Purpose To explore the efficacy and safety of intravenous immunoglobulin (IVIG) in the treatment of severe Japanese encephalitis (JE). Methods A retrospective study of 124 children diagnosed with the severe or very severe form of JE was undertaken. There were 62 cases in the IVIG group and control group. The efficacy, safety, and tolerability of IVIG were evaluated 3 days, as well as 1, 2, and 3 weeks after IVIG, respectively, and the prognosis was assessed at 6 months. Results Cox regression survival analysis suggested that the IVIG group reached the criteria for efficacious treatment faster than that in the control group. The duration of unconsciousness and the number of days of seizures, a dull response to light, the pyramidal sign, and meningeal-irritation sign in the IVIG group were shorter than those in the control group (p < 0.05). The number of complications occurring in the IVIG group (including gastrointestinal bleeding and pneumonia) was less than that in the control group (p < 0.05). Cox regression survival analysis suggested that age (p = 0.003) and imaging abnormalities (p = 0.042) had an effect on the efficacy of IVIG treatment. The Liverpool Outcome Score at 6 months showed that the prognosis of the IVIG group was better than that of the control group (p < 0.05). IVIG treatment was safe and tolerable. Conclusion IVIG showed good efficacy, safety, and tolerance for treatment of the severe form of JE. The age and imaging abnormalities of patients affect the efficacy of IVIG treatment.

The descending serotonergic pathway, which originates in various populations of brainstem neurons, plays an important role in generating the rhythmic motor pattern associated with locomotor movement. Although the development of its innervation has been studied in rodent spinal cord, it has not been clearly identified how the projection of serotonergic pathway is related to its function. Here, we evaluated the pattern of serotonergic innervation on the lumbar spinal cord from embryonic day 14.5 (E14.5) to adulthood. Before birth, we found that 5-hydroxytryptamine (5-HT) fibers invade the lumbar cord as early as E14.5, penetrate into the gray matter from lateral funiculus by E16.5, and then mainly occupied the ventral horn by E18.5 before localizing in the dorsal horn. After birth, we found that 5-HT invasion of both dorsal horn and ventral horn were present by the 7th postnatal day (P7). Additionally, the 5-HT innervation of these two areas evolved progressively from a diffuse network to a more restricted pattern, particularly at the ventral horn within the motoneuron area from P21 to adulthood. This 5-HT innervation pattern in the lumbar cord provides anatomical evidence that serotonergic fibers establish direct connections with lumbar motoneurons, which offers us a solid foundation that enhancing the plasticity of serotonergic pathway following SCI may facilitate locomotor functional recovery. Therefore, we employed treadmill training to activate serotonergic plasticity after SCI. We found that mice which underwent treadmill training exhibited a better locomotor functional recovery. Meanwhile, the density of 5-HT fibers in the ventral horn was significantly increased and the synaptic formation of 5-HT fibers with lumbar motoneurons was also significantly rescued in the training group mice after SCI. These findings demonstrate that the descending serotonergic projection is a robust and flexible parallel pathway for modulating spinal locomotor function.

The aim of this study was to investigate the pathogenesis of autophagy and apoptosis mediated by Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signal pathway after the onset of acute spinal cord injury (ASCI). A total of 45 Sprague-Dawley adult rats of either sex were selected for this study. The age of rats ranged from 8 to 10 weeks, and the average weight was 245 g. These rats were randomly divided into three groups, i.e. sham-operated group, model group, and the AG-490 intervention group (AG-490 is an inhibitor of JAK2). Each group contained 15 rats. Models were prepared using the modified Allen method. Five rats in each group were sacrificed at 6, 12 and 24 h, respectively, and the expression levels of p-JAK2 and p-STAT3 were detected in spinal cord tissue via western blot analysis. The levels of proinflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected via ELISA, positive expression of light chain 3 (LC3)-II of microtubule-associated protein 1 via immunofluorescence labeling method, and mRNA expression levels of caspase-3 and Bax/Bcl-2 via RT-PCR. In the model group, the expression levels of p-JAK2, p-STAT3, IL-6, TNF-α and LC3-II, and the mRNA expression levels of caspase-3 and Bax/Bcl-2 at all time-points were significantly higher than those in the AG-490 intervention group, and the levels in the sham-operated group were the lowest (p<0.05). In the model group, peak levels of p-JAK2 and p-STAT3 were attained at 12 h, but a decline was seen at 24 h; while increasing trend was seen in other indicators. In conclusion, JAK2/STAT3 signal pathway can mediate the activity of autophagy and apoptosis in an early stage after the onset of ASCI of rat.

