| Objective:This article through the quantitative assessment of the effects of oxidant on antigen-antibody binding activity,to explore the toxicity of oxidative stress on the immune system,at the same time comparative observation of oxidative stress and DNA damage,oxidative stress and blood coagulation function change,etc.Further comprehensive assessment of the biological effects of oxidative stress,in-depth understanding of the impact of oxidative stress on the body.Methods:1.Adopted double diffusion test of precipitation reaction,by observing the clear degree of precipitation line formed in the AGAR gel plate,to represent the effects of potassium permanganate,iodine and hydrogen peroxide on antigen-antibody binding activity in the reaction.Polyclonal antibodies were determined with the precipitation reaction,not the monoclonal antibodies.2.Adopted B erythrocytes and O-serum from healthy people,agglutination reaction by B antigen on the B erythrocytes surface and the anti-B antibodies in the O-serum,the area with aggregation and the corresponding oxidant concentration were determined as a measure of the effect of oxidant on the agglutination reaction and the activity of antibody response intensity.IgM antibodies were measured with the agglutination test.3.The enzyme-linked immunosorbent assay of enzyme immunoassay technology was used to detect hepatitis B surface antibody,the OD value can be determined through enzyme standard instrument to represent the size of the activities of three kinds of oxidants to IgG antibody in ELISA test.4.Calculated ID50 concentrations which had an obvious inhibitory effect on the activity of antibody were added to serum,then used the method of direct determination of redox potential determine the serum total antioxidant capacity,in order to measure the impact of this concentration of oxidant on the serum total antioxidant power.5.This article selected polymerase chain reaction（PCR）,its DNA template was oxidized treatment,through the observation of the clear degree of agarose gel amplification bands,to analyze the effect of theoxidant on DNA damage.6.In this paper,used the quantitative blood clot disposable suction tube,automatic blood coagulation analyzer and blood coagulation reagent to detect blood samples after dealing with the oxidant of coagulation four indicators,for exploring the oxidant effects on the blood coagulation function.Results:1.In this study,the double diffusion test showed that potassium permanganate,iodine and hydrogen peroxide had an obvious inhibitory effect on the activity of the antibody,and the ID50 concentrations were 1.9×10-3 mol/L,2.96×10-3 mol/L and15.9×10-3 mol/L,respectively.2.In this study,a simple agglutination reaction was used to determine serum IgM antibody activity.The results demonstrated that potassium permanganate,iodine and hydrogen peroxide had obvious inhibition activity on the antibody,and the ID50 concentrations were 2.58×10-3 mol/L,3.83×10-3 mol/L and 22.3×10-3 mol/L,respectively.3.In this study,an ELISA kit was used to determine hepatitis B surface antibody activity,and was found to be more sensitive,and resulted in better quantitative analysis and accuracy than the precipitation and agglutination techniques.The study also found that potassium permanganate,iodine and hydrogen peroxide had an obvious inhibitoryeffect on the antibody activity,and the ID50 concentrations were 2.40×10-3 mol/L,3.06×10-3 mol/L and 13.5×10-3 mol/L,respectively.4.Calculated ID50 concentrations were added to serum to detect the serum total antioxidant capacity,and it was observed that the concentration of oxidant had no significant effect on serum total antioxidant capacity.5.In this paper,through the HLA-GAPDH and internal CRP in agarose gel electrophoresis of PCR products amplification result diagram can be seen that with the increase of concentration of the oxidant,GAPDH and CRP amplification bands gradually dim,after reaching a certain concentration,stripe disappear.The study also found that potassium permanganate,iodine and hydrogen peroxide had an obvious damage on the DNA,and the ID50 concentrations were 0.625×10-3mol/L,1.25×10-3mol/L,6.25×10-3mol/L,respectively.6.In the detection of coagulation,we found that with the increase of concentration of the oxidant,the PT,APTT and TT values increased gradually,but the Fib values decreased gradually.Conclutions:1.Oxidants had no significant effect on the content of antibody,but had a marked influence on the activity of the antigen-antibody reaction.2.Oxidative stress can cause DNA damage and the sensitivity of the DNA to the oxidants was higher than the activity of antibody.3.Oxidative stress can damage the stability of coagulation factors and the structure of fibrinogen in plasma,result in coagulation function abnormalities. |
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