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The mechanisms of many diseases, including central nervous system disorders, are regulated by circadian rhythms. The development of brain disorders such as depression, autism, and stroke is strongly associated with circadian cycles. Previous studies have shown that cerebral infarct volume is smaller at night (active phase) than during the day (inactive phase) in ischemic stroke rodent models. However, the underlying mechanisms remain unclear. Increasing evidence suggests that glutamate systems and autophagy play important roles in the pathogenesis of stroke. Here, we report that GluA1 expression was decreased and autophagic activity was increased in active-phase male mouse models of stroke compared with the inactive-phase models. In the active-phase model, induction of autophagy decreased the infarct volume, whereas inhibition of autophagy increased the infarct volume. Meanwhile, GluA1 expression was decreased following activation of autophagy and increased following inhibition of autophagy. We used Tat-GluA1 to uncouple p62, an autophagic adapter, from GluA1 and found that this blocked the degradation of GluA1, an effect similar to that of inhibition of autophagy in the active-phase model. We also demonstrated that knock-out of the circadian rhythm genePer1abolished the circadian rhythmicity of the volume of infarction and also abolished GluA1 expression and autophagic activity in wild-type (WT) mice. Our results suggest an underlying mechanism by which the circadian rhythm participates in the autophagy-dependent regulation of GluA1 expression, which influences the volume of infarction in stroke.SIGNIFICANCE STATEMENTCircadian rhythms affect the pathophysiological mechanisms of disease. Previous studies suggested that circadian rhythms affect the infarct volume in stroke, but the underlying mechanisms remain largely unknown. Here, we demonstrate that the smaller infarct volume after middle cerebral artery occlusion/reperfusion (MCAO/R) during the active phase is related to lower GluA1 expression and activation of autophagy. The decrease in GluA1 expression during the active phase is mediated by the p62–GluA1 interaction, followed by direct autophagic degradation. In short, GluA1 is the substrate of autophagic degradation, which mainly occurs after MCAO/R during the active phase but not the inactive phase.

Previous studies demonstrated that diabetic stroke patients had a poor prognosis and excess complement system activation in the peripheral blood. In this study, the association of serum complement levels with the prognosis of diabetic stroke was examined. Patients with acute ischemic stroke were recruited and were divided into two groups according to their history of diabetes. Baseline data on the admission, including C3 and C4 were collected. Neurologic function at discharge was the primary outcome and was quantified by the National Institutes of Health Stroke Scale (NIHSS). A total of 426 patients with acute ischemic stroke (116 diabetic strokes and 310 non-diabetic strokes) were recruited in this study. There were significant differences between the two groups in hypertension, coronary disease, triglyceride, high-density lipoprotein cholesterol, fasting blood sugar, C4, and mortality rates. Furthermore, the values of complement protein levels were divided into tertiles. In the diabetic stroke group, serum C4 level at the acute phase in the upper third was independently associated with NIHSS score at discharge and concurrent infection. These associations were not significant in non-diabetic stroke. High serum C4 level at admission, as a unique significant predictor, was associated with unfavorable clinical outcomes in the diabetic stroke, independently of traditional risk factors.

Central post-stroke pain (CPSP) is a disabling condition in stroke patients. It is a type of neuropathic pain for which the mechanism and relevant drug pathways remain unknown. Inflammatory response and central disinhibition have been suggested recently. Our previous research has shown targeting P2X4 receptors (P2X4R) may be effective in the treatment of CPSP, but the downstream pathway of the P2X4R has not been studied. In this study, we found the increase in tumor necrosis factor alpha (TNF-α) level and endocytosis of surface gamma-aminobutyric acid a receptors (GABAaR) in CPSP, and these effects were inhibited by blocking P2X4R. Furthermore, antagonizing TNF-α can increase surface GABAaR expression and mechanical pain threshold. Meanwhile, knocking down TNFR1 but not TNFR2 reversed the endocytosis of surface GABAaR and alleviated mechanical allodynia. Thus, the neuropathic pain was mediated, in part, through P2X4R/TNF-α/TNFR1/GABAaR signaling, which was induced after stroke. PERSPECTIVE: P2X4R regulates the pathophysiological mechanism of CPSP through central disinhibition mediated by TNF-α/TNFR1. Our results suggest that modulation of P2X4R-TNF-α/TNFR1-GABAaR signaling could provide a new therapeutic strategy to treat CPSP.

