| Objective : The purpose of this study was to study the clinical characteristics of Brucellosis patients in Hohhot area,the cytokines related to Th1/Th2/ Th17 cells in peripheral blood,and the expression characteristics of virulence genes and drug resistance genes of isolated strains.To further understand the correlation between Th1/Th2/ Th17 cell-related cytokines and disease progression in patients with brucellosis and the distribution of human brucellosis virulence genes and drug resistance genes,so as to provide experimental basis for the diagnosis and treatment of brucellosis.Methods:1.The selection in May 2019 to May 2020,in Inner Mongolia medical university affiliated hospital diagnosis of brucellosis patients as the research object,set the blood culture positive for group(31 cases blood culture positive brucella patients),blood culture negative group(blood culture negative brucella patients 32cases)and control group(31 cases)healthy check-up crowd.At the same time,the relevant clinical data of each group were collected for comparative analysis.2.From May 2018 to May 2020,39 Brucellosis strains were isolated from the whole blood samples of 39 patients with brucellosis,and virulence genes and drug resistance genes were detected.3.Flow cytometry was used to detect the levels of cytokines related to Th1/Th2/ Th17 cells in peripheral blood of patients with brucellosis and healthy subjects,including IL-2,IL-4,IL-6,IL-10,IL-17 A,IFN-γ and TNF-α.4.PCR method was used to identify seven virulence genes(sensory protein BVRS,regulatory protein BVRR,virulence factor B4,heat shock protein 20,ABC type transporter(ABC)),Brucella protein A(Baca,chaperone protein BP(HFQ))and erythromycin resistance gene(ERM)in blood samples isolated from patients with Brucellosis Detect and analyze the distribution condition.Results:1.Changes of blood system: Among 63 patients diagnosed with brucellosis,35 patients(55.6%)presented anemia,16 patients(25.4%)presented thrombocytopenia,10 patients(15.9%)presented leukopenia,24 patients(38.1%)presented elevated transaminase,and 39 patients(61.9%)presented elevated CRP.Among the 31 patients with blood cultur-positive brucellosis,20(64.5%)had anemia,10(32.2%)had thrombocytopenia,6(19.4%)had leucopenia,18(58.1%)had elevated transaminase,and 28(90.3%)had elevated CRP.The levels of CRP,ALT and AST in peripheral blood in blood culture-positive group were significantly higher than those in blood culture-negative group,and the two were significantly higher than those in control group,the difference was statistically significant(P<0.05).The hemoglobin and platelet in the blood culture-positive group were significantly lower than those in the blood culture-negative group,and they were significantly lower than those in the control group,with statistical significance(P<0.05).There was no significant difference in the WBC count between the positive group and the negative group(P>0.05).They were significantly lower than the control group,and the difference was statistically significant(P<0.05).2.Th1/Th2/ Th17 cell related cytokines: Serum IFN-γ concentrations(15.86(6.18,24.84)),IL-10 concentrations(4.98(3.92,7.23)),IL-6 concentrations(18.03(11.29,28.52)),and IL-17 A concentrations(5.03(2.73,7.50))in the blood culture-positive group;Serum IFN-γconcentrations(4.51(3.39,8.50)),IL-10 concentrations(3.97(3.31,5.87)),IL-6concentrations(11.17(6.46,31.74)),and IL-17 A concentrations(4.53(3.39,7.06))in the blood culture-negative group;Serum IFN-γ concentrations(3.38(2.99,3.61)),IL-10 concentrations(3.21(2.93,3.94)),IL-6 concentrations(4.32(3.65,4.32)),and IL-17 A concentrations(3.26(2.21,4.81)in the control group;The concentrations of IL-6 and IL-17 A were increased in both the positive and negative blood culture groups,and there was no significant difference between the two groups,but they were significantly higher than the control group,with statistical significance(P<0.05).The serum concentrations of IFN-γ and IL-10 in the blood culture-positive group were significantly higher than those in the blood culture-negative group,and they were significantly higher than those in the control group,with statistical significance(P<0.05).Of the 31 patients with positive blood culture,12.9%(4/31)had negative detection of Brucella antibody.3.Distribution characteristics of virulence genes and drug resistance genes: 39 strains of brucellosis were isolated from the whole blood samples of 39 patients with brucellosis.The PCR results showed that the positive rates of sensory protein BVRS,regulatory protein BVRR,virulence factor B4,heat shock protein 20 and ABC type transporter(ABC)genes were 100%(39/39).The genes encoding Brucella protein A(Bac A)were not amplified in the two strains.One strain did not amplify the chaperone BP(HFQ)coding gene.Conclusion : 1.The number of peripheral blood cells,the concentration of serum inflammatory factors and the indexes of liver function are often changed in patients with brucellosis.2.Serum IL-10 and IFN-γ concentrations changed significantly in patients with Brucella,and there were significant differences between patients in acute and chronic stages.3.The combined detection method of blood culture,serum antibody and cytokines of brucellosis can maximize the diagnostic rate of brucellosis;4.The overall carrying rate of virulence genes in human Brucella was high,and the distribution characteristics of virulence genes among different strains were similar. |
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