To realize the automatic identification of electroencephalogram (EEG) signals from patients with schizophrenia, this study calculated graph-theoretic features derived from event-related potentials (ERP) recorded during a Go/NoGo task. The ERP data were collected from 42 individuals diagnosed with schizophrenia and 29 healthy controls. Different functional networks of single-frequency band and all band of these EEG data, were constructed by phase locking value method. Their graph-theoretic features were extracted. Three machine learning classifiers were used to automatically distinguish the patients and health controls. The classification accuracies of different models with graph-theoretic features of different networks were compared. The results revealed that F-score combined with support vector machine (SVM) model achieved the highest accuracy. This model with graph-theoretic features of all-band network and theta band networks resulted in the highest classification accuracy, which was 95.45

BackgroundPreviously we showed that elevated cerebrospinal fluid (CSF) levels of kininogen during the acute phase of encephalitis are associated with symptomatic epilepsy. However, the functional role of kininogen in epileptogenesis remains unexplored.ObjectiveThis study investigated the brain expression of kininogen and its influence on seizure susceptibility. Additionally, we examined the effects of bradykinin, a nonapeptide derived from kininogen, as a potential downstream mediator of kininogen's effect on seizure susceptibility and the underlying circuitry mechanisms.MethodsWe analyzed brain mRNA expression of kininogen using publicly available single-cell RNA-sequencing (scRNA-seq) datasets and assessed protein expression through immunofluorescence in various brain regions. Immunoblot was conducted following pilocarpine-induced status epilepticus (Pilo-SE) to understand post-seizure kininogen dynamics. Next, to understand its functional role, kininogen was overexpressed in the hippocampal CA1 area via its AAV-mediated gene delivery, and bradykinin was administered through the fourth ventricle in mice. The effects on seizure susceptibility were evaluated using a pentylenetetrazole-induced seizure susceptibility test. Furthermore, two-photon in vivo calcium imaging of cortical layer 2/3 glutamatergic neurons and GABAergic parvalbumin-positive neurons was performed to explore the potential circuitry mechanisms.ResultsWhile scRNA-seq analyses found kininogen gene expressions in various cell types across the brain, immunofluorescence revealed its preferential localization in neurons but not in glia. Pilo-SE decreased intact kininogen levels in the hippocampus and increased cleaved to intact kininogen (cHK / iHK) ratio. Overexpression of kininogen and exogenous bradykinin administration significantly increased pentylenetetrazole-induced seizure susceptibility in mice. Mechanistically, bradykinin was found to enhance calcium activities in cortical glutamatergic excitatory neurons in Thy1-GCaMP mice when they were treated with a subthreshold dose of pentylenetetrazole. In contrast, calcium activities in GABAergic parvalbumin-positive inhibitory neurons were reduced by bradykinin in PVCre-GCaMP mice, suggesting potential circuitry mechanisms by which kininogen may render increased seizure susceptibility.ConclusionOur findings reveal a novel pathological role of kininogen in seizure occurrence, explaining why kininogen might be elevated in the CSF of epilepsy-susceptible patients and suggest its potential mechanisms where it might regulate the activities of glutamatergic and GABAergic neurons through the downstream release of bradykinin. Altogether, we propose kininogen as a potential target for developing therapeutics for epilepsy intervention.

AIMS:To explore the role of voltage-gated calcium channels (VGCC) in 5-HT2A/2C receptor agonist 2,5-dimethoxy-4-iodophenyl-2-aminopropane hydrochloride's improvement of spinal cord injury (SCI) induced detrusor sphincter dyssynergia and the expressions of the 5-hydroxy tryptamine (5-HT) 2A receptors and VGCCs in lumbosacral cord after SCI. METHODS:Female Sprague-Dawley rats were randomized into normal control group and SCI group (N = 15 each). Cystometrogram (CMG), simultaneous CMG, and external urethral sphincter electromyography (EUS-EMG) were conducted in all groups under urethane anesthesia. Drugs were administered intrathecally during CMG and EUS-EMG. Rats were euthanized and L6-S1 spinal cord were acquired for immunofluorescence. RESULTS:In SCI rats, intrathecal administration of 2,5-dimethoxy-4-iodophenyl-2-aminopropane hydrochloride or L-type VGCC blocker, nifedipine, could significantly increase voiding volume, voiding efficiency, and the number of high-frequency oscillations. They could also prolong EUS bursting activity duration on EUS-EMG. Moreover, the effect of 2,5-dimethoxy-4-iodophenyl-2-aminopropane hydrochloride can be eliminated with the combined administration of L-type VGCC agonist, (±)-Bay K 8644. No significant differences were observed in CMG after intrathecal administration of T-type VGCC blocker TTA-P2. Additionally, immunofluorescence of the lumbosacral cord in control and SCI rats showed that the 5-HT2A receptor and Cav1.2 immunolabeling-positive neurons in the anterior horn of the lumbosacral cord were increased in SCI rats. CONCLUSIONS:Our study demonstrated that 5-HT2A/2C agonist 2,5-dimethoxy-4-iodophenyl-2-aminopropane hydrochloride may improve SCI-induced DSD by inhibiting the L-type voltage-gated calcium channel in lumbosacral cord motoneurons.