As the main organelles for the clearance of damaged proteins and damaged organelles, the function of lysosomes is crucial for maintaining the intracellular homeostasis of long-lived neurons. A stable acidic environment is essential for lysosomes to perform their functions. TMEM175 has been identified as a new K+channel that is responsible for regulating lysosomal membrane potential and pH stability in neurons. This study aimed to understand the role of TMEM175 in lysosomal function of neurons and neuronal injury following cerebral ischemia-reperfusion (I/R). A middle-cerebral-artery occlusion/reperfusion (MCAO/R) model was established in adult male Sprague-Dawley rats in vivo, and cultured neurons were exposed to oxygen-glucose deprivation/reoxygenation (OGD/R) to mimic ischemia-reperfusion (I/R) injury in vitro. We found that the protein level of TMEM175 decreased after cerebral I/R injury and that TMEM175 overexpression ameliorated MCAO/R-induced brain-cell death and neurobehavioral deficits in vivo. Furthermore, these results were recapitulated in cultured neurons. Acridine orange (AO) staining, as well as LysoSensor Green DND-189, cathepsin-B (CTSB), and cathepsin-D (CTSD) activities, showed that TMEM175 deficiency inhibited the hydrolytic function of lysosomes by affecting lysosomal pH. In contrast, TMEM175 upregulation reversed OGD/R-induced lysosomal dysfunction and impaired mitochondrial accumulation in cultured neurons. TMEM175 deficiency induced by cerebral I/R injury leads to compromised lysosomal pH stability, thus inhibiting the hydrolytic function of lysosomes. Consequently, lysosomal-dependent degradation of damaged mitochondria is suppressed and thereby exacerbates brain damage. Exogenous up-regulation of TMEM175 protein level could reverse the neuronal lysosomal dysfunction after ischemia-reperfusion.

D-Serine is thought to be involved in N-methyl-D-aspartate (NMDA)-type glutamate receptor-mediated neurotoxicity and plays a pathophysiologic role in stroke. D-Serine is synthesized by serine racemase (SR), which directly converts L-serine into D-serine. The deletion of SR has been reported to protect against cerebral ischemia damage. Additionally, SR catalytic activity is physiologically regulated by its binding to stargazin. However, whether the stargazin-SR interaction affects the level of stroke damage remains elusive. We showed that cerebral ischemia increased the interaction of stargazin and SR and decreased the levels of D-serine. Disrupting the stargazin-SR interaction by knocking down stargazin aggravated cerebral ischemic insults. We found that cerebral ischemia decreased the phosphorylation of stargazin at the Thr-321 residue, which was phosphorylated by cAMP-dependent protein kinase A (PKA). Treatment with the PKA inhibitor H89 blocked stargazin T321 phosphorylation, augmented the stargazin-SR interaction, decreased D-serine levels, and alleviated focal cerebral ischemic damage in rats subjected to middle cerebral artery occlusion and reperfusion (MCAO/R). Thus, the stargazin-SR interaction is a promising strategy in the treatment of stroke.

BackgroundRecent studies on cluster headache suggested that headache includes three phases.Oral zolmitriptan is effective in the treatment of acute headache attack.We aim to describe the clinical characteristics of different phases of episodic cluster headache(eCH) and observe the efficacy and safety of oral zolmitriptan in preictal phase of eCH attacks.MethodsForty-eight patients with eCH were enrolled in this study. We divided the CH attack into three phases: preictal, ictal and postictal phase and analyzed the clinical features of each phase. We collected the data of twenty-three patients who took zolmitriptan in the preictal phase of three consecutive CH attacks. The Visual Analogue Scale (VAS) score and duration of headache after three consecutive treatments were compared with the baseline respectively. Chi square test, t test, Mann Whitney U test, Pearson linear correlation analysis, paired t-test and paired non-parametric rank-sum test were used for statistical analysis.ResultsForty-five patients (93.8%) reported the presence of preictal symptoms. The most frequent preictal symptom was mild to moderate head discomfort in the area of the subsequent serious headache, accounting for 87.5%. The duration of preictal period was positively correlated with the duration of headache. The Pearson correlation coefficient was 0.313 (P=0.018). Among the twenty-three patients taking oral zolmitriptan in the preictal phase, the VAS max score was significantly lower than that of baseline headache (PConclusions Preictal symptoms were common in CH. Mild to moderate discomfort in the subsequent serious headache was the most common symptom in the preictal phase. Oral administration of zolmitriptan in the preictal period could reduce the severity of headache and shorten the duration of headache in a cluster period without serious adverse reactions.

