Corticosteroids (CS), first-line therapeutics for Crohn’s disease (CD) with moderate or severe disease activity, were found to restore intestinal permeability in CD patients, whereas the underlying ...molecular events are still largely unknown. This study aimed to investigate the effect and mechanisms of CS prednisolone on epithelial barrier using CD patient-derived intestinal organoids. 3D intestinal organoids were generated from colon biopsies of inactive CD patients. To mimic the inflammatory microenvironment, a mixture of cytokines containing TNF-α, IFN-γ, and IL-1β were added to the organoid culture with or without pre-incubation of prednisolone or mifepristone. Epithelial permeability of the organoids was assessed by FITC-D4 flux from the basal to luminal compartment using confocal microscopy. Expression of junctional components were analyzed by qRT-PCR, immunofluorescence staining, and western blot. Activity of signaling pathways were analyzed using western blot. Exposure of the cytokines significantly disrupted epithelial barrier of the intestinal organoids, which was partially restored by prednisolone. On the molecular level, the cytokine mixture resulted in a significant reduction in E-cadherin and ILDR-1, an increase in CLDN-2, MLCK, and STAT1 phosphorylation, whereas prednisolone ameliorated the abovementioned effects induced by the cytokine mixture. This study demonstrates that prednisolone confers a direct effect in tightening the epithelial barrier, identifies novel junctional targets regulated by prednisolone, and underscores intestinal barrier restoration as a potential mechanism that contributes to the clinical efficacy of prednisolone in CD patients.
Key messages
Prednisolone confers a direct preventive effect against cytokine-induced barrier dysfunction.
Prednisolone regulates the expression of CLDN-2, E-cadherin, and ILDR-1.
The effect of prednisolone is GR-, MLCK-, and STAT1-dependent.
Upconversion (UC) luminescent lanthanide nanoparticles (LNPs) are expected to play an important role in imaging and photodynamic therapy (PDT) in vitro and in vivo. However, with the absorption of UC ...emissions by photosensitizers (PSs) to generate singlet oxygen (1O2) for PDT, the imaging signals from LNPs are significantly weakened. It is important to activate another imaging route to track the location of the LNPs during PDT process. In this work, Nd3+-sensitized LNPs with dual-band visible and near-infrared (NIR) emissions under single 808 nm excitation were reported to address this issue. The UC emissions in green could trigger covalently linked rose bengal (RB) molecules for efficient PDT, and NIR emissions deriving from Yb3+ and magnetic resonance imaging (MRI) were used for imaging simultaneously. Notably, the designed therapeutic platform could further effectively avoid the overheating effect induced by the laser irradiation, due to the minimized absorption of biological media at around 808 nm. TdT-mediated dUTP nick end labeling (TUNEL) assay showed serious cell apoptosis in the tumor after PDT for 2 weeks, leading to an effective tumor inhibition rate of 67%. Benefit from the PDT, the tumor growth-induced liver and spleen burdens were largely attenuated, and the liver injury was also alleviated. More importantly, pulmonary and hepatic tumor metastases were significantly reduced after PDT. The Nd3+-sensitized LNPs provide a multifunctional nanoplatform for NIR light-assisted PDT with minimized heating effect and an effective inhibition of tumor growth and metastasis.
The autophagic ubiquitin‐like protein LC3 functions through interactions with LC3‐interaction regions (LIRs) of other autophagy proteins, including autophagy receptors, which stands out as a ...promising protein–protein interaction (PPI) target for the intervention of autophagy. Post‐translational modifications like acetylation of Lys49 on the LIR‐interacting surface could disrupt the interaction, offering an opportunity to design covalent small molecules interfering with the interface. Through screening covalent compounds, we discovered a small molecule modulator of LC3A/B that covalently modifies LC3A/B protein at Lys49. Activity‐based protein profiling (ABPP) based evaluations reveal that a derivative molecule DC‐LC3in‐D5 exhibits a potent covalent reactivity and selectivity to LC3A/B in HeLa cells. DC‐LC3in‐D5 compromises LC3B lipidation in vitro and in HeLa cells, leading to deficiency in the formation of autophagic structures and autophagic substrate degradation. DC‐LC3in‐D5 could serve as a powerful tool for autophagy research as well as for therapeutic interventions.
A potent probe, DC‐LC3in‐D5, was discovered which demonstrates high selectivity to LC3A/B in the proteome. DC‐LC3in‐D5 inhibits autophagy by attenuating LC3B lipidation, which subsequently reduces the extent of autophagic structure formation and later substrate degradation.