N6-methyladenosine (m6A) modification controls cell fate determination. Here, we show that liquid-liquid phase separation (LLPS) of YTH N6-methyladenosine RNA binding protein 1 (YTHDF1), a pivotal m6A "reader" protein, promotes the transdifferentiation of spermatogonial stem cells (SSCs) into neural stem cell-like cells by activating the IκB-nuclear factor κB (NF-κB)-CCND1 axis. The inhibition of IκBα/β mRNA translation mediated by YTHDF1 LLPS is the key to the activation of the IκB-NF-κB-CCND1 axis. Disrupting either YTHDF1 LLPS or NF-κB activation inhibits transdifferentiation efficiency. Moreover, overexpression of the YTH domain of YTHDF1 inhibits the activation of the IκB-NF-κB-CCND1 axis by promoting IκBα/β mRNA translation. Overexpression of the tau-YTH fusion protein reactivates IκB-NF-κB-CCND1 axis by inhibiting the translation of IκBα/β mRNAs, and tau LLPS is observed, which can restore transdifferentiation efficiency. Our findings demonstrate that the protein-RNA LLPS plays essential roles in cell fate transition and provide insights into translational medicine and the therapy of neurological diseases.

Alzheimer’s disease (AD) is the most common neurodegenerative disease characterized by the accumulation of amyloid β peptides (Aβ) and impaired glucose metabolism in the brain. Osteocalcin (OCN), an osteoblast-derived protein, has been shown to modulate brain functions but whether it has any effect on AD is undetermined. In this study, daily intraperitoneal injection of OCN for 4 weeks ameliorated the anxiety-like behaviors and cognitive dysfunctions in the APP/PS1 transgenic AD mice model, as shown in the increased entries into the central area in open field test, the increased time and entries into open arms in elevated plus maze test, the increased time spent in the light chamber in light-dark transition test, as well as the reduced escape latency and the increased preference for target quadrant in Morris water maze test. Aβ burden in the hippocampus and cortex of AD mice was ameliorated by OCN. Besides, OCN improved the neural network function of the brain, mainly in the enhanced power of high gamma band in the medial prefrontal cortex of AD mice. The proliferation of astrocytes in the hippocampus in AD mice was also inhibited by OCN as demonstrated by immunofluorescence. Furthermore, OCN enhanced glycolysis in astrocytes and microglia, as evidenced by elevated glucose consumption, lactate production, and increased extracellular acidification rate. Such an effect was abolished when the receptor of OCN – Gpr158 was knockdown in astrocytes. Our study revealed OCN as a novel therapeutic factor for AD potentially through reducing Aβ burden and upregulation of glycolysis in neuroglia.

Persistent stress increases the probability for developing depression significantly thereafter. Repeated social defeat stress is a widely used model to investigate depressive-like behavior in preclinical models. Hence, the repeated social defeat stress model provided an ideal animal model, through which the hypotheses of prevention and treatment can be investigated. We have successfully induced depressive-like behavior for male C57BL/6J mice with this model. Here, we reported that certain level of during-stress social interactions with single female or multiple male peer(s) exerted a positive role in preventing the development of depressive-like behavior induced by repeated social defeat stress. Our data suggested that the stress-susceptible mice may benefit from positive social interaction, which reduces the chance for depressive-like behavior development. Since numerous studies indicate that the metabotropic glutamate receptor 5 (mGluR5) plays an important role in various cognitive functions, we further investigate the treatment effect of 3-cyano-N-(1,3-diphenyl-1H-pyrazol-5-yl) benzamide (CDPPB) on the depressive-like behavior induced by repeated social defeat stress. Most importantly, robust anti-depressant effects have been achieved through modulating the mGluR5 function. We found that single oral dose administration of CDPPB (20 mg/kg), to some extent, alleviated the social avoidance behaviors for the stress-susceptible mice. Our data implies that the CDPPB, a positive allosteric modulator of mGluR5, is a promising anti-depressant candidate with limited side effect.

