We report a case of a Chinese girl who presented with multiple seizure types of epilepsy, followed by motor and intellectual regression, vision impairment, and cerebral and cerebellar atrophy. She carries an unreported compound heterozygous variant of the ASAH1 gene and is diagnosed with spinal muscular atrophy associated with progressive myoclonic epilepsy (SMA-PME), a disorder in which ceramide accumulation in lysosomes due to a decrease in acid ceramidase activity. This case suggests attention to this rare class of deceases involving both the central and peripheral nervous systems.

AIM:Drug resistance is an intractable issue urgently needed to be overcome for improving efficiency of antiepileptic drugs in treating refractory epilepsy. microRNAs (miRNAs) have been proved as key regulators and therapeutic targets in epilepsy. Accordingly, the aim of the present study was to identify a novel differentially expressed miRNA which could improve the efficiency of antiepileptic drugs during the treatment of refractory epilepsy.METHODS AND RESULTS:Serum samples were collected from children with refractory epilepsy. An in vivo refractory epilepsy model was developed in SD rats by electrical amygdala kindling. We identified that miR-139-5p was decreased and multidrug resistance-associated protein 1 (MRP1) was remarkably upregulated in the serum samples from children with refractory epilepsy and the brain tissues from rat models of refractory epilepsy. After phenobarbitone injection in rat models of refractory epilepsy, the after discharging threshold in kindled amygdala was detected to screen out drug-resistant rats. Dual-luciferase reporter gene assay demonstrated that MRP1 was a target of miR-139-5p. In order to evaluate the effect of miR-139-5p/MRP1 axis on drug resistance of refractory epilepsy, we transfected plasmids into the hippocampus of drug-resistant rats to alter the expression of miR-139-5p and MRP1. TUNEL staining and Nissl staining showed that miR-139-5p overexpression or MRP1 downregulation could reduce the apoptosis and promote survival of neurons, accompanied by alleviated neuronal damage.CONCLUSION:Collectively, these results suggest an important role of miR-139-5p/MRP1 axis in reducing the resistance of refractory epilepsy to antiepileptic drugs.

OBJECTIVE:To explore the role of miR-181b in alterations of apoptosis and autophagy in the kainic acid (KA)-induced epileptic juvenile rats via modulating TLR4 and P38/JNK signaling pathway.METHODS:Dual-luciferase reporter assay was performed to testify the targeting relationship between miR-181b and TLR4. After intracerebroventricular injection (i.c.v.) of KA, rats were injected with miR-181b agomir and TLR4 inhibitor (TAK-242). The TLR-4 activator lipopolysaccharide (LPS) was also administered into rats immediately after injection with miR-181b agomir. Quantitative real-time-polymerase chain reaction (qRT-PCR) was used for detections of miR-181b and TLR4 expressions, hematoxylin-eosin (HE) and Nissl staining for observation of the hippocampus morphological changes, and TUNEL staining for apoptosis analysis. Moreover, western blot was determined to detect TLR4 and P38/JNK pathway proteins, as well as autophagy- and apoptosis-related proteins.RESULTS:TLR4 was identified as a direct target of miR-181b using Dual-luciferase reporter assay. KA rats injected with miR-181b agomir or TAK-242 had improved learning and memory abilities, reduced seizure severity of Racine's scale, and lessened neuron injury. Additionally, miR-181b agomir or TAK-242 could significantly inhibit P38/JNK signaling, decrease LC3II/I, Beclin-1, ATG5, ATG7, ATG12, Bax, and cleaved caspases-3, but increase p62 and Bcl-2 expression. No significances were found between KA group and KA + miR-181b + LPS group.CONCLUSION:MiR-181b could inhibit P38/JNK signaling pathway via targeting TLR4, thereby exerting protective roles in attenuating autophagy and apoptosis of KA-induced epileptic juvenile rats.

