Histopathology Serial sections of tumor tissue were processed for routine histological examination. The specimens were 3-Methyladenine in vivo washed with PBS to remove blood, fixed with formaldehyde, dehydrated with a graded alcohol series, and embedded in paraffin. Hematoxylin eosin staining (H&E) was performed on the specimens, for histopathologic evaluation of hemorrhage, necrosis, and inflammation. Statistical Analysis Statistical analyses were performed by the SPSS 13.0 software package (SPSS, Inc,

Chicago, IL). All values were expressed as mean ± SD. Analysis of variance with t test and analysis of variance AZD6738 purchase (ANOVA) test were used to determine the significance of the difference in a multiple comparison. If the ANOVA was significant, the

Tukey’s procedure was used as a post hoc test. Differences with a P value of less than 0.05 were considered to be statistically significant. Results Identification of pCMV-LUC by Restriction Enzymes Digestion After double-enzyme cutting by Bam HI and Hind III, the restriction enzymes digestion results showed Selleck Alvespimycin that the objective fragment of the pCMV-LUC plasmid could be detected at around 2000 bp, which was exactly coincidence with the size of the designed DNA (Figure 1). Figure 1 Identification of pCMV-LUC by restriction enzymes digestion. 1, Marker, 1 kb ladder; 2, pCMV-LUC; 3, pCMV-LUC/Hind III + Bam HI; the plasmid pCMV-LUC or with the restriction enzymes digestion showed two bands after electrophoresis. A correct insertion was

showed a band of 2000 bp (as arrowhead indicated) cut off by Hind III and Bam HI. Gel Retardation Analysis of PEI/DNA Complexes Agarose gel electrophoresis analysis showed Decitabine molecular weight that (Figure 2), with the increase of N/P ratio of PEI/DNA complexes, the plasmid DNA migrated more slowly. When N/P ≥ 3, the plasmid DNA migration could not be observed, and the PEI/DNA complexes with positive charge remained in the hole. PEI could effectively condensate the plasmid DNA, and the electrophoresis analyses of both plasmids were similar (Figure 2A-B). According to the results of electrophoresis, the N/P ratio was chose for 6 in this study and used in the following experiments. Figure 2 Electrophoretic patterns of plasmid DNA complexes prepared with PEI at various N/P ratios: N/P ratio = PEI nitrogen/DNA phosphate; (A) pCMV-LUC, (B) pSIREN-S. Lanes 1-10: the N/P molar ratios of 1/4, 1/2, 1, 3/2, 2, 5/2, 3, 4, 6, and 8. Enhanced RFP Expression in Transplanted Tumors by Combination of UTMD and PEI Regardless of ultrasound irradiation, there was no obvious RFP expression in Group A and B (Figure 3A-B). Without ultrasound irradiation, there were only a few cells expressing RFP in pSIREN-C/SonoVue group and red fluorescent signal was weak in the majority of samples (Figure 3C).