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Objective@#To construct a Tat-dependent MazF expression vector pcDNA3.1-GFP-HA-MazF-U3TAR.@*Methods@#HIV-1 U3TAR and MazF gene were amplified from pCR2.1-U3TAR and pET28a-MazF, respectively. Two gene fragments were linked together to form U3TAR-MazF by an overlapping PCR, and then cloned into pMD18T. An N-terminal HA tag was added to MazF to form U3TAR-MazF-HA. After double enzyme digestion using EcoR I and Hind Ⅲ, U3TAR-MazF-HA was reversely inserted into pcDNA 3.1 to form pcDNA3.1-HA-MazF-U3TAR. Then, a fluorescent reporter gene GFP was inserted into the downstream of U3TAR to form pcDNA3.1-GFP-HA-MazF-U3TAR.@*Results@#The co-transfection experiments with pcDNA3.1-tat-flag showed that pcDNA3.1-GFP-HA-MazF-U3TAR is Tat dependent. MazF was expressed only under the presence of Tat. In addition, compared with the cells transfected with pcDNA3.1-GFP-HA-MazF-U3TAR, less green fluorescent signal was observed in the cells co-transfected with pcDNA3.1-GFP-HA-MazF-U3TAR and pcDNA3.1-tat-flag, indicating that expressed MazF down-regulated the expression of GFP.@*Conclusions@#The expression vector pcDNA3.1-GFP-HA-MazF-U3TAR will provide an important tool for the development of MazF-based AIDS gene therapy strategies.
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Objective To study the relationship of serum pulmonary surfactant protein and vascular endothelial growth factor re‐lated indexes to lung cancer .Methods Totally 56 patients with lung cancer in our hospital from September 2014 to December 2015 were selected as the observation group ,56 healthy personnel at the same period were selected as the control group ,then the serum pulmonary surfactant protein and vascular endothelial growth factor related indexes of control group and observation group ,obser‐vation group with different stages and pathological classifications were compared ,and the relationship between serum pulmonary surfactant protein and vascular endothelial growth factor related indexes and lung cancer were satisfacted with Logistic analysis .Re‐sults The serum pulmonary surfactant protein and vascular endothelial growth factor related indexes of observation group were all higher than those of control group ,and the serum pulmonary surfactant protein and vascular endothelial grow th factor related inde‐xes of observation group with higher stages were all higher than those of patients with lower stages ,and the levels of patients with adenocarcinoma were all higher than those of patients with squamous cell carcinoma ,and the serum pulmonary surfactant protein and vascular endothelial growth factor related indexes all had close relationship to the lung cancer (all P<0 .05) .Conclusion The serum pulmonary surfactant protein and vascular endothelial growth factor related indexes all have close relationship to the lung cancer ,and those indexes of patients with lung cancer should be paid more attention .
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Objective To analyze the expression of excision repair cross-complementation group 1 (ERCC1),MutS protein homolog 2 (MSH2) and poly ADP-ribose polymerase 1 (PARP1) in non-small cell lung cancer (NSCLC) and their prognostic value for patients receiving platinum-based postoperative adjuvant chemotherapy.Methods Immunohistochemistry was applied to detect the expression of ERCC1,MSH2 and PARP1 in 111 cases of NSCLC paraffin embedded surgical specimens.Through OG-Rank survival analysis,the prognostic value of the ERCC1,MSH2,PARP1 and the related clinic pathological factors were evaluated.Cox regression analysis was used to determine whether ERCC1,MSH2 and PARP1 were independent prognostic factors or not.Results Among the 111 NSCLC patients,the positive expression rates of ERCC1,MSH2 and RARP1 were 33.3 ％(37/111),36.9 ％(41/111) and 55.9 ％(62/111),respectively.Besides,a total of 72 patients who received platinum-based chemotherapy had complete follow-up data.ERCC1 (P＜ 0.001) and PARP1 (P=0.033) were found to be correlated with the survival time while there was no correlation for MSH2 (P =0.298).Patients with both ERCC1 and PARP1 negative had significant longer survival time than those with ERCC1 (P=0.042) or PARP1 (P=0.027) positive alone.Similarly,the survival time of patients with both ERCC1 and PARP1 positive was shorter than those with ERCC1 (P=0.048) or PARP1 (P=0.010) positive alone.Conclusions The NSCLC patients with ERCC1 and (or) PARP1 negative may benefit from platinumbased postoperative adjuvant chemotherapy.The detection of ERCC1 and PARP1 can be used as an important method to assess the prognosis of patients with NSCLC.