Objective:To compare the expression and immunizing efficiency of the hCGβ-C3d3 fusion protein between pCMV4 and pcDNA3.Methods:The plasmid pCMV4-hCGβ-C3d3 was constructed from pcDNA3-hCGβ-C3d3.COS-7 cells were transfected with pCMV4-hCGβ-C3d3 and pcDNA3-hCGβ-C3d3,and the expression efficiency in vitro was compared between them.BALB/C mice of 6 weeks old were immunized intramuscularly by pcDNA3,pcDNA3-hCGβ-C3d3,pCMV4-hCGβ-C3d3 ,respectively.The anti-hCGβ antibody titers were determined by indirect ELISA in 6 weeks of last immunization.Results:There was a significant difference of expression efficiency in vitro in COS-7 cells between the pcDNA3-hCGβ-C3d3 and pCMV4-hCGβ-C3d3( P 0.01).The mice produced significantly higher anti-hCGβ antibody level after DNA immunization with pcDNA3-hCGβ-C3d3 and pCMV4-hCGβ-C3d3, than that with pcDNA3-hCGβ-C3d3( P 0.05).Conclusion:The expression efficiency of pCMV4 was much higher than that of pcDNA3,and pCMV4 can induce stronger immune response than pcDNA3 because of its high expression efficiency.
Objective To investigate the effects of TIMP-2 gene expressed by host cells on the ability of growth and invasion in vitro in cultured aortic smooth muscle cells (SMCs).Methods Human TIMP-2 gene was transferred into SMCs by adenoviral Adeasy-1 to build TIMP-2-transferred cell line.Reverse-transcriptase polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA) were applied to detect the expression of TIMP-2.Growth activity was explored using direct cell counting method in vitro.Cell invasion ability was also examined by Boyden chamber.Results RT-PCR revealed that the genome of TIMP-2-transferred cells contained a 590 bp specific fragment of TIMP-2 gene.It had been certified that TIMP-2 was expressed stably in transferred cell line by ELISA method.The ability of growth of cells transferred by TIMP-2 gene were inhibited markedly as compared with control cells.The cells were markedly reduced in TIMP-2 transferred group as compared with controls (21.38± 12.81 vs 53.64± 11.27, 44.23± 12.75, P 0.05).Conclusion Recombinant adenovirus mediated gene transfer of TIMP-2 gene can inhibit growth and proliferation of aortic SMCs effectively.
The purpose of this study was to investigate differentially expressed long noncoding RNAs (lncRNAs) in pulmonary adenocarcinoma tissue and adjacent noncancerous tissue from Chinese patients using lncRNA expression microarray and preliminary analysis.RNA extracted from three paired pulmonary adenocarcinoma tissue and adjacent noncancerous tissue specimens was used to synthesize double-stranded complementary DNA after labeling and hybridization. The complementary DNA was labeled and hybridized to the lncRNA expression microarray, and array data were analyzed for hierarchical clustering. Gene coexpression networks were constructed to identify interactions among genes. To validate the microarray findings, we measured the relative expression levels of four random differentially expressed lncRNAs in the same tissue used for microarray using real-time quantitative polymerase chain reaction. The expression level of one lncRNA, AK124939, in the paired pulmonary adenocarcinoma/adjacent noncancerous tissue of another 30 patients was measured using real-time quantitative polymerase chain reaction. The experimental data were further analyzed and compared with clinical features.Of 39,000 lncRNAs investigated, 704 were differentially expressed in pulmonary adenocarcinoma tissue; 385 were upregulated and 319 were downregulated compared with those in the adjacent noncancerous tissue (fold change ≥2 and ≤-2, P<0.05). AK124939 expression levels in poorly differentiated adenocarcinoma tissue were lower than those found in well to moderately differentiated adenocarcinoma tissue (P=0.05).There are significant differences in the lncRNA expression profiles in Chinese patients with pulmonary adenocarcinoma. LncRNAs such as AK124939 may be anticancer factors related to the progression of pulmonary adenocarcinoma.
Objective To observe the protective effectiveness of Shuganning injection on hepatic injury caused by chemotherapy drug. Methods Total of 46 patients with tumors who had slight hepatic injury after chemotherapy were randomly divided into treatment group and control group. In treatment group, Shuganning was additionally added by intravenous infusion to 26 patients. However, the other 20 patients in control group were given routine liver-protective drugs only. The change of ALT was researched before and after therapy to test the effectiveness of Shuganning injection. Results The serum level of ALT in treatment group was obviously lower than that of control group, the difference was significant (P 0.05). Conclusions Shuganning injection has noticeable effect to relieve hepatic injury caused by chemotherapy.
Objective:In the past study, a plasmid of pcDNA3 hCGβ C3d3 had been constructed. It was suggested that the fusing protein be expressed by the pcDNA3 hCGβ C3d3 plasmid both in the transient expression system in COS 7 cells, and the stable expression system in CHO cells. In the present research, it would be testified that the C3d molecular adjuvant could improve the immunogenicity of the hCGβ DNA immunization or not. Methods:BALB/C mice of 6 weeks old were immunized intramuscularly two times at interval of 3 weeks by the plasmid pcDNA3, pcDNA3 hCGβ, pcDNA3 hCGβ C3d3 at dosage of 5, 10, 20 pmol, respectively. The anti hCGβ antibody titers were determined by indirect ELISA in 6 weeks of last immunization. Results:The result showed that the C3d molecular adjuvant could increase significantly the titer of anti hCGβ antibody in a dose dependent manner after DNA immunization.Conclusion:The C3d molecular adjuvant does improve the humoral immunity of the hCGβ DNA immunization, which would be helpful for technical progress and clinical trial of the contraceptive vaccine. [
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