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Currently, there is increasing attention on the regulatory effects of cannabidiol (CBD) on the inflammatory response and the immune system. However, the mechanisms have not yet been completely revealed. Mitofusin 2 (Mfn2) is a mitochondrial fusion protein involved in the inflammatory response. Here, we investigated whether Mfn2 confers the anti-inflammatory effects of CBD. We found that treatment with CBD decreased the levels of tumor necrosis factor α, interleukin 6, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and ionized calcium-binding adaptor molecule-1 (Iba1) in lipopolysaccharide (LPS)-challenged microglia. CBD also significantly suppressed the increase in reactive oxygen species (ROS) and the decline of mitochondrial membrane potential in BV-2 cells subjected to LPS. Interestingly, CBD treatment increased the expression of Mfn2, while knockdown of Mfn2 blocked the effect of CBD. By contrast, overexpression of Mfn2 reversed the increase in the levels of iNOS, COX-2, and Iba1 induced by Mfn2 small interfering RNA. In mice challenged with LPS, we found that CBD ameliorated the anxiety responses and cognitive deficits, increased the level of Mfn2, and decreased the expression of Iba1. Since neuro-inflammation and microglial activation are the common events that are observed in the experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis, we treated EAE mice with CBD. Mice that received CBD showed amelioration of clinical signs, reduced inflammatory response, and increased myelin basic protein level. Most importantly, the adeno-associated virus delivery of short hairpin RNA against Mfn2 reversed the protective effects of CBD. Altogether, these results indicate that Mfn2 is an essential immunomodulator conferring the anti-inflammatory effects of CBD. Our results also shed new light on the mechanisms underlying the protective effects of CBD against inflammatory diseases including multiple sclerosis.

A super-large ensemble simulation dataset with 110 members has been produced by the fully coupled model FGOALS-g3 developed by researchers at the Institute of Atmospheric Physics, Chinese Academy of Sciences. This is the first dataset of large ensemble simulations with a climate system model developed by a Chinese modeling center. The simulation has the largest realizations up to now worldwide in terms of single-model initial-condition large ensembles. Each member includes a historical experiment (1850–2014) and an experiment (2015–99) under the very high greenhouse gas emissions Shared Socioeconomic Pathway scenario (SSP5-8.5). The dataset includes monthly and daily temperature, precipitation, and other variables, requiring storage of 275 TB. Additionally, the surface air temperature (SAT) and land precipitation simulated by the FGOALS-g3 super-large ensemble have been validated and projected. The ensemble can capture the response of SAT and land precipitation to external forcings well, and the internal variabilities can be quantified. The availability of more than 100 realizations will help researchers to study rare events and improve the understanding of the impact of internal variability on forced climate changes.

Neuromyelitis optica spectrum disorder (NMOSD) is an inflammatory demyelinating disease of the central nervous system (CNS). Our previous study indicated that neutrophil-related chemokine CXCL7 is elevated in the cerebrospinal fluid (CSF) of NMOSD patients. To study the potential function of CXCL7 during NMOSD, we measured the chemokines level in CSF of follow-up patients, and established three NMOSD mouse models by injecting aquaporin4 (AQP4)-IgG. Astrocytes loss, inflammatory infiltration, and myelin sheath damage were detected by western blot or immunofluorescence analysis. We found CXCL7 was significantly increased in the serum and CSF of model mice, and exogenous CXCL7 caused serious astrocyte injury, obvious microglia activation, and increased infiltration of neutrophils and macrophages, resulting in secondary demyelination. Consistently, knocking down the CXCL7 reversed the loss of AQP4, and attenuated the inflammatory response. Collectively, our data indicates that CXCL7 aggravates NMOSD-like pathological damage to astrocytes and myelin sheath mainly via promoting neuroinflammatory response.

