RESUMO
SARS-CoV-2 and its induced COVID-19 remains as a global health calamity. Severe symptoms and high mortality, caused by cytokine storm and acute respiratory distress syndrome in the lower respiratory airway, are always associated with elderly individuals and those with comorbidities; whereas mild or moderate COVID-19 patients have limited upper respiratory flu-like symptoms. There is an urgent need to investigate SARS-CoV-2 and other coronaviruses replication and immune responses in human respiratory systems. The human reconstituted airway epithelial air-liquid interface (ALI) models are the most physiologically relevant model for the investigation of coronavirus infection and virus-triggered innate immune signatures. We established ALI models representing both the upper and the lower respiratory airway to characterize the coronavirus infection kinetics, tissue pathophysiology, and innate immune signatures from upper and lower respiratory tract perspective. Our data suggested these in vitro ALI models maintain high physiological relevance with human airway tissues. The coronavirus induced immune response observed in these upper and lower respiratory airway models are similar to what has been reported in COVID-19 patients. The antiviral efficacy results of a few promising anti-coronavirus drugs in these models were consistent with previous reports and could be valuable for the human dose prediction. Taken together, our study demonstrates the importance of 3D airway epithelial ALI model for the understanding of coronavirus pathogenesis and the discovery and development of anti-coronavirus drugs.
RESUMO
Objective This exploratory study aimed to assess effectiveness with ethylene oxide treatment for removing DNA contamination. Methods 98 different spiked samples such as saliva, dander, skin cell, hair, blood and cartilage were conducted with ethylene oxide treatment. After extraction of samples, the dna was amplified and then the STR analysis was performed with 3130xl or 3500xl. Results A 6h EO treatment results showed that two saliva stains of 44 samples STR profile were detected; Just one hair of 54 samples treated with ethylene oxide was detected contaminating DNA with EO treatment for 8 hours. Conclusion This work suggested that it was more successful to reduce DNA contamination by using ethylene oxide treatment.
RESUMO
Objective This study aimed to assess the efficiency and purification of the Trace DNA extraction with a quantified method for the magnetic bead-based DNA extraction as performed on the Tecan Automated systems with TE-MAGS magnetic separator. Methods Serial dilutions of standard commercial DNA 9947A were used with the total DNA contents, 0.1ng, 0.2ng,0.3ng, 0.4ng,0.5ng, 0.6ng, 0.7ng, 0.8ng, 0.9ng,1ng,diluted progressively and a 1ng DNA (standard commercial DNA 9947A) admixed with 6 common DNA-PCR inhibitors were extracted on the Automated systems and then performed via Fluorogenic probe quantitative PCR and STR genotype for the quantification analysis of recovery and purification. Results The recovery rate of standard 9947A DNA diluted with 0.1~1ng was 38.92~60.01%, and 0.3ng and more DNA could observed the full STR profiles. For the different PCR inhibitors, above 94.5% of bile acid, collagen and urea were efficient removal during the purification process, and the hemoglobin, melanin and humic acid removal efficiency were about 97.5%, 97.85%, 82.14%, respectively.Conclusion Our results suggested that The TE-MAGS magnetic bead-based DNA extraction was suitable for the extraction of trace DNA with high recovery efficiency and purification ability.