Abstract Studies have shown that manual and electrical acupuncture have significant effects on brain functions. However, these treatments often evoke pain and fear. Non-invasive acupuncture therapies, such as mechanical (acupressure) and thermal (moxibustion), have been developed and shown to effectively relieve pain and treat various conditions. However, little is known about their influence on brain network function. The current study explored the effects of mechanical, thermal, and combined thermal and mechanical stimulations on acupoint Large Intestine 4 (LI4, Hegu) on EEG in fifty-two healthy participants. A 30-minute combined thermal and mechanical stimulation on LI4 enhanced the powers of the β and γ bands and reduced the ratio of θ/β in both the parietal and temporal lobes when compared with the same stimulation on the control point. In contrast, neither the mechanical nor thermal stimulation alone had such effects. The distinct impact of combined thermal and mechanical stimulation, as opposed to either method alone, underscores the importance of joint activation of thermosensory, touch, and pain-sensory fibers for effective acupoint stimulation. Furthermore, our findings provide insights into the use of combined thermal plus mechanical stimulation as an effective non-invasive alternative therapy.

Considerable evidence has revealed that essential oils and their main constituents possess anti-depressant and anxiolytic properties.In the current study,we report the effect of β-citronellol,the main component of rose essential oil,on depressive-like and anxiety-like behaviors in chronic restraint stress(CRS)mice.We found that chronic inhalation of β-citronellol for 14 days could increase locomotor activity in the open field test,de-crease the percentage of immobility duration in the forced swimming test,and increase open arms exploration in elevated plus-maze test in CRS mice.Western blot experiment shows that chronic β-citronellol inhalation res-cues parvalbumin(PV)expression loss in the prefrontal cortex(PFC)of CRS mice.Correlation analysis reveals a strong relationship between the PV expression in PFC and the percentage of sucrose preference of the mice.These findings indicate the relationships between the PV gene expression of PFC and the effects ofβ-citronellol inhalation.

BackgroundParkinson's disease (PD) is a neurodegenerative disorder with no absolute cure. The evidence of the involvement of gut microbiota in PD pathogenesis suggests the need to identify certain molecule(s) derived from the gut microbiota, which has the potential to manage PD. Osteocalcin (OCN), an osteoblast-secreted protein, has been shown to modulate brain function. Thus, it is of interest to investigate whether OCN could exert protective effect on PD and, if yes, whether the underlying mechanism lies in the subsequent changes in gut microbiota.ResultsThe intraperitoneal injection of OCN can effectively ameliorate the motor deficits and dopaminergic neuronal loss in a 6-hydroxydopamine-induced PD mouse model. The further antibiotics treatment and fecal microbiota transplantation experiments confirmed that the gut microbiota was required for OCN-induced protection in PD mice. OCN elevated Bacteroidetes and depleted Firmicutes phyla in the gut microbiota of PD mice with elevated potential of microbial propionate production and was confirmed by fecal propionate levels. Two months of orally administered propionate successfully rescued motor deficits and dopaminergic neuronal loss in PD mice. Furthermore, AR420626, the agonist of FFAR3, which is the receptor of propionate, mimicked the neuroprotective effects of propionate and the ablation of enteric neurons blocked the prevention of dopaminergic neuronal loss by propionate in PD mice.ConclusionsTogether, our results demonstrate that OCN ameliorates motor deficits and dopaminergic neuronal loss in PD mice, modulating gut microbiome and increasing propionate level might be an underlying mechanism responsible for the neuroprotective effects of OCN on PD, and the FFAR3, expressed in enteric nervous system, might be the main action site of propionate.

With the fast-accelerating pace of life, most of the population is troubled by anxious feelings, or they even developed anxiety disorders, which affect their life qualities viciously. The current treatment is not without drawbacks, such as exhibiting slow or acute effects only. Music therapy has shown promising outcomes in significantly reducing tension and anxiety in many studies. To explore the underlying mechanism, we recruited 12 undergraduate student volunteers from Shanghai Jiao Tong University with habits of music listening. We explored the changes in the electroencephalogram (EEG) power spectrum both during and after listening to self-select favorite music, and the relation between EEG signals and anxiety-relieving during music listening. The results showed that the power of the β band in the right parietal area was significantly higher in those who were self-report able to relax during music listening compared with those who could not get relaxed. We observed power increased in the right parietal area of self-reports, not able to relax while music playing. Also, a significant difference in θ band power in the left frontal area and γ band in the right frontal area before and after music listening was observed between high anxiety group and low anxiety group. These results imply that β band activity in the left frontal area is associated with the anxiety-relieving capability of anxiety, which provides evidence for further elucidating the mechanism underlying anxiety-relieving during music listening.

