duration of surgery was 195 (163–275) minutes in group TIVA and 247 (174–276) in CT99021 molecular weight group INHALATION. Blood samples were collected in tubes coated with ethylenediaminetetraacetic acid (EDTA), 7.2 mg EDTA per 4.5 ml blood. After centrifugation for removal of cells, the samples were frozen within 30 min and stored at −80 °C. The blood samples provided data on the levels of complement split products (C3a and SC5b-9), pro-inflammatory cytokines (TNF-α, IL-1β, IL-6 and IL-8) and anti-inflammatory cytokines (IL-4 and IL-10). The levels of TNF-α, IL-1β, IL-4, IL-6, IL-8 and IL-10 were obtained with SearchLight method (Pierce Biotechnology, Woburn, MA, USA), which is a multiplex sandwich enzyme-linked immunosorbent assay (ELISA) in a planar, plate-based array format,
for the quantitative measurement of secreted proteins in different biological materials. Diluted samples and controls were incubated for one hour on the arrayed plates. All incubations were performed at room temperature selleck chemicals llc with shaking at 200 rpm. The plates were decanted and washed six times before adding a cocktail of biotinylated detection antibodies to each well. After incubating with detection antibodies for 30 min, the plates were washed three times and incubated for 30 min with streptavidin–horseradish peroxidase. The plates were again washed before adding SuperSignal Femto Chemiluminescent substrate. The plates were immediately imaged using the SearchLight Black Ice imaging system, and data were analysed using Array Analyst software (Auchon Digestive enzyme Biosystems, Billerica, MA, USA). All results were in the range of the standard curve. Differences in age, duration of anaesthesia and surgery, blood loss, American Society of Anesthesiologists (ASA) physical status classification scores and length of stay at hospital post-operatively were tested between the treatment groups using Mann–Whitney U-tests. Chi-squared tests were used to compare proportions. Mean values of each inflammatory marker for each anaesthetic treatment group at each measurement point were inspected graphically. The repeated measurements were
then analysed using linear mixed models with an unstructured covariance structure and maximum likelihood estimates for both all patients and those without inflammatory bowel disease (IBD). All exploratory and formal statistical tests were carried out using SPSS for Windows (Version 18; SPSS Inc, Chicago, IL, USA). All P-values were two-tailed, and P-values <0.05 were considered statistically significant. There were no significant differences between the anaesthesia groups regarding clinical parameters. The parameters are given in Table 1. Complement and interleukin determinations are given in Table 2 and in Figs. 1–6. The C3a levels were increased during surgery in both groups compared with baseline (P < 0.001).