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ABSTRACT Respiratory tract infections (RTIs) caused by viruses are prevalent and significant conditions in clinical settings. Accurate and effective detection is of paramount importance in the diagnosis, treatment, and prevention of viral RTIs. With technological advancements, multiplex real-time reverse transcription polymerase chain reaction (rRT-PCR) assays have been developed and extensively adopted for the diagnosis of viral RTIs. Given the potential challenges in the detection performance of multiplex assays, this study evaluated the analytical sensitivity and competitive interference of the six most commonly used multiplex rRT-PCR kits for detection of respiratory viruses in China. The results revealed that the limits of detection were variable across the viruses and kits. Most of the evaluated multiplex kits demonstrated comparable or enhanced analytical sensitivity compared with singleplex kits for clinically significant viruses, including human adenovirus (HAdV)-3, HAdV-7, Omicron BA.5, H1N1pdm09, H3N2, B/Victoria, respiratory syncytial virus subtype A, and respiratory syncytial virus subtype B, whereas multiplex kits showed relatively less analytical sensitivity for human rhinovirus-B72, human metapneumovirus-A2, parainfluenza virus (PIV)-1, and PIV-3. In addition, most multiplex kits successfully identified co-infections when one analyte was present at a low concentration and another analyte was present at a high concentration.IMPORTANCEThe complexity and severity of viral respiratory tract infections (RTIs) emphasize the pivotal role of precise diagnosis for viral RTIs in guiding effective public health responses and ensuring appropriate medical interventions, given the substantial population at risk. This study highlights the necessity and importance of evaluating the analytical validity of multiplex real-time reverse transcription polymerase chain reaction assays, offering valuable insights into their optimization and application.

The real-time input of ground motions can effectively assess earthquake disasters in building structures. In this study, we proposed a nearby real-time simulation method that can be applied to assess regional earthquake disasters using structural monitoring recordings based on an urban earthquake simulation system. Real-time accelerations recorded during the 5.1 magnitude Tangshan earthquake were used to update and modify the computing models. El Centro waves were calibrated to 400 cm/s2 for structural seismic resilience analyses. The results indicated that approximately 70% of the structural functions were lost during the rare earthquake. Regional numerical models of 216 buildings were constructed and timeously updated using a geographic information system and measured data. The inputting ground motions recorded in the 5.1 magnitude Tangshan earthquake and typical El Centro waves were selected to analyze the structural seismic response, in which the damage indexes were computed, and the damage predictions of the regional 216 buildings were also simulated in different levels of earthquakes by being combined with the simplified principles of structural damage estimation. Finally, the evaluation results were visualized in three dimensions using ParaView software. The simulated results of earthquake disasters at an urban scale will promote the prediction abilities of local earthquake administration agencies and have considerable potential to provide essential information for emergency responses and regional disaster mitigation.

Abstract Targeted panel-based tumor mutation burden (TMB) assays are widely employed to guide immunotherapy for patients with solid tumors. However, the accuracy and consistency of this method can be compromised due to the variability in technical details across different laboratories, particularly in terms of panel size, somatic mutation detection and TMB calculation rules. Currently, systematic evaluations of the impact of these technical factors on existing assays and best practice recommendations remain lacking. We assessed the performance of 50 participating panel-based TMB assays involving 38 unique methods using cell line samples. In silico experiments utilizing TCGA MC3 datasets were performed to further dissect the impact of technical factors. Here we show that the panel sizes beyond 1.04 Mb and 389 genes are necessary for the basic discrete accuracy, as determined by over 40,000 synthetic panels. The somatic mutation detection should maintain a reciprocal gap of recall and precision less than 0.179 for reliable psTMB calculation results. The inclusion of synonymous, nonsense and hotspot mutations could enhance the accuracy of panel-based TMB assay. A 5% variant allele frequency cut-off is suitable for TMB assays using tumor samples with at least 20% tumor purity. In conclusion, this multicenter study elucidates the major technical factors as sources of variability in panel-based TMB assays and proposed comprehensive recommendations for the enhancement of accuracy and consistency. These findings will assist clinical laboratories in optimizing the methodological details through bioinformatic experiments to enhance the reliability of panel-based methods.

Nontypeable Haemophilus influenzae (NTHi) is the dominant pathogen in several infectious diseases. Currently the use of antibiotics is the main intervention to prevent NTHi infections, however with the emergence of drug resistant strains, it has compromised the treatment of respiratory infections with antibiotics. Therefore there is an urgent need to develop a safe and effective vaccine to prevent NTHi infections. We investigate the potential of C-HapS-P6 fusion protein as a vaccine for treating NTHi in murine models. PGEX-6P2/C-HapS-P6 fusion gene was constructed using overlap extension polymerase chain reaction. The recombined plasmid was transformed into Escherichia coli for protein expression. The mice were subjected to intraperitoneal immunization using purified antigens. Immunoglobulin (Ig) G in serum samples and IgA in nasal and lung lavage fluids were analyzed using enzyme-linked immunosorbent assay. Cytokine release and proliferation capacity of splenic lymphocytes in response to antigens were measured in vitro. The protective effect of the C-HapS-P6 protein against NTHi infection was evaluated by NTHi count and histological examination. The data showed that the C-HapS-P6 fusion protein increased significantly the levels of serum IgG and nasal and lung IgA, and promoted the release of interleukin (IL)-2, interferon-ϒ, IL-4, IL-5, and IL-17 and the proliferation of splenic lymphocytes compared with C-HapS or P6 protein treatment alone. Moreover, C-HapS-P6 effectively reduced the NTHi colonization in the nasopharynx and lungs of mice. In conclusion, our results demonstrated that the C-HapS-P6 fusion protein vaccine can significantly enhance humoral and cell immune responses and effectively prevent against NTHi infection in the respiratory tract in murine models.

Abstract Transverse cracking is a serious problem for semi-rigid base asphalt pavement. The shrinkage of the base course and the surface course, as well as reflective cracks, are key factors for transverse cracking in asphalt pavement. Crack spacing can directly reflect the degree of transverse cracking in pavements. Therefore, this study aims to calculate the transverse crack spacing and discuss its affecting factors. To this end, a calculation model of transverse crack spacing for the semi-rigid base asphalt pavement was first established. Then, the transverse crack spacings of different composite structures were calculated, and the influences of the shrinkage coefficient, the structural layer thickness, and the pavement tensile strength on transverse crack spacing were expounded. Finally, the transverse crack spacing of the pavement after the appearance of the reflective was calculated. The results show that the lower lime and fly ash content and skeleton gap gradation can be adopted during the design of the base course. Meanwhile, the lower lime and fly ash content in the macadam base, the skeleton gap gradation and asphalt concrete with a larger particle size in the surface layer can be used during the design of surface layer. In addition, the transverse crack spacing of the semi-rigid base asphalt pavement could be increased by reducing the shrinkage coefficient, increasing the thicknesses of the surface course and the base course, and improving the tensile strength of the pavement. After the appearance of reflective cracks, the transverse crack spacing of the surface layer ranged between 32.8 m and 66.5 m. 15fp-AC25, 15fp-AC20, 15df-AC25, and 17fp-AC25 were found to be the best semi-rigid base asphalt pavement structures to reduce transverse cracking. Finally, transverse cracking in pavement composite structures under different bonding conditions needs to be analyzed in the follow-up work.