The impact of COVID-19 on a Southern Chinese cohort with neuromyelitis optica spectrum disorders.

BACKGROUND: There are few studies on risk factors for coronavirus disease 2019 (COVID-19) infection in patients with Neuromyelitis Optica Spectrum Disorders (NMOSD). The relationship between NMOSD relapse and COVID-19 needs to be evaluated. The objective of our study is to identify the risk factors of COVID-19 infection and NMOSD relapse among NMOSD patients with COVID-19. METHOD: A total of 379 NMOSD patients registered in a NMOSD database were included in this case-control study after the end of the COVID-19 quarantine and restriction policies on December 6, 2022 in China. Data were obtained from the database. Additional information was obtained by questionnaires and the Neurology out-patient clinic. The clinical characteristics of NMOSD patients with COVID-19 were described. Risk factors associated with COVID-19 infection and outcome among patients with NMOSD were analyzed. Risk factors associated with relapse in NMOSD patients with COVID-19 were also identified. RESULTS: 239 (63.1%) NMOSD patients were infected with COVID-19. Patients with NMOSD who were infected with COVID-19, in comparison to those without COVID-19, were younger at the time of interview (median [IQR] age: 43.00 [32.00-55.00] vs 49.50 [35.25-56.00] years, P = 0.029), younger at NMOSD onset (median [IQR] age: 38.00 [27.00-51.00] vs 45.00 [32.00-52.75] years, P = 0.013), had abnormal visual evoked potentials before infection (73.4% vs 54.3% P = 0.029), had lower baseline Activities of Daily Living Scale (ADL) scores (median [IQR] ADL: 14.00 [14.00-16.00] vs 14.00 [14.00-19.00], P = 0.014) or lower baseline modified Rankin Scale (mRS) scores (1.12±0.749 vs 1.33±0.991, P = 0.037), and were less frequently treated with more than 10 mg prednisone or 8 mg methylprednisolone (25.0% vs 36.0%,p = 0.026). All 9 NMOSD patients who had symptomatic cerebral syndrome developed moderate/severe COVID-19. A higher percentage of patients with moderate/severe COVID-19 experienced more than one core clinical NMOSD symptoms (61.5% vs 55.1%, p = 0.044), compared to patients with mild COVID-19. Higher risk of NMOSD relapse among NMOSD patients with COVID-19 was associated with higher Expanded Disability Status Scale (EDSS) scores (median[IQR] EDSS: 2.00 [1.00-3.00] vs 1.50 [1.00-2.25], P = 0.037) and drug treatments disruption (21.6% vs 5.0% P<0.001). CONCLUSIONS: NMOSD patients with younger age, lower baseline ADL or mRS had higher incidence of being diagnosed with COVID-19 during pandemic. Glucocorticoid use may decrease the risk of COVID-19. NMOSD patients with symptomatic cerebral syndrome before the COVID-19 pandemic are associated with worse COVID-19 outcomes. Drug treatment disruption may result in relapse among NMOSD patients with COVID-19.

Inhibiting Caveolin-1-Related Akt/mTOR Signaling Pathway Protects Against N-methyl-D-Aspartate Receptor Activation-Mediated Dysfunction of Blood-Brain Barrier in vitro.

BACKGROUND: The aim of this study was to further explore the role of caveolin-1 (Cav-1) related Akt/mTOR signaling pathway in blood brain barrier (BBB) dysfunction caused by NMDAR activation. METHODS: The cell localization of NMDAR GluN1 subunit and Cav-1 was observed on human brain microvascular HBEC-5i cells after immunofluorescence double staining. The transendothelial resistance (TEER) of BBB in vitro was measured by Millicell-ERS cell resistance meter. Sodium fluorescein (SF) was used to measure the permeability of BBB in vitro. A stable Cav-1-silenced HBEC-5i cell line was established by infecting the cells with a lentivirus encoding Cav-1 shRNA. The changes of the protein and mRNA of MMP9 and Occludin induced by NMDA were detected by Western blot (WB) and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), respectively. The phosphorylated proteins of Cav-1, Akt, and mTOR were detected by WB. RESULTS: NMDAR GluN1 was expressed in the cytoplasm and part of the cell membrane of the HBEC-5i cell line. NMDAR activation decreased TEER and increased the SF of BBB in vitro. HBEC-5i cells incubated with NMDA enhanced the phosphorylation of Cav-1, Akt, and mTOR, also promoting the expression of MMP9 along with the degradation of Occludin. These effects could be reversed by pretreatment with NMDAR antagonist (MK801) or Cav-1 antagonist (Daidzein), or Akt antagonist (LY294002), respectively. Further silencing Cav-1 with LV-Cav-1-RNAi also played a similar protective effect. CONCLUSION: Caveolin-1 (Cav-1) related Akt/mTOR signaling probably contributes to BBB dysfunction by activating NMDAR on human brain microvascular cells.

Clinical Characteristics of Myasthenia Gravis Patients with COVID-19 in Guangxi, China: A Case-Control Study.