目的:通过测试子代雄性SD大鼠成年后的焦虑样行为,阐明婴幼期声音干预对大鼠成年后焦虑样行为的影响.方法:6只孕14～15 d的SD大鼠随机分为正常噪声环境对照组(NN组)、莫扎特D大调双钢琴奏鸣曲K448组(MK组)和大自然声音组合组(NS组).NN组大鼠不给予任何特殊的声音刺激;MK组大鼠每天给予莫扎特D大调双钢琴奏鸣曲K448声音刺激2 h,音量为50～70 dB,持续30 d;NS组大鼠每天给予大自然声音组合刺激2 h,音量为65～75 dB,持续30 d.刺激结束后5周,选取体质量相近的NN组11只、MK组8只和NS组11只雄性子代大鼠进行旷场实验、明暗箱实验和高架十字迷宫实验.旷场实验检测大鼠总运动路程、站立次数、进入中央区域次数和进入中央区域的时间,明暗箱实验检测大鼠在明箱停留时间百分率、穿梭次数和总运动路程,高架十字迷宫实验检测大鼠在开臂的时间百分率、进入次数和运动路程.结果:旷场实验中3组大鼠的总运动路程、站立次数、进入中央区域次数和进入中央区域的时间比较差异均无统计学意义(P>0.05).明暗箱实验,与NN组比较,NS组大鼠在明箱停留时间百分率明显升高(P<0.05);与NN组和MK组比较,NS组大鼠在明暗箱之间的穿梭次数和总运动路程明显增加(P<0.05或P<0.01).高架十字迷宫实验,与NN组比较,NS组大鼠在开臂的时间百分率明显升高(P<0.05).结论:婴幼期大自然声音的干预可减少雄性SD大鼠成年后的焦虑样行为.

Acupuncture treatment is based on acupoint stimulation; however, the biological basis is not understood. We stimulated one acupoint with catgut embedding for 8 weeks and then used isobaric tags for relative and absolute quantitation to screen proteins with altered expression in adjacent acupoints of Sprague Dawley rats. We found that kininogen expression was significantly upregulated in the stimulated and the nonstimulated adjacent acupoints along the same meridian. The enhanced kininogen expression was meridian dependent and was most apparent among small vessels in the subcutaneous layer. Enhanced signals of nitric oxide synthases, cGMP-dependent protein kinase, and myosin light chain were also observed at the nonstimulated adjacent acupoints along the same meridian. These findings uncover biological changes at acupoints and suggest the critical role of the kininogen-nitric oxide signaling pathway in acupoint activation.

Objective::The asparagine-linked glycosylation 13 homolog ( Alg13) has been identified as causative for congenital disorders of glycosylation type I with epilepsy. The aim of this study was to determine whether mice carrying a mutated version of Alg13 could be used as a model for epileptic encephalopathies or congenital disorders of glycosylation type I. Methods::A model of epileptic encephalopathy was established in C57BL/6 mice by introducing mutations in Alg13 via the clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9 (CRISPR/Cas9) system. All surgical procedures were approved by the Institutional Animal Care and Use Committee of Shanghai Jiao Tong University (A2016084) on October 8, 2016. Results::Mice with 3 different mutations, Alg13 -54nt, Alg13 -5nt and Alg13 -4nt, all of which are located in Alg13 transcript variant 1, were created. The Alg13 -5nt mice exhibited spontaneous seizures similar to patients with Alg13 mutations, suggesting that they could be used as a model for epilepsy. Western blot analysis demonstrated that Alg13 -5nt mice had lower levels of Alg13 expression than wild-type mice. Video observations showed that two of the 17 Alg13 -5nt mice had stage 5 seizures involving jumping and falling, while 12 had stage 3 seizures with head nodding. Conclusion::The Alg13 mouse model provides an outstanding tool for studying epileptic encephalopathies and investigating different aspects of defects in glycosylation or other post-translational modification that cannot be assessed in patients or cell culture systems.

