The Study Of Correlation Between Japanese Encephalitis Syndromes And Human Genetic Polymorphisms, Associated Cytokine

Objective:JE cases which collected in2011and2012, using double-antibodysandwich ELISA to measure control group JE dusunnaoluo group、duxianxinbao group expression in blood IL-10, TNF-α and IFN-γ in level,whether the statistical difference was observed.To explore (1) JE possiblemechanism of IL-10、TNF-α and IFN-γ in the process of brain tissue damage;(2) correlation between different JE syndromes and IL-10、TNF-α and IFN-γ;(3) explore the relevance of the different stages of the symptoms ofencephalitis and cytokines, revealing its essence and material basis. Toprovide an objective basis for the future of Japanese encephalitis andtreatment，and to provide new ideas for TCM standardization process.The collection in2011in Children’s Hospital of Chongqing MedicalUniversity of Japanese encephalitis patients (JE) specimens collected braincollateral damage poison, poison trap pericardial syndrome clinical datacollection and testing physical and chemical indicators, screening syndromespredisposing genes associated test specimen. Using Taqman assay genotyping,to find statistical polymorphism among different groups, aims to revealrelevance between the IL-10promoter region position-592and IFN-γpromoter region prone position-1616and sensibility、type and outcome ofpatients with encephalitis. Methods:All84patients are Children have been hospitalized in Children’s Hospitalof Chongqing Medical University, Department of Infectious Diseases fromJune2011to September2012. Study group that is toxic brain collateraldamage group of30patients (17male,13female), mean age6.62±4.01years old; poison trap pericardium group of30patients (18men,12women),mean age5.16±2.79years old. Study group is composed of the twogroups.Patients in the control group of24patients (14male,10female) overthe same period in outpatient examination, the average age of5.58±3.20years old. All subjects in the admission or the next morning after fastingvenous blood4ml (with EDTA anticoagulant), after centrifugationconditions under3000r/min separated serum aliquots into the upper800℃freezer saved. Double antibody sandwich ELISA assay in human peripheralblood IL-10、TNF-α levels and IFN-γ, human interleukin10ELISA kit、human tumor necrosis factor α ELISA kit and man interferon γ ELISA kitprovided by Boster. Detected in the First Hospital of Wuhan BiologicalLaboratory reagents according to the instructions to complete. All data are inform of±S said count data were compared using chi-square test tocompare measurement data mining t-test and analysis of variance.All the study subjects (n:297) of JE epidemic season in June2011toSeptember2012JE patient informed consent. Of these,66cases of poisoningpatients with brain collateral damage (based on clinical diagnosis and IgMantibody positive diagnosis),39cases of patients with pericardial poison trap(based on clinical diagnosis and IgM antibody positive diagnosis), Japaneseencephalitis patients from Chongqing Medical University Department ofInfectious Diseases, Children’s Hospital inpatient children, in addition to192cases for the control group of patients, all from the children’s Hospital ofChongqing Medical University medical clinic for children. All subjects in the admission or the next morning after fasting venous blood4ml (with EDTAanticoagulant) at-80℃cryopreservation. Use Kangwei century universalpillar genome extraction kit to extract genomic DNA in a hospital in Wuhan,biological laboratories, measuring genotyping PCR using TaqMan probefluorescence methods, primers and fluorescent probes designed by Goldenintellectualism synthesis. Sequencing results were analyzed, the data isdetermined by chi-square test (χ2) using standard statistical software, p <0.05considered statistically significant. Genotype frequency is determined byHardy-Weinberg equilibrium statistical test method. Use PLINK software tocalculate the value of χ2value of genotype and allele frequencies, odds ratiosand95%confidence intervals.Results:①Brain collateral damage poison, toxic levels of depression-10ILpericardial contrast with the control group serum brain collateral damagepoison, poison trap pericardium in serum IL-10levels were significantlyhigher in comparison with a significant difference between the two (P <0.05).Poison trap pericardial serum levels of IL-10was significantly higher than thegroup of brain collateral damage poison, there is a significant differencebetween the two.②Brain collateral damage poison, poison trap pericardium and controlgroups in serum IFN-γ levels of contrast, brain collateral damage poison,poison trap pericardium group serum IFN-γ levels were significantly higher incomparison with a significant difference between the two (P <0.05). Poisontrap pericardium group serum IFN-γ levels were significantly higher thanpoison brain collateral damage group, a significant difference between thetwo groups.③Poison trap obstacle awareness pericardium group was significantly higherthan the severity of brain collateral damage toxin group, a significant difference between the two.④Poison trap pericardium group was significantly higher than the duration ofconsciousness toxic brain collateral damage group, a significant differencebetween the two groups.⑤Pearson rank correlation test will be the extent of60cases ofunconsciousness and IL10-level correlation coefficient was0.276, P=0.032<0.05significantly correlated.⑥The extent of60cases of unconsciousness and IFN-γ levels Pearson rankcorrelation test, correlation coefficient was0.424, P=0.001<0.01significantly correlated.⑦Between IFN-γ promoter region-1616loci TT homozygote frequency andthe frequency of TC and CC plus the JE group (ie, all Japanese encephalitispatients, including brain collateral damage poison and poison traps setpericardium group) and control group.frequency distribution was statisticallysignificant (TT vs TC+CC: Χ2=4.79, P=0.0287**).⑧IL-10promoter region-592CC homozygotes in the frequency and thefrequency and JE CA plus AA group (ie all encephalitis patients, includingbrain collateral damage poison and poison traps set pericardium group)between the control group and the frequency distribution was statisticallysignificant (CC vs CA+AA: Χ2=3.93, P=0.0475**), in addition, thefrequency of a allele C ratio in the JE group (ie, all Japanese encephalitispatients, including toxic brain damage contact between the group and thepoison trap pericardium group) and control group was statistically significantfrequency distribution (C vs A: Χ2=5.344, P=0.021**, OR=0.638,95%CI=0.44-0.94).⑨C allele frequency than the frequency of A in the brain collateral damageamong drug group and control group was statistically significant frequencydistribution (C vs A: Χ2=4.045, P=0.044**, OR=0.634,95%CI= 0.41-0.99).Conclusion:1. IL-10, IFN-γ levels in different syndromes content is statisticallysignificant, suggesting that IL-10, IFN-γ can be used as a basis for clinicalclassification for blood gas business in medical and public health provide thebasis for mass change, and as the extent of the infection, one used in clinicalbrain injury indicators.2. IL-10inhibition of inflammatory cytokines play, prevent excessive damagecell function, and therefore IL10-levels can indirectly reflect the severity ofinflammation, the present study reflects the degree of IL10-levelconsciousness significant correlation can be seen IL-10can reflect the degreeof brain tissue damage and inflammation severity, it can be used as a clinicalsyndrome type basis.3. The experiment reflects the IFN-γ levels and the degree of consciousnessthere is a significant correlation, we can see that IFN-γ can reflect the extentof brain damage, can also prompt evil blood into the camp, the disturbance ofmind, it can be used as a clinical syndrome type basis.4. IFN-γ in-1616TT genotype homozygous loci not more susceptible toJapanese encephalitis;5. IL-10-592A allele subjects more susceptible to encephalitis, nothomozygous CC genotype is more susceptible to encephalitis. In the era ofpersonalized medicine, gene mutations and susceptibility to understand thelink between infectious diseases is necessary, this information may helpdoctors understand the prognosis of patients with the disease, the treatmentmay take appropriate measures in advance.