Background: The previously approved botulinum toxin and nowadays promising calcitonin gene-related peptide (CGRP) monoclonal antibody have shown efficacy for preventing chronic migraine (CM). However, there is no direct evidence for their relative effectiveness and safety. In this study, we conducted an indirect treatment comparison to compare the efficacy and safety of CGRP monoclonal antibody with botulinum toxin for the preventive treatment of chronic migraine.Methods: Up to August 31, 2020, we systematically searched PubMed, Embase, and Cochrane Library Central Register of Controlled Trials (Central). Weighted mean difference (WMD) and relative risk (RR) were used to evaluate clinical outcomes. Indirect treatment comparison (ITC) software was used to conduct indirect treatment comparison.Results: Ten studies were pooled with 6,325 patients in our meta-analysis. Both botulinum toxin and CGRP monoclonal antibody demonstrated favorable efficacy in the change of migraine days, headache days, HIT-6 score, and 50% migraine responder rate compared with placebo. In indirect treatment comparison, CGRP monoclonal antibody was superior to botulinum toxin in the frequency of acute analgesics intake (WMD = −1.31, 95% CI: −3.394 to 0.774, p = 0.02113), the rate of treatment-related adverse events (AEs) (RR = 0.664, 95% CI: 0.469 to 0.939, p = 0.04047), and the rate of treatment-related serious adverse events (RR = 0.505, 95% CI: 0.005 to 46.98, p < 0.001).Conclusion: For chronic migraine patients, CGRP monoclonal antibody was slightly better than botulinum toxin in terms of efficacy and safety. In the future, head-to-head trials would be better to evaluate the efficacy and safety between different medications in the prevention of chronic migraine.

Stroke is the leading cause of disability and death in the world. Central post-stroke pain (CPSP), a central neuropathic pain syndrome occurring after cerebral stroke, is a serious problem. But on account of the lack of reliable animal models, the mechanisms underlying CPSP remains poorly understood. To better understand of the pathophysiological basis of CPSP, we developed and characterized a new rat model of CPSP. This model is based on a hemorrhagic stroke lesion with intra-thalamic autologous blood (ITAB) injection in the ventral posterolateral nucleus of the thalamus. Behavioral analysis demonstrated that the animals displayed a significant decrease in mechanical allodynia threshold. We found a significant increase in P2 × 4 receptor expression in microglia in thalamic peri-lesion tissues post-hemorrhage. The mechanical allodynia in rats with CPSP were reversed by blocking P2 × 4 receptors. A significant alleviation of mechanical allodynia was achieved following the administration of adrenergic antidepressants and antiepileptics. Meanwhile, we found a significant decrease in P2 × 4 receptor expression after treatment with these drugs. Taken together, our results suggest that targeting P2 × 4 receptor may be effective in the treatment of CPSP.

Central poststroke pain (CPSP) is a neuropathic pain syndrome that usually occurs after cerebrovascular accidents. Currently, the pathogenesis of CPSP is not fully understood. Purinergic P2X4 receptor (P2X4R) is implicated in neuropathic pain including CPSP. Herein, we demonstrated that the levels of microRNA-133b-3p (miR-133b-3p), which targets P2X4R transcripts, were significantly downregulated in the ventral posterolateral nucleus of the thalamus (VPL), cerebrospinal fluid (CSF), and plasma of CPSP rats. The expression levels of miR-133b-3p negatively correlated with the severity of allodynia. Genetic knockdown of P2X4R in the VPL protected CPSP rats against allodynia. Similarly, genetic overexpression of miR-133b-3p in the VPL reversed the allodynia established in CPSP rats via downregulation of P2X4R expression. Treatment using gabapentin in CPSP rats significantly restored the decreased miR-133b-3p expression in the VPL, CSF, and plasma and blocked allodynia in CPSP rats. The administration of an miR-133b-3p inhibitor into the VPL abolished the antiallodynic activity of gabapentin. This mechanism was associated with P2X4R expression and involved the endogenous opioid system. Human patients with CPSP showed decreased plasma levels of miR-133b-3p compared with those of control participants. Logistic regression analysis of our patient cohort showed that determining plasma levels of miR-133b-3p may be useful for CPSP diagnosis and treatment.

