MDH2 is an RNA binding protein involved in downregulation of sodium channel Scn1a expression under seizure condition.

Voltage-gated sodium channel α-subunit type I (NaV1.1, encoded by SCN1A gene) plays a critical role in the excitability of brain. Downregulation of SCN1A expression is associated with epilepsy, a common neurological disorder characterized by recurrent seizures. Here we reveal a novel role of malate dehydrogenase 2 (MDH2) in the posttranscriptional regulation of SCN1A expression under seizure condition. We identified that MDH2 was an RNA binding protein that could bind two of the four conserved regions in the 3' UTRs of SCN1A. We further showed that knockdown of MDH2 or inactivation of MDH2 activity in HEK-293 cells increased the reporter gene expression through the 3' UTR of SCN1A, and MDH2 overexpression decreased gene expression by affecting mRNA stability. In the hippocampus of seizure mice, the upregulation of MDH2 expression contributed to the decrease of the NaV1.1 levels at posttranscriptional level. In addition, we showed that the H2O2 levels increased in the hippocampus of the seizure mice, and H2O2 could promote the binding of MDH2 to the binding sites of Scn1a gene, whereas ß-mercaptoethanol decreased the binding capability, indicating an important effect of the seizure-induced oxidation on the MDH2-mediated downregulation of Scn1a expression. Taken together, these data suggest that MDH2, functioning as an RNA-binding protein, is involved in the posttranscriptional downregulation of SCN1A expression under seizure condition.

Effects of Embryo Microbial Contamination on ART and Neonatal Outcomes.

Purpose: This study investigated the incidence and microbial etiology of embryo contamination in assisted reproductive technology (ART), and its influence on embryo development, pregnancy and neonatal outcomes. Methods: A retrospective analysis was conducted on embryo contamination at the Reproductive Centre of the Third Affiliated Hospital of Guangzhou Medical University, between 2018 and 2021. Results: In the period from 2018 to 2021, the average incidence of embryo contamination was 0.12%. Bacterial growth was observed in 39 cases, with a preponderance of Escherichia coli (20, 51.28%), Streptococcus agalactiae (7, 17.95%). The fertilization rate of contaminated embryos was 18.18% (Klebsiella pneumoniae) to 94.79% (S. agalactiae), the cleavage rate was 9.09% (Enterobacter cloacae) to 98.90% (S. agalactiae), and the available embryo rate of Day 3 was 0 (Klebsiella pneumoniae, Enterobacter cloacae) to 63.33% (S. agalactiae). Blastocyst formation rate was 3.23% (Proteus mirabilis) to 64.29% (Streptococcus mitis). E. coli contamination occurred mostly on Day 1, and S. agalactiae on Days 3 and 5. After rinsing and rescuing treatment, six healthy male babies were born. Conclusion: E. coli and S. agalactiae were the most common bacterial embryo contaminants. Most microbial contamination can significantly decrease the fertilization rate. Embryo transfer after rinsing and continuing culture had no negative effect on neonatal outcomes, but there was an increased risk of early abortion due to E. coli contamination.

Epigenetic Downregulation of Scn3a Expression by Valproate: a Possible Role in Its Anticonvulsant Activity.

Upregulation of sodium channel SCN3A expression in epileptic tissues is known to contribute to enhancing neuronal excitability and the development of epilepsy. Therefore, certain strategies to reduce SCN3A expression may be helpful for seizure control. Here, we reveal a novel role of valproate (VPA) in the epigenetic downregulation of Scn3a expression. We found that VPA, instead of carbamazepine (CBZ) and lamotrigine (LTG), could significantly downregulate Scn3a expression in mouse Neuro-2a cells. Luciferase assays and CpG methylation analyses showed that VPA induced the methylation at the -39C site in Scn3a promoter which decreased the promoter activity. We further showed that VPA downregulated the expression of methyl-CpG-binding domain protein 2 (MBD2) at the posttranscriptional level and knockdown of MBD2 increased Scn3a expression. In addition, we found that VPA induced the expression of fat mass and obesity-associated (FTO) protein and FTO knockdown abolished the repressive effects of VPA on MBD2 and Nav1.3 expressions. Furthermore, VPA, instead of other two anticonvulsant drugs, induced the expressions of Scn3a and Mbd2 and reduced Fto expression in the hippocampus of VPA-treated seizure mice. Taken together, this study suggests an epigenetic pathway for the VPA-induced downregulation of Scn3a expression, which provides a possible role of this pathway in the anticonvulsant action of VPA.