Image1_Construction of ceRNA network and identification of hub genes in aniridia-associated keratopathy using bioinformatics analysis.JPEG

Aniridia-associated keratopathy (AAK) is characteristic at ocular surface of aniridia caused by haploinsufficiency of PAX6. Competing endogenous RNA (ceRNA) has been reported to play an important role in various diseases, whereas its function on AAK is unclear. The microarray data of 20 AAK patients and 20 healthy people were downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed lncRNAs, miRNAs, and mRNAs were analyzed using “limma” packages and weighted gene co-expression network analysis (WGCNA). A ceRNA network was constructed by Cytoscape 3.9.1, and miR-224-5p, miR-30a-5p, and miR-204-5p were at the center of the network. CIBERSORTx algorithm and ssGSEA analyses revealed that AAK was associated with immune cell infiltration, showing that activated Mast cells increased while resting Mast cells decreased and NK cells decreased in AAK. Type II INF Response, CCR, parainflammation, T cell co-stimulation, and APC co-stimulation of AAK patients differed from healthy individuals. Additionally, the ROC curve of five genes, MITF(AUC = 0.988), RHOB(AUC = 0.973), JUN(AUC = 0.953), PLAUR (AUC = 0.925), and ARG2 (AUC = 0.915) with high confidence in predicting AAK were identified. Gene set enrichment analysis (GSEA) analysis of hub genes enriched in the IL-17 signaling pathway.</p

Western blot and densitometric analyses of NLRP3, procaspase-1,and caspase-1 protein expressions in the eyes of controls, NSSDE, and SSDE subjects.

<p>Expression of 120-kDa NLRP3 increased in SSDE versus controls. Expression of 45-kDa procaspase-1 and 20-kDa caspase-1 showed increased protein expressions in SSDE and NSSDE compared with controls.</p

Upregulation of IL-1β expression in the tear and CIC samples of dry eye patients.

<p>(<b>A</b>) Quantitative RT-PCR for IL-1β mRNA expression in CIC specimens showed significant upregulation in eyes with SSDE and NSSDE (***<i>p</i>≤0.001, **<i>p</i>≤0.01, compared with the controls, Kruskal-Wallis test). (<b>B</b>) Tear IL-1β protein expressions via ELISA demonstrated higher levels in SSDE group compared with controls. (**<i>p</i>≤0.01, compared with the controls, Kruskal-Wallis test).</p

Semipinacol Rearrangement of Iododifluorohomoallyl Alcohols and Its Application in the Allylic C–H Esterification Reactions

We herein present a study on the Ag(I)-mediated semipinacol rearrangement of iododifluorohomoallyl alcohols, the resulting allylic difluoromethyl ketones underwent oxidative allylic C–H esterification under palladium catalysis in the absence of external ligand. This process yielded a range of difluoromethyl ketones derived from allyl esters in a single operation. The reaction features broad scope of o-nitrobenzoic acids and homoallylic iododifluoroalcohols affording the targeted molecules in synthetically useful yields. Control experiments illustrated that the silver salt acted as not only a Lewis acid to promote the cleavage of a C–I bond and furnish the semipinacol rearrangement but also a co-oxidant in the catalytic cycle for the allylic C–H esterification

Sequence Data for Gene Amplification in Quantitative RT-PCR.

<p>Sequence Data for Gene Amplification in Quantitative RT-PCR.</p

Upregulation of IL-18 expression in the tear and CIC samples of dry eye patients.

<p>(<b>A</b>) Significant elevation of IL-18 mRNA level was found in SSDE and NSSDE groups using quantitative RT-PCR.(***<i>p</i>≤0.001, compared with the controls, Kruskal-Wallis test). (<b>B</b>) ELISA analyses of IL-18 protein expression demonstrated tear IL-18 increased significantly in SSDE and NSSDE groups. (*<i>p</i><0.05, ***<i>p</i>≤0.001, compared with the controls, Kruskal-Wallis test).</p

Upregulation of NLRP3 mRNA expression in the CIC samples of dry eye patients.

<p>Quantitative analysis of mRNA expression of NLRP3 showed significantly upregulation in SSDE. (*<i>p</i><0.05, compared with the controls, Kruskal-Wallis test).</p

Summary of Diagnostic Tests for Study Groups.

<p>Abbreviations: SSDE = Sjögren's syndrome dry eye; NSSDE = non-Sjögren's syndrome dry eye; TBUT = tear break-up time; FS = fluorescein score;</p><p>^a<i>p <0</i>.<i>05</i>, compared with normal controls, Kruskal-Wallis test.</p><p>^b<i>p<0</i>.<i>05</i>, compared with NSSDE patients, Kruskal-Wallis test.</p><p>Summary of Diagnostic Tests for Study Groups.</p

Additional file 4: of CpG site methylation in CRYAA promoter affect transcription factor Sp1 binding in human lens epithelial cells

Row data of qRT-PCR for CRYAA after Zebularine treatment, Experiment 3. It is the rata data of the third experiment described above. (XLS 2630 kb

Decoding Molecular Mechanism Underlying Human Olfactory Receptor OR8D1 Activation by Sotolone Enantiomers

Sotolone, a chiral compound, plays an important role in the food industry. Herein, (R)-/(S)-sotolone were separated to determine their odor characteristics and thresholds in air (R-form: smoky, burned, herb, and green aroma, 0.0514 μg/m3; S-form: sweet, milk, acid, and nutty aroma, 0.0048 μg/m3). OR8D1 responses to (R)-/(S)-sotolone were detected in a HEK293 cell-based luminescence assay. (S)-Sotolone was a more potent agonist than (R)-sotolone (EC50 values of 84.98 ± 1.05 and 167.20 ± 0.25 μmol/L, respectively). Molecular dynamics simulations and molecular mechanics Poisson–Boltzmann surface area analyses confirmed that the combination of (S)-sotolone and OR8D1 was more stable than that of (R)-sotolone. Odorant docking, multiple sequence alignments, site-directed mutagenesis, and functional studies with recombinant odorant receptors (ORs) in a cell-based luminescence assay identified 11 amino-acid residues that influence the enantioselectivity of OR8D1 toward sotolone significantly and that N2065.46 was indispensable to the activation of OR8D1 by (S)-sotolone