目的 探讨不同剂量拉莫三嗪对戊四氮所致癫痫大鼠的行为学、海马组织形态学及Caspase-3和热休克蛋白70(HSP70)表达的影响.方法 选取清洁级健康6周龄雄性Sprague-Dawley大鼠100只,随机分为正常对照组、模型组和拉莫三嗪低、中、高剂量组,每组20只.正常对照组大鼠腹腔注射生理盐水(3.5 mL·kg-1),其余各组大鼠均腹腔注射戊四氮(35 mg·kg-1)并观察其行为学变化.以连续出现3次Ⅳ级及其以上发作作为点燃标准.全部点燃后,模型组和拉莫三嗪低、中、高剂量组大鼠给予戊四氮35 mg·kg-1每日1次腹腔注射维持点燃,拉莫三嗪低、中、高剂量大鼠组分别给予拉莫三嗪20、30、60 mg·kg-1·d-1灌胃,连续2周.在末次灌胃24 h内,对各组大鼠进行心脏灌注固定取海马组织,采用免疫组织化学法检测海马CA3区Caspase-3和HSP70蛋白表达.结果 除正常对照组外,其余各组大鼠均有痫性发作.正常对照组大鼠海马结构完整,其他组大鼠海马神经元体积缩小,与周围组织有脱离,细胞核深染,有核分裂及核溶解,具有凋亡特征,多数细胞质呈空泡状,海马区神经元细胞排列散乱,无层次感,尼氏体减少,部分神经元丢失,细胞密度小.与正常对照组比较,模型组大鼠海马CA3区Caspase-3和HSP70阳性细胞数增多(P<0.01);拉莫三嗪低、中、高剂量组大鼠海马CA3区Caspase-3阳性细胞数均少于模型组,HSP70阳性细胞数均多于模型组(P<0.05);拉莫三嗪低、中、高剂量组大鼠海马CA3区Caspase-3阳性细胞数随着剂量的增加而减少,HSP70阳性细胞数随着剂量的增加而增加,拉莫三嗪低、中、高剂量组大鼠海马CA3区Caspase-3、HSP70阳性细胞数两两比较差异均有统计学意义(P<0.05).与正常对照组比较,模型组大鼠海马CA3区Caspase-3及HSP70蛋白表达均增加(P<0.05).拉莫三嗪低、中、高剂量组Caspase-3蛋白表达低于模型组(P<0.05),HSP70蛋白表达高于模型组(P<0.05).拉莫三嗪低、中、高剂量组大鼠海马CA3区Caspase-3蛋白表达随着剂量的增加而降低,HSP70蛋白表达随着剂量的增加而增加,拉莫三嗪低、中、高剂量组大鼠海马CA3区Caspase-3、HSP70蛋白表达两两比较差异均有统计学意义(P<0.05).结论 拉莫三嗪对戊四氮所致癫痫大鼠的癫痫发作有保护作用,可以减轻大鼠癫痫发作的严重程度,其机制可能是通过降低海马组织中Caspase-3表达、增加HSP70表达来抑制癫痫过程中的神经元凋亡.

目的 探讨拉莫三嗪对戊四氮致癫痫大鼠认知功能及海马组织中Bax、Bcl-2蛋白表达的影响.方法 将健康雄性Sprague-Dawley大鼠100只随机分为正常组、模型组及拉莫三嗪低、中、高剂量组,每组20只.模型组及拉莫三嗪低、中、高剂量组大鼠每天腹腔注射戊四氮35 mg·kg-1进行造模;正常组大鼠腹腔注射同体积生理盐水.造模成功后,正常组大鼠继续每日腹腔注射生理盐水;其余4组大鼠继续腹腔注射戊四氮,在此基础上,拉莫三嗪低、中、高剂量组大鼠分别给予拉莫三嗪20、30、60 mg·kg-1·d-1灌胃,模型组大鼠给予生理盐水,共2周.用Morris水迷宫实验评价大鼠的认知功能,免疫组织化学方法检测大鼠海马组织中Bax、Bcl-2蛋白表达.结果 与正常组比较,模型组大鼠各时间点逃避潜伏期显著延长(P＜0.05);拉莫三嗪低、中、高剂量组大鼠各时间点逃避潜伏期均短于模型组(P＜0.05,P＜0.01);拉莫三嗪低、中、高剂量组大鼠逃避潜伏期两两比较差异均有统计学意义(P＜0.05).空间探索实验结果显示,模型组大鼠记忆探索正确时间短于正常组(P＜0.05);与模型组比较,拉莫三嗪低、中、高剂量组大鼠记忆探索正确时间显著增加(P＜0.05);拉莫三嗪低、中、高剂量组大鼠记忆探索正确时间两两比较差异均有统计学意义(P＜0.05).与正常组比较,模型组大鼠海马组织中Bax蛋白表达增加,Bcl-2蛋白表达减少(P＜0.01);拉莫三嗪低、中、高剂量组大鼠海马组织中Bax蛋白表达均低于模型组,Bcl-2蛋白表达均高于模型组(P＜0.05);拉莫三嗪低、中、高剂量组之间大鼠海马组织中Bax、Bcl-2蛋白表达两两比较差异均有统计学意义(P＜0.05).结论 拉莫三嗪对戊四氮致癫痫大鼠海马神经元有保护作用,并能提高大鼠的学习、记忆和判断能力.