Objective To study the effect of N-Methyl-D-asparticacid ( NMDA ) on the intracellular free calcium concentration ( [ Ca2+] i ) in primary cultured rat calvaria osteoblasts. Methods A calcium imaging technique was applied to observe [ Ca2+] i changes in primary cultured rat calvaria osteoblasts after stimulating by NMDA with various concentrations or pretreated with NMDA receptor noncompetitive antagonism MK801 ( Dizocilpin) . Results Different concentrations of NMDA caused [ Ca2+] i increases in varying degrees and by different ways. NMDA could evoke transient increase and secondary change in [ Ca2+] i including calcium oscillation or steady increase. MK801 inhibited NMDA-induced [ Ca2+] i increase in varying degrees. Conclusion These results indicated that there are abundant functional NMDA receptors expressed in primary cultured rat calvaria osteoblasts, showing different forms and varying degrees of [ Ca2+] i increases in response to different concentrations of NMDA. The characters of the blocking effect of MK801 to NMDA-induced [ Ca2+] i increasing, indicated that the NMDA receptors expressed in primary cultured rat calvaria osteoblasts differ in channel properties from those in central nervous system.

Purpose: Epilepsy is a highly disabling neurological disorder. Brain insult is the most critical cause of epilepsy in adults. This study aimed to find reliable and efficient biomarkers for predicting secondary epilepsy. Materials and methods: The LiCl-pilocarpine (LiCl-Pilo) chronic epilepsy rat model was used, and rat cerebrospinal fluid (CSF) was collected 5 days after status epilepticus (SE). The CSF was analyzed using the label-free LC-ESI-Q-TOF-MS/MS. Differential expression of proteins was confirmed using enzyme-linked immunosorbent assay (ELISA) and Western blotting. The corresponding protein level in the CSF of patients with encephalitis in the postacute phase was determined using ELISA and compared between patients with and without symptomatic epilepsy after encephalitis during a 2-year follow-up. Results: The proteomics and ELISA results showed that the protein level of kininogen (KNG) was obviously elevated in both CSF and hippocampus, but not in serum, 5 days after the onset of SE in LiCl-Pilo chronic epilepsy model rats. In patients with encephalitis, the protein level of KNG in the CSF in the postacute phase was significantly elevated in patients with a recurrent epileptic seizure during a 2-year follow-up than in patients without a recurrent seizure. Conclusion: KNG in the CSF may serve as a potential biomarker for predicting epileptogenesis in patients with encephalitis.

Neonatal seizures are different from adult seizures, and many antiepileptic drugs that are effective in adults often fail to treat neonates. Here, we report that gluconate inhibits neonatal seizure by inhibiting CLC-3 chloride channels. We detect a voltage-dependent outward rectifying Cl − current mediated by CLC-3 Cl − channels in early developing brains but not adult mouse brains. Blocking CLC-3 Cl − channels by gluconate inhibits seizure activity both in neonatal brain slices and in neonatal animals with in vivo EEG recordings. Consistently, neonatal neurons of CLC-3 knockout mice lack the outward rectifying Cl − current and show reduced epileptiform activity upon stimulation. Mechanistically, we demonstrate that activation of CLC-3 Cl − channels alters intracellular Cl − homeostasis and enhances GABA excitatory activity. Our studies suggest that gluconate can suppress neonatal seizure activities through inhibiting CLC-3 Cl − channels in developing brains.

It is now generally accepted that the extra-skeleton functionalities of bone are multifaceted. Its endocrine functions came first to light when it was realized that osteoblasts, the bone forming cells, maintain energy homeostasis by improving glucose metabolism, insulin sensitivity and energy expenditure through osteocalcin, a multipurpose osteokine secreted by osteoblasts. Recently, the emerging knowledge on the functional aspects of this osteokine expanded to properties including adult and maternal regulation of cognitive functions. Therapeutic potential of this osteokine has also been recently reported in experimental Parkinson's disease models. This review highlights such findings on the functions of osteocalcin in the brain and emphasizes on exploring and analyzing much more in-depth basic and clinical studies.

The level of nicotinamide adenine dinucleotide (NAD) decreases in Parkinson's disease (PD), and its reduction has been reported to be involved in many age-associated neurodegenerative pathologies. Thus, we investigated whether NAD replenishment is beneficial in a 6-hydroxydopamine (6-OHDA)-induced mouse model of PD. Preinjection with NAD in the striatum ameliorated motor deficits and dopaminergic neuronal damage in the substantia nigra and striatum of a mouse model of PD. Moreover, preincubation with NAD protected PC12 cells against the loss of cell viability, morphological damage, oxidative stress and mitochondrial dysfunction caused by 6-OHDA. These results add credence to the beneficial role of NAD against parkinsonian neurodegeneration in mouse models of PD, provide evidence for the potential of NAD for the prevention of PD, and suggest that NAD prevents pathological changes in PD via decreasing mitochondrial dysfunctions.