Objective: Previous studies demonstrated that diabetic stroke patient had a poor prognosis and excess activation of the complement system in the peripheral blood. In this study, the association of serum complement levels with prognosis of diabetic stroke was examined. Methods: Patients with acute ischemic stroke were recruited and were divided into two groups according to the history of diabetes. Baseline data on the admission including C3 and C4 were collected. Neurologic function at discharge was the primary outcome and was quantified by the National Institutes of Health Stroke Scale (NIHSS). Results: A total of 426 patients with acute ischemic stroke (116 diabetic stroke and 310 non-diabetic stroke) were recruited in this study. There were significant differences on hypertension, CHD, TG, HDL, FGB, C4, and mortality rate between two groups. Furthermore, the values of complement protein levels were divided into tertiles. In diabetic stroke group, serum C4 level at acute phase in the upper third was independently associated with NIHSS score at discharge and concurrent infection. This associations were not significant in non-diabetic stroke. Conclusion: High serum C4 level at admission, as an unique significant predictor, was associated with unfavorable clinical outcome in the diabetic stroke, independently of traditional risk factors.

Background Immune factors are found to be involved in the pathophysiological process of ischemic stroke. However, the clinical role of the immune system in stroke remains unclear. Our study was designed to investigate the correlation between immunocytes (T cell, B cell, NK cell), complements and stroke severity. Methods 236 patients with first-ever ischemic stroke were included in our study and divided to mild, moderate and severe groups according to NHISS score at stroke onset. Modified Rankin Scale (mRS) was used to assess short-term prognosis 3 months after stroke. We also collected clinical data and test circulating T cells, CD8 + T cells, CD4 + T cells, B cells, NK cells, C3, C4, complement factor B (CFB) of each patient. Results We found NK cell(p = 0. 015), CFB (p = 0. 007), C3 (p = 0. 035) were significantly elevated in groups with higher NHISS score. CD4 + T cell (p = 0. 009), T cell (p = 0. 001) were decreased in groups with higher NHISS score. In multivariate logistic regression, CFB and C3 were independently associated with severity of stroke. CD4 + T cell, NK cell, T cell were independently associated with severe ischemic stroke. NK cell (p = 0.049), CFB (p = 0.035), C4 (p = 0.028) are were significantly higher in groups with higher mRS score. In binary analyses, CFB (p = 0.009, OR: 1.056, 95%CI: 1.014–1.099), C4 (p = 0.011, OR: 134.444, 95%CI: 3.115-5803.504) were independently associated with prognosis at 3 months after stroke adjusting onset. Conclusions Elevated circulating CFB was correlated with both severe symptom at stroke onset and worse prognosis. It indicates that CFB may be a predictor of stroke severity and prognosis. Reduced T cell counts, elevated C3 and NK cell were associated with severe symptom at stroke onset. C4 was associated with short-term prognosis.

As a very common risk of adverse outcomes of the ischemic stroke patients, sarcopenia is associated with infectious complications and higher mortality. The goal of this retrospective study is to explore the predictive value of serum Cr/CysC ratio in acute ischemic stroke patients receiving nutritional intervention. We reviewed adult patients with AIS from December 2019 to February 2020. Patients with acute kidney injury were excluded and all patients received nutritional intervention during a 3-month follow-up period. We collected baseline data at admission including creatinine and cystatin C. The primary poor outcome was major disability (modified Rankin Scale score ≥ 4) at 3 months after AIS. A total of 217 patients with AIS were identified for this study. Serum Cr/CysC ratio was significantly correlated with NIHSS at discharge, 1-month modified Rankin Scale score, and 3-month modified Rankin Scale score. During 3 months, 34 (15.70%) patients had a poor outcome after AIS and 11 (5.10%) patients died within 30 days. In multivariable logistic regression analyses, serum Cr/CysC ratio at admission was independently associated with 3-month poor outcomes (OR: 0.953, 95% CI: 0.921–0.986, p =.006) and 30-day mortality (OR: 0.953, 95% CI: 0.921–0.986, p =.006). As a blood biochemical indexes reflecting the muscle mass and aiding in risk stratification, Cr/CysC ratio at admission could be used as a predictor of 30-day mortality and long-term poor prognosis in AIS patients.

