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Supplemental material, sj-pdf-2-eso-10.1177_23969873231173285 for Individual trigger factors for hemorrhagic stroke: Evidence from case-crossover and self-controlled case series studies by Junyu Liu, Chun Luo, Yuxin Guo, Fang Cao and Junxia Yan in European Stroke Journal

Supplemental material, sj-docx-1-eso-10.1177_23969873231173285 for Individual trigger factors for hemorrhagic stroke: Evidence from case-crossover and self-controlled case series studies by Junyu Liu, Chun Luo, Yuxin Guo, Fang Cao and Junxia Yan in European Stroke Journal

Supplemental material, sj-docx-1-eso-10.1177_23969873231173285 for Individual trigger factors for hemorrhagic stroke: Evidence from case-crossover and self-controlled case series studies by Junyu Liu, Chun Luo, Yuxin Guo, Fang Cao and Junxia Yan in European Stroke Journal

Supplemental material, sj-pdf-2-eso-10.1177_23969873231173285 for Individual trigger factors for hemorrhagic stroke: Evidence from case-crossover and self-controlled case series studies by Junyu Liu, Chun Luo, Yuxin Guo, Fang Cao and Junxia Yan in European Stroke Journal

BackgroundDiabetic retinopathy (DR) is a common and serious microvascular complication of diabetes mellitus (DM), but its pathological mechanism, especially the formation mechanism of new blood vessels remains unclear. Thrombospondin-1 (THBS1) is a potent endogenous inhibitor of angiogenesis and it was found over expressed in DR in our previous study. Our study aimed to determine whether overexpression of THBS1 is associated with its promoter methylation level, and whether methylation of THBS1 is regulated by genetic variants in DR.MethodsPatients diagnosed with DR and DM patients without retinal problems were included in the case-control study. DNA methylation detection of THBS1 by bisulfite sequencing and genotyping of specific SNPs by MassARRAY analysis were performed in the patients recruited from 2019-2020. Real time quantitative PCR was performed to obtain mRNA expression of THBS1 in the patients recruited from August to October 2022. The differentially methylated CpG loci of THBS1 were identified by logistic regression, and associations between 13 SNPs and methylation levels of CpG loci were tested by methylation quantitative trait loci (meQTLs) analysis. Mediation analysis was applied to determine whether CpG loci were intermediate factors between meQTLs and DR.Results150 patients diagnosed with DR and 150 DM patients without retinal complications were enrolled in the first recruitment, seven DR patients and seven DM patients were enrolled in the second recruitment. The patients with DR showed promoter hypomethylation of THBS1 (P value = 0.002), and six out of thirty-nine CpG sites within two CpG islands (CGIs) showed hypomethylation(P value < 0.05). THBS1 mRNA expression in peripheral blood was significantly higher in DR patients than in DM patients. Five out of thirteen cis-meQTLs were identified to be associated with CpG sites: rs13329154, rs34973764 and rs5812091 were associated with cis-meQTLs of CpG-4 (P value=0.0145, 0.0095, 0.0158), rs11070177 and rs1847663 were associated with cis-meQTLs of CpG-2 and CpG-3 respectively (P value=0.0201, 0.0275). CpG-4 methylation significantly mediated the effect of the polymorphism rs34973764 on DR (B=0.0535, Boot 95%CI: 0.004~0.1336).ConclusionTHBS1 overexpression is related to THBS1 hypomethylation in patients with DR. DNA methylation may be genetically controlled in DR.

Using the closed chamber technique daily changes of net ecosystem carbon dioxide (CO2) exchange (NEE) and ecosystem respiration (Re) were determined in a ryegrass ecosystem on eight separated days at half hour interval from morning to afternoon in Taiyuan basin, Shanxi province of China. The gross primary production (GPP) was calculated as the difference of NEE and Re. The objective of the study was to examine the relationship between ecosystem C fluxes and canopy temperature (Tc), air temperature (Ta) and soil temperature (Ts), and finally to explore the possibility that Tc could substitute Ta or Ts for estimate of ecosystem carbon fluxes. The results showed on diurnal scales that the coefficients of determination of both NEE and GPP with Tc were significantly higher than those of the GPP and NEE with Ta, and that the Re was also closely related with both Ta and Tc on all measurement dates, but that significant correlation of Re with Ts existed only on 7 out of 8 measurement dates. The determination coefficients between Re and Ts were lower than those between Re and both Ta and Tc on diurnal scales. These results suggested that at plot scale Tc also is a good factor for ecosystem carbon fluxes estimate as both Ta and Ts factors. This gives some implications for linking remotely sensed infrared temperature of canopy to ecosystem carbon flux estimate. However, further experiments at ecosystem scale are still needed.