Preparation of purified proteins from fresh Pheretima and their inhibitory effect against pulmonary fibrosis in mice / 南方医科大学学报

OBJECTIVE@#To develop a convenient method for rapid purification of fresh Pheretima proteins and assess the inhibitory effect of these proteins against pulmonary fibrosis.@*METHODS@#The crude extract of fresh Pheretima was obtained by freeze-drying method and then purified by size exclusion chromatography. The composition of the purified proteins was analyzed by mass spectrometry. MRC-5 cells were treated with 5 ng/mL TGF-β1 alone (model group) or in combination with SB431542 (2 μmol/L) or the purified proteins (13.125 μg/mL), and the cytotoxicity of purified proteins and their inhibitory effects on cell proliferation were detected with CCK8 assay. Flow cytometry was used to detect the changes in cell apoptosis, and the cellular expressions of α-SMA, Vimentin, E-cadherin, collagen I, Smad2/3 and P-Smad2/3 were detected using RT-PCR and Western blotting. In the animal experiment, adult male C57BL/6 mice were subjected to intratracheal instillation of bleomycin followed by treatment with the purified proteins (5 mg/mL) for 21 days, after which HE and Masson staining was used to observe the pathological changes in the lung tissue of the mice.@*RESULTS@#We successfully obtained purified proteins from fresh Pheretima protein by size exclusion chromatography. Treatment with the purified proteins significantly inhibited TGF-β1-induced proliferation of MRC-5 cells (P < 0.01), reduced the cellular expressions of α-SMA, Vimentin and collagen I (P < 0.001 or P < 0.01), increased the expression of E-cadherin (P < 0.01), and inhibited the expressions of Smad2/3 and P-Smad2/3 (P < 0.001 or P < 0.01). In male C57BL/6 mice models of bleomycin-induced pulmonary fibrosis, treatment with the purified proteins obviously reduced the number of inflammatory cells and fibrotic area in the lungs.@*CONCLUSION@#The purified proteins from fresh Pheretima obtained by size exclusion chromatography can inhibit pulmonary fibrosis in mice by regulating the TGF-β/ Smad pathway.

Application evaluation and analysis of clinical case discussion in CPPT course / 中华医学教育探索杂志

Objective:To explore and optimize the teaching content of clinical case discussion course based on clinical pathophysiology and therapy (CPPT), and to improve the teaching quality of this course.Methods:Self-compiled evaluation form was sent to all the students who participated in this course in 2016-2017 school year, and the teaching satisfaction and teaching characteristics were evaluated. The differences of evaluation results from students with different educational programs or teachers with different seniority were compared, and the correlation between evaluation results and teachers' seniority was analyzed.Results:The students had a high evaluation on the course, but students from different programs had significant different comments on thinking inspiring, speech encouraging, questions guiding, process holding and effective feedback, as well as the self-evaluation of mastery degree. For teachers with different seniority, there were significant differences in scores in all aspects except speech encouraging, but no differences in mastery degree. Spearman analysis showed that there was a certain correlation between scores and teachers' seniority.Conclusion:The overall situation of the CPPT case discussion course in our school is worthy of recognition, but it needs to be improved in many aspects.

Blood biochemical indicators, IgM and IgG antibodies in 56 confirmed and suspected COVID-19 cases / 上海预防医学

Objective To provide reliable indicators for effective prevention and control of COVID-19, we examined the biochemical indicators as well as anti-SARS-CoV-2 IgM and IgG antibodies in confirmed and suspected COVID-19 patients. Methods A total of 56 confirmed and suspected COVID-19 cases quarantined during January-March, 2020 in Gansu Provincial People′s Hospital and People′s Hospital of Xigu District, Gansu Province were included.Based on the results of nucleic acid testing and CT scan finding, they were divided into three groups: positive in both nucleic acid testing and CT scan finding; positive in nucleic acid testing but negative in CT scan finding; negative in both nucleic acid testing and CT scan finding.COVID-19 viral nucleic acid was detected and chest CT scan was performed.The following biochemical indicators were examined: total protein, albumin, total bilirubin, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, γ-glutamyltranspeptidase, creatine kinase and lactate dehydrogenase.SPSS 22.0 statistical software was used to analyze the differences in the biochemical indicators among these three groups, as well as the temporal trend of IgM and IgG antibodies at different points of time. Results There were significant differences in the mean values of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and lactate dehydrogenase between these three groups (P < 0.05), whereas no significant difference in the mean value of total protein and albumin (P>0.05).ROC curve indicated that aspartate aminotransferase had the largest maximum area under the curve (AUC, 0.834), whereas alanine aminotransferase had the highest sensitivity (1.0) and total bilirubin had the highest specificity (0.927).Thus, aspartate transaminase provided the best prediction for the diagnosis of COVID-19, with sensitivity of 0.786, specificity of 0.854, and the maximum AUC of 0.834.In 12 of 16 confirmed COVID-19 patients tested IgG positive after 10 days of diagnosis, and 10 of 10 confirmed COVID-19 patients tested IgG positive after 14 days of diagnosis. Conclusion Aspartate aminotransferase may be the most useful indicator in the diagnosis of COVID-19.

Effects of Bruton's tyrosine kinase on the proliferation and differentiation of osteoclasts / 华西口腔医学杂志

OBJECTIVE@#To observe the effect of Bruton's tyrosine kinase (BTK) on the proliferation and differentiation of osteoclasts and to explore the mechanism of BTK on bone destruction in periapical periodontitis.@*METHODS@#After RAW264.7 cells induced with 100 ng·L⁻¹ receptor activator for nuclear factor-κB ligand (RANKL) for 5 days, osteoclast induction was confirmed by light microscopy, tartrate-resistant acid phosphatase (TRAP) staining, and quantitative real-time PCR (RT-qPCR). Then, BTK-small interfering RNA (BTK-siRNA) was transfected into cells induced for 5 days. After 24 h, the expression of TRAP mRNA was measured using RT-qPCR, and the proliferation and differentiation of osteoclasts were detected using CCK-8 and TRAP activity assay. Statistical analysis was performed.@*RESULTS@#After RAW264.7 was induced with RANKL for 5 days, a large number of round, ellipse, irregularly protuberant, and TRAP-positive macrophages were observed under light microscopy. The expression of TRAP mRNA significantly reduced after 24 h of BTK-siRNA transfection (P<0.05). The detection of CCK-8 and TRAP activities showed that the proliferation and differentiation of osteoclasts significantly decreased (P<0.05).@*CONCLUSIONS@#Silencing of BTK can inhibit the proliferation and differentiation of osteoclasts. BTK can be used as a new target for the inhibition of osteoclasts.