Purpose: With the adjustment of prevention strategies in December 2022, coronavirus disease 2019 (COVID-19) became widely prevalent in China. This study is aimed to describe the clinical characteristics of myasthenia gravis (MG) patients with COVID-19 and identify risk factors of exacerbation in MG patients with COVID-19 in Guangxi. Patients and Methods: A total of 489 MG patients and 587 control subjects in Guangxi during the COVID-19 pandemic were enrolled in this case-control study. After contacting the participants, the clinical data of MG patients and the control group were analyzed. The clinical characteristics of MG patients with COVID-19 were described. Multivariable logistic regression analysis was used for discovering independent risk factors of MG exacerbation in the patients with MG and COVID-19. Results: A total of 311 (75.30%) MG patients and 428 (72.91%) control subjects were infected with COVID-19, and 64.31% of MG patients with COVID-19 were women. The median age at the time of interview was 41 (IQR: 28, 54) years old, and median onset age was 36 (IQR: 24, 51), both of which were lower than those in MG patients without COVID-19. MG duration was 24 (IQR: 9, 72) months. About 44.69% of patients were generalized MG (GMG). About 11.90% of MG patients with COVID-19 showed severe COVID-19 symptoms and the duration of symptomatic COVID-19 was 9.57 ± 6.79 days, higher than those in the control group. About 35.69% MG patients with immunosuppressive drugs were infected with COVID-19, which is higher than those in the non-infected MG patients (21.57%). A total of 120 (38.59%) MG patients with COVID-19 had comorbidities. About 21 (20.19%) of the 104 MG patients without vaccination showed severe COVID-19 symptoms. Multivariable logistic regression analysis showed that baseline MG activities of daily living profile (MG-ADL, OR 1.280, 95% CI: 1.010-1.621, p = 0.041), duration of COVID-19 (OR 1.158, 95% CI: 1.100-1.220, p < 0.001), GMG (OR 2.331, 95% CI: 1.228, 4.426, p = 0.010), and lack of COVID vaccination (OR 2.075, 95% CI: 1.152, 3.738, p = 0.015) were independent factors of exacerbation in MG patients with COVID-19. Conclusion: MG patients with immunosuppressive drugs, younger onset, longer MG duration, or comorbidities are more susceptible to COVID-19. The baseline MG-ADL, duration of symptomatic COVID-19, GMG, and lack of COVID-19 vaccination are independent risk factors of exacerbation in MG patients with COVID-19.

Risk Factors and Prognosis in Anti-NMDA Receptor Encephalitis Patients with Disturbance of Consciousness.

Purpose: Disturbance of consciousness is common in patients with severe anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis. However, little is known about it. This study aimed to analyze the clinical manifestations and prognostic factors of anti-NMDAR encephalitis with disturbance of consciousness. Methods: In this retrospective study, the clinical features, treatment results, and long-term outcomes of anti-NMDAR encephalitis patients with disturbance of consciousness were analyzed, and multivariate logistic regression was used to analyze the factors affecting their prognosis. Results: In the group with disturbance of consciousness, the incidences of seizures, involuntary movements, pulmonary infection, mechanical ventilation, intensive care unit (ICU) admission, neutrophil-lymphocyte ratio (NLR), abnormal cerebrospinal fluid index, plasma exchange, and immunosuppressive therapy were higher than those in the group without disturbance of consciousness (all P<0.05). During the follow-up period (median: 36 months, range: 12-78 months), the modified Rankin scale (mRS) score, the maximum mRS score during hospitalization, the mRS score at discharge, and the mRS score at 12 months after discharge were higher in the disturbance of consciousness group (all P < 0.001). However, there was no significant difference in long-term outcomes and recurrence between the two groups. Multivariate logistic regression analysis showed that mechanical ventilation, elevated IgG index, and delayed immunotherapy were independent risk factors for poor outcomes in patients with anti-NMDAR encephalitis with disturbance of consciousness at 12 months (odds ratio: 22.591, 39.868, 1.195). The receiver operating characteristics (ROC) curve analysis showed that the area under the curve (AUC) of mechanical ventilation, elevated IgG index, and delayed immunotherapy was 0.971 (95% CI=0.934-1.000, P<0.001). Conclusion: Mechanical ventilation, elevated IgG index, and delayed immunotherapy may be the influencing factors of poor prognosis of anti-NMDAR encephalitis patients with disturbance of consciousness. Although their condition is relatively serious, most patients with anti-NMDAR encephalitis with disturbance of consciousness will achieve favorable long-term outcomes after long-term treatment.

Inhibiting PI3K/Akt-Signaling Pathway Improves Neurobehavior Changes in Anti-NMDAR Encephalitis Mice by Ameliorating Blood-Brain Barrier Disruption and Neuronal Damage.

