Decreased Expression of the Host Long-Noncoding RNA-GM Facilitates Viral Escape by Inhibiting the Kinase activity TBK1 via S-glutathionylation.

Viruses have evolved multiple strategies to evade elimination by the immune system. Here we examined the contribution of host long noncoding RNAs (lncRNAs) in viral immune evasion. By functional screening of lncRNAs whose expression decreased upon viral infection of macrophages, we identified a lncRNA (lncRNA-GM, Gene Symbol: AK189470.1) that promoted type I interferon (IFN-I) production and inhibited viral replication. Deficiency of lncRNA-GM in mice increased susceptibility to viral infection and impaired IFN-I production. Mechanistically, lncRNA-GM bound to glutathione S-transferase M1 (GSTM1) and blocked GSTM1 interaction with the kinase TBK1, reducing GSTM1-mediated S-glutathionylation of TBK1. Decreased S-glutathionylation enhanced TBK1 activity and downstream production of antiviral mediators. Viral infection reprogrammed intracellular glutathione metabolism and furthermore, an oxidized glutathione mimetic could inhibit TBK1 activity and promote viral replication. Our findings reveal regulation of TBK1 by S-glutathionylation and provide insight into the viral mediated metabolic changes that impact innate immunity and viral evasion.

CCR7 Chemokine Receptor-Inducible lnc-Dpf3 Restrains Dendritic Cell Migration by Inhibiting HIF-1α-Mediated Glycolysis.

CCR7 chemokine receptor stimulation induces rapid but transient dendritic cell (DC) migration toward draining lymph nodes, which is critical for the initiation of protective immunity and maintenance of immune homeostasis. The mechanisms for terminating CCR7-mediated DC migration remain incompletely understood. Here we have identified a long non-coding RNA lnc-Dpf3 whose feedback restrained CCR7-mediated DC migration. CCR7 stimulation upregulated lnc-Dpf3 via removing N6-methyladenosine (m6A) modification to prevent RNA degradation. DC-specific lnc-Dpf3 deficiency increased CCR7-mediated DC migration, leading to exaggerated adaptive immune responses and inflammatory injuries. Mechanistically, CCR7 stimulation activated the HIF-1α transcription factor pathway in DCs, leading to metabolic reprogramming toward glycolysis for DC migration. lnc-Dpf3 directly bound to HIF-1α and suppressed HIF-1α-dependent transcription of the glycolytic gene Ldha, thus inhibiting DC glycolytic metabolism and migratory capacity. We demonstrate a critical role for CCR7-inducible lnc-Dpf3 in coupling epigenetic and metabolic pathways to feedback-control DC migration and inflammatory responses.

Adult Connective Tissue-Resident Mast Cells Originate from Late Erythro-Myeloid Progenitors.

Tissue-resident mast cells are associated with many inflammatory and physiological processes. Although mast cells arise from the yolk sac, the exact ontogeny of adult mast cells remains unclear. Here we have investigated the hematopoietic origin of mast cells using fate-mapping systems. We have shown that early erythro-myeloid progenitors (EMPs), late EMPs, and definitive hematopoietic stem cells (HSCs) each gave rise to mast cells in succession via an intermediate integrin ß7+ progenitor. From late embryogenesis to adult, early EMP-derived mast cells were largely replaced by late EMP-derived cells in most connective tissues except adipose and pleural cavity. Thus, mast cells with distinct origin displayed tissue-location preferences: early EMP-derived cells were limited to adipose and pleural cavity and late EMP-derived cells dominated most connective tissues, while HSC-derived cells were a main group in mucosa. Therefore, embryonic origin shapes the heterogeneity of adult mast cells, with diverse functions in immunity and development.

Rhbdd3 controls autoimmunity by suppressing the production of IL-6 by dendritic cells via K27-linked ubiquitination of the regulator NEMO.

