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.
Objective To explore the immunopathologic mechanism underlying the inflammatory response after severe acute respiratory syndrome（SARS） invasion.Methods Pathway focused cDNA microarrays were employed for comparision of the gene expression patterns in 16HBEs treated with interferon-γ(IFN-γ) or the S protein of SARS-CoV.The S proteins were administered to BALB/c mice and the pathological changes of lung and spleen were observed.Results S protein activated JAK/STAT signal pathway in the 16HBEs with inducible protein 10（IP-10） gene expression，and the specific inhibitors of the JAK/STAT signal pathway were able to downregulate the induction of IP-10.The mice instilled intracheally with S proteins revealed obvious acute diffuse damage and increased IP-10 expression and CD68+ macrophages infiltration in both lung and spleen tissues.In contrast，the treatment with JAK3 inhibitors attenuated lung and spleen injury in the mice.Conclusion Our findings support that the activation of JAK/STAT pathway induced by SARS-CoV S protein plays a key role in promotion of an IFN -γ inducible chemokine cascade，which can help in the development of novel drug and therapeutics for prevention and treatment of SARS.
Objective To explore whether epithelial to mesenchymal transition ( EMT) occurs in bleomycin( BLM) induced pulmonary fibrosis, and the involvement of bronchial epithelial cells( BECs) in the EMT. Methods BLM-induced peribronchial fibrosis in an α-smooth muscle actin-Cre transgenic mouse( α-SMACre /R26R) was examined by pulmonary βgal staining and α-SMA immunofluorescence staining. Results BLMtreated mice showed significantly enhanced βgal staining in subepithelial areas in bronchi, terminal bronchioles and walls of pulmonary vessels. Some alveolar epithelial cells( AECs) in certain peribronchial areas or even a small subset of BECs were also positively stained, as confirmed by α-SMA immunostaining. Conclusions EMT occurs in BLM-induced peribronchial fibrosis mice. BECs, like AECs, have the capacity to undergo EMT and to contribute to mesenchymal expansion in pulmonary fibrosis.
Objective To investigate the expression of high mobility group protein-B1( HMGB1)and α-smooth muscle actin( α-SMA) in Bleomycin induced pulmonary fibrosis in mice. Methods Twenty C57BL/ 6 male mice were randomly divided into a Bleomycin group and a control group. The Bleomycin group was treated with Bleomycin( 3 mg/kg) by endotracheally injection to induce pulmonary fibrosis. The control group were treated with normal saline( NS) . Then they were sacrificed by abdominal aortic bleeding 10 days after the injection. The right lung was stained with hematoxylin-eosin and Masson trichrome respectively for pathological examination. Immunohistochemistry and RT-PCR were performed to identify the protein and mRNA levels of α-SMA and HMGB1 respectively. Results The mRNA( 0. 89 ±0. 12, 0. 61 ±0. 08) and protein( 13. 66 ±1. 01, 13. 12 ±1. 33) expressions of α-SMA and HMGB1 in the Bleomycin group were all significantly higher than those of the control group( mRNA: 0. 60 ±0. 07, 0. 15 ±0. 02; protein: 8. 18 ±1. 33,7. 92 ±1. 10; all P lt; 0. 01) . Conclusions The expressions of HMGB1 and α-SMA are increased in Bleomycin induced pulmonary fibrosis. HMGB1 participates in the pathological process of pulmonary fibrosis probably by activation of the α-SMA expression.
Objective To investigate the feasibility of detection of epidermal growth factor receptor ( EGFR) exon 19 deletions and exon 21 L858R mutations in pleural effusion fromnon-small-cell lung cancer ( NSCLC) patients by mutant enriched PCR assay. Methods The mutations of exon 19 and 21 of EGFR gene in pleural samples fromthirty NSCLC patients were analyzed using both the mutant-enriched PCR assay and the non-enriched PCR assay. Results Ten ( 33. 3% , 10/ 30) exon 19 deletions and five ( 16. 7% , 5/30) exon 21 L858R mutation were detected by the mutant-enriched PCR assay, while only 6 cases ( 20. 0% ) and 1 case ( 3. 3% ) were detected by the non-enriched PCR assay respectively. The difference of mutation detection rate of EGFR gene between the two methods was statistically significant ( P = 0. 032) . Mutations were detected in all of partial responders ( 2 /4) among the four patients who received gefitinib therapy. Conclusions Mutant-enriched PCR assay can detect EGFR exon 19 deletions and exon 21 L858R mutation in pleural effusion from NSCLC patients effectively, economically and accurately. It may be a valuable biomarker for gefitinib therapy in advanced NSCLC.
