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Objective To analyze the value of serum Treg cell related factors and chemokines in patients with tuberculous pleurisy .Methods From July 2015 to December 2016 ,92 cases of tuberculous pleurisy in our hospital were selected as the observation group ,and 92 healthy persons at the same time were selected as the control group .The levels of Treg cell related factors[monocyte chemoattractant protein (MCP)-1 ,IP-10 ,CCL-3 and CCL-16] and IL-10 ,TGF-βand IL-35] were detected and compared in the two groups ,and the levels of these indexes were compared in different classifications and stages of tuberculous pleuritis .Results The ser-um Treg cell related factors and chemokine levels in the observation group were significantly higher than those in the control group (P<0 .05) .The expression level of tuberculous empyema was higher than that of dry pleuritis and exudative pleuritis ,the patients with exudative pleuritis were higher than those of dry pleuritis , and the patients with multiple pleuritis were higher than those with idiopathic and concomitant pleuritis ,the difference was statistically significant (P<0 .05) .Conclusion The serum Treg cell related factors and chemo-kines in patients with tuberculous pleurisy are highly expressed ,and the classification and staging of the dis-ease have great influence on the expression ,and the above indexes have high detection value in the patients with tuberculous pleurisy .
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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.
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<p><b>OBJECTIVE</b>To investigate the cytochrome P450 2E1 (CYP2E1) RsaI/PstI and DraI polymorphisms in workers exposed to benzene.</p><p><b>METHODS</b>A cross-sectional survey was carried out. A total of 71 workers exposed to benzene were included in observation group and the same number of people without occupational benzene exposure were included in control group. Blood samples from the two groups were collected and genotyping for CYP2E1 RsaI/PstI and DraI were conducted using the polymerase chain reaction-restriction fragment length polymorphism.</p><p><b>RESULTS</b>There were no significant differences in CYP2E1 DraI genotype and allele distributions between the observation group and the control group (χ² = 2.374, P > 0.05; χ² = 2.113, P > 0.05). Significant differences in CYP2E1 RsaI/PstI genotype and allele distributions between the two groups were observed (χ² = 9.129, P < 0.01; χ² = 6.028, P < 0.05).</p><p><b>CONCLUSION</b>Mutations at CYP2E1 RsaI/PstI can enhance the expression of CYP2E1 and this suggests individuals with the mutated gene have increased susceptibility to chronic benzene poisoning.</p>
Subject(s)
Humans , Alleles , Benzene , Poisoning , Cross-Sectional Studies , Cytochrome P-450 CYP2E1 , Genetics , Metabolism , Genetic Predisposition to Disease , Genotype , Poisoning , Genetics , Polymerase Chain Reaction , Polymorphism, Genetic , Genetics , Polymorphism, Restriction Fragment LengthABSTRACT
In order to express the gene of LEN-5 β-lactamase from a Klebsiella pneumoniae strain,plasmids in the strain were extracted and an 879bp product of LEN-5 gene was obtained with PCR.After being digested with Nde I and Xho I,LEN-5 gene was cloned into pET-26b (+) vector.Then it was confirmed by digestion and DNA sequencing in recombinant plasmid before transformed into E.coli BL21 (DE3).After inducing by IPTG,LEN-5 β-lactamase was expressed.Protein extraction was processed by ultrasonic and protein activity was detected by nitrocefin.The isoelectric focusing electrophoresis showed a pI of 7.6.These results indicated that the LEN-5 gene has been cloned and expressed in prokaryote cell successfully.