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Objective·To explore plasma immune and inflammatory proteins that could serve as potential screening markers for Alzheimer's disease (AD).Methods·Healthy controls (n=19) and AD patients (n=19) were enrolled.Plasma samples were collected and 70 kinds of immune and inflammatory proteins were detected.The immune and inflammatory proteins associated with AD were screened by Mann-Whitney U test and partial correlation analysis.Discriminant analysis was used to develop multi-protein combined algorithm to distinguish plasma samples of AD patients from those of healthy controls.Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficacy for the multi-protein combined algorithm.Results·Among the 70 proteins analyzed,23 were significantly higher in AD patients (P＜0.05),among which 19 were strongly correlated with AD (P＜0.05).These 19 proteins were analyzed with Wilks' lambda stepwise analysis to develop discriminant algorithm for detecting plasma samples of AD.Finally,the discriminant algorithm established by 11 plasma immune and inflammatory proteins (EGF,GRO,MDC,MCP-1,MCP-2,MCP-4,TARC,SCF,TRAIL,CTACK,GCP-2) was found to have an optimal diagnostic efficacy (AUC=0.994).The optimal cutoff value of the algorithm was-0.609.When the optimal cutoff value was obtained,the sensitivity of the equation could reach 100％ and the specificity could reach 94.7％.Conclusion·The discriminant equation composed of the above 11 plasma immune and inflammatory proteins has the potential to assist AD screening.
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Objective To investigate the toxic effect of the carboxyl-terminal peptide of β-amyloid precursor protein (APPC31) on Neuro2a cells as well as its role in the toxic process in Neuro2a cells induced by Aβ42 in vitro.Methods The plasmid vector and the APPC31 construct were transiently transfected into Neuro2a cells respectively by lipofectamine 2000.The viability of the cells was measured by the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay at 48 h after transfection,and their morphocytology was observed by 4', 6-diamidino-2-phenylindole (DAPI) nucleus staining.Afterword different constructs including vector, WTAPP695, APP( D664A), the amino-terminal peptide of β-amyloid precursor protein (APP△C31) and APPC31 were transiently transfected into Neuro2a cells respectively via the same method.At 24 h after transfection Aβ42 was added into the culture medium of Neuro2a cells with the desired concentration for another 24 h for cell studies.The viabihty and morphocytology of the cells were measured by using the MTT assay and DAPI nucleus staining, respectively.Results When incubated in the absence of Aβ42, the viability of cells transfected with vector and APPC31construct were 0.81 ±0.10 and 0.88 ±0.12 respectively, and accordingly there was no significant difference between the these two groups (t = - 0.78, P = 0.48 ); meanwhile no obvious cell nuclear morphological changes of apoptosis or death occurred.However when incubated in the presence of Aβ42, the viability of cells transfected with vector, WTAPP695, APP( D664A), APP△C31 and APPC31 constructs were 0.82 ±0.01, 0.78 ±0.03, 0.55 ±0.04, 0.81 ±0.04, 0.78 ±0.02 and 0.54 ±0.02 respectively.The viability of cells transfected with WTAPP construct and APPC31 construct decreased significantly ( F = 47.53, P ＜0.05) compared with the control group, meanwhile cells displayed condensed nuclear and even nuclear fragmentation.Conclusions In vitro, over-expression of a certain level of APPC31 in Neuro2a cells fails in causing cell death, but this short peptide enhances cytotoxicity induced by Aβ42 in Neuro2a cells.Thus,these results provide the experimental basis for the further explication of the pathogenesis of Alzheimer's disease.
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Objective To observe the change of invasion and migration of the pancreatic carcinoma cell line SW1990 transfeeted with EEF1A2 gene.Methods Pancreatic carcinoma cell line SW1990 was transfected with EEF1A2 by recombinant adenovirus vector.The alteration of motility、invasion and adhesion property of SW1990 was evaluated by wound healing assay,transwell With or without Matrigel basement membrane and adhesion assay.Results Wound healing assay revealed that EEF1A2 enhanced cell motility and transwell assay with Matrigel indicated that the average numbers of transwell cells with EEFlA2 was increased from 23.25±5.23 to 65.42±8.24(P＜0.05).The adhesive rate was substantially increased in EEF1A2 transfected SW1990 cells compared with control cells.Conclusions EEF1A2 gene can promote the migration.invasion and adhesion ability of pancreatic cancer cell in vitro.It is indicated that EEF1A2 may involve in the development of human pancreatic cancer by influencing cell biological characteristics.
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Objective To investigate the effects of EEF1 A2 on growth and proliferation of the human pancreatic cancer cell line SW1990. MethodsThe EEF1 A2 gene was introduced into pancreatic cancer cell line SW1990 by adenovirus vector. The effects of EEF1 A2 on the growth of human pancreatic cancer cell were measured by MTT. Cell cycle was detected by flow cytometry and cell growth rate was examined by soft agar cloning formation test. ResultsAfter EEF1A2 induction, the expression of EEFA1 A 2 mRNA in pancreatic cancer cell line SW1990 increased, value of A750 at 72 h was 1. 2996 ±0. 2091, number of cells was 81250, cloning efficiency at 14 d was 82%, all of these parameters were significantly higher than those in the groups of blank adenoviras vector and PBS groups (P <0.05 ) ; the percentage of G1 phase cell was 28.5%, which was significantly lower than those in the groups of blank adenovirus vector and PBS groups; the percentage of Sphase ceil was 60.9%, which was significantly higher than those in the groups of blank adenovirus vector and PBS groups (P < 0.05 ). ConclusionsEEF1 A2 gene significantly enhanced cell growth and proliferation in human pancreatic cancer in vitro.
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We selected defective retrovirus LXSN approved by the recombinant DNA Advisory Comittee, NIH, USA as a vector to study the transduction of human TIL with TNF gene, on the basis of which, we engaged in the clinical application. The results showed that the logarithmic growth phase was the optimal transducing time when the TIL were isolated and cultured from 9 to 25 days. It also showed that the transduction efficiency was correlated to the MOI value, the more MOI value, the more TIL being transduced. Successful gene inserted and expressing were confirmed by the polymerase chain reaction and the L929 cell test. The clinical trial of treatment for metastastic hepatic cancer suggested that the better therapeutic effectiveness and less side effect depend on the specificity of the TIL, the proper infusion pathway and the dose of the transduced TIL.