ABSTRACT
Objective To investigate the value of combined detection of viral capsid antibody(VCA -IgA), early antigen antibody(EA -IgG),and Rta protein antibody IgG(Rta -IgG)in the diagnosis of EB virus associated nasopharyngeal carcinoma(NPC).Methods A total of 47 nasopharynx cancer patients,45 patients with benign rhinitis and 45 healthy controls were recruited.The serum levels of VCA -IgA,EA -IgA and Rta -IgG were tested by enzyme -linked immunosorbent assay (ELISA).Specificity and sensitivity of the indicators alone and combined detection were compared using the clinical diagnosis as the gold standard.Results The expression levels of serum VCA -IgA,EA -IgA and Rta -IgG in the rhinitis group were (0.82 ±0.25),(0.74 ±0.13),(0.89 ±0.27),the levels in the NPC group were (2.16 ±0.39),(1.26 ±0.24),(3.95 ±0.76),and the levels in the healthy control group were (0.65 ±0.14),(0.51 ±0.11),(0.41 ±0.16)respectively.The levels of VCA -IgA,EA -IgA and Rta-IgG in the rhinitis group,NPC group and healthy group were significantly different (F =400.065,232.803, 740.215,P =0.000,0.000,0.000).The levels of VCA -IgA,EA -IgA and Rta -IgG in the NPC patients with different TNMstages were significantly different(F =195.679,30.878,38.561,P =0.000,0.000,0.000),and the trend of each antibody was increased with the severity of the disease.The sensitivity of VCA -IgA was the highest (85.11%),and the specificity of EA -IgA was the highest(95.56%).The sensitivity of the combined assay was 95.74%,which was higher than that of the other three combinations.Conclusion The combination of VCA -IgA, EA -IgA and Rta -IgG can reflect the expression of EBV -associated antigen to a greater extent,and it is superior to single or two combined detection in the diagnosis of EB -associated NPC,with a high clinical value.
ABSTRACT
Objective To construct a lentiviral vector carrying rat sirt1 gene and observe the expression of sirt1 in retinal ganglion cell (RGC) of rat.Methods Rat sirt1 cDNA was inserted into pLV5 vector.After identification by sequencing analysis and PCR,the recombinant sirt1expressinglentivirus vector was packaged by cotransfecting 293T cells with packaged plasmid.Then pLV5-sirt1 was used to infect the cultured Sprague-Dawley rat RGC cell in vitro.The expressions of sirt1 protein and mRNA in infected rat RGC were detected by quantitative real-time PCR and Western blot.Results The sirt1 expression vector pLV5 was successful constructed and sequence was proved to be correct.The expression of sirt1 protein and mRNA in RGC was significantly increased than that in cells infected with control lentiviruses (P ＜ 0.05).Conclusion We have successful constructed a sirt1 expression lentivirus vector pLV5-sirt1 and it can increase the expression of sirt1 protein and mRNA in the rat retinal ganglion cells.
ABSTRACT
<p><b>OBJECTIVE</b>To establish a canine cell line with p53 gene knockdown by lentivirus- mediated RNA interference (RNAi).</p><p><b>METHODS</b>Four pairs of oligonucleotide sequences of p53 gene were synthesized and cloned into the pGMLV-SC5 RNAi vector. Following confirmation by PCR and DNA sequencing, the recombinant pGMLV-p53 plasmids and packaging mix vectors were packaged into mature lentivirus to infect 293T cells. The supernatant of the infected cells was harvested to infect WRD cells, in which p53 expression was detected using Western blotting and real-time PCR. The lentivirus with the strongest p53⁻ silencing effect was packaged for infecting WRD cells at the optimal multiplicity of infection (MOI) to establish a stably infected cell line (WRD/p53⁻) screened using puromycin.</p><p><b>RESULTS</b>The lentivirus carrying p53 shRNA was constructed successfully and a virus titer of 1 × 10⁹ TU/ml. The 4 pGMLV-p53 plasmids all produced strong p53 interference affects, and pGMLV- p53A1 with the strongest effect. The stably infected cells line WRD/p53⁻ was established successfully using pGMLV-p53A1 plasmid.</p><p><b>CONCLUSION</b>The canine cell line WRD/p53⁻ with stable lentivirus-mediated p53 silencing has been established successfully.</p>