OBJECTIVE To investigate the possible effects of meteorological and environmental factors on IL-4 gene specific DNA methylation levels in CD4+T cells of children with AR.METHODS Thirty five pediatric AR patients(6-12 years) followed up for one year from the Department of Otolaryngology, Shanghai Children Medical Center from Jan, 1, 2012 to Dec. 31, 2012 were included in this study. Data on daily particulate matter of diameter smaller than 10 micrometer(PM10) and particulate matter of diameter smaller than 2.5 micrometer(PM2.5) was available as average values derived from the data of 6 state-controlled monitoring stations distributed across Pudong District, Shanghai. We quantified IL-4(interleukin-4) gene specific DNA methylation levels in CD4+T cells from 35 patients with AR and 30 healthy controls. mRNA levels of IL-4 gene were measured by real-time reverse transcriptase-PCR. Methods of personal exposure assessment of PM2.5 and PM10 were measured.RESULTS Compared with controls, IL-4 promoter region was hypomethylated in AR CD4+T cells(P=0.038). Of all observed CpG sites in IL-4 promoter region, there was significant differences in CpG-48, CpG+54(P=0.041, 0.032). IL-4 mRNA expression was significantly increased in CD4+T cells(P=0.039). The level of IL-4 mRNA expression was negatively correlated to the mean level of methylation in IL-4 promoter region. After adjusting, level of PM10 exposure was negatively correlated with level of methylation in IL-4 promoter region(r2=0.419,β=-0.470,SD=0.781,P=0.045). CONCLUSION Level of methylation in IL-4 promoter region may be affected by PM10 exposure.

Objective:To establish autoverification rules for coagulation tests in multicenter cooperative units, in order to reduce workload for manual review of suspected results and shorten turnaround time (TAT) of test reports, while ensure the accuracy of results.Methods:A total of 14 394 blood samples were collected from fourteen hospitals during December 2019 to March 2020. These samples included: Rules Establishment Group 11 230 cases, including 1 182 cases for Delta check rules; Rules Validation Group 3 164 cases, including 487cases for Delta check; Clinical Application Trial Group 77 269 cases. Samples were analyzed for coagulation tests using Sysmex CS series automatic coagulation analyzers, and the clinical information, instrument parameters, test results, clinical diagnosis, medication history of anticoagulant and other relative results such as HCT, TG, TBIL, DBIL were summarized; on the basis of historical data, the 2.5 and 97.5 percentile of all data arranged from low to high were initially accumulated; on the basis of clinical suggestions, critical values and specific drug use as well as relative guidelines, autoverification rules and limits were established.The rules were then input into middleware, in which Stage I/Stage II validation was done. Positive coincidence, negative coincidence, false negative, false positive, autoverification pass rate, passing accuracy (coincidence of autoverification and manual verification) were calculated. Autoverification rules underwent trial application in coagulation results reports.Results:(1) The autoverification algorisms involve 33 rules regarding PT/INR, APTT, FBG, D-dimer, FDP,Delta check, reaction curve and sample abnormalities; (2)Autoverification Establishment Group showed autoverification pass rate was 68.42% (7 684/11 230), the false negative rate was 0%(0/11230), coincidence of autoverification and manual verification was 98.51%(11 063/11 230), in which positive coincidence and negative coincidence were respectively 30.09% (3 379/11 230) and 68.42%(7 684/11 230); Autoverification Validation Group showed autoverification pass rate was 60.37%(1 910/3 164), the false negative rate was 0%(0/11 230), coincidence of autoverification and manual verification was 97.79%(3 094/3 164), in which positive coincidence and negative coincidence were respectively 37.42%(1 184/3 164) and 60.37%(1 910/3 164); (3) Trialed implementation of these autoverification rules on 77 269 coagulation samples showed that the average TAT shortened by 8.5 min-83.1 min.Conclusions:This study established 33 autoverification rules in coagulation tests. Validation showedthese rules could ensure test quality while shortening TAT and lighten manual workload.

Objective:To investigate the possible effects of meteorological and environmental factors on AR of children and IFN-γgene specific DNA methylation levels in CD4⁺ T cells of patients with AR. Method:Undergoing follow-up on 35 pediatric AR patients (6-12 years). Data on daily sulfur dioxide (SO₂), nitrogen dioxide (NO₂), particulate matter of diameter smaller than 10 micrometer (PM-10) and particulate matter of diameter smaller than 2.5 micrometer (PM2.5), the average of ozone (O₃) per 8 hours was available as average values derived from the data of 6 state controlled monitoring stations distributed across Pudong district, Shanghai. We quantified IFN-γ (interferon-γ) gene specific DNA methylation levels in CD4⁺ T cells from 35 patients with AR and 30 healthy controls. mRNA levels of IFN-γ gene were measured by real-time reverse transcriptase-PCR. Methods of personal exposure assessment of PM2.5 and PM10 were measured. Result:Compared with control, IFN-γ promoter region was hypermethylated in AR CD4⁺ T cells (P<0.05). Of all observed CpG sites in IFN-γ promoter region, there were significant differences in CpG⁻²⁹⁹, CpG⁺¹¹⁹, CpG⁺¹⁶⁸ (P=0.004, P=0.029, P=0.035). IFN-γ mRNA expression was significantly increase in CD4⁺ T cells (P<0.05). The level of IFN-γ mRNA expression was negatively correlated to mean level of methylation in IFN-γ promoter region. After adjusting, level of long exposure PM2.5 was positively correlated with level of methylation in IFN-γ promoter region. Conclusion:Level of methylation in IFN-γ promoter region may be affected by long exposure PM2.5.