Vestibular migraine (VM) is a multidisciplinary disease under exploration. Multiple temporal patterns of vertigo and migraine make it difficult to diagnose VM, and their effect on the clinical features of VM is still unclear. Here we investigated the clinical features of VM under three temporal patterns. 172 VM patients were enrolled in this study and divided into three groups: 86 patients in group A had an earlier onset of migraine than vertigo, 35 patients in group B had an earlier onset of vertigo than migraine, and 51 patients in group C had concurrent vertigo and migraine. No significant difference was found among three groups regarding types, intensity and accompanying symptoms of the vestibular attack. Patients in group C presented higher frequency and longer duration of vertigo than group A and B, while patients in group A presented lower frequency and shorter duration of headaches than group B and C. Additionally, the frequency, duration, intensity and accompanying symptoms of headache in group A decreased significantly after the onset of vertigo, especially in women around menopause. We hypothesized that vestibular stimulation could inhibit the trigeminal pain pathway, while painful trigeminal stimulation could excite the vestibular system. Our findings may contribute to the clinical identification of VM and further clarification of its pathogenesis.

Additional file 1. Supplementary material.

The aim of this study was to investigate the alterations in regional homogeneity assessed by fMRI in patients with migraine without aura (MWoA). Fifty-six eligible MWoA patients and 32 matched healthy volunteers were enrolled in this study. MWoA patients were divided into three groups according to the headache days per month within 3 months: infrequent episodic migraine (IEM) group, frequent episodic migraine (FEM) group, and chronic migraine (CM) group. Data collection and rest-state fMRI examination were performed in all cases. The ReHo method was used to analyze the blood oxygen level dependent (BLOD) signals of the adjacent voxels in the brain regions of each patient, and the consistency of their fluctuations in the sequences of same time. Compared with normal controls, ReHo values of bilateral thalami, right insula and right middle temporal gyrus increased and both precentral gyri decreased in the IEM group; ReHo values of bilateral thalami and the right middle temporal gyrus increased; ReHo values of both anterior cingulate cortex, precentral gyri and putamen decreased in the FEM group. Compared with control group, ReHo values of left olfactory cortex, right hippocampus, parahippocampal gyrus, suboccipital gyrus and precuneus increased, both precentral gyri, precuneus, putamen and anterior cingulate cortex decreased in the CM group. Compared with IEM group, ReHo values of both putamen, left middle frontal gyrus, right superior frontal gyrus increased, and the left precuneus decreased in the FEM group. Compared with FEM group, ReHo values of left olfactory and left precuneus increased, and the right superior frontal gyrus, insula, middle temporal gyrus, thalami, both superior temporal gyri decreased in the CM group. In the IEM group, the changes of function focus on the regions associated with coding, conduction and regulation of pain signals. In the FEM group, functional alterations mainly concentrated on the regions associated with pain regulation and emotion cognition. In the CM group, the changes focus on the regions related to spatial attention and cognition, affective disorders and pain feedback, which may be associated with migraine production, development and chronification.

High-flow oxygen inhalation is one of the most effective acute treatments for cluster headache. The therapy was first described for the treatment of cluster headache in 1952 by Horton, and has exhibited some advantages and efficacy compared to other acute medicines. The mechanism is not very clear, but some evidence has demonstrated its relationship to the trigeminovascular system and neuroinflammation. High-flow oxygen inhalation via a non-rebreather mask during cluster headache attacks has been widely recommended. Patients with frequent attacks and/or intolerance to drugs may prefer the oxygen treatment.