The disruption of the blood-brain barrier (BBB) is hypothesized to be involved in the progression of anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis, but its mechanism is still unclear. Recently, the phosphatidylinositol 3-kinase (PI3K)/threonine kinase (Akt) pathway is involved in the regulation of the BBB in various diseases. This study is aimed to investigate the mechanism of BBB damage and neurobehavior changes in anti-NMDAR encephalitis mice. Female C57BL/6J mice were actively immunized to establish an anti-NMDAR encephalitis mouse model and evaluate the neurobehavior changes of mice. To study its potential mechanism, LY294002 (PI3K inhibitor, 8 mg/kg) and Recilisib (PI3K agonist, 10 mg/kg) were treated by intraperitoneal injection, respectively. Anti-NMDAR encephalitis mice showed neurological deficits, increased BBB permeability, open endothelial tight junctions (TJs), and decreased expression of TJ-related proteins zonula occludens (ZO)-1 and Claudin-5. However, administration of PI3K inhibitor significantly reduced the expression of p-PI3K and p-Akt, improved neurobehavior function, decreased BBB permeability, and upregulated the expressions of ZO-1 and Claudin-5. Furthermore, PI3K inhibition reversed the decline of NMDAR NR1 in the membranes of hippocampal neurons, which reduced the loss of neuron-specific nucleoprotein (NeuN) and microtubule-associated protein 2 (MAP2). In contrast, administration of the PI3K agonist Recilisib showed a tendency to exacerbate BBB breakdown and neurological deficits. Our results showed that the activation of PI3K/Akt, along with the changes in TJ-related proteins ZO-1 and Claudin-5, may be closely related to BBB damage and neurobehavior changes in anti-NMDAR encephalitis mice. PI3K inhibition attenuates BBB disruption and neuronal damage in mice, thereby improving neurobehavior.

DCAF12 and HSPA1A May Serve as Potential Diagnostic Biomarkers for Myasthenia Gravis.

Background: Myasthenia gravis (MG) is an autoimmune disease that severely affects the life quality of patients. This study explores the differences in immune cell types between MG and healthy control and the role of immune-related genes in the diagnosis of MG. Methods: The GSE85452 dataset was downloaded from the Gene Expression Omnibus (GEO) database and analyzed using the limma package to determine differentially expressed genes (DEGs) between patients with MG and the control group. Differentially expressed immune cells were analyzed using single-sample gene set enrichment analysis (GSEA), while immune cell-associated modules were identified by weighted gene coexpression network analysis (WGCNA). Then, the expression of the identified hub genes was confirmed by RT-PCR in peripheral blood mononuclear cells (PBMCs) of MG patients. The R package pROC was used to plot the receiver operating characteristics (ROC) curves. Results: The modules related to CD56bright natural killer cells were identified by GSEA and WGCNA. The proportion of CD56bright natural killer cells in the peripheral blood of MG patients is low. The results of RT-PCR showed that the levels of DDB1- and CUL4-associated factor 12 (DCAF12) and heat shock protein family A member 1A (HSPA1A) were significantly decreased in peripheral blood mononuclear cells of MG patients compared with healthy controls. The ROC curve results of DCAF12 and HSPA1A mRNA in MG diagnosis were 0.780 and 0.830, respectively. Conclusions: CD56bright NK cell is lower in MG patients and may affect MG occurrence. DCAF12 and HSPA1A are lowly expressed in PBMCs of MG patients and may serve as the diagnostic biomarkers of MG.

N-methyl-D-aspartic acid increases tight junction protein destruction in brain endothelial cell via caveolin-1-associated ERK1/2 signaling.

N-methyl-D-aspartic acid (NMDA), a glutamate analog, can activate N-Methyl-D-Aspartate receptor (NMDAR) to induce vascular endothelial cell injury but the mechanisms are not fully understood. The present study intended to evaluate the role of caveolin-1 (Cav-1) in NMDA-induced dysfunction of human brain microvascular endothelial cells (HBEC-5i), and verify that endothelial NMDAR activation mediates the disruption of tight junction barrier integrity via extracellular signal-regulated kinase (ERK)1/2 pathway. The expression of NMDAR NR1 were confirmed firstly in HBEC-5i and compared with that in mouse brain by Western blot. To study the role of Cav-1 in NMDA mediated reduction of tight junction protein zonula occludens- (ZO) 1 expression, HBEC-5i were transduced with Cav-1 shRNA or Control shRNA, and the Cav-1 knockdown rate tested with qRT-PCR and Western blot is 99.98% or 87.5%, respectively. NMDA exposure decreased mRNA and protein levels of tight junction protein ZO-1 and suppressed transendothelial electrical resistance (TEER) values in HBEC-5i but blocked by NMDAR antagonist MK801. In addition, NMDA induced Cav-1 and ERK1/2 sequential phosphorylation，but these effects were attenuated by silencing the Cav-1 gene with shRNA and ERK1/2 inhibitor U0126, respectively. These results show that the functional presence of NMDAR NR1 in HBEC-5i. Endothelial NMDAR NR1 activation regulate the maintenance of HBEC-5i-constructed tight junction barrier integrity via the caveolin-1-associated ERK1/2 signaling pathway.