Purpose This study aimed to reveal a new pathogenic gene locus linked to mitochondrial diseases. Methods The patient's clinical data were collected; peripheral blood specimens were obtained from him and his family. Variants were identified using exome sequencing and confirmed through Sanger sequencing and pedigree analysis. Functional validation of the variant locus was accomplished via in vitro vector construction, cell transfection, mRNA and protein level detection, nonsense-mediated mRNA decay (NMD), and guanosine triphosphatase levels. Results The proband presented with recurrent convulsive seizures, hyperlactatemia, and developmental delay. Brain magnetic resonance imaging showed symmetrical abnormal signals in the bilateral thalamus. Genetic testing revealed a compound heterozygous variant of GTPBP3: NC_000019.10(NM_133644.3):c.43_52del (var1) and NC_000019.10(NM_133644.4):c.872A>G (var2), derived from the father and mother (both heterozygous), respectively, and consistent with an autosomal recessive disorder. Both the proband and his parents had significantly lower GTPBP3 mRNA levels than those of healthy controls (p < 0.05). In vitro experiments revealed that var1 leads to decreased GTPBP3 expression through the NMD pathway; var2 affects oxidative phosphorylation by reducing guanosine triphosphatase activity. Conclusion Identification of a novel compound heterozygous variant of GTPBP3 related to mitochondrial diseases offers guidance for patient diagnosis and genetic counseling.

A ketogenic diet (KD) has demonstrated significant therapeutic efficacy in drug-resistant epilepsy. The molecular mechanisms through which KDs exert therapeutic effects on temporal lobe epilepsy (TLE) are not yet fully understood. Recent studies suggest that ferroptosis, a cell death pathway driven by iron-dependent lipid peroxidation, plays a role in the pathophysiological progression of epilepsy. This research revealed that lithium-pilocarpine (LI-PILO)-induced status epilepticus in TLE models triggered pronounced ferroptosis in the rat hippocampus and that KDs inhibited neuronal ferroptosis in the hippocampus, as evidenced by elevated levels of the antioxidant factors, glutathione (GSH) and catalase (CAT), and decreased levels of 4-HNE, Fe2+ and the lipid peroxidation product malondialdehyde (MDA). We also observed ferroptosis-related mitochondrial abnormalities, including reduced mitochondrial volume, disrupted cristae, and the outright disappearance of cristae, in the epilepsy model group. These morphological alterations were markedly attenuated following KD intervention. Furthermore, KDs alleviated both neuronal loss and cognitive impairment in TLE rats. However, the neuroprotective effects of KDs were completely abolished by the ferroptosis inducer erastin. In addition, treatment with the ferroptosis inhibitor ferrostatin-1 (Fer-1) not only reduced hippocampal neuronal damage, as confirmed by Nissl staining and immunofluorescence but also improved cognitive performance in TLE rats, as evidenced by better outcomes in the Morris water maze and novel object recognition tests. With respect to the underlying mechanism, multiomics analysis revealed that KDs alter circulating metabolite profiles. Notably, we revealed that deoxycholyl-L-dopa may be a key metabolite for targeting Keap1, xCT and HO-1. Western blot and qPCR results revealed that KDs activated the Nrf2/HO-1/GPX4 signaling axis and upregulated the expressions of Nrf2, HO-1, FTH1, xCT and GPX4. Our findings identify ferroptosis inhibition as a mechanism underlying the efficacy of KDs in epilepsy.

The objective of this study was to elucidate the molecular mechanisms by which cAMP-regulated transcription coactivator1 (CRTC1) regulates autophagy and GluA2 expression in patients with epilepsy. Methods: We initially established a magnesium-free epilepsy cell model and recorded cellular discharges using the whole-cell patch clamp technique. Next, we experimentally activated autophagy and identified effective methods for silencing the CRTC1 gene using RNA interference technology. Furthermore, we developed an animal models of status epilepticus and employed immunofluorescence and Western Blot to elucidate CRTC1’s role in regulating autophagy-related genes and GluA2 expression in epilepsy. Results: We observed mouse hippocampal neurons under magnesium-free extracellular conditions. Treatment with an autophagy activator decreased GluA2 expression; however, CRTC1 was not dephosphorylated. CRTC1 siRNA suppressed LC3 and PSD95 expression, whereas CRTC1 siRNA intervention restored GluA2 expression. Conclusion: CRTC1 indirectly influences the expression of synaptic-related proteins and GluA2 by directly modulating autophagy during the pathological process of epilepsy. The findings of this study reveal novel molecular targets for the treatment of epilepsy.

