Objective To design a novel efficient CEA vaccine of fission epitopes with the vehicle of HPV16L1,and construct the recombinant baculovirus in Bac-to-Bac baculovirus expression system.Methods The recombinant baculovirus expression vector pFastBac1-V was constructed.HPV16L1 and CEA-PADRE synthesis primers were inserted into bacmid in E.coli DH10Bac with the help of Tn7 transposition system.Then the recombinant baculovirus was got by infecting insect cell sf9.Results HPV16L1 and CEA-PADRE synthesis primers that were cloned to the baculovirus transfer vector pFastBac1 were assessed by restriction analysis and direct sequencing.The recombinant baculovirus of the novel vaccine had been constructed.After infected sf9,the correct fragment was amplified by PCR.Conclusion The baculovirus of CEA vaccine of fussion epitopes with the vehicle of HPV16L1 was constructed.

Objective To analyze the differential expressions of miR-146a,miR-206,miR-223 and let-7c-1,such as cell differentiation-related miRNAs,in CXCR4-positive and CXCR4-negative subsets of the Lewis lung cancer cell lines(LLC).Methods CXCR4-positive and CXCR4-negative subsets were isolated from LLC by immunomagnetic beads sorting,and then their total cellular RNA were extracted by Trizol,expression of 4 miRNAs were detected by real-time fluorescence quantitative PCR(TaqMan probe),and potential target genes of miRNA whose differential expression was the most significant were predicted.Immunohistochemistry was carried out to confirm differential expression of the key molecule of certain research value within CXCR4-positive and CXCR4-negative subsets growing tumor tissue,and a BLAST search was performed to identify homologies of its 3′UTR.Results Compared to CXCR4-negative subsets,the expression of 4 miRNAs were lower in CXCR4-positive subsets,and expression of miR-223 had the most significant difference(Fold change=8.26).By softwares forecasting,miR-223 had potential target sites of IGF1R,IGFBP5,Pik3cb,ELK-1 and E2F1 mRNA,such as key molecular of IGF1R signaling pathway.The expression of IGF1R of CXCR4-positive subsets growing tumor tissue was significantly higher than that of CXCR4-negative subsets.Conclusion miR-223 is lowly expressed in CXCR4 positive cells from Lewis lung carcinoma cell lines.Position 238～244 nt and 688～695 nt in target sequences of 3′UTR of IGF1R mRNA was highly homologous by screening.Close correlation is found between miR-223 and IGF1R signaling pathway.The mechanisms underlying this biologically important finding need to be further explored.

0.05).Tumor formation was found in subgroups of positive CXCR4 cells(7 ? 103 ).Negative and positive CXCR cells were inoculated into 3 mice respectively at the concentration of 5 ? 105 and 2 ? 104 cells.No metastasis occurred in the former group.However,lung metastasis was found in 2 mice and ear metastasis was observed in 1mouse of the latter group.Conclusion Subgroups of positive CXCR4 cells in LLC are characterized by certain properties of cancer metastatic stem cells and have the ability to renew themselves and metabolize.

Objective To investigate the clinical value of combined detection of serum carcinoembryonic an-tigen(CEA) ,carbohydrate antigen-125 (CA125) ,cytokeratin 19 fragment antigen (CYFRA21-1) ,squamous cell carcinoma(SCC) antigen ,neuron-specific enolase(NSE) and plasma progastrin-releasing peptid(ProGRP) in the diagnosis ,pathological typing and clinical staging in lung cancer .Methods The serum CEA ,CA125 , CYFRA21-1 ,SCC ,NSE levels and plasma ProGRP level were detected by adopting the chemiluminescent mi-croparticle immunoassay method in 378 cases of lung cancer ,200 cases of benign lung diseases and 200 people undergoing healthy physical examination .Results The levels and positive rates of CEA ,CA125 ,CYFRA21-1 , SCC ,NSE and ProGRP in the patients with lung cancer were significantly higher than those in the patients with benign lung diseases and healthy control group ,the difference was statistically significant (P<0 .05);in the single index detection ,the biomarkers of highest positive rate in adenocarcinoma ,squamous cell carcinoma and small cell lung carcinoma were CEA ,CYFRA21-1 and SCC ,NSE and ProGRP respectively .The sensitivi-ty ,specificity ,accuracy ,negative predictive value and positive predictive value of combined detection of these 6 indexes were 92 .86％ ,85 .00％ ,88 .17％ ,92 .64％ and 85 .40％ respectively ,except the specificity and positive predictive value were slightly decreased ,the levels of other indexes were higher than those of single index de-tection .The aresa under the receiver operating characteristic (ROC) curves of CEA ,CA125 ,CYFRA21-1 ,SCC ,NSE and ProGRP were 0 .775 ,0 .778 ,0 .891 ,0 .585 ,0 .710 and 0 .620 respectively .The area under ROC curves of the combined detection was 0 .950 .The positive rates of CA125 ,CYFRA21-1 ,NSE and the combined detection in the patients with stage Ⅲ ,Ⅳof lung cancer were obviously higher those in the patients with stageⅠand Ⅱof lung cancer ,the difference was statistically significant (P<0 .05);the combined detection obviously improved the positive rates for the diagnosis in the patients with different stages of lung cancer .Conclusion The combined detection of CEA ,CA125 ,CYFRA21-1 ,SCC ,NSE and ProGRP is conducive to improve the di-agnosis performance and early detection rate for lung cancer ,differentially diagnosing different pathological types of lung cancer and judge the clinical stage of lung cancer .The combined detection of 6 tumor biomarkers is an ideal diagnosis index of lung cancer .