Reducing excessive inflammation is beneficial for the recovery from intracerebral hemorrhage (ICH). Here, the roles and mechanisms of A20 (TNFAIP3), an important endogenous anti-inflammatory factor, are examined in ICH. A20 expression in the PBMCs of ICH patients and an ICH mouse model was detected, and the correlation between A20 expression and neurologic deficits was analyzed. A20 expression was increased in PBMCs and was negatively related to the modified Rankin Scale score. A20 expression was also increased in mouse perihematomal tissues. A20−/− and A20-overexpressing mice were generated to further analyze A20 function. Compared with wild-type (WT) mice, A20−/− and A20-overexpressing mice showed significant increases and decreases, respectively, in hematoma volume, neurologic deficit score, mortality, neuronal degeneration, and proinflammatory factors. Moreover, WT-A20−/− parabiosis was established to explore the role of A20 in peripheral blood in ICH injury. ICH-induced damage, including brain edema, neurologic deficit score, proinflammatory factors, and neuronal apoptosis, was reduced in A20−/− parabionts compared with A20−/− mice. Finally, the interactions between TRAF6 and Ubc13 and UbcH5c were increased in A20−/− mice compared with WT mice; the opposite occurred in A20-overexpressing mice. Enhanced IκBα degradation and NF-κB activation were observed in A20−/− mice, but the results were reversed in A20-overexpressing mice. These results suggested that A20 is involved in regulating ICH-induced inflammatory injury in both the central and peripheral system and that A20 reduces ICH-induced inflammation by regulating TRAF6 polyubiquitination. Targeting A20 may thus be a promising therapeutic strategy for ICH.

BackgroundBlood–brain barrier (BBB) disruption aggravates brain injury induced by intracerebral hemorrhage (ICH); however, the mechanisms of BBB damage caused by ICH remain elusive. Mfsd2a (major facilitator superfamily domain containing 2a) has been known to play an essential role in BBB formation and function. In this study, we investigated the role and underlying mechanisms of Mfsd2a in BBB permeability regulation after ICH.Methods and ResultsUsing ICH models, we found that Mfsd2a protein expression in perihematomal brain tissues was significantly decreased after ICH. Knockdown and knockout of Mfsd2a in mice markedly increased BBB permeability, neurological deficit score, and brain water contents after ICH, and these were rescued by overexpressing Mfsd2a in perihematomas. Moreover, we found that Mfsd2a regulation of BBB permeability after ICH correlated with changes in vesicle number. Expression profiling of tight junction proteins showed no differences in Mfsd2a knockdown, Mfsd2a knockout, and Mfsd2a overexpression mice. However, using electron microscopy following ICH, we observed a significant increase in pinocytotic vesicle number in Mfsd2a knockout mice and decreased the number of pinocytotic vesicles in mouse brains with Mfsd2a overexpression. Finally, using multiple reaction monitoring, we screened out 3 vesicle trafficking–related proteins (Srgap2, Stx7, and Sec22b) from 31 vesicle trafficking‐related proteins that were markedly upregulated in Mfsd2a knockout mice compared with controls after ICH.ConclusionsIn summary, our results suggest that Mfsd2a may protect against BBB injury by inhibiting vesicular transcytosis following ICH.