BACKGROUND AND PURPOSE:This study investigated the predictive value of clinical characteristics and cerebrospinal fluid (CSF) metabolites for nusinersen efficacy in children with spinal muscular atrophy (SMA). METHODS:In this study, clinical data and CSF samples were collected. We used liquid chromatography-tandem mass spectrometry to analyze CSF metabolites from 42 patients with type II and type III SMA. RESULTS:Although clinical indicators, such as age at treatment initiation and disease duration, did not predict the clinical efficacy of nusinersen, we identified 47 differentially expressed metabolites between effective- and ineffective-treatment patients with type II disease and 109 metabolites in patients with type III disease. Additionally, KEGG-enriched pathway analysis revealed differences in several pathways between the effective- and ineffective-treatment groups for both types II and III. N-myristoyl arginine and 1,1,1,2,2,2-Pentafluoro-7-phenylheptan-3-one were negatively associated with Hammersmith Functional Motor Scale Expanded changes in patients with type III and type III SMA. Furthermore, multivariate receiver operating characteristic curve analysis indicated that differential metabolites have some accuracy in predicting SMA treatment efficacy. CONCLUSION:This study identified CSF metabolites that are predictive of nusinersen efficacy. The results of this study may guide the development of adjunctive therapies for improving the efficacy of nusinersen.

OBJECTIVE:To investigate the clinical efficacy and safety of the combination of lacosamide (LCM) and oxcarbazepine (OXC) in children with focal epilepsy, either as a dual therapy or in combination with other anti-seizure medications. METHODS:We collected clinical data retrospectively from pediatric epilepsy patients treated with both LCM and OXC for at least 6 months at the Department of Pediatrics, Second Affiliated Hospital of Xi'an Jiaotong University, between January 2021 and July 2023. Efficacy and adverse events were investigated. RESULTS:Collectively, 69 pediatric patients were included, with ages ranging from 1 to 18 years. The age at onset of epilepsy varied from 15 days to 14.8 years. Disease duration ranged from 1 month to 14 years. All pediatric patients exhibited focal seizures, with the maximum number of concurrent combined medications being six. The maintenance dose of LCM for all pediatric patients was between 3.8 and 10 mg/kg, while that of OXC was between 15 and 40 mg/kg. Over half of the patients showed a greater than 50 % reduction in seizure frequency during long-term follow-up. The most frequently reported adverse reactions included hyponatremia, headache, dizziness, somnolence, and nausea; no patients experienced rash as an adverse reaction. CONCLUSION:The combination of LCM and OXC has shown promising efficacy and tolerability in pediatric patients with focal epilepsy. This therapeutic combination is a worthwhile option to consider for patients with focal epilepsy, especially those with underlying structural abnormalities.

Objective:This study aimed to observe the neurophysiological characteristics of type II and type III 5q spinal muscular atrophy (SMA) patients and the changes in peripheral motor nerve electrophysiology after Nusinersen treatment, as well as the influencing factors.Methods:This single-center retrospective case-control study collected clinical data and peripheral motor nerve CMAP parameters from 42 5qSMA patients and 42 healthy controls at the Second Affiliated Hospital of Xi'an Jiaotong University (January 2021 to December 2022). It evaluated changes in motor function and CMAP amplitude before and after Nusinersen treatment.Results:Our investigation encompassed all symptomatic and genetically confirmed SMA patients, consisting of 32 type II and 10 type III cases, with a median age of 57 months (29.5 to 96 months). Comparative analysis with healthy controls revealed substantial reductions in CMAP amplitudes across various nerves in both type II and type III patients. Despite the administration of Nusinersen treatment for 6 or 14 months to the entire cohort, discernible alterations in motor nerve amplitudes were not observed, except for a significant improvement in younger patients (≤36 months) at the 14-month mark. Further scrutiny within the type II subgroup unveiled that individuals with a disease duration ≤12 months experienced a noteworthy upswing in femoral nerve amplitude, a statistically significant difference when compared to those with >12 months of disease duration.Conclusion:Motor nerve amplitudes were significantly decreased in type II and type III 5q SMA patients compared to healthy controls. Nusinersen treatment showed better improvement in motor nerve amplitudes in younger age groups and those with shorter disease duration, indicating a treatment-time dependence.

