OBJECTIVE: To explore the biomechanical proteins different between low myopic corneas and moderate to high myopic corneas.
METHODS: A total of 27 myopic corneas were used for the Tandem Mass Tag (TMT) proteomics analysis. Differentially expressed proteins (DEPs) were clustered with fold changes > 1.20 or < 0.83 and p < 0.05. Proteins and Proteins Interactions (PPIs) were conducted to find hub proteins; Uniprot database was to screen proteins with biomechanical functions, and Parallel Reaction Monitoring (PRM) was performed to verify the TMT results. Pearson analysis was used to reveal the correlations between myopic degrees and biomechanical proteins. The Immunofluorescence (IF) staining was used to observe the protein distributions.
RESULTS: In total, 34 DEPs were observed between moderate myopic corneas and low myopic corneas; 103 DEPs were observed between high myopic corneas and low myopic corneas, 20 proteins overlapped. The PPIs analysis showed keratin 2, keratins 10 and PRSS1 were hub proteins. The Uniprot function analysis suggested keratin 2 and keratin 10 exhibited biomechanical functions. The PRM demonstrated keratin 2 and keratin 10 levels were significantly lower in moderate and high myopic corneas, which was consistent with the TMT proteomics results. IF staining also demonstrated keratin 2 and keratin 10 were less distributed in moderate and high myopic corneas than in low myopic corneas.
CONCLUSIONS: The levels of biomechanical proteins keratin 2 and keratin 10 are significantly lower in moderate and high myopic corneas than in low myopic corneas.

The infection and replication of avian influenza virus (AIV) in host cells is a complex biological process that involves the transport of viral genes through the host cell\'s transport systems. Actin, microtubules and vimentin are known to facilitate transport of endosomes to the perinuclear region, but the biological role of Keratin, another intermediate filament, in viral transport during AIV replication is not well understood. In this study, the viral NS2 protein was used as the target protein to identify the potential interacting proteins following GST-Pulldown method and protein mass spectrometry. It was discovered that Keratin10 interacted with NS2. Subsequently, it was found AIV infection did not affect the gene level or protein level of keratin10 in HeLa cells, but when Keratin10 was knocked down, the expressions of viral NP mRNA and protein were reduced, and the generation of offspring virus also was also decreased. Furthermore, in early viral infection, Keratin10 could aggregate and co-localize with NP proteins, suggesting that Keratin10 might be connected to early viral transport. Additionally, it was demonstrated that Keratin10 co-localized with Lamp1 and that AIV particles were trapped in late endosomes/Lysosomes after Keratin10 was knocked down. Finally, it was discovered that the knocking down Keratin10 in HeLa cells led to an increase in the acidic pH of endosomes and lysosomes, which prevented AIV from undergoing fusion and uncoating, and then inhibited the process of the viral infection. Overall, the results suggested that Keratin10 might play the critical role in the release of vRNPs from LEs/Ls and can affect the generation of offspring virus. The study provides the novel insights into the role of Keratin10 in the process of AIV infection and transmission, which may have implications for developing new strategies to against AIV infections.

BACKGROUND: Ichthyosis with confetti (IWC) is an extremely rare autosomal-dominant genodermatosis characterized by erythroderma with numerous confetti-like pale spots. IWC is caused by mutations in KRT10 (IWC-I) or KRT1 (IWC-II) which affect their tail domains. In IWC-I, the mutations lead to replacement of glycine/serine-rich keratin 10 (K10) tail with arginine- or alanine-rich frameshift motifs, causing K10 mis-localization which might trigger loss of the mutant KRT10 allele via mitotic recombination, leading to genetic reversion.
OBJECTIVE: To investigate mutations in five IWC-I patients and their functional consequences.
METHODS: We performed Sanger sequencing of KRT1 and KRT10 in peripheral blood samples of five patients, with highly polymorphic KRT10 SNPs genotyped to confirm loss-of-heterozygosity in the epidermis of pale spots. K10 expression pattern was examined in both patient skin biopsies and HaCaT cells overexpressing mutant KRT10-enhanced green fluorescence protein fusion.
RESULTS: Four novel and one recurrent KRT10 mutations were identified in patient peripheral blood samples but not in the corresponding pale spot epidermis. Two of the mutations, c.1696_1699dupCACA and c.1676dupG, affected residues close to K10 carboxyl terminus and encoded only 3 and 6 arginine residues, which were far fewer than reported previously. Interestingly, imaging analyses for K10 in HaCaT cells overexpressing either of these two mutations and in the corresponding patients\' affected skin, showed a remarkably lower level of K10 mis-localization compared to that of other mutations reported in this study.
CONCLUSIONS: Our findings suggest that the number of arginine residues in the mutant tail may correlate with the level of K10 mis-localization in IWC-I keratinocytes. These results expand the genotypic and phenotypic spectrum of IWC-I.