Increasing evidence indicates that immune system dysfunction affects anti‐N‐methyl‐D‐aspartate receptor (NMDAR) encephalitis. This study aims to investigate the relationship between adhesion molecules and the pathophysiology in anti‐NMDAR encephalitis. Soluble forms of Intercellular adhesion molecule‐1 (sICAM‐1), vascular adhesion molecule‐1 (sVCAM‐1), and L‐selectin (sL‐selectin), were measured in the CSF and serum of 26 participants with anti‐NMDAR encephalitis, 11 patients with schizophrenia and 22 patients with noninflammatory disorders. CSF levels of sICAM‐1, sVCAM‐1 and sL‐selectin were significantly elevated in the anti‐NMDAR encephalitis group. sVCAM‐1 levels were positively associated with modified Rankin scale score in anti‐NMDAR encephalitis patients at the onset and 3‐month follow‐up.

Background: There is conflicting data regarding transcatheter aortic valve replacement in patients with paradoxical low-flow, low-gradient aortic stenosis (pLFLG-AS). The authors aimed to examine the hemodynamic and clinical benefit of transcatheter aortic valve replacement in symptomatic patients with pLFLG-AS compared with high-gradient severe AS. Methods: A single-center retrospective analysis of patients who underwent transcatheter aortic valve replacement using a transfemoral approach was performed. Patients with a mean gradient ≥40 mm Hg were included in the high-gradient aortic stenosis (HG-AS) group (n=217). Patients with mean gradient 2 were included in the pLFLG-AS group (n=73). Clinical end points including treatment futility, survival, changes in functional status and quality of life, and echocardiographic outcomes of left ventricular reverse remodeling were evaluated. Two primary composite outcomes were used for treatment futility. The first as a composite outcome defined as death or New York Heart Association class III/IV at 1 year. The second was a composite of death or Kansas City Cardiomyopathy Questionnaire 12 score of ≤25 at 1 year. Results: There were no differences in mortality, the Kansas City Cardiomyopathy Questionnaire-based treatment futility outcome (HG-AS: 8.8% versus pLFLG-AS: 6.1%; P =0.482), or New York Heart Association-based treatment futility outcome (HG-AS: 9.1% versus pLFLG-AS: 11.6%; P =0.546) at 1 year. Both groups had similar improvement in mean Kansas City Cardiomyopathy Questionnaire 12 score when compared to baseline (HG-AS: 88.56±12.38% versus pLFLG-AS: 137.24±31.75%; P =0.382). There was a similar relative interval decrease in the proportion of patients with New York Heart Association class III/IV (HG-AS: 53.6% versus pLFLG-AS: 55.1%; P =0.838). Echocardiographic evidence of reverse remodeling of the left ventricle in pLFLG-AS group was documented by improvements in global longitudinal strain (pre: −13.95±0.36% versus post: −14.83±0.38%, P =0.004) and stroke volume index at 30 days. Conclusions: pLFLG-AS did not confer a worse prognosis. Mortality rate and functional class improvement at 1 year was similar in both groups. Left ventricular reverse remodeling in the pLFLG-AS group was apparent.

Additional file 1. Supplementary Methods.

BackgroundIDO1 is the initial and rate-limiting enzyme that metabolizes tryptophan (TRP) to kynurenine (KYN). IDO1-dependent neurotoxic KYN metabolism plays a crucial role in pathogenesis of many neurodegenerative disorders. However, the function of IDO1 in epilepsy is still unclear. MethodsPatients with epilepsy and controls were enrolled. Male C57BL/6 mice and IDO1 knockout (KO) mice were subjected to intraperitoneal injection of lithium and pilocarpine to induce epilepsy. The level of IDO1 and concentrations of TRP and KYN in the patients with epilepsy and epileptic mice were evaluated by enzyme-linked immunosorbent assay (ELISA) and liquid chromatography-mass spectrometry (LC-MS) respectively. Then, SRS and neuronal damage was compared between KO and wild-type (WT) mice in lithium-pilocarpine-induced epilepsy. To explore underlying pathways involved in IDO1 deficiency, concentrations of kynurenic acid (KYNA) and quinolinic acid (QUIN), glial cells activation, major pro-inflammatory cytokines, and antioxidant enzymes activity were measured by LC-MS, immunohistochemistry and ELISA.ResultsIn this study, IDO1 level and KYN/TRP ratio were increased in the patients with epilepsy and epileptic mice. IDO1 deficiency attenuated the frequency, duration and severity of SRS and improved neuronal survival. Additionally, IDO1-/- epileptic mice showed a progressive decline in QUIN production, glial cells activation and pro-inflammatory cytokines and enhanced antioxidant enzymes activity.ConclusionsIDO1 deletion alleviated SRS and neuronal damage in the chronic period after SE through a reduction in IDO1-dependent neurotoxic metabolites, which finally inhibited pro-inflammatory cytokine production and glial cells activation and improved antioxidant enzymes activity. Our study demonstrates that IDO1 may be involved in the pathogenesis of epilepsy and has potential to be a therapeutic target for the treatment of epilepsy.

Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a recently discovered autoimmune syndrome associated with psychosis, dyskinesia, and seizures. However, the underlying mechanisms of this disease remain unclear, in part because of a lack of suitable animal models. This study describes a novel mouse model of anti-NMDAR encephalitis that was induced by active immunization against NMDARs using amino-terminal domain peptides. After 12 weeks of immunization, the mice showed significant behavioral disorders and memory loss. Furthermore, antibodies from the cerebrospinal fluid of immunized mice lowered the surface NMDAR cluster density in hippocampal neurons. Immunization also impaired long-term potentiation at Schaffer collateral–CA1 synapses and reduced NMDAR-induced calcium influx. This novel mouse model may allow further research into the pathogenesis of anti-NMDAR encephalitis and aid in the development of new therapies for this disease.

Anti-N-methyl-D-aspartate receptor (NMDA) receptor encephalitis is a recently discovered autoimmune syndrome associated with psychosis, dyskinesias, and seizures. Ectopic expression of NMDA receptors associated with ovarian teratoma is thought to mediate the initial autoimmune response against NMDA receptor encephalitis. Due to the lack of suitable animal models, the underlying mechanism of the disease remains unclear. This study described a new mice model of active immunization against the NMDA receptor with amino-terminal domain (ATD) peptides. After 12 weeks of immunization, mice were showed significant behavioral disorders and memory loss. Antibodies from CSF of immunized mice decreased surface NMDAR cluster density on hippocampus neurons. It also impaired the LTP induced at the Schaffer collateral to CA1 synapse and reduced NMDA receptors-induced calcium influx. The new model may help further research into the pathogenesis of the disease and the development of potential new therapies.

Background Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a recently discovered autoimmune syndrome associated with psychosis, dyskinesia, and seizures. However, the underlying mechanisms of this disease remain unclear, in part because of a lack of suitable animal models. Methods This study describes a novel female C57BL/6 mouse model of anti-NMDAR encephalitis that was induced by active immunization against NMDARs using an amino terminal domain (ATD) peptide from the GluN1 subunit (GluN1356–385). Results Twelve weeks after immunization, the immunized mice showed significant memory loss. Furthermore, antibodies from the cerebrospinal fluid of immunized mice decreased the surface NMDAR cluster density in hippocampal neurons which was similar to the effect induced by the anti-NMDAR encephalitis patients’ antibodies. Immunization also impaired long-term potentiation at Schaffer collateral–CA1 synapses and reduced NMDAR-induced calcium influx. Conclusion We established a novel anti-NMDAR encephalitis model using active immunization with peptide GluN1356–385 targeting the ATD of GluN1. This novel model may allow further research into the pathogenesis of anti-NMDAR encephalitis and aid in the development of new therapies for this disease.