Abstract ID 127906Poster Board 336Neuropsychiatric disorders present significant challenges due to their complex etiology, involving inflammatory responses, neurotransmitter dysregulation, and altered gut-brain interactions. Recent interdisciplinary research uncovers the gut-brain axis (GBA), connecting the central and enteric nervous systems through various pathways. The intestinal microbiota, crucial to the GBA, impacts brain function via bacterial metabolic products like butyrate. This study addresses these complexities by examining the role of butyrate in modulating the influence of proinflammatory responses and oxidative stress on the transport of neurotransmitter precursor amino acids tyrosine and tryptophan and gene expression of these transporters.Fibroblasts derived from human skin obtained from a healthy control were used as model cells for the blood-brain barrier in this research project. Fibroblasts were treated with proinflammatory cytokines (IL-1β, IL-6, IFN-γ, and TNF- α) and oxidative stress (hydrogen peroxide) to assess the uptake of 14C tyrosine. Additionally, fibroblasts were treated with oxidative stress and different concentrations of butyrate to measure the transport of 3H tryptophan across the cell membranes. Furthermore, gene expression profiles in response to oxidative stress and butyrate treatment of tyrosine and tryptophan amino acid transporters were analyzed.Proinflammatory cytokines and oxidative stress were found to significantly decrease tyrosine transport. Oxidative stress significantly decreased tryptophan uptake, while butyrate counteracted this effect. However, treatment of oxidative stressed and control cells with different concentrations of butyrate differentially regulated the gene expression of large amino acid transporters 1 and 2, which are the major transporters of tyrosine and tryptophan.In conclusion, this research project elucidated the pathophysiological mechanisms of proinflammatory responses in neuropsychiatric disorders. Moreover, it highlights the therapeutic potential of butyrate and emphasizes the significance of gut-brain interactions in these conditions. By shedding light on these aspects, this research contributes to the development of innovative and personalized treatment strategies for enhanced management of neuropsychiatric disorders.

PURPOSE:To draw clinical attention to rashes caused by lacosamide. METHODS:This retrospective analysis included patients admitted to the Department of Pediatrics, Second Affiliated Hospital of Xi'an Jiaotong University between January 2021 and September 2023. We focused on patients who developed rashes after lacosamide treatment and analyzed all patients who exhibited rashes after lacosamide treatment to analyze the risk factors. RESULTS:In total, 190 patients received lacosamide, of whom four developed allergies (2.1 %). Three patients had severe rashes, and two patients had high fever. All of these adverse events improved after the withdrawal of lacosamide. Of the 13 patients reported to date, including the four patients in this study, eight used various antiseizure medicines, including seven patients who used four or more antiseizure medicines. Four patients underwent testing for HLA-B*1502, and two patients were positive. Patients developed rashes within 1-10 days after treatment initiation, and the duration of the rash ranged 2-37 days. CONCLUSIONS:Lacosamide-induced rash was detected in 2.1 % of patients in our cohort. Rashes are potentially serious, and prompt evaluation is required. Rashes are more likely to occur when multiple antiseizure medicines are used simultaneously, typically within 10 days of treatment initiation.

AIMS:Classification of spinal muscular atrophy (SMA) is associated with the clinical prognosis; however, objective classification markers are scarce. This study aimed to identify metabolic markers in the cerebrospinal fluid (CSF) of children with SMA types II and III.METHODS:CSF samples were collected from 40 patients with SMA (27 with type II and 13 with type III) and analyzed for metabolites.RESULTS:We identified 135 metabolites associated with SMA types II and III. These were associated with lysine degradation and arginine, proline, and tyrosine metabolism. We identified seven metabolites associated with the Hammersmith Functional Motor Scale: 4-chlorophenylacetic acid, adb-chminaca,(+/-)-, dodecyl benzenesulfonic acid, norethindrone acetate, 4-(undecan-5-yl) benzene-1-sulfonic acid, dihydromaleimide beta-d-glucoside, and cinobufagin. Potential typing biomarkers, N-cyclohexylformamide, cinobufagin, cotinine glucuronide, N-myristoyl arginine, 4-chlorophenylacetic acid, geranic acid, 4-(undecan-5-yl) benzene, and 7,8-diamino pelargonate, showed good predictive performance. Among these, N-myristoyl arginine was unaffected by the gene phenotype.CONCLUSION:This study identified metabolic markers are promising candidate prognostic factors for SMA. We also identified the metabolic pathways associated with the severity of SMA. These assessments can help predict the outcomes of screening SMA classification biomarkers.