Epilepsy is a chronic disease of the nervous system which is commonly contracted by children.The cognitive abilities of these affected children are mostly in an average level ,but still lower as a whole than those of the normal children .The cognitive impairment caused by epilepsy is mainly reflected in the children′s decreasing ability of integrating the external information .The extent of the damage caused by this impairment to the integrating ability can be decided by the epilepsy′s seizure type ,age of onset ,affection process,seizure frequency,disease duration,lesion focus,basic diseases/causes that may lead to epilepsy, social psychology,and so on.For a long time,the most important objective of epilepsy treatments has been to take control of epilepsy seizures .A great deal of current experiments and researches has demonstrated that the theory of newly-invented anti-epilepsy medicines is to contain epilepsy seizures through neurotransmission ,ion channels,second messenger and other links .In this way,the cognitive impairment is relieved and the patient′s life quality improved .However , one disadvantage of this treatment is its harmfulness to the children′s cognitive functions in some ways.Antiepileptic drugs through the neurotransmitter ,ion channels,the second messenger ,and other links to adjust control seizures .Thus reduce cognitive impairment of epilepsy , improve the quality of life of patients ,but also affect children′s cognitive function in some ways .

Objective To investigate the effect of hydrogen sulfide(H2S) on expressions of interleukin-1β(IL-1β)and IL-18 in hippocampus of rats with recurrent febrile seizures(FS).Methods Sprague-Dawlery rats were randomly divided into 4 groups: control group,FS group,FS+ sodium hydrosulfide(NaHS) group and FS+hydroxylamine(HA) group.The FS rats were bathed in warm water 10 times,once every 3-5 minutes;the pathological changes in hippocampus was observed by Toluidine blue staining;the protein expressions of IL-1β,IL-18 were detected by immunohistochemistry;and the expressions of IL-1β and IL-18 mRNA were measured by reverse transcription polymerase chain reaction.Results The expressions of IL-1β and IL-18 proteins and mRNA in FS group were significant higher than those in control group.Toluidine blue staining showed that the hippocampal neurons′s arrangement was indiscriminate,cell nucleus size was different,part cell showed vacuolar degeneration,concentration,and syneresis.The levels of IL-1β proteins and mRNA in FS+NaHS group at different points were significant lower as compared with those in FS group,the expressions of IL-18 proteins and mRNA in the hippocampus in FS +NaHS group at post-seizures 1 d,3 d,7 d at different points were significant lower as compared with those in FS group.About the expressions of IL-1β proteins and mRNA,FS+HA group of 6 h,3 d,7 d were significantly higher than those in FS group.The levels of IL-18 in FS+HA group increased significantly compared with those in FS group at post-seizures 6 h.The levels of IL-1β and IL-18 in FS+NaHS group at different points were significant lower compared with those in HA group.Conclusion H2S can protect the brain injury which induced by recurrent FS,through inhibiting the expressions of IL-1β and IL-18.