A visible light‐mediated difluoroacetylation of cinnamamides with ethyl bromodifluoroacetate as a CF2 radical precursor is described. The reaction incorporates tandem cyclization or ...cyclization/dearomatization processes. The latter process occurs when there is a p‐RO substituent on the aniline. This protocol affords straightforward new routes to separately synthesize quinoline‐2‐ones and spiro4.5decanes in moderate to good yields.
Ocean temperature prediction plays crucial roles in the ocean-related fields. The graph neural networks (GNNs) show advantages for modeling complex environmental issues. However, the prior methods ...typically focus on node features to predict ocean temperature. Note that the edge features are essential, especially for 3-D ocean temperature prediction. In this letter, a GNN with optimized attention mechanisms (GNN-OAMs) model is proposed to predict 3-D ocean temperature. The GNN-OAM introduces a random forest (RF) module to capture the nonstationary temporal dependencies. Especially, the OAMs, via combination of multiple adjacency matrices construct edge features, are presented to capture the dynamic spatial dependencies. The prediction performance of GNN-OAM model is evaluated in the 3-D ocean temperature experimental dataset, and the results show that the GNN-OAM achieves ocean temperature prediction with high accuracy, and the MAEs are 0.146 and 0.26, when predicting temperatures in next one day and five days.
Hydrogels with high mechanical strength are essential for its most industrial applications. In this work, the hydrophobic SiO2 nanoparticles (M‐SiO2 NPs) modified with octadecyltrimethoxysilane were ...used as physical crosslinker to toughen the mechanical properties of hydrogels. The solution of monomers containing M‐SiO2 NPs, N‐hydroxymethyl acrylamide (NHAM), acrylic acid (AA), 2‐acrylamido‐2‐methylpropanesulfonic acid (AMPS), was polymerized to prepare a P(NHAM/AA/AMPS)‐based composite hydrogel without any chemical cross‐linker. The composition and microstructure of the hydrogel were carefully studied with Fourier transform infrared spectroscopy and scanning electron microscopy. The mechanical properties of the hydrogel were investigated through compressive and tensile tests. Dynamic swelling tests were carried out at different pH values (1.0–12.0) and salinity (2000–20,000 mg/L) to study the acid resistance, alkali resistance, and salt resistance of the hydrogel. The obtained results showed that the hydrogel exhibited excellent mechanical performance, which was attributed to the unique 3D network formed by the hydrogen bond between M‐SiO2 NPs and polymer as well as the hydrophobic association effect between the hydrophobic chains of M‐SiO2 NPs. Furthermore, the hydrophobic association effect was enhanced when contacting with salt, acid, and alkali solutions, thus endowing the hydrogel with intensified salt resistance and wider acid–base adaptability range.
Paeonia lactiflora Pall. (Paeonia) has aroused many concerns due to its extensive medicinal value, in which monoterpene glucoside paeoniflorin and its derivatives are the active chemical components. ...However, little is known in the molecular mechanism of monoterpenoids biosynthesis, and the regulation network between small RNAs and mRNAs in monoterpenoids biosynthesis has not been investigated yet. Herein, we attempted to reveal the tissue-specific regulation network of miRNAs and their targets related to paeoniaflorin and monoterpenoids biosynthesis in Paeonia by combining mRNA and miRNA expression data with degradome analysis. In all, 289 miRNAs and 30177 unigenes were identified, of which nine miRNAs from seven miRNA families including miR396, miR393, miR835, miR1144, miR3638, miR5794 and miR9555 were verified as monoterpenoids biosynthesis-related miRNAs by degradome sequencing. Moreover, the co-expression network analysis showed that four monoterpenoids-regulating TFs, namely AP2, MYBC1, SPL12 and TCP2, were putatively regulated by five miRNAs including miR172, miR828, miR858, miR156 and miR319, respectively. The present study will improve our knowledge of the molecular mechanisms of the paeoniaflorin and monoterpenoids biosynthesis mediated by miRNA to a new level, and provide a valuable resource for further study on Paeonia.
The properties of high theoretical capacity, low cost, and large potential of metallic sodium (Na) has strongly promoted the development of rechargeable sodium‐based batteries. However, the issues of ...infinite volume variation, unstable solid electrolyte interphase (SEI), and dendritic sodium causes a rapid decline in performance and notorious safety hazards. Herein, a highly reversible encapsulation‐based sodium storage by designing a functional hollow carbon nanotube with Zn single atom sites embedded in the carbon shell (ZnSA‐HCNT) is achieved. The appropriate tube space can encapsulate bulk sodium inside; the inner enriched ZnSA sites provide abundant sodiophilic sites, which can evidently reduce the nucleation barrier of Na deposition. Moreover, the carbon shell derived from ZIF‐8 provides geometric constraints and excellent ion/electron transport channels for the rapid transfer of Na+ due to its pore‐rich shell, which can be revealed by in situ transmission electron microscopy (TEM). As expected, Na@ZnSA‐HCNT anodes present steady long‐term performance in symmetrical battery (>900 h at 10 mA cm−2). Moreover, superior electrochemical performance of Na@ZnSA‐HCNT||PB full cells can be delivered. This work develops a new strategy based on carbon nanotube encapsulation of metallic sodium, which improves the safety and cycling performance of sodium metal anode.