Purpose Previous studies have demonstrated that fibronectin (FN) levels are increased in brain tissues from patients and animals with epilepsy. This study aimed to assess FN levels in cerebrospinal fluid (CSF) and serum samples from patients with epilepsy. Methods Fibronectin levels were assessed in CSF and serum samples from 56 patients with epilepsy (27 and 29 individuals with intractable epilepsy and nonintractable epilepsy, respectively) and 25 healthy controls, using sandwich enzyme-linked immunosorbent assays (ELISA). Results CSF-FN levels were higher in patients with epilepsy (8.07 ± 1.51 mg/l versus 6.20 ± 1.18 mg/l, p < 0.05) than in the control group. In addition, serum-FN levels in the group with epilepsy and in the control group were 236.96 ± 65.7 mg/l and 181.43 ± 72.82 mg/l, respectively, indicating a statistically significant difference (p = 0.01). Interestingly, serum- and CSF-FN levels in individuals with epilepsy were not affected by antiepileptic drug and duration of epilepsy. Of note, the increase of CSF- and serum-FN levels was more pronounced in subjects with intractable epilepsy than in patients with nonintractable epilepsy. Conclusion Serum- and CSF-FN levels constitute a potential clinical diagnostic biomarker for epilepsy and could also be used for differential diagnosis.

Mounting evidence supports the involvement of brain inflammation and the associated blood–brain barrier damage from which spontaneous and recurrent seizures originate. Detection of the soluble form of adhesion molecules (AM) has also been proven to predict outcomes in central nervous system (CNS) disorders. A recent study has shown that expression of AM in brain vessels was upregulated 24 h after kainic acid (KA) induced seizures. The aim of the present study was to investigate soluble AM levels in the cerebrospinal fluid (CSF) and serum of epilepsy patients. Paired CSF and serum samples were analyzed by sandwich enzyme-linked immunosorbent assay (ELISA) to determine the concentrations of soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-1 (sICAM-1). Increased serum concentrations of sICAM-1 were present in epileptic patients (41 localization-related of unknown etiology, 19 idiopathic generalized). Serum sICAM-1 level in drug-refractory epilepsy was elevated as compared to new diagnosis epilepsy and drug-responsive epilepsy. CSF sVCAM-1 and serum sVCAM-1 concentrations in the epilepsy group were higher as compared to the neurosis group. Moreover, CSF sVCAM-1 and serum sVCAM-1 concentrations in drug-refractory epilepsy were raised as compared to drug-responsive epilepsy and new diagnosis epilepsy. However, there were no significant differences in concentrations of CSF sICAM-1 between the epilepsy and neurosis groups. Our results suggest that sVCAM-1 and sICAM-1 could play an important role in the drug-refractory epilepsy.

Our previous study on proteomic analysis has shown that clusterin (CLU) is significantly decreased in the cerebrospinal fluid (CSF) of patients with epilepsy. Therefore, the present study aimed to confirm CLU concentration reduction in the CSF of patients with drug-resistant epilepsy and drug-responsive epilepsy. Fifty-two patients with epilepsy (23 drug resistance and 29 drug effectivity) and 20 control individuals were recruited. The concentrations of CSF and serum CLU were detected. Moreover, alteration of CLU was detected in the rat hippocampus over time after pilocarpine-induced status epilepticus (SE). Our results showed that human CSF-CLU levels were decreased in patients with both drug-resistant epilepsy and drug-responsive epilepsy compared to controls, and concentration of CSF-CLU was obviously lower in drug-resistant epilepsy than in drug-responsive epilepsy. In the pilocarpine-induced seizure rats, expression of neuronal CLU was gradually decreased in a time-dependent manner from acute phase to chronic phase after the onset of SE. In conclusion, CLU level is decreased in the CSF of human epilepsy and the similar alteration is confirmed in a rat model with epilepsy. Therefore, CLU might contribute to the development of epilepsy and be a potential CSF biomarker for resistant epilepsy.