Excessive activation of dendritic cells (DCs) leads to the development of autoimmune and inflammatory diseases, which has prompted a search for regulators of DC activation. Here we report that Rhbdd3, a member of the rhomboid family of proteases, suppressed the activation of DCs and production of interleukin 6 (IL-6) triggered by Toll-like receptors (TLRs). Rhbdd3-deficient mice spontaneously developed autoimmune diseases characterized by an increased abundance of the TH17 subset of helper T cells and decreased number of regulatory T cells due to the increase in IL-6 from DCs. Rhbdd3 directly bound to Lys27 (K27)-linked polyubiquitin chains on Lys302 of the modulator NEMO (IKKγ) via the ubiquitin-binding-association (UBA) domain in endosomes. Rhbdd3 further recruited the deubiquitinase A20 via K27-linked polyubiquitin chains on Lys268 to inhibit K63-linked polyubiquitination of NEMO and thus suppressed activation of the transcription factor NF-κB in DCs. Our data identify Rhbdd3 as a critical regulator of DC activation and indicate K27-linked polyubiquitination is a potent ubiquitin-linked pattern involved in the control of autoimmunity.

Unspliced XBP1 Counteracts ß-Catenin to Inhibit Vascular Calcification.

BACKGROUND: Vascular calcification is a prevalent complication in chronic kidney disease and contributes to increased cardiovascular morbidity and mortality. XBP1 (X-box binding protein 1), existing as the XBP1u (unspliced XBP1) and XBP1s (spliced XBP1) forms, is a key component of the endoplasmic reticulum stress involved in vascular diseases. However, whether XBP1u participates in the development of vascular calcification remains unclear. METHODS: We aim to investigate the role of XBP1u in vascular calcification. XBP1u protein levels were reduced in high phosphate-induced calcified vascular smooth muscle cells, calcified aortas from mice with adenine diet-induced chronic renal failure, and calcified radial arteries from patients with chronic renal failure. RESULTS: Inhibition of XBP1u rather than XBP1s upregulated in the expression of the osteogenic markers Runx2 (runt-related transcription factor 2) and Msx2 (msh homeobox 2), and exacerbated high phosphate-induced vascular smooth muscle cell calcification, as verified by calcium deposition and Alizarin red S staining. In contrast, XBP1u overexpression in high phosphate-induced vascular smooth muscle cells significantly inhibited osteogenic differentiation and calcification. Consistently, smooth muscle cell-specific XBP1 deficiency in mice markedly aggravated the adenine diet- and 5/6 nephrectomy-induced vascular calcification compared with that in the control littermates. Further interactome analysis revealed that XBP1u is bound directly to ß-catenin, a key regulator of vascular calcification, via amino acid (aa) 205-230 in its C-terminal degradation domain. XBP1u interacted with ß-catenin to promote its ubiquitin-proteasomal degradation and thus inhibited ß-catenin/TCF (T-cell factor)-mediated Runx2 and Msx2 transcription. Knockdown of ß-catenin abolished the effect of XBP1u deficiency on vascular smooth muscle cell calcification, suggesting a ß-catenin-mediated mechanism. Moreover, the degradation of ß-catenin promoted by XBP1u was independent of GSK-3ß (glycogen synthase kinase 3ß)-involved destruction complex. CONCLUSIONS: Our study identified XBP1u as a novel endogenous inhibitor of vascular calcification by counteracting ß-catenin and promoting its ubiquitin-proteasomal degradation, which represents a new regulatory pathway of ß-catenin and a promising target for vascular calcification treatment.

Urate-Lowering Therapy Inhibits Thoracic Aortic Aneurysm and Dissection Formation in Mice.