Hydrogen sulfide (H2S) is recognized to be a novel mediator after carbon monoxide and nitric oxide in the organism. It can be produced in various mammalian tissues and exert many physiological effects in many systems including the cardiovascular system. A great amount of recent studies have demonstrated that endogenous H2S and exogenous H2S-releasing compounds (such as NaHS, Na2S, and GYY4137) provide protection in many cardiovascular diseases, such as ischemia/reperfusion injury, heart failure, cardiac hypertrophy, and atherosclerosis. In recent years, many mechanisms have been proposed and verified the protective role exhibited by H2S against myocardial ischemia/reperfusion injury, and this review is to demonstrate the protective role of exogenous and endogenous H2S on myocardial ischemia/reperfusion injury.

Although recent studies have implicated serotonin 5-HT1B/D receptors in the nociceptive sensitivity of primary afferent neurons, the underlying molecular and cellular mechanisms remain unclear. In this study, we identified a novel functional role of the 5-HT1D receptor subtype in regulating A-type potassium (K(+)) currents (IA) as well as membrane excitability in small trigeminal ganglion (TG) neurons. We found that the selective activation of 5-HT1D, rather than 5-HT1B, receptors reversibly increased IA, while the sustained delayed rectifier K(+) current was unaffected. The 5-HT1D-mediated IA increase was associated with a depolarizing shift in the voltage dependence of inactivation. Blocking G-protein signaling with pertussis toxin or by intracellular application of a selective antibody raised against Gαo or Gβ abolished the 5-HT1D effect on IA. Inhibition of protein kinase A (PKA), but not of phosphatidylinositol 3-kinase or protein kinase C, abolished the 5-HT1D-mediated IA increase. Analysis of phospho-p38 (p-p38) revealed that activation of 5-HT1D, but not 5-HT1B, receptors significantly activated p38, while p-ERK and p-JNK were unaffected. The p38 MAPK inhibitor SB203580, but not its inactive analogue SB202474, and inhibition of B-Raf blocked the 5-HT1D-mediated IA response. Functionally, we observed a significantly decreased action potential firing rate induced by the 5-HT1D receptors; pretreatment with 4-aminopyridine abolished this effect. Taken together, these results suggest that the activation of 5-HT1D receptors selectively enhanced IA via the Gβγ of the Go-protein, PKA, and the sequential B-Raf-dependent p38 MAPK signaling cascade. This 5-HT1D receptor effect may contribute to neuronal hypoexcitability in small TG neurons.

Objective The research aim was to investigate the effects of dl-3-n-butylphthalide (NBP) on the level of circulating endothelial progenitor cells (EPCs) and clinical outcome in patients with acute ischemic stroke (AIS). Materials and Methods A total of 170 patients were included and randomly assigned to NBP group and control group. All patients were administrated a basic antiplatelet and lipid-lowering therapy. Among the patients, 86 received additional NBP administration for 30 days, whereas 84 received only basic therapy (the control). The level of circulating EPCs (marked with CD34 + /CD133 + /KDR + ) was determined by flow cytometry at baseline and days 7, 14, and 30 after therapy. Impairment of neurological function was evaluated by the National Institutes of Health Stroke Scale (NIHSS) on days 7, 14, 30, and 90 after therapy. The association between the increased level of circulating EPCs and improvement of NIHSS score was evaluated by Pearson analysis. The clinical outcome was evaluated by modified Rankin Scale (mRS) on day 90. During the observation period, any adverse events related to drugs were reported. Results The levels of circulating EPCs on days 14 and 30 were significantly higher in the NBP group than in the control group. In contrast, NIHSS score was notably lower in NBP group on day 14, 30 and day 90. Pearson correlation analysis revealed a significant association between the increased level of EPCs and improvement of NIHSS score. Also, the mRS score in the NBP group was lower on day 90. Importantly, the reported adverse events in the 2 groups were comparable. Conclusion NBP significantly increases the circulating level and improves clinical outcome in patients with AIS.