PurposeOur previous study using proteomic analysis showed that superoxide dismutase 1 (SOD1) was significantly decreased in cerebrospinal fluid (CSF) of patients with epilepsy. However, the relevance of CSF-SOD1 alterations for the pathophysiology of epilepsy is currently unknown. The present study was intended to add to our understanding of this issue by measuring SOD1 levels in the CSF of patients with resistant epilepsy and non-resistant epilepsy.MethodsA total of 52 patients with epilepsy were recruited. 29 were non-resistant, 23 drug-resistant. 20 individuals with no evidence of any neurological diseases were used as control. The concentration of CSF and serum SOD1 was measured by enzyme-linked immunosorbent assay.ResultsThe concentration of CSF-SOD1 was decreased in both the drug-resistant (0.13 ± 0.12 ng/ml) and the non-resistant epilepsy subgroups (0.29 ± 0.23 ng/ml) compared to the control group (0.40 ± 0.35 ng/ml). SOD1 was significantly lower in the drug-resistant than the non-resistant epilepsy subgroup (P < 0.05).ConclusionSOD1 levels are decreased in the CSF of patients with epilepsy, especially of patients with intractable epilepsy. Low CSF-SOD1 levels may be a predictor of antiepileptic drug resistance in patients with epilepsy.

Recent studies suggest that angiogenesis and vascular endothelial growth factor (VEGF) are involved in the pathophysiology of epilepsy. However, relatively little data are available linking placenta growth factor (PIGF) with epilepsy. In this study, we assessed concentrations of PIGF in cerebrospinal fluid (CSF) of 60 epileptic patients and 24 non-seizure subjects using sandwich enzyme-linked immunosorbent assays. Epileptic patients in general had higher concentration of CSF-PIGF than controls (7.95 ± 0.88 ng/l vs. 5.87 ± 0.79 ng/l, P < 0.01). CSF-PIGF level in secondary epileptic patients (8.59 ± 1.26 ng/l) was higher than that in idiopathic epileptic patients (7.62 ± 0.20 ng/l) (P < 0.05). In idiopathic epilepsy, CSF-PIGF level in patients with high seizure frequency was higher than those in patients with low seizure frequency and seizure-free in recent 3 years (7.78 ± 0.23 ng/l vs. 7.49 ± 0.09 ng/l and 7.59 ± 0.10 ng/l, P < 0.05). Concentration of CSF-PIGF in patients with a disease duration of >5 years was higher than those in patients with durations of 1–5 years and <1 year (7.72 ± 0.20 ng/l vs. 7.52 ± 0.09 ng/l and 7.41 ± 0.07 ng/l, P < 0.05). These results indicate that preexisting brain damage, seizure frequency and disease duration are important factors contributing to elevated PIGF.