BACKGROUND: Thoracic aortic aneurysm and dissection (TAAD) is a highly lethal vascular disease without effective drug therapy. Whether elevated serum concentrations of uric acid are involved in TAAD development remains unclear. METHODS: Serum uric acid levels were detected in different TAAD mouse models and patients. The urate-lowering drug allopurinol was administered in the drinking water of TAAD mice. Adenine diet-induced mice were established to investigate the role of hyperuricemia in TAAD formation and RNA-sequencing of thoracic aortas from these mice was performed. RESULTS: We found serum uric acid levels were elevated in various mouse TAAD models, including mice fed a ß-aminopropionitrile diet, Marfan mice with fibrillin-1 haploinsufficiency (Fbn1C1041G/+), and ApoE-/- mice infused with Ang II (angiotensin II), as well as in patients with TAAD. Administration of urate-lowering drug allopurinol in the drinking water significantly alleviated TAAD formation in ß-aminopropionitrile-treated mice, Fbn1C1041G/+ mice, and Ang II-infused ApoE-/- mice. Moreover, an adenine diet was used to induce hyperuricemia in mice. Intriguingly, a 4-week adenine diet feeding directly induced TAAD formation characterized by increased maximal thoracic aortic diameters and severe elastin degradation, which were ameliorated by allopurinol. Unbiased RNA-sequencing in mouse thoracic aortas suggested that FcγR (Fc gamma receptor) was upregulated upon adenine diet, but reciprocally repressed by allopurinol. Mechanistically, hyperuricemia activated FcγR-mediated ERK1/2 (extracellular signal-regulated kinase 1/2) phosphorylation to induce macrophage inflammation and TAAD development, which was abrogated by allopurinol or FcγR deficiency. CONCLUSIONS: This study uncovered an important and previously unrecognized role of hyperuricemia in mediating the pathogenesis of TAAD, and uric acid-lowering drug may represent a promising therapeutic approach for TAAD.

Tet2 promotes pathogen infection-induced myelopoiesis through mRNA oxidation.

Varieties of RNA modification form the epitranscriptome for post-transcriptional regulation. 5-Methylcytosine (5-mC) is a sparse RNA modification in messenger RNA (mRNA) under physiological conditions. The function of RNA 5-hydroxymethylcytosine (5-hmC) oxidized by ten-eleven translocation (Tet) proteins in Drosophila has been revealed more recently. However, the turnover and function of 5-mC in mammalian mRNA have been largely unknown. Tet2 suppresses myeloid malignancies mostly in an enzymatic activity-dependent manner, and is important in resolving inflammatory response in an enzymatic activity-independent way. Myelopoiesis is a common host immune response in acute and chronic infections; however, its epigenetic mechanism needs to be identified. Here we demonstrate that Tet2 promotes infection-induced myelopoiesis in an mRNA oxidation-dependent manner through Adar1-mediated repression of Socs3 expression at the post-transcription level. Tet2 promotes both abdominal sepsis-induced emergency myelopoiesis and parasite-induced mast cell expansion through decreasing mRNA levels of Socs3, a key negative regulator of the JAK-STAT pathway that is critical for cytokine-induced myelopoiesis. Tet2 represses Socs3 expression through Adar1, which binds and destabilizes Socs3 mRNA in a RNA editing-independent manner. For the underlying mechanism of Tet2 regulation at the mRNA level, Tet2 mediates oxidation of 5-mC in mRNA. Tet2 deficiency leads to the transcriptome-wide appearance of methylated cytosines, including ones in the 3' untranslated region of Socs3, which influences double-stranded RNA formation for Adar1 binding, probably through cytosine methylation-specific readers, such as RNA helicases. Our study reveals a previously unknown regulatory role of Tet2 at the epitranscriptomic level, promoting myelopoiesis during infection in the mammalian system by decreasing 5-mCs in mRNAs. Moreover, the inhibitory function of cytosine methylation on double-stranded RNA formation and Adar1 binding in mRNA reveals its new physiological role in the mammalian system.

Evaluating the microcirculation of the dome-shaped macula and its complications in adults with highly myopic eyes by swept-source optical coherence tomography angiography.