Designing a functional encapsulation‐based sodium storage void with abundant pore structure and enriched Zn single atom sites in the inner wall is presented here. Zn single atoms provide abundant selective sodiophilic nucleation sites. Due to the rich pore structure, ZIF‐derived carbon shells can provide geometric constraints and excellent ion/electron transport channels to maintain fast electron/ion contact, benefiting for encapsulating large amounts of metallic sodium.
NOD-like receptors affect multiple stages of cancer progression in many malignancies. NACHT, LRR, and PYD domain-containing protein 7 (NLRP7) is a member of the NOD-like receptor family, although its ...role in tumorigenesis remains unclear. By analyzing clinical samples, we found that NLRP7 protein levels were upregulated in colorectal cancer (CRC). We proposed the hypothesis that a high level of NLRP7 in CRC may promote tumor progression. Here, we further investigated the role of NLRP7 in CRC and the underlying mechanism.
NLRP7 expression in human CRC and adjacent non-tumorous tissues was examined by quantitative real-time polymerase chain reaction (qRT-PCR), western blotting, and immunohistochemistry. The effect of NLRP7 in CRC progression was investigated in vitro and in vivo. Proteins interacting with NLRP7 were identified by immunoprecipitation and mass spectrometry analysis while immunofluorescence staining revealed the cellular location of the proteins. Cellular ubiquitination and protein stability assays were applied to demonstrate the ubiquitination effect on NLRP7. Cloning and mutagenesis were used to identify a lysine acceptor site that mediates NLRP7 ubiquitination. Cytokines/chemokines affected by NLRP7 were identified by RNA sequencing, qRT-PCR, and enzyme-linked immunosorbent assay. Macrophage phenotypes were determined using qRT-PCR, flow cytometry, and immunohistochemistry.
NLRP7 protein levels, but not mRNA levels, were upregulated in CRC, and increased NLRP7 protein expression was associated with poor survival. NLRP7 promoted tumor cell proliferation and metastasis in vivo and in vitro and interacted with ubiquitin-specific protease 10, which catalyzed its deubiquitination in CRC cells. NLRP7 stability and protein levels in CRC cells were modulated by ubiquitination and deubiquitination, and NLRP7 was involved in the ubiquitin-specific protease 10 promotion of tumor progression and metastasis in CRC. K379 was an important lysine acceptor site that mediates NLRP7 ubiquitination in CRC cells. In CRC, NLRP7 promoted the polarization of pro-tumor M2-like macrophages by inducing the secretion of C-C motif chemokine ligand 2. Furthermore, NLRP7 promoted NF-κB nuclear translocation and activation of C-C motif chemokine ligand 2 transcription.
We showed that NLRP7 promotes CRC progression and revealed an as-yet-unidentified mechanism by which NLRP7 induces the polarization of pro-tumor M2-like macrophages. These results suggest that NLRP7 could serve as a biomarker and novel therapeutic target for the treatment of CRC.
Sweetpotato (Ipomoea batatas L. Lam.) is an important food crop. However, the genetic information of the nuclear genome of this species is difficult to determine accurately because of its large ...genome and complex genetic background. This drawback has limited studies on the origin, evolution, genetic diversity and other relevant studies on sweetpotato.
The chloroplast genomes of 107 sweetpotato cultivars were sequenced, assembled and annotated. The resulting chloroplast genomes were comparatively analysed with the published chloroplast genomes of wild species of sweetpotato. High similarity and certain specificity were found among the chloroplast genomes of Ipomoea spp. Phylogenetic analysis could clearly distinguish wild species from cultivars. Ipomoea trifida and Ipomoea tabascana showed the closest relationship with the cultivars, and different haplotypes of ycf1 could be used to distinguish the cultivars from their wild relatives. The genetic structure was analyzed using variations in the chloroplast genome. Compared with traditional nuclear markers, the chloroplast markers designed based on the InDels on the chloroplast genome showed significant advantages.
Comparative analysis of chloroplast genomes of 107 cultivars and several wild species of sweetpotato was performed to help analyze the evolution, genetic structure and the development of chloroplast DNA markers of sweetpotato.
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