The discovery and application of genome editing introduce a new era of plant breeding, giving researchers efficient tools for the precise engineering of crop genomes 1 . Here, we demonstrate the power of genome editing for engineering broad-spectrum disease resistance in rice (Oryza sativa). We first isolated a lesion mimic mutant (LMM) from a mutagenized rice population, demonstrated that a 29-bp deletion in a gene we named RESISTANCE TO BLAST1 (RBL1) caused this phenotype and showed that this mutation caused a ca. 20-fold reduction in yield. RBL1encodes a cytidine diphosphate diacylglycerol (CDP-DAG) synthase required for phospholipid biosynthesis 2 . Mutation of RBL1 results in reduced levels of phosphatidylinositol (PI) and its derivative PI(4,5)P2. Rice PI(4,5)P2 is enriched in cellular structures specifically associated with effector secretion and fungal infection, suggesting a role as a disease susceptibility factor 3 . Using targeted mutagenesis, we obtained an allele of RBL1, named RBL12, which confers broad-spectrum resistance but does not decrease yield in a model rice variety as assessed in small-scale field trials. Our study has demonstrated the usefulness of editing of an LMM gene, a strategy relevant to diverse LMM genes and crops. Main textGenome editing has been widely used in functional studies of genes but its potential for crop improvement has not yet been broadly utilized 4 . Plant diseases cause severe losses in agriculture, threatening global food security 5 . Rice blast alone, caused by the fungal pathogen Magnaporthe oryzae, results in annual yield losses that are sufficient to feed more than 60 million people worldwide 6 . Given this cost, cultivating crops with resistance to diseases, particularly broad-spectrum resistance, is highly desirable 7 . Despite the importance of this goal, only a limited number of broad-spectrum resistance genes have been cloned and used in the field, such as rice Xa21 (ref. 8 ), bsr-d1 (ref. 9 ), Pigm 10 , IPA1 (ref. 11 ), ROD1 (ref. 12 ), UMP1 (ref. 13 ), wheat Lr34 (ref. 14 ) and PsIPK1 (ref. 15 ), and barley mlo 16,17 .LMMs form hypersensitive response-like lesions (a form of programmed cell death) in the absence of pathogens 18 . LMMs often confer durable and broad-spectrum resistance, representing a potential source for breeding resistance to diseases. However, LMMs are usually associated with reduced yield, and therefore the use of the genes conferring LMM phenotypes (hereafter referred to as LMM genes) has not been fully exploited in plant breeding due to the lack of useful alleles.Phospholipids are essential components of biological membranes and are involved in various biological processes, including development and response to biotic and abiotic stress 19 . In phospholipid biosynthesis (Extended Data Fig. 1), phosphatidic acid (PA) and cytidine triphosphate (CTP) are converted to CDP-DAG by CDP-DAG synthases (CDSs). CDP-DAG and Myo-inositol are used to produce PI by phosphatidylinositol synthases (PISs) 20 . PI is added to by a varied number of p...

BackgroundSpinal cord injury (SCI) is a devastating disease that lacks effective treatment. Interestingly, recent studies indicated that vagus nerve stimulation (VNS), neuromodulation that is widely used in a variety of central nervous system (CNS) diseases, improved motor function recovery after SCI. But the exact underlying mechanism of how VNS ameliorates SCI is unclear. This study aimed to confirm the efficacy and further explore the potential therapeutic mechanism of VNS in SCI.MethodA T10 spinal cord compression model was established in adult female Sprague-Dawley rats. Then the stimulation electrode was placed in the left cervical vagus nerve (forming Sham-VNS, VNS, and VNS-MLA groups). Basso-Beattie-Bresnahan (BBB) behavioral scores and Motor evoked potentials (MEPs) analysis were used to detect motor function. A combination of histological and molecular methods was used to clarify the relevant mechanism.ResultsCompared with the Sham-VNS group, the VNS group exhibited better functional recovery, reduced scar formation (both glial and fibrotic scars), tissue damage, and dark neurons, but these beneficial effects of VNS were diminished after alpha 7 nicotinic acetylcholine receptor (α7nAchR) blockade. Specifically, VNS inhibited the pro-inflammatory factors TNF-α, IL-1β, and IL-6 and increased the expression of the anti-inflammatory factors IL-10. Furthermore, we found that VNS promotes the shift of M1-polarized Iba-1+/CD86+ microglia to M2-polarized Iba-1+/CD206+ microglia via upregulating α7nAchR to alleviate neuroinflammation after SCI.ConclusionOur results demonstrated that VNS promotes microglial M2 polarization through upregulating α7nAChR to reduce neuroinflammation, thus improving motor function recovery after SCI. These findings indicate VNS might be a promising neuromodulation strategy for SCI.

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