PURPOSE: The aim of this study was to investigate the microcirculatory characteristics of the dome-shaped macula (DSM), its complications in highly myopic eyes and to explore the factors associated with a DSM. METHODS: This cross-sectional case-control study included a total of 98 subjects (98 eyes): 49 eyes with DSM and 49 eyes without DSM. The axial length (AL) of the myopic eyes was matched 1:1 to eliminate the effect of AL differences on the results. Choroidal (CT) and scleral thickness (ST) and other structural parameters were assessed by swept-source optical coherence tomography (SS-OCT). OCT angiography was used to measure microcirculatory parameters in highly myopic eyes. RESULTS: Subjects with DSM had thinner subfoveal choroidal thickness (46.01 ± 13.25 vs. 81.62 ± 48.26 µm; p < 0.001), thicker subfoveal scleral thickness (SFST; 331.93 ± 79.87 vs. 238.74 ± 70.96 µm; p < 0.001) and thinner foveal CT (66.86 ± 24.65 vs. 107.85 ± 52.65 µm; p < 0.001) compared to subjects without DSM. The foveal choroidal perfusion area (0.72 ± 0.04 vs. 0.76 ± 0.04 mm2; p < 0.001) and foveal choroidal vascularity index (0.15 ± 0.04 vs. 0.33 ± 0.14; p < 0.001) were significantly lower in eyes with DSM. Retinoschisis (81.6% vs. 38.8%; p < 0.001) was more common in eyes with DSM. Eyes with horizontal DSM had worse best-corrected logMAR visual acuity than eyes with round DSM (0.34 ± 0.22 vs. 0.23 ± 0.22; p = 0.03). DSM height (98.95 ± 65.17 vs. 104.63 ± 44.62 µm; p = 0.05) was lower in the horizontal DSM. SFST (OR = 1.06, p = 0.04) and foveal choroidal vascularity index (OR = 0.711, p = 0.02) were significantly associated with DSM. DSM width (p < 0.001), foveal choroidal perfusion area (p = 0.01), foveal choriocapillaris perfusion area (p = 0.02) and parafoveal choroidal vascularity index (p = 0.03) were the most significantly associated factors with DSM height. CONCLUSIONS: The microcirculatory characteristics of eyes with DSM differed from those without DSM. Microcirculatory abnormalities were significantly associated with a DSM. The height of the DSM was associated with decreased blood perfusion.

Targeting endothelial tight junctions to predict and protect thoracic aortic aneurysm and dissection.

AIMS: Whether changes in endothelial tight junctions (TJs) lead to the formation of thoracic aortic aneurysm and dissection (TAAD) and serve as an early indicator and therapeutic target remains elusive. METHODS AND RESULTS: Single-cell RNA sequencing analysis showed aberrant endothelial TJ expressions in the thoracic aortas of patients with TAAD. In a ß-aminopropionitrile (BAPN)-induced TAAD mouse model, endothelial TJ function was disrupted in the thoracic aortas at an early stage (5 and 10 days) as observed by a vascular permeability assay, while the intercellular distribution of crucial TJ components was significantly decreased by en face staining. For the non-invasive detection of endothelial TJ function, two dextrans of molecular weights 4 and 70 kDa were conjugated with the magnetic resonance imaging (MRI) contrast agent Gd-DOTA to synthesize FITC-dextran-DOTA-Gd and rhodamine B-dextran-DOTA-Gd. MRI images showed that both probes accumulated in the thoracic aortas of the BAPN-fed mice. Particularly, the mice with increased accumulated signals from 5 to 10 days developed TAAD at 14 days, whereas the mice with similar signals between the two time points did not. Furthermore, the protease-activated receptor 2 inhibitor AT-1001, which seals TJs, alleviated the BAPN-induced impairment of endothelial TJ function and expression and subsequently reduced TAAD incidence. Notably, endothelial-targeted ZO-1 conditional knockout increased TAAD incidence. Mechanistically, vascular inflammation and edema were observed in the thoracic aortas of the BAPN-fed mice, whereas these phenomena were attenuated by AT-1001. CONCLUSION: The disruption of endothelial TJ function is an early event prior to TAAD formation, herein serving as a potential indicator and a promising target for TAAD.

Prediction model for asymptomatic carotid atherosclerosis using retinal microvascular intelligent analysis: A retrospective study.

