomography and Histopathology of Connective Tissue Diseases associated Interstitial Lung Disease in Chinese Population: Avoid Lung Biopsy in Those Patients? Publication History: Received: February 16, 2017 Accepted: June 04, 2017 Published: June 06, 2017

Objective: Pirfenidone is a new, anti-fibrotic drug used for the treatmentof idiopathic pulmonary fibrosis (IPF). This study was designed to verify the effect of pirfenidone administrated in connective tissue diseases related interstitial lung disease (CTD-ILD). Methods: Twenty two patients diagnosed with CTD-ILD were administrated with a 6-months treatment with or without pirfenidone. At baseline and at months 3rd and 6th, pulmonary function tests, six minute walk distance (6MWD) and HRCT scores were performed. Results: PFD group presented a significant improvement in TLC in the 3rd and 6th month points and DLco was greatly increased in 6th month point, which were all adjusted by the baseline levels, compared with control group (p=0.043, 0.048 and 0.043, respectively). During the follow-up period, TLC was also improved compared to the initial time point and DLco increased obviously in the 6th month point in PFD group (p=0.005, 0.004 and p <0.001 respectively). A lower ground glass score and fibrotic score in the PFD group approached statistical significance after 6-months treatment with pirfenidone when compared with control group (p=0.037 and 0.018, respectively) and decreased greatly in the 6th month point when compared to the former time points in PFD group (p=0.006 and 0.013, respectively), whereas no such changes were observed in the control group. Conclusion: These results suggest that pirfenidone could improve the pulmonary function tests and HRCT scores of connective tissue diseases related interstitial lung disease. It is a potential therapeutic candidate for treatment.

AIM:To investigate the effect of pirfenidone on transforming growth factor-β1 (TGF-β1)-induced fibroblast-to-myofibroblast transition in vitro. METHODS:The cell viability was measured by MTT assay. The prolifera-tion of human lung fibroblasts ( HLFs) was detected by EdU incorporation. Migratory and invasive abilities were measured by Boyden chamber assay. The α-smooth muscle actin (α-SMA) protein expression was determined by Western blot and immunofluorescence. The mRNA expression of α-SMA and type Ⅰ and Ⅲ collagens was evaluated by RT-qPCR. RE-SULTS:Pirfenidone at different concentrations (0. 1, 0. 2, 0. 3, 0. 5 and 0. 8 mg/L) had no cytotoxic effect on the HLFs, and pirfenidone at 0. 2 mg/L was used for the intervention. Pretreatment of the HLFs with 0. 2 mg/L pirfenidone prior to TGF-β1 not only markedly suppressed the changes of proliferation, migration, invasion and reorganization of actin cytoskeleton in the HLFs (P<0. 01), but also down-regulated the expression ofα-SMA and typeⅠandⅢcollagens trig-gered by TGF-β1 ( P<0. 05 ) . CONCLUSION: Pirfenidone has an inhibitory effect on TGF-β1-induced activated cell functions and fibroblast-to-myofibroblast transition in HLFs.

结缔组织病相关间质性肺病(connective tissue disease-associated interstitial lung disease,CTD-ILD)是结缔组织病较为严重的并发症,预后极差,目前治疗手段有限,因此早期诊断、早期干预极为关键.CTD-ILD生物标志物检测简单易行,对于疾病的早期诊断及评估有着重要的意义.目前对CTD-ILD生物标志物的研究主要集中在:肺泡上皮蛋白、自身抗体、基质金属蛋白酶(matrix metalloproteinases,MMPs)及基质金属蛋白酶组织抑制因子(tissue inhibitor metalloproteinases,TIMPs)、趋化因子CCL家族和CXCL家族、白细胞介素家族、赖氨酰氧化酶、小窝蛋白-1、S100A8/A9等.本文就CTD-ILD的生物标志物最新进展进行综述.

OBJECTIVETo study the influence of hepatitis B virus (HBV) replication and expressions of different viral genes on CDC37 level in hepatocytes.METHODSWe amplified and cloned 6 HBV genes (P, preS1, preS2, S, C and X) into pCMV expression vectors, which were transfected in Huh7 and HepG2 hepatoma cell lines, and CDC37 expression level in the cells was detected using Western blotting. Wealso cloned the promoter sequence of CDC37 into pGL3 vector, and co-transfected pGL3 with pCMV recombinant plasmids into Huh7 and HepG2 cells and the fluorescent signals were detected. To study the influence of HBV replication on CDC37 expression, we constructed 1.28-copy overlength genomes of HBV genotypes B, C, D and CD recombinant. The overlength HBV genomes were transformed into Adeasier-1 cells for recombination and into 293 cells for packaging. Huh7 and HepG2 cell lines infected with the packaged HBV recombinant adenoviruses were examined for CDC37 expression with Western blotting.RESULTSWestern blotting showed that the expression of different HBV genes did not obviously affect the protein level of CDC37 in the hepatocytes. The protein expression of HBV genes had no effect on the activity of CDC37 promoter. Huh7 and HepG2 cells infected with 1.28-copy HBV replicon showed no significant changes in the expression level of CDC37.CONCLUSIONHBV replication and its gene expression have no effect on the level of CDC37 in hepatocytes in vitro.