SiteFinding-PCR: a simple and efficient PCR method for chromosome walking

In this paper, we present a novel PCR method, termed SiteFinding-PCR, for gene or chromosome walking. The PCR was primed by a SiteFinder at a low temperature, and then the target molecules were amplified exponentially with gene-specific and SiteFinder primers, and screened out by another gene-specific primer and a vector primer. However, non-target molecules could not be amplified exponentially owing to the suppression effect of stem–loop structure and could not be screened out. This simple method proved to be efficient, reliable, inexpensive and time-saving, and may be suitable for the molecules for which gene-specific primers are available. More importantly, large DNA fragments can be obtained easily using this method. To demonstrate the feasibility and efficiency of SiteFinding-PCR, we employed this method to do chromosome walking and obtained 16 positive results from 17 samples

Role of neuromedin B and its receptor in the innate immune responses against influenza A virus infection in vitro and in vivo

International audienceAbstractThe peptide neuromedin B (NMB) and its receptor (NMBR) represent a system (NMB/NMBR) of neuromodulation. Here, it was demonstrated that the expression of NMBR in cells or murine lung tissues was clearly upregulated in response to H1N1/PR8 influenza A virus infection. Furthermore, the in vitro and in vivo activities of NMB/NMBR during PR8 infection were investigated. It was observed that A549 cells lacking endogenous NMBR were more susceptible to virus infection than control cells, as evidenced by the increased virus production in the cells. Interestingly, a significant decrease in IFN-α and increased IL-6 expression were observed in these cells. The role of this system in innate immunity against PR8 infection was probed by treating mice with NMB. The NMB-treated mice were less susceptible to virus challenge, as evidenced by increased survival, increased body weight, and decreased viral NP expression compared with the control animals. Additionally, the results showed that exogenous NMB not only enhanced IFN-α expression but also appeared to inhibit the expression of NP and IL-6 in PR8-infected cells and animals. As expected, opposing effects were observed in the NMBR antagonist-treated cells and mice, which further confirmed the effects of NMB. Together, these data suggest that NMB/NMBR may be an important component of the host defence against influenza A virus infection. Thus, these proteins may serve as promising candidates for the development of novel antiviral drugs

Analysis of the correlation between the pavement structure combinations and pavement performance using experimental sections

The new generation of pavement technology with the goal of longevity is an important supporting technology that can promote the achievement of sustainable development of high-speed roadways. To further investigate the evolution trend of long-life pavement performance, this paper paved experimental sections to explore the correlations between pavement structure combinations and pavement performances. This paper presented four experimental sections with different pavement structures, asphalt concrete layer thicknesses, and pavement materials. Then, this paper analyzed the effects of the seasonal factors, pavement structure, and lanes on the deflection value and rut depth from three dimensions by the Pearson correlation coefficient (PCC). Finally, this paper used the analysis of variance (ANOVA) to analyze the relationships between the layer thickness of various materials in the pavement structure and the pavement performances, including the deflection value, international roughness index (IRI), texture depth (TD), British Pendulum Number (BPN), sideway force coefficient (SFC), rut depth, and disease area. The results showed that the seasonal factors significantly affected the deflection values of pavement structures with PCCs of 0.61, 0.72, 0.53, and 0.78. The high temperatures increased the average deflection values by 22.85%, 72.88%, 77.61%, and 88.13%, respectively. Under the influence of high temperature in summer and traffic loads, the increased ranges of average rut depth were −0.2%, 4.89%, 9.56%, and 7.31%, respectively. The results of ANOVA showed that the pavement structure type and thickness of each structural layer significantly affected the deflection value, and there also was a strong correlation between the pavement structure type, thickness, BPN, and SFC with p-values less than 0.05. Increasing the thickness of the asphalt surface was beneficial for reducing the area of defects, while laying the semi-rigid base layer was beneficial for maintaining the deflection value and rut depth at a lower level

Safety and tolerability of a single dose T0001 in Chinese healthy adult volunteers: a first-in-human ascending dose study

T0001 is the first mutant of etanercept with a higher affinity to tumor necrosis factor α (TNF-α) than etanercept. In order to investigate the safety and tolerability of T0001, a study was carried out in healthy Chinese subjects. A first-in-human, dose escalation study was conducted in healthy Chinese subjects. Fifty-six subjects were divided into six dose cohorts (10 mg, 20 mg, 35 mg, 50 mg, 65 mg and 75 mg) to receive a single subcutaneous injection of T0001. Safety and tolerability assessment were based on the records of vital signs, physical examinations, clinical laboratory tests, 12-lead electrocardiograms and adverse events (AEs). All subjects were in good compliance and none withdraw due to AEs. No serious AEs occurred. A total of twenty-three AEs in sixteen subjects were recorded, and eighteen of these AEs were believed to be related to T0001. The most frequently reported AEs were injection site reactions and white blood cell count increase. All these AEs were of mild to moderate intensity and most of them recovered spontaneously within 14 days. In this study, no dose-limiting toxicity was observed, and the maximum tolerated dose was identified as 75 mg. T0001 was considered safe and generally well tolerated at doses up to 75 mg in healthy Chinese volunteers

Preliminary discussion on the research and application of univariate linear weighted regression in analytical chemistry

The paper mainly expounds the principles， differences and selection principles of ordinary least squares and weighted least squares in univariate linear regression， and introduces the application in four aspects， including heteroscedasticity test， optimal weight function selection， regression equation test and comparison of fitting quality. The application and existing problems of the weighted least squares method at home and abroad is analyzed. Suggestions on how to promote the application research of regression technology in analytical chemistry to provide reference and inspiration for researchers are put forward