IMPORTANCE: Early detection and timely diagnosis of asymptomatic carotid atherosclerosis significantly assist in the prevention of ischemic stroke for them. OBJECTIVE: This observational study aimed to develop and validate a novel prediction model to assist in the early diagnosis of carotid atherosclerosis based on new characteristic variables screened by retinal microvascular intelligence analysis. MAIN OUTCOME(S) AND METHOD (S): The least absolute shrinkage and selection operator (LASSO) combined with 10-fold cross-validation were screened for characteristic variables, and nomograms were plotted to demonstrate the prediction model. Receiver operating characteristic (ROC) curves and area under the curve (AUC), calibration plots and brier score (BS), and decision curve analysis (DCA) were used to evaluate the risk model's discrimination, calibration, and clinical applicability. RESULTS: Age, gender, diabetes mellitus (DM), drinking history, vascular branching angle, mean vascular diameter within 0.5-1.0 papillary diameter (PD), curvature tortuosity arteriole in the inferior region of the optic disc, and vascular density in the nasal region of the optic disc were identified as characteristic variables for carotid atherosclerosis with retinal microvascular intelligence analysis. The predictive nomogram model presented good discrimination with AUCs of 0.790 (0.774-0.806), and the calibration curve displayed high consistency between predicted and actual probability. The DCA demonstrated that this nomogram model led to net benefits in a threshold probability range of 20 %-94 % and could be adapted for clinical decision-making. The results of the 100-bootstrap resampling strategy for internal validation also show that the risk model is well discriminated with an AUC of 0.789 and excellent calibration. External validation showed good discrimination with AUCs of 0.703 (0.627 - 0.779) and good calibration, the risk threshold is 10 %-92 % in terms of DCA. CONCLUSIONS AND RELEVANCE: The novel prediction model based on retinal microvascular intelligence analysis constructed in this study could be effective prognoses for predicting the risk of asymptomatic carotid atherosclerosis in a Chinese screening population.

High-fibre diets regulate antioxidative capacity and promote intestinal health by regulating bacterial microbiota in growing pigs.

This experiment was conducted to investigate the effects of a high-fibre diet on growth performance, nutrients digestibility, intestinal health, and intestinal microbiota composition of growing pigs. Twelve healthy "Duroc × Landrace × Yorkshire" castrates (49 ± 1.35 kg) were randomly divided into two groups with six replicates and one pig per replicate. The two diet treatments were fed the basal diet (CON) based on corn and soybean meal and high fibre diet (HF) respectively. The nutritional levels of the two treatments were the same. The experiment lasted 28 days. The results showed that the addition of 16% wheat bran fibre to the diet of growing pigs did not affect growth performance (p > 0.05). Compared with the CON, contents of isobutyric and butyric acid, GSH-PX and T-AOC in serum were increased in the HF. It decreased the gross energy digestibility and acetic acid content in feces of growing pigs (p < 0.05), the contents of GSH-PX and T-AOC in serum. It decreased the gross energy digestibility and acetic acid content in feces of growing pigs (p < 0.05). Compared with the CON, the Shannon, and Chao1 indexes of the HF were increased (p < 0.05). At the phylum level, the abundance of g_Lactobacillus increased in the HF (p < 0.05). Correlation analysis showed that a total of 18 microbial genera were correlated with antioxidant capacity and volatile fatty acid levels (p < 0.05). In summary, this study showed that adding 16% wheat bran to the diet of growing pigs had no effect on growth performance but helped to improve the richness and stability of intestinal microbiota, promote posterior intestinal fermentation and increase serum antioxidant capacity.

Metabolic enzyme Suclg2 maintains tolerogenicity of regulatory dendritic cells diffDCs by suppressing Lactb succinylation.

Metabolic reprogramming plays a pivotal role in the differentiation and function of immune cells including dendritic cells (DCs). Regulatory DCs can be generated in regional tissue niches like splenic stroma and act as an important part of stromal control of immune response for the maintenance of immune tolerance. However, the metabolic alterations during splenic stroma-driven regulatory DCs differentiation and the metabolic enzyme involved in regulatory DCs function remain poorly understood. By combining metabolomic, transcriptomic, and functional investigations of mature DCs (maDCs) and diffDCs (regulatory DCs differentiated from activated mature DCs through coculturing with splenic stroma), here we identified succinate-CoA ligase subunit beta Suclg2 as a key metabolic enzyme that reprograms the proinflammatory status of mature DCs into a tolerogenic phenotype via preventing NF-κB signaling activation. diffDCs downregulate succinic acid levels and increase the Suclg2 expression along with their differentiation from mature DCs. Suclg2-interference impaired the tolerogenic function of diffDCs in inducing T cell apoptosis and enhanced activation of NF-κB signaling and expression of inflammatory genes CD40, Ccl5, and Il12b in diffDCs. Furthermore, we identified Lactb as a new positive regulator of NF-κB signaling in diffDCs whose succinylation at the lysine 288 residue was inhibited by Suclg2. Our study reveals that the metabolic enzyme Suclg2 is required to maintain the immunoregulatory function of diffDCs, adding mechanistic insights into the metabolic regulation of DC-based immunity and tolerance.