Background Neuromyelitis optica (NMO), multiple sclerosis (MS) and autoimmune glial fibrillary acidic protein (GFAP) astrocytopathy belong to autoimmune diseases incentral nervous system mainly manifestate encephalomyelitis. The glycocalyx (GLX), consists of several membrane-bound macromolecules, is located on the luminal side of the endothelium and mediates the blood and the vessel interaction. Until now, there is still lacking a holistic understanding of the GLX degradation in autoimmune encephalomyelitis. Aim This study aimed to detect the shedding levels of GLX components, heparan sulfate (HS) and hyaluronic acid (HA) in serum and cerebrospinal fluid (CSF), correlate them with the severity and assess the diagnostic value of them, and evaluate their correlations with pro-inflammatory cytokines. Methods Serum and CSF samples were obtained from 24 NMO patients, 15 MS patients, 10 autoimmune GFAP astrocytopathy patients, and 18 controls without non-inflammatory neurological diseases. Soluble HS, HA and IFN-γ, IL-17A, matrix metalloproteinase (MMP)-1 were detected by enzyme linked immunosorbent assay ELISA. Results Besides levels of serum and CSF levels of HS, HA and related cytokines were significantly elevated in these diseases. Notably, HS, HA in NMO, MS patients, and autoimmune GFAP astrocytopathy diseases were widely correlated with EDSS scores. Importantly, the ROC curve analysis suggested a potential diagnostic role of HS or HA . Conclusions The results here suggested the GLX degradation and inflammation in NMO, MS and autoimmune GFAP astrocytopathy. Moreover, increased shedding of HS or HA may indicate worse clinical situation. Importantly, CSF HS and HA may be informative diagnostic biomarkers for telling autoimmune encephalomyelitis from the non-inflammatory neurological controls. Furthermore, therapeutic strategy for protecting GLX may be effective to these diseases.

Ningaloo Nino, an interannual sea surface temperature (SST) warming phenomenon off western Australia, has a significant impact on the regional ecosystem and arouses economic damage. Using two coupled models of FGOALS-g3 and FGOALS-f3-L, we evaluated their simulation ability for Ningaloo Nino. The results show that the two models can reproduce the basic characteristics of Ningaloo Nino well, including the spatial pattern and phase-locking feature. The amplitude of Ningaloo Nino is underestimated in FGOALS-g3 but is well duplicated in FGOALS-f3-L. We found that the difference in cloud shortwave radiation-SST feedback between the two models is the key contributor to the different Ningaloo Nino amplitudes between the two models. A positive stratus cloud shortwave radiation-SST regime exhibits in the FGOALS-f3-L, which is similar to that operates in the observation; while FGOALS-g3 unsuccessfully simulates such shortwave radiation-SST regime, which may be traced back to the model parameterization schemes in the atmospheric component of FGOALS-g3. In the observation, the development of Ningaloo Nino involves two different modes (local mode and non-local mode). Our results show that the two modes can be reproduced well in FGOALS-g3, whereas only one single mode (say, local mode) is manifested in FGOALS-f3-L. From the statistical perspective, both the observation and FGOALS-g3 show that there is a significant connection between tropical ENSO signals and the Ningaloo Nino/Nina to some extent; while the FGOALS-f3-L does not have such a significant statistical relationship. Thus, it is argued that the failure in representing the inter-basin connection in the FGOALS-f3-L may be responsible for the absent non-local mode.

The NOD-like receptor family pyrin domain-containing protein 3 (NLRP3) is a protein complex which mediates pro-inflammatory response and pyroptosis. The activation of NLRP3 inflammasome triggers pyroptosis via cleavage of the pore-forming protein gasdermin D. Previous studies have demonstrated that NLRP3 inflammasome activation in microglial contributes to the pathogenesis of EAE, while Inhibition of microglial NLRP3 inflammasome and the subsequent pyroptosis attenuates the severity of EAE. In addition, Interleukin-1 receptor-associated kinase (IRAK)-M, mainly expressed in microglial in the central nervous system, negatively regulates the TLR4/IL-1R signaling pathway through inhibiting the phosphorylation of IRAK1 and dissociation of IRAK1 from MyD88/IRAK-4/IRAK-1 complex as well as the formation of IRAK-1 and TRAF6 complex. Our recent study suggests the protective effect of microglial IRAK-M in EAE mice. Therefore, we hypothesize that IRAK-M may alleviate IRAK1-induced NLRP3 inflammasome activation and pyroptosis through inhibiting dissociation of IRAK1 from MyD88/IRAK-4/IRAK-1 complex and the following association of IRAK1 and TRAF6. Here, we used IRAK-M knockout mice and their microglia to clarify the role of IRAK-M in EAE. We demonstrated that IRAK-M decreased the incidence rate and improved the clinical symptoms accompanied by mild demyelination in EAE mice, and which was parallel to the inhibition of microglial NLRP3 inflammasome activation, pyroptosis, and IRAK1/TRAF6 formation in EAE. Our findings fill the gap in knowledge of IRAK-M on activation of NLRP3 inflammasome and pyroptosis and provide a potential novel target for the therapeutic strategy of multiple sclerosis and NLRP3-related diseases.