Although the ketogenic diet (KD) is known to control seizures and improve cognition function in patients with drug-refractory epilepsy, the underlying mechanism remains unknown. In the present study, using pentylenetetrazol (PTZ)-induced and kindled rats, we found that KD significantly improved the impaired spatial reference memory of PTZ-kindled rats in the Morris water maze. To explore the mechanism underlying the action of KD in PTZ-kindled rats, quantitative real-time PCR (qRT-PCR) and immunohistochemical analysis were used to detect the expression of GluR1 and NR2B. The results showed that both the mRNA and protein expression of GluR1 and NR2B were significantly downregulated in the hippocampus of PTZ-kindled rats, while KD could observably improve both the mRNA and protein expression of GluR1 and NR2B in the hippocampus of PTZ-kindled rats. Additionally, KD improved the over-activated MAPK in PTZ-kindled rats, but not CAMKII, as detected by enzyme-linked immuno sorbent assay (ELISA), suggesting that the MAPK signaling pathway might be involved in the memory improvement of KD in PTZ-kindled rats. In conclusion, these results demonstrate that KD can indeed improve impaired spatial reference memory in PTZ-kindled rats, and KD can improve the expression of NR2B and GluR1.

Background Ischemic heart disease (IHD) is a common cardiovascular disorder associated with inadequate blood supply to the myocardium. Chronic coronary ischemia leads to ischemic cardiomyopathy (ICM). Despite their rising prevalence and morbidity, few studies have discussed the lipids alterations in these patients. Methods In this cross-sectional study, we analyzed serum lipids profile in IHD and ICM patients using a lipidomics approach. Consecutive consenting patients admitted to the hospital for IHD and ICM were enrolled. Serum samples were obtained after overnight fasting. Non-targeted metabolomics was applied to demonstrate lipids metabolic profile in control, IHD and ICM patients. Results A total of 63 and 62 lipids were detected in negative and positive ion mode respectively. Among them, 16:0 Lyso PI, 18:1 Lyso PI in negative ion mode, and 19:0 Lyso PC, 12:0 SM d18:1/12:0, 15:0 Lyso PC, 17:0 PC, 18:1-18:0 PC in positive ion mode were significantly altered both in IHD and ICM as compared to control. 13:0 Lyso PI, 18:0 Lyso PI, 16:0 PE, 14:0 PC DMPC, 16:0 ceramide, 18:0 ceramide in negative ion mode, and 17:0 PE, 19:0 PC, 14:0 Lyso PC, 20:0 Lyso PC, 18:0 PC DSPC, 18:0-22:6 PC in positive ion mode were significantly altered only in ICM as compared to IHD and control. Conclusion Using non-targeted lipidomics profiling, we have successfully identified a group of circulating lipids that were significantly altered in IHD and ICM. The lipids metabolic signatures shed light on potential new biomarkers and therapeutics for preventing and treating ICM.

Role of butyrate in counteracting the influence of oxidative stress on amino acid transport implicated in neuropsychiatric disorders

OBJECTIVE:To study the effect of cyclooxygenase -2 selective inhibitor celecoxib on the expression of major vault protein ( MVP) in the brain of rats with status epilepticus and its possible roles in the treatment of refractory epilepsy.METHODS:Sixty adult male Sprague-Dawley rats were randomly assigned to blank control (n=16), epilepsy model (n=22) and celecoxib treatment groups (n=22). After the status epilepticus was induced in rats by injecting lithium and pilocarpine, each group had 16 rats enrolled as subjects. Immunohistochemical method and Western blot method were used to detect the expression of MVP in the frontal cortex and hippocampus.RESULTS:The expression of MVP was significantly higher in the epilepsy model group than in the control group (P<0.01). The expression of MVP in the celecoxib treatment group was significantly decreased compared with the epilepsy model group, but it was still higher than in the control group (P<0.01).CONCLUSIONS:Celecoxib could decrease the expression of MVP in brain tissue of rats with status epilepticus, suggesting that it is promising for the treatment of intractable epilepsy.