Theory of all-coupling angulon for molecules rotating in many-body environment.

The formation of angulon, stemming from the rotor (molecule or impurity), rotating in the quantum many-body field, adds a new member to the quasi-particles' family and has aroused intense interest in multiple research fields. However, the analysis of the coupling strength between the rotor and its hosting environment remains a challenging task, both in theory and experiment. Here, we develop the all-coupling theory of the angulon by introducing a unitary transformation, where the renormalization of the rotational constants for different molecules in the helium nanodroplets is reproduced, getting excellent agreement with the experimental data collected during the past decades. Moreover, the strength of molecule-helium coupling and the effective radius of the solvation shell co-rotating along with the molecular rotor could be estimated qualitatively. This model not only provides significant enlightenment for analyzing the rotational spectroscopy of molecules in the phononic environment, but also provides a new method to study the transfer of the phonon angular momentum in the angulon frame.

FEATURES OF VASCULATURE IN PATIENTS WITH RETINA ARTERIAL MACROANEURYSM USING OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY.

PURPOSE: To assess morphologic characteristics of retinal arterial macroaneurysm (RAM) and their vascular changes using optical coherence tomography angiography (OCTA). METHODS: This observational study included 31 eyes of 29 participants diagnosed with RAM based on fundus fluorescein angiography in Tianjin Medical University Eye Hospital. Multimodal imaging modalities, including fundus photography, fluorescein angiography, and OCTA, were used to examine RAMs. The demographic and clinical characteristics of the RAMs were recorded. RESULTS: Depending on the fundus fluorescein angiography examination, 40 cases of RAM were confirmed in 29 patients. Twenty-three patients were female (79%), and six patients were male (21%). Two patients had binocular RAM, and four eyes had more than one RAMs. Relying on the OCTA technology, RAMs have four different vascular morphology types (i.e., distended, meshed, malformed, and occult types). In the distended type, round or encircled thrombi caused asymmetrical or symmetrical distention of retinal arteriolar, leading to separate true lumen and false thrombus lumen in RAM. In the meshed type, the meshed or dendritic vascular network around the RAM was likely to be the neovascularization due to the ischemia and hypoxia of the arteriolar wall. Finally, in the malformed and occult type, the RAM usually regressed, and the retinal arterioles were remodeled to distorted or normal arterioles accompanied by capillary degradation. CONCLUSION: Relying on the OCTA technology, we found that the RAMs have four different types of vascular morphology. Each group of RAM has different vascular features. The application of OCTA in patients with RAM furthers our understanding of the vasculature of RAMs.

CRISPR-Cas9-Mediated Mutation of Methyltransferase METTL4 Results in Embryonic Defects in Silkworm Bombyx mori.

DNA N6-methyladenine (6mA) has recently been found to play regulatory roles in gene expression that links to various biological processes in eukaryotic species. The functional identification of 6mA methyltransferase will be important for understanding the underlying molecular mechanism of epigenetic 6mA methylation. It has been reported that the methyltransferase METTL4 can catalyze the methylation of 6mA; however, the function of METTL4 remains largely unknown. In this study, we aim to investigate the role of the Bombyx mori homolog METTL4 (BmMETTL4) in silkworm, a lepidopteran model insect. By using CRISPR-Cas9 system, we somatically mutated BmMETTL4 in silkworm individuates and found that disruption of BmMETTL4 caused the developmental defect of late silkworm embryo and subsequent lethality. We performed RNA-Seq and identified that there were 3192 differentially expressed genes in BmMETTL4 mutant including 1743 up-regulated and 1449 down-regulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that genes involved in molecular structure, chitin binding, and serine hydrolase activity were significantly affected by BmMETTL4 mutation. We further found that the expression of cuticular protein genes and collagens were clearly decreased while collagenases were highly increased, which had great contributions to the abnormal embryo and decreased hatchability of silkworm. Taken together, these results demonstrated a critical role of 6mA methyltransferase BmMETTL4 in regulating embryonic development of silkworm.

