Effect of desflurane on expression of Bcl-2, c-IAP1 and SOD activity.

<p>Western blot analysis showing concentration of Bcl-2 (A), c-IAP1 (B). Equal loading was confirmed by western blot with an anti-β-actin or anti-tubulin antibody. The Bcl-2 and c-IAP1protein concentrations were calculated by averaging the results obtained from five independent experiments. The SOD activity was measured by WST-1 assay (C). Data are presented as mean±SD. *<i>P</i><0.05 versus CON group, ^#<i>P</i><0.05 versus DES+A/R group.</p

Desflurane preconditioning induced oscillation of NF-κB between nucleus and cytoplasm.

<p>Broken arrow refers to unconfirmed part of current study.</p

Experiment protocols.

<p>HUVECs submitted to the anoxia and reoxygenation (A/R) and pretreated with and without 1.0 minimum alveolar concentration (MAC) desflurane or 5 µM BAY11-7082 (BAY). For cell viability, apoptosis, SOD activity assays, the cell samples were collected at the end of experiments. For western blot analysis, cell samples were collected at the end of reoxygenation (samples were collected at 45 min after baseline in CON group and 15 min after desflurane exposure in DES group).</p

Eeffect of desflurane on SMAC release and capase-3 cleavage.

<p>Western blot analysis showing cytosolic SAMC (A) and total caspase-3 (B). Equal loading was confirmed by western blot with an anti-β-actin antibody. The cytosolic SMAC and total caspsase-3 concentrations were calculated by averaging the results obtained from five independent experiments. Data are presented as mean±SD. *<i>P</i><0.05 versus CON group, ^#<i>P</i><0.05 versus DES+A/R group.</p

HUVECs immunofluorescence stained with anti-factor VIII related antibody.

<p>Column 1 is the image of factor VIII (green) in cytoplasm, column 2 is the image of nuclei (blue) and column 3 is the merged image. Magnification ×40.</p

Effect of desflurane on NF-κB p65 nuclear translocation.

<p>Immunofluorescence assay showing NF-κB p65 subunit (green) translocated to nuclei (blue) after exposure to desflurane or A/R. Arrowheads signify some positive nuclei. Magnification ×200.</p

Effect of desflurane preconditioning on HUVECs submitted to A/R.

<p>Cells were submitted to A/R with or without pretreated with desflurane for 30 min or 5 µM BAY11-7082 (BAY). The viability was measured by MTT assay (A). The apoptosis rate was measured by FACSan flow cytometry (B). Data are presented as mean±SD. n = 6/group. *<i>P</i><0.05 versus CON group,^#<i>P</i><0.05 versus DES+A/R group.</p

Data_Sheet_1_Circulating vitamin levels mediate the causal relationship between gut microbiota and cholecystitis: a two-step bidirectional Mendelian randomization study.docx

BackgroundThe relationship between gut microbiota and the occurrence of cholecystitis remains unclear. Existing research lacks a clear understanding of how circulating vitamin levels modulate this relationship. Therefore, our study aims to investigate whether circulating vitamin levels mediate the causal relationship between gut microbiota and cholecystitis using a two-step bidirectional Mendelian randomization approach.MethodsIn this study, we initially employed Linkage Disequilibrium Score Regression (LDSC) analysis to assess the genetic correlation of five circulating vitamin level genome-wide association study (GWAS) summary datasets, thereby avoiding potential sample overlap. Subsequently, we conducted a two-step analysis to investigate the causal effects between gut microbiota and cholecystitis. In the second step, we explored the causal relationship between circulating vitamin levels and cholecystitis and identified the mediating role of vitamin D. The primary method used for causal analysis was the inverse variance-weighted approach. We performed additional sensitivity analyses to ensure result robustness, including the cML-MA method and reverse Mendelian randomization (MR) analysis.ResultsAn increment of one standard deviation in RuminococcaceaeUCG003 was associated with a 25% increased risk of cholecystitis (OR = 1.25, 95%CI = 1.01–1.54, p = 0.04), along with a 3% decrease in 25-hydroxyvitamin D levels (OR = 0.97, 95%CI = 0.944–0.998, p = 0.04). However, following the rigorous Bonferroni correction, every one standard deviation decrease in circulating vitamin D levels was associated with a 33% increased risk of cholecystitis (OR = 0.67, 95%CI = 0.49–0.90, p = 0.008, Padjust = 0.04). Thus, the potential link between gut microbiota and cholecystitis risk might be mediated by circulating vitamin D levels (proportion mediated = 5.5%). Sensitivity analyses provided no evidence of pleiotropy.ConclusionOur study results suggest that an elevated abundance of specific gut microbiota is associated with an increased susceptibility to cholecystitis, with the causal relationship being mediated by circulating vitamin D levels. Further large-scale randomized controlled trials are necessary to validate the causal effects of gut microbiota on cholecystitis risk. This study provides novel insights into cholecystitis prevention through the regulation of gut microbiota.</p

Effects of propofol on CoCl₂ induced the destruction of intracellular Ca²⁺ homeostasis and the phosphorylation of CAMKIIα, ERK, NF-κB.

<p>A and B, CoCl₂-increased intracellular Ca²⁺ concentration was attenuated by propofol. C, D, E, F, G and H, CoCl₂-induced the phosphorylation of CAMKIIα, ERK and NF-κB were attenuated by propofol. (* p < 0.05 vs. control, # p < 0.05 vs. CoCl₂ treatement, n = 5, Data were shown as mean ± SD).</p

Effects of propofol, BAPTA-AM, KN93, U0126 on CoCl₂ induced the phosphorylation of CAMKIIα, ERK, NF-κB.

<p>CoCl₂-induced the phosphorylation of CAMKIIα, ERK and NF-κB were attenuated by propofol, BAPTA-AM, KN93 and U0126. (* p < 0.05 vs. control, # p < 0.05 vs. CoCl₂ treatement, n = 5, Data were shown as mean ± SD).</p