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OBJECTIVE: To investigate the effect of nickel sulfate on cell survival rate and apoptosis of normal human liver L02 cells. METHODS: i) L02 cells in logarithmic growth phase were divided into 9 groups, each with 6 wells. L02 cells in each group were treated with 0, 100, 200, 300, 400, 500, 600, 700 and 800 μmol/L nickel sulfate. The survival rate of L02 cells was determined by CCK-8 assay after cells were treated for 0, 6, 12, 24, 48 and 72 hours. The nickel sulfate exposure dose and exposure time for subsequent experiments were selected based on the results of CCK-8 assay. ii) L02 cells in logarithmic growth phase were divided into control group, 100 and 300 μmol/L dose groups, and were exposed to 0, 100 and 300 μmol/L nickel sulfate for 12 hours, respectively. Western blot was used to detect the relative protein expression of B cell lymphoma/leukemia 2(BCL-2), Bcl-2 related protein X(BAX), caspase-3, phosphorylated RNA-dependent protein kinase-like endoplasmic reticulum kinase(p-PERK), phosphorylated eukaryotic translation initiation factor 2α(p-eIF2α), CCAAT/enhancer-binding protein homologous protein(CHOP) and glucose regulatory protein 78(GRP78). RESULTS: i) After treatment with nickel sulfate, the survival rate of cells decreased with the increase of dose and the prolongation of exposure time(all P values were <0.01). According to the half inhibitory concentration of nickel sulfate on L02 cells, the nickel sulfate exposure time in subsequent experiments was selected as 12 hours, and the exposure concentration was 100 and 300 μmol/L. ii) Compared with the control group, the relative expression of BCL-2 protein in L02 cells in the 100 and 300 μmol/L dose groups decreased(all P values were <0.05), while the relative protein expression of BAX, caspase-3 protein and ratio BAX/BCL-2 increased(all P values were <0.05). Compared with 100 μmol/L dose group, the relative expression of BCL-2 protein in L02 cells of 300 μmol/L dose group decreased(P<0.05), while the relative expression of BAX and caspase-3 protein and the ratio of BAX/BCL-2 increased(all P values were <0.05). Compared with the control group, the relative expression levels of p-PERK, p-eIF2α, CHOP and GRP78 protein in L02 cells were increased in 100 and 300 μmol/L dose groups(all P values were P<0.05). Compared with 100 μmol/L dose group, the relative expression levels of p-eIF2α, CHOP and GRP78 protein in 300 μmol/L dose group were increased(all P values were<0.05).CONCLUSION: Nickel sulfate can regulate the expression of apoptosis related proteins and PERK signaling pathway related proteins in L02 cells, aggravate apoptosis of L02 cells and decrease the cell survival rate.
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Objective To detect serum brucellosis in high risk population,three methods were used and their advantages and disadvantages was compared.Methods In accordance with the surveillance standard for brucello-sis(GB 16885 -1997)and brucellosis diagnostic criteria(WS269 -2007)in the prescribed method,rose -bengal plate agglutination test (RBPT),standard -tube agglutination test (SAT)and enzyme linked immunoassay assay (ELISA)were used to detect brucellosis and analysis of its diagnostic significance in the high risk population of sheep farm of Guoyang county in Anhui province.Results The positive rates of RBPT,SAT and ELISA were 19.1%,12.1% and 16.3% in 257 blood samples,respectively.Compared to SAT,the sensitivity,specificity,accura-cy and the area under the ROC curve of RBPT were 88.5%,91.8%,91.4%,0.81,respectively,which of ELISA were 93.9%,95.1%,94.9%,0.88.Conclusion The sensitivity,specificity,accuracy and the area under the ROC curve of ELISA were higher than those of other methods.Proper method,early surveillance and effective technology can help to control the occurrence and epidemic of brucellosis in the actual test work promotion.
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Objective To establish a magnetic nanoparticles separation-based quantitative real-time PCR(RT-PCR)assay for fast and accurate detection of Plasmodium falciparum and providing a technical support for improving the control and preven-tion of imported malaria. Methods According to the conserved sequences of the P. falciparum genome 18SrRNA,the species-specific primers and probe were designed and synthetized. The RT-PCR was established by constructing the plasmid standard , fitting the standard curve and using magnetic nanoparticles separation. The sensitivity and specificity of the assay were evaluat-ed. Results The relationship between the threshold cycle(Ct)and logarithm of initial templates copies was linear over a range of 2.5×101 to 2.5×108 copies/μl(R2=0.999). Among 13 subjects of entry frontier,a P. falciparum carrier with low load was de-tected by using the assay and none was detected with the conventional examinations(microscopic examinations and rapid tests). Conclusion This assay shows a high sensitivity in detection of P. falciparum,with rapid and accurate characteristics,and is especially useful in diagnosis of P. falciparum infectors with low parasitaemia at entry-exit frontier ports.