ABSTRACT
Objective To analyze changes of DNA demethylation analysis of FOXP3 TSDR at the beginning of the secondary pulmonary tuberculosis patientsby utilizing real-time PCR technology.Methods To select 47 patients of secondary pulmona-ry tuberculosis as a research group from June 2014 to May 2015 and 40 healthy donors as a control group.The peripheral blood mononuclear cells (PBMC)of research group and control group were isolated.CD4+CD25+T cells were isolated from PBMC.Genomic DNA was isolated from CD4+CD25+T cells.PCR was performed in a final reaction volume containing dem-ethylation-specific primers.Plasmid standard was generated by PCR products were enzyme digestion,TOPO TA cloning,and recycling and purification.A real-time PCR system was established by quantitatively analyzing the specificity of FOXP3 TS-DR demethylation to treg (regulatory T-cell).Treg numbers of control group at week 0 and research group treated at week 0,week 2,week 4 and week 8 by using real-time PCR assay of the FOXP3 TSDR demethylation.The experimental data was analyzed by using SPSS 1 6.0 software.Results The M.tuberculosis in sputum of research group were positive by smear mi-croscopy,however the results of control group were negative.The treg frequency of control group,2+ group and 3+ group respectively was 1.63%±0.70%,1.96%±0.10% and 0.86%±0.21%,respectively.The difference between the treg fre-quency of control group and that of 2+ group by smear microscopy had not statistical significance,however which of 3+group was opposite.The average treg frequency of research group treated at week 0,2,4 and 8 respectively was at 1.05%, 2.04%,3.44% and 2.79%,range of which respectively was 0.32%~2.03%,0.95%~3.95%,2.35%~4.95% and 1.02%~4.27%,95% confidence interval of which respectively was (0.93%,1.18%),(1.85%,2.24%),(3.27%,3.61%) and (2.60%,2.98%).The treg frequency of difference between control group and research group at week 0 had statistical significance (t=4.669,P<0.05).The treg frequency was influenced by time of therapy,using One-Way ANOVA analysis (F=347.2,P<0.001,df=3,within-subjects Contrasts:F=407.4,P<0.001,df=3).Test of the treatment time and group interaction effect was linear (F=678.2,P<0.001,df=1).Conclusion DNA demethylation analysis of FOXP3 TSDR was high sensitivity and specificity in monitoring changes of treg at the beginning of the secondary pulmonary tuberculosis pa-tients.
ABSTRACT
Objective To understand Shenzhen Longgang,guangming and longhua new district four district hospital ICU pa-tients with secondary pulmonary tuberculosis merger lower respiratory infection of pathogenic bacteria distribution and drug resistance status of provide a reference for clinical diagnosis and rational use of antibiotics therapy.Methods Random selec-tion from February 2013 to October 2015 in the three district hospital ICU diagnosis of secondary pulmonary tuberculosis patients with lower respiratory infection in 593 cases of sputum specimen pathogenic bacteria culture and drug susceptibility results were retrospectively analyzed.Results 593 cases of ICU secondary pulmonary tuberculosis patients with respiratory tract infection of the communist party of China isolated 617 strains of pathogenic bacteria,fungi accounted for 49.6% (306/617),gram negative bacilli accounted for 40.4% (249/617),gram positive cocci accounted for 10.0% (62/617).Fungal in-fection main pathogens for white smooth candida yeast and candida yeast,respectively accounted for 44.2% (273/617)and 4.5% (28/617),gram negative bacillus mainlyKlebsiellaPneumoniae,Pseudomonasaeruginosa,and H.influenzae,respec-tively accounted for 16.7% (103/617),12.0% (74/617)and 7.3% (45/617),gram-positive cocci mainly for Saphylococcus aureus and Epidermisstaphylococcus and Hemolyticstaphylococci,respectively accounted for 4.5% (28/617),3.2% (20/617)and 0.9% (5/617).Pathogenicbacteria isolated from the multiple drug resistant bacteria,present different levels of resistance to commonly used antimicrobial agents.Conclusion ICU patients with secondary pulmonary tuberculosis merger of lower respiratory tract infection pathogens to fungi and gram-negative bacilli,the most commonWhite candida,Klebsiella pneumoniae,and Pseudomonasaeruginosa,and different levels of resistance to commonly used antimicrobial agents.