Primers and probes sequences. (DOC 73 kb)

Influenza virus infections are the main contagious respiratory disease with high levels of morbidity and mortality worldwide. Antiviral drugs are indispensable for the prophylaxis and treatment of influenza and other respiratory viral infections. In this study, the Arbidol hydrochloride (ARB), which has been licensed in Russia and China, is used to investigate its anti-viral and anti-inflammatory efficacy in vitro and in vivo. The antiviral results in vitro showed that ARB had a better inhibition on Influenza virus A/PR/8/34 (H1N1), A/Guangdong/GIRD07/09 (H1N1), A/Aichi/2/68 (H3N2), A/HK/Y280/97 (H9N2) with IC50 ranging from 4.4 to 12.1μM. The further mechanisms study demonstrated that ARB is able to inhibit hemagglutinin-mediated hemolysis at concentration of 3.91-15.63μg/mL. The anti-inflammatory efficacy in vitro indicated that IL-6, IP-10, MCP-1, RANTES and TNF-α levels were diminished by ARB at concentrations of 22.6 and 18.8μM. The in vivo results in mice model displayed that the survival rates of mice administered 25mg/mL and 45mg/mL ARB were 40% and 50% respectively. And also, ARB can inhibit the decrease of body weight at 45mg/mL and inhibit the increase of mice lung index at 25mg/mL and 45mg/mL comparing to virus group. In ferret model, the ARB-treated ferrets showed a fever that peaked at 2 dpi and gradually decreased beginning at 3 dpi while relatively high temperatures were observed until 4 dpi in the virus group. The ARB-treated group scored 0-1 in the activity level at 2 dpi and 3 dpi at all time points. The transcription levels of cytokines in the respiratory tract of ferrets were detected at 3 dpi. Several proinflammatory cytokines induced by influenza (IL-10, TNF-α, IL-8 and IL-6) were down-regulated by post-treatment with ARB. The histopathological results of ferret lung displayed that ARB can alleviate the influenza virus induced lung lesions. Our results clarified the activity of ARB in both suppressing virus propagation and modulating the expression of inflammatory cytokines in vitro and in vivo, it can be as an effective drug to treat the influenza virus infection.

Background: Lianhuaqingwen Capsule (LH-C) is a traditional Chinese medicine (TCM) formula used to treat respiratory tract infectious diseases in Chinese. The aim of this study was to determine the antiviral activity of LH-C and its immunomodulatory effects on viral infection. Method: The in vitro cytotoxicity and antiviral activity of LH-C was determined by MTT and Plaque reduction assays. Time course study under single-cycle virus growth conditions were used to determine which stage of viral replication was blocked. The effect of LH-C on the nuclear export of the viral nucleoprotein was examined using an indirect immunofluorescence assay. The regulation to different signaling transduction events and cytokine/chemokine expression of LH-C was evaluated using Western blotting and real-time RT-PCR. After virus inoculation, BALB/c mice were administered with LH-C of different concentrations for 5 days. Body-weight, viral titers and lung pathology of the mice were measured, the level of inflammatory cytokines were also examined using real-time RT-PCR. Results: LH-C inhibited the proliferation of influenza viruses of various strain in vitro, with the 50% inhibitory concentration (IC50) ranging from 0.35 to 2 mg/mL. LH-C blocked the early stages (0–2 h) of virus infection, it also suppressed virus-induced NF-kB activation and alleviated virus-induced gene expression of IL-6, IL-8, TNF-a, IP-10, and MCP-1 in a dose-dependent manner. LH-C treatment efficiently impaired the nuclear export of the viral RNP. A decrease of the viral titers in the lungs of mice were observed in groups administered with LH-C. The level of inflammatory cytokines were also decreased in the early stages of infection. Conclusions: LH-C, as a TCM prescription, exerts broad-spectrum effects on a series of influenza viruses, including the newly emerged H7N9, and particularly regulates the immune response of virus infection. Thus, LH-C might be a promising option for treating influenza virus infection.