Chemokine C-X3-C motif ligand 1 (CX3CL1, alias fractalkine), is highly expressed in the central nervous system and participates in inflammatory responses. Recent studies indicated that inflammatory processes within the brain constitute a common and crucial mechanism in the pathophysiological characteristics of epilepsy. This study investigated the expression pattern of CX3CL1 in epilepsy and its relationship with neuronal loss. Double immunolabeling, MC, and immunoblotting results showed that CX3CL1 expression was upregulated in the temporal neocortex of patients with temporal lobe epilepsy. In a rat model of epilepsy, CX3CL1 up-regulation began 6 hours after epilepsy, with relatively high expression for 60 days. In addition, ELISA revealed that the concentrations of CX3CL1 in cerebrospinal fluid and serum were higher in epileptic patients than in patients with neurosis but lower than in patients with inflammatory neurological diseases. Moreover, H&E staining demonstrated significant neuronal loss in the brains of epileptic patients and in the rat model. Finally, the expression of tumor necrosis factor related apoptosis-inducing ligand was significantly increased in both patients and the animal model, suggesting that tumor necrosis factor related apoptosis-inducing ligand may play a role in CX3CL1-induced cell death. Thus, our results indicate that CX3CL1 may serve as a possible biomarker of brain inflammation in epileptic patients. (Am J Pathol 2012, 180:1950-1962; DOI: 10.1016/j.ajpath.2012.01.024)

Down syndrome cell adhesion molecule (Dscam) is a neural adhesion molecule that plays an essential role in the establishment of neural circuits. Considerable evidence suggests that Dscam is required for axon guidance and dendritic arborization. Our aim was to investigate the expression of Dscam in the temporal lobes of patients with intractable epilepsy (IE) and of experimental animals. In this study, we used immunohistochemistry, immunofluorescence, and western blotting to examine Dscam expression in thirty‐five surgical samples from brains of IE patients and 15 control brain samples. We also measured the levels of Dscam during the entire epileptic process in a rat model of temporal lobe epilepsy. Dscam expression in IE patients was significantly higher compared with that in the controls. In addition, Dscam was also highly expressed in the rat brain during the different phases of the epileptic process. It is the first time to find abnormal expression of Dscam in the brain tissues in patients with IE. And this finding provides an experimental evidence for the study of neuronal circuit remodeling and synaptic plasticity in IE, furthermore, our results also suggest that Dscam may be involved in the generation and the development of IE. Synapse, 2011. © 2011 Wiley‐Liss, Inc.

Gephyrin, which is a postsynaptic scaffolding protein participated in clustering GABAA receptors at inhibitory synapses, has been reported to be involved in temporal lobe epilepsy (TLE) recently. Here, we investigate gephyrin protein expression in the temporal lobe epileptic foci in epileptic patients and experimental animals in order to explore the probable relationship between gephyrin expression and TLE. Using immunohistochemistry, immunofluorescence, and western blot analysis, gephyrin expression was examined in 30 human temporal neocortex samples from patients who underwent surgery to treat drug‐refractory TLE and 10 histological normal temporal neocortex from the controls. Meanwhile, we investigated the gephyrin expression in the hippocampus and adjacent neocortex from experimental rats on 24 h, 48 h, 1 week, 2 weeks, 1 month, and 2 months postseizure and from control rats. Gephyrin protein was mainly expressed in the membrane and cytoplasm of neurons in temporal lobe epileptic foci in humans and experimental rats. Gephyrin expression was significantly lower in the temporal neocortex of TLE patients compared to the controls. In experimental rats, the expression of gephyrin in temporal lobe was downregulated in epileptic groups compared to the control group. Gephyrin expression gradually decreased during the acute period and the latent period, but then began to increase below the levels seen in controls during the chronic phase. Our findings suggest that gephyrin may be involved in the development of TLE. Synapse, 2011. © 2011 Wiley‐Liss, Inc.

Background: Tetranectin (TN) is a plasminogen kringle 4 binding protein and regulates fibrinolysis and proteolytic processes via binding to plasminogen. A previous proteomics study identified TN in the cerebrospinal fluid (CSF) of epileptic patients but not in healthy controls. We determined the concentrations of TN in CSF and serum of epileptic patients to evaluate the changes in TN levels after epileptic attack.Methods: We detected TN in the CSF and serum of 64 epileptic patients and 26 healthy subjects using sandwich enzyme-linked immunosorbent assays.Results: Compared with the control group, CSF-TN levels increased in epileptic patients while serum-TN levels decreased. These differences were statistically significant. The decrease of serum-TN in patients with drug-refractory epilepsy was the most striking.Conclusion: CSF-TN and serum-TN are potential biomarkers in epilepsy and drug-refractory epilepsy and would be useful for diagnosis. (C) 2010 Elsevier B.V. All rights reserved