Effects of different starch diets on growth performance, intestinal health and faecal microbiota of growing pigs.

This experiment was conducted to investigate the effects of different starch source diets on growth performance, intestinal health, and, microbiota of growing pigs. Eighteen healthy "Duroc × Landrace × Yorkshire" pigs (50 ± 0.61 kg, Castrated boar) were randomly divided into three groups with six replicates and one pig per replicate. The pigs in the three treatments were fed diets prepared with cassava flour (CF), rice bran (RB) and sorghum flour (SF), respectively, and the nutritional levels of the three treatments were the same. The experiment lasted for 28 days. The results showed that pigs in the RB group had significantly increased average daily gain (ADG, p < 0.05) compared with pigs in CF and SF groups. Compared with pigs in the CF group, the final body weight (FBW) of growing pigs in the RB group was increased and the ratio of feed to gain (F: G) was decreased (p < 0.05). There was no significant difference between FBW and F: G between the SF group and the other two groups. Compared with the CF group, the RB group significantly increased the jejunum amylase activity (p < 0.05), and there was no significant difference between the SF group and the other two groups. Compared with growing pigs in the CF group and SF group, the duodenal villus height and villus height/crypt depth ratio of growing pigs in the RB group were significantly increased (p < 0.05). The concentrations of acetic acid, propionic acid, and total VFA in the colon and caecum of piglets in the SF group were significantly increased (p < 0.05) compared to piglets in CF and RB groups, and there was no significant difference between the CF group and RB group. Compared with the RB group, caecal butyric acid concentration was significantly increased in SF and CF groups (p < 0.05). Seven dominant phyla were identified at the phylum level, among which Firmicutes, Bacteroidota and Spirochaetota were dominant phyla, accounting for 74.18%, 14.87% and 6.56% of the RB group respectively. Cassava flour group accounted for 80.22%, 9.64% and 3.71%; Accounting for 65.33%, 17.34% and 13.07% of the SF group. Through the comparative analysis of microbial differences among the treatment groups, it was found that at the phylum level, compared with the SF group, the abundance of Synergistota in the diet of the CF group and the diet of the RB group was significantly increased (p < 0.05). The abundance decreased significantly (p < 0.05). The quantity of Desulfobacterota in the RB group was significantly higher than that in the CF group (p < 0.05). In conclusion, compared with sorghum starch and cassava starch, RB starch can improve the activity of digestive enzymes and villus height in the small intestine of growing pigs and promote the growth of pigs by protecting the intestinal health of growing pigs.

Matricellular proteins: Potential biomarkers and mechanistic factors in aortic aneurysms.

Aortic aneurysms, including thoracic aortic aneurysms and abdominal aortic aneurysms, are life-threatening macrovascular diseases with high mortality. The already known key mechanisms implicated in aortic aneurysm pathogenesis involve the remodeling of the extracellular matrix and a set of cellular responses, such as inflammation, oxidative stress and vascular smooth muscle cell dysfunction. Matricellular proteins constitute a group of nonstructural extracellular proteins that link the interaction between cells and their extracellular microenvironment and have been widely reported in different diseases, including aortic aneurysms. In the present review, we summarize the role of various matricellular proteins in the pathogenesis and progression of aortic aneurysms, as well as address the possibility of using these proteins as biomarkers and pathogenic factors, to highlight their clinical significance in aortic aneurysms.

Histone methyltransferase Ash1l suppresses interleukin-6 production and inflammatory autoimmune diseases by inducing the ubiquitin-editing enzyme A20.

