Spss release 17.0 for windows

The percentages of apple wounds infected with P. expansum were converted to Bliss angular values (arcsine square root) before analysis of variance. Data were subjected to analysis of variance (ANOVA, SPSS release 17.0 for Windows; SPSS Inc., Chicago, IL, United States). All the means were compared by using Duncan’s multiple range test (P < 0.05). All the experiments were performed twice.

The characteristics of the subjects are expressed as the median and range unless otherwise stated. A one-sample Kolmogorov-Smirnov test, calculated to assess normality, showed non-normal distribution of the parameters studied. Between-group differences were analysed by the Mann-Whitney test and within-group differences by the Wilcoxon signed rank test. Differences were considered significant at a p value of ≤0.05. Multivariate logistic regression was used to analyse the independent association between asthma severity and the type of agent involved. All variables that were related to the quantities of interest and/or factors previously reported in the literature were considered as potential confounders. Results were reported using odds ratios (OR) and 95% confidence intervals (CI). SPSS release 17.0 for Windows (SPSS; Chicago, IL) and GraphPad InStat4 (GraphPad Software Inc; San Diego, CA) were used for the statistical analyses.

Each three RNA samples were extracted by the Yeast RNAiso Kit (Code No. 9751, TAKARA, Japan) according to the manufacturer’s instructions. The RNA concentration and purity was detected by Nanodrop2000 (Thermo scientific, USA). One μg of RNA was used to form cDNA by reverse transcription with PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (Takara, Japan) followed manufacturer’s instructions. The primers were designed by Primer 5.0. All the primers are shown in supplementary Table S1. The qRT-PCR was conducted using an ABI 7300 Real-Time PCR System (Applied Biosystems, USA). The qRT-PCR system was 20 μl which contained 10 μl SYBR Premix Ex Taq II (Tli RNaseH Plus)(10×), each 0.8 μl Forward primer (10 μΜ) and Reverse primer (10 μΜ), 0.4 μl Reference Rox Dye (50×), 2 μl cDNA and distilled water up to 20 μl. The conditions were as follows: 95 °C for 30 s, 40 cycles of 95 °C for 5 s, 60 °C for 31 s, and a dissociation curve of 95 °Cfor 15 s, 60 °C for 1 min, 95 °C for 15 s followed the amplification cycle was added to determine the reliability of the quantitative results. Each sample have three technical replicates. Data were subjected to analysis of variance by using t test (SPSS release 17.0 for Windows; SPSS Inc., Chicago, Illinois, USA).