We read with great interest the case reports about patients with COVID-19 with Miller Fisher syndrome and polyneuritis cranialis by Gutierrez-Ortiz et al.,1 which provided a link between these diseases and the symptom spectrum of SARS-CoV-2 infection. As we know, the Miller Fisher syndrome and polyneuritis cranialis are an autoimmune neuropathy triggered by autoantibodies specific for the polysialogangliosides GQ1b and GT1a in axonal terminals and causes the inflammation and demyelination of the peripheral and cranial nerves.2 A recent article also proposed that autoimmune injury may be involved in the mechanisms of nervous system symptoms of COVID-19.3 In addition, Zika virus—another coronavirus—is also associated with autoimmune peripheral neuropathy.4,5 These thus support the connection of Miller Fisher syndrome and polyneuritis cranialis with SARS-CoV-2 infection. We wonder whether the authors examined the presence of SARS-CoV-2 RNA and antibodies for SARS-CoV-2 in CSF and ruled out the infection by other common viruses in 2 patients with COVID-19? This will provide more convincing evidence to support SARS-CoV-2 infection-induced autoimmune injury on peripheral nerves, and immunoglobulin therapy may be considered for some patients with COVID-19.

Since the power transformers are indispensable in power systems, their reliable operation are of significance. Winding deformation is one of the most common failures within transformers. At present, many methodologies have been proposed to detect these faults, and frequency response analysis (FRA) technique is most frequently used because of its low cost, convenience, simplicity and effectiveness. However, there is no standard and reliable code to interpret the mechanical deformations from FRA traces as so far. In this study, an actual transformer experimental platform was used to simulate variable winding deformation faults in order to standardize the interpretation of winding fault degree from FRA data. The measured FRA raw data are processed from the perspective of statistics, and some numerical indices are found to be more suitable for the diagnosis of transformer winding deformation degree.

In this study, we aimed to investigate the expression of miR-145 before and after hASCs osteogenic differentiation. We also intended to explore the influence of the target relationship between miR-145 and FoxO1 on osteogenic differentiation. Dual-luciferase reporter gene assay and real-time PCR were used to confirm the target relationship between miR-145 and FoxO1. Furthermore, the modulatory effects of miR-145 and FoxO1 on hASCs osteoinductive differentiation were measured by real-time PCR , Western blot, ALP staining, ARS staining, and cell immunofluorescence assay. After osteogenic differentiation, miR-145 was gradually down-regulated, while FoxO1 was up-regulated in hASCs. MiR-145 could directly target FoxO1 3′UTR. FoxO1 was negatively regulated by miR-145. After osteoinductive differentiation, BSP, Ocn, and OPN expression was lowered with the overexpression of miR-145 or the knockdown of FoxO1. Furthermore, ALP and ARS staining assay results showed weakened ALP activity and extracellular matrix calcification. When overexpressing miR-145 and FoxO1 simultaneously, no obvious change in ALP activity and extracellular matrix calcification was seen. MiR-145 could suppress hASCs osteoinductive differentiation by suppressing FoxO1 directly. Impact statement Researching on ASCs was a promising strategy to study osteogenic differentiation. The regulatory role of miR-145 on hASCs osteogenic differentiation remained partially explored. Our study revealed a novel mechanism of the osteogenic differentiation process and suggested that miR-145 and its target gene FoxO1 may be potential targets for the therapy of human osteogenic-related disorders.

Pseudorabies (PR) has been prevalent in Chinese swine breeding farms since the outbreak at the end of 2011. For investigating current prevalence of PR, a nationwide surveillance has been performed in this study. The swine serum samples were collected from 93, 100, 92, and 91 swine farms in China during 2013–2016, respectively. Since the extensive use of gE-deleted pseudorabies virus (PRV) vaccine, we could apply the PRV-gE antibody for determining wild-type virus infection and the PRV-gB antibody for evaluating vaccine immunization. The results were concluded as follows: (1) Nationally, the positive rate of PRV-gB was maintained at a high level (> 90%), while the positive rate of PRV-gE continued to decrease (from 22.17 to 13.14%). (2) The positive rates of PRV-gE were greatly varied in different geographical regions and swine farms (0~100%), while the positive rate of PRV-gB was generally high (> 90%). (3) The number of imported PRV attenuated vaccines were about twice that of domestic PRV attenuated vaccines, while the positive rate of PRV-gB was not significantly different (P > 0.05). (4) The performance of PR eradication developing or developed farms was better than the performance of common farms, with higher positive rate of PRV-gB (> 90%) and much lower positive rate of PRV-gE (nearly 0%).