Objective To investigate the effects and mechanisms of lactic acid bacteria on MAPK signaling in immune response of dust mite sensitized mice. Methods Forty C57BL/6 mice in Group M, P and L, were sensitized and challenged with mite extract while then the animals in Group N were treated with saline as control. The mice in Group L and P were fed with Lactococcus lactis or Lactobacillus respectively.Three days after the last challenge, all mice were sacrificed for lung pathological examination. IL-10 level in culture supernatant of splenocytes stimulated with mite extract was detected by ELISA. The expression of IL-4/ IFN-γon CD3 +CD4 + cells was detected by flow cytometry. Western blot were performed for detection of MAPK signaling ( P38, ERK, and JNK) from mice’s spleen cells stimulated with mite extract. Results The mice fed with Lactococcus lactis ( Group L) had lower rate of eosinophilic airway inflammation and higher level of IL-10 in the culture supernatant of splenocytes than Group P. Meanwhile, the number of CD4 + T cell with IL-4 expression was decreased revealed by the analysis of flow cytometry. P38 signaling inspleen cells was activated in the mice of Group M, similarly in the mice of Group P, but not of Group L.Conclusion Oral treatment of Lactococcus lactis can induce an immune tolerance in response to mite by up-regulating the level of Tr cells secreting IL-10, thus inhibiting activation of P38 signaling.
Objective To evaluate the clinical significance of epidermal growth factor receptor EGFR) mutations in the treatment of non-small cell lung cancer ( NSCLC) . Methods Plasma DNAs solated fromblood specimens of 170 NSCLC patients, who were admitted in the First Affiliated Hospital of uangzhou Medical College from December 2005 to December 2007, were subjected to the test of EGFR utant-enriched PCR. The correlation of mutant detection with clinical characteristics was analyzed as well.Results Out of the total 170 patients, EGFR mutations were identified in 77 cases ( 77 /170, 45. 3% ) .EGFR mutations were more frequent in the patients with adenocarcinoma ( P lt; 0. 001) and in the nonsmokers P =0. 001) . In the 33 patients treated with gefitinib, those with mutations ( + ) showed a higher esponse rate and prolonged progression-free survival after the treatment compared with those with mutations( - ) ( P =0. 001 and 0. 001, respectively) . Conclusions EGFR active mutations can be specifically and ensitively detected by EGFR mutant enriched PCR assay. Plasma EGFR mutants detection is valuable in uiding clinical decision.
ObjectiveTo research on the correlation between modified shock index (MSI), traditional vital sign assessment indexes and the proportion of patients entering resuscitation room through emergency triage, and to discuss its significance and advantages for emergency triage. MethodA total of 22 153 emergency patients between January 1 and May 31, 2009 were retrospectively analyzed. We counted the shock index (SI), mean arterial pressure (MAP), MSI, and evaluated the reference range of MSI, based on which, the patients were divided into groups, and the proportion of patients entering resuscitation room in each group was compared. Based on pulse, systolic blood pressure (SBP), SI, MAP and MSI, the patients were again grouped for comparing the proportion of patients entering resuscitation room, and the positive predictive value, negative predictive value, and odds ratio (OR) were also analyzed. ResultsReference value of MSI ranged from 0.34 to 1.70 times/(min·mm Hg) (1 mm Hg=0.133 kPa). Positive predictive values:MSI (16.239%) >MAP (6.115%) >pulse (5.746%) >SBP (5.321%) >SI (3.689%). The negative predictive values were all at high levels and similar with each other. OR:MSI (6.138) >MAP (2.498) >pulse (2.431) >SBP (2.117) >SI (1.361). ConclusionsPulse, SBP, SI, MAP, and MSI are correlated with the proportion of patients entering resuscitation room, and can be regarded as guide for emergency triage, among which MSI may be superior to all other indexes.
The three-dimensional (3D) liver and tumor segmentation of liver computed tomography (CT) has very important clinical value for assisting doctors in diagnosis and prognosis. This paper proposes a tumor 3D conditional generation confrontation segmentation network (T3scGAN) based on conditional generation confrontation network (cGAN), and at the same time, a coarse-to-fine 3D automatic segmentation framework is used to accurately segment liver and tumor area. This paper uses 130 cases in the 2017 Liver and Tumor Segmentation Challenge (LiTS) public data set to train, verify and test the T3scGAN model. Finally, the average Dice coefficients of the validation set and test set segmented in the 3D liver regions were 0.963 and 0.961, respectively, while the average Dice coefficients of the validation set and test set segmented in the 3D tumor regions were 0.819 and 0.796, respectively. Experimental results show that the proposed T3scGAN model can effectively segment the 3D liver and its tumor regions, so it can better assist doctors in the accurate diagnosis and treatment of liver cancer.