Histone modifications play important roles in multiple physiological processes by regulating gene expression. However, the roles of histone modifications in immunity remain poorly understood. Here we report that Ash1l, a H3K4 methyltransferase, suppressed interleukin-6 (IL-6), and tumor necrosis factor (TNF) production in Toll-like receptor (TLR)-triggered macrophages, protecting mice from sepsis. Ash1l-silenced mice were more susceptible to autoimmune disease as a result of enhanced IL-6 production. Ash1l enhanced A20 expression through induction of H3K4 modification at the Tnfaip3 promoter via H3K4 methyltransferase activity of Ash1l SET (Su[var]3-9, E[z] and trithorax) domain. Ash1l suppressed NF-κB, mitogen-activated protein kinase (MAPK) pathways, and subsequent IL-6 production via facilitating A20-mediated NF-κB signal modulator NEMO and transducer TRAF6 deubiquitination. Therefore, Ash1l-mediated H3K4 methylation at the Tnfaip3 promoter is required for controlling innate IL-6 production and suppressing inflammatory autoimmune diseases, providing mechanistic insight into epigenetic modulation of immune responses and inflammation.

Two-way comparison of brain perfusion image processing software for patients with acute ischemic strokes in real-world.

PURPOSE: Perfusion imaging generates multimaps of ischemic tissues and is a proven decision-making tool in patients with acute ischemic stroke. However, the reliability of perfusion post-processing outcomes has been debated, given disparate results of various software applications, especially for patients with small ischemic core volume. This study was undertaken to compare ischemic volume estimates determined by imSTROKE (a software with new imaging protocol) and RAPID computer applications, respectively. METHODS: A total of 611 patients qualified for study, each having met inclusion and exclusion criteria of the Multicenter Randomized Clinical Trial of Endovascular Treatment for Acute Ischemic Stroke in the Netherlands (MR CLEAN trial). Subjects were examined by computed tomography perfusion (CTP) imaging (n = 349) or perfusion-weighted (PWI) and diffusion-weighted (DWI) imaging (n = 262). Ischemic volumes estimated by imSTROKE and RAPID applications were then compared. We used Bland-Altman analysis and intraclass correlation coefficients (ICCs) to ascertain agreement between applications. Accuracies of estimated core infarct and penumbra volumes were tested at specific thresholds (core: 25 mL, 50 mL, and 70 mL; penumbra: 45 mL, 90 mL, and 125 mL). RESULTS: Median core infarct volumes by imSTROKE and RAPID were 29.18 mL and 29.53 mL, respectively (ICC = 0.9880, 95% confidence interval [CI]: 0.9860-0.9898). Median penumbra volumes by imSTROKE and RAPID were 68.20 mL and 68.55 mL, respectively (ICC = 0.9885, 95% CI: 0.9865-0.9902). CONCLUSION: In estimating core infarct and penumbra volumes, imSTROKE and RAPID applications showed high-level agreement. For patients with small ischemic core volume, compared with RAPID, imSTROKE may have better sensitivity.

Novel mutations of the USH2A gene cause Usher syndrome in five Chinese families.

BACKGROUND: Usher syndrome (USH) is a leading disorder of deaf-blindness. The phenotypic and genetic heterogeneity of USH makes the diagnosis of this disorder difficult. However, diagnosis can be facilitated by employing molecular approaches, especially for diseases without pronounced pathognomonic symptoms. Therefore, this study aimed to reveal the genetic defects in five USH patients using clinical targeted exome sequencing (TES). METHODS: USH patients and their family members from five unrelated Chinese USH families were recruited and subjected to TES. Ophthalmic information was obtained for all patients to ensure a meaningful interpretation. The TES data were analysed using an established bioinformatics pipeline to identify causative mutations. Further verification by Sanger sequencing and cosegregation analysis were performed on available family members. RESULTS: We identified genetic mutations in five USH patients using TES. Seven mutations, four of which were novel, were identified in the USH2A gene. One proband (F1-II-3) was found to have a homozygous mutation inherited from nonconsanguineous parents, and another proband (F5-III-1) was found to carry three USH2A gene mutations. CONCLUSION: In conclusion, the study revealed the importance of TES in the clinical diagnosis of USH patients with variable phenotypes. The correlation between USH2A gene mutations and clinical phenotypes will help to refine the clinical diagnosis of USH.