Hepatocellular carcinoma (HCC) represents the most common primary malignancy of the liver with a worldwide increasing incidence. Although the risk factors for HCC are well characterized, the molecular mechanisms responsible for malignant transformation of hepatocytes are not well understood. In this study, a case–control study including 291 HCC patients and 294 healthy controls was conducted to investigate the association between HCC susceptibility and with a 4-bp insertion/deletion polymorphism (rs66465034) in the proximal promoter of CD3G. Logistic regression analysis showed that the heterozygote and the homozygote 4-bp ins/ins confer a significantly increased risk of HCC after controlling for other covariates (adjusted odds ratio [OR]=1.51, 95% confidence interval [C.I.] 1.01–2.27, p=0.040; OR=1.71, 95% C.I. 1.07–2.89, p=0.025, respectively). Carriage of the 4-bp insertion allele was associated with a greatly increased risk of developing the disease (OR=1.30, 95% C.I. 1.02–1.64, p=0.027). Moreover, hepatitis B virus (HBV) stratification analysis showed that the differences between cases and controls were more obvious in HBV-positive than in the HBV-negative population, suggesting a possible role of this polymorphism in the immune regulation during HBV infection. Further, luciferase-based transient transfection assays revealed that rs66465034 can affect promoter activity of CD3G, indicating its possible functional significance. Our data suggested that common genetic polymorphisms in CD3G may influence HCC risk in Chinese population. Considering the relative small sample size, replication in other populations with larger sample size and further functional analysis are required for fully understanding the roles of CD3G polymorphisms in predisposition for HCC.

Highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) initially emerged in China and currently prevails in other Asian countries as well, resulting in immense economic losses. HP-PRRS virus (HP-PRRSV) has undergone rapid evolution since its first recognition in 2006. To analyze the genomic and pathogenic characteristics of 2010 HP-PRRSV, we tested 919 clinical samples collected from China, Laos and Vietnam, sequenced 29 complete genomes of HP-PRRSV isolates, and determined the pathogenicity of seven HP-PRRS viruses isolated from 2006 to 2010. HP-PRRSV was detected from 45.2% (415/919) samples, while only 0.1% (1/919) was classical PRRSV, indicating that HP-PRRSV isolates with a unique discontinuous deletion of 30 amino acids (aa) in non-structural protein 2 (Nsp2) are still the predominant viruses. 2010 HP-PRRSV together with 2009 HP-PRRSV isolates form a new evolutionary branch based on phylogenetic analyses. The numbers of potential N-glycosylation sites are variable in major glycoprotein GP5 but are conserved in minor glycoproteins GP2, GP3 and GP4. Pathogenicity studies showed that HP-PRRS viruses isolated from 2006 to 2010 maintain similar level of high pathogenicity, which caused high fever (>41 °C for at least four days), 100% morbidity, and 40–100% mortality in 4–10 weeks old pigs. Real time monitoring information from this study could help to understand the genetic and pathogenic evolution of HP-PRRSV and assist in the control of HP-PRRS in Asia.

Porcine circovirus type 2 (PCV2) is continuously evolving through point mutation and genome recombination. In the present study, genetic affiliations of 40 PCV2 strains were determined by amplification, sequencing and phylogenetic analyses during the nationwide molecular epidemiology investigation from 2009 to 2010 in mainland China. The results revealed that PCV2b was the predominant genotype in mainland China from 2009 to 2010 and PCV2b-1C within PCV-2b genotype was an emerging predominant subtype. More interestingly, five strains (09HaiN-1, 09HaiN-2, 10AH, 10GX and 10QH) were classified into a novel cluster along with the two main PCV2 genotypes PCV2a and PCV2b. Further analyses revealed that this novel cluster arose from recombination between PCV2a and PCV2b stains within the ORF2 gene. Moreover, BLAST search on NCBI website revealed that PCV2 strains of the novel cluster also emerged in Thailand, Indonesia and Laos, indicating that the novel cluster of PCV2 has also been circulating in some other Asian countries. This study is the first time to perform comprehensive analyses to demonstrate a cluster of PCV2 strains derived from the same type of inter-genotypic recombination pattern. Our findings provide valuable information on PCV2 evolution.