Objectives: Talin 2 plays an important role in cell adhesion and recycling of synaptic vesicles. To explore the possible role of talin 2 in epilepsy, we designed current study to quantitatively evaluate the changes in talin 2 levels in different epilepsy patients.Design and methods: We measured talin 2 levels in CSF and the serum of postencephalitic epilepsy (PEE) patients (subdivided into drug-refractory PEE-RPEE, and drug-effective PEE-EPEE groups) and acute encephalitis patients (subdivided into acute encephalitis with secondary seizures-AESS, and acute encephalitis without seizures-AE) by enzyme-linked immunosorbent assay.Results: We found that, in RPEE patients, interictal CSF-talin 2 concentrations significantly increased, whereas serum-talin 2 significantly decreased when compared with the AE, AESS, and EPEE groups.Conclusions: This result suggested that the differential concentration of CSF and serum-talin 2 in the RPEE group is an intractability-related phenomenon that might be involved in the development of RPEE. (C) 2010 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

PURPOSE:In epilepsy, many studies were focus on apoE gene polymorphism and found APOE epsilon4 to be associated with earlier onset of temporal lobe epilepsy. There is not any study about apoE in cerebrospinal fluid (CSF) of epileptic patients, so we detect the CSF-apoE and determine whether it is changed after seizure. METHODS:A total of 60 epileptic patients and 28 subjects with no evidence of any neurological diseases were studied. The concentrations of CSF-apoE were detected with enzyme-linked immunosorbent assay. RESULTS:The CSF-apoE levels in epilepsy and control group were 5.78+/-2.15 mg/l and 13.60+/-12.11 mg/l, and there were statistical difference. In epilepsy group, the CSF-apoE concentration was 6.53+/-2.55 mg/l in male patients, and 4.98+/-1.21 mg/l in female. In secondary epilepsy group was 5.06+/-1.31 mg/l, and in idiopathic epilepsy was 6.04+/-2.34 mg/l. In different seizure types groups, including complex partial seizure (CPS), secondarily generalized tonic-clonic seizure (SGTC), generalized tonic-clonic seizure (GTCS), and absence seizure (AS), the mean concentrations of CSF-apoE were 6.62+/-3.13 mg/l, 5.21+/-1.22 mg/l, 5.00+/-1.09 mg/l and 7.25+/-1.88 mg/l, respectively. CONCLUSIONS:CSF-apoE concentration decreases after seizures, correlated with the gender, etiological factor and seizure types.

Repellent guidance molecules provide targeting information to outgrowing axons along predetermined pathways during development. These molecules may also play a role in synaptic reorganization in the adult brain and thereby promote epileptogenesis. Our aim was to investigate the expression of Slit2, one of repellent guidance molecules, in temporal lobe epileptic foci from epileptic patients and experimental animals. Thirty-five temporal neocortex tissue samples from patients with intractable temporal lobe epilepsy (TLE) and fifteen histological normal temporal lobes from controls were selected. Fifty-four Sprague-Dawley rats were divided randomly into six groups, including five groups with epilepsy induced by lithium-pilocarpine administration and one control group. Temporal lobe tissue samples were taken from rats at 1, 7, 14, 30, and 60 days post-seizure and from controls. Expression of Slit2 was assessed by immunohistochemistry, immunofluorescence, and Western blot analysis. Slit2 was mainly expressed in neurons in human controls and in both neurons and astrocytes in TLE patients. Slit2 expression was significantly higher in TLE patients as compared with the controls. Slit2-positive cells were mainly neurons in the rat temporal lobe tissues of the control group, the acute period group, and the latent period group, while the Slit2-positive cells were mainly astrocytes in chronic phase. Compared with controls, Slit2 expression in animals in the TLE group gradually decreased from days 1 to 14 post-seizure, but then increased over the levels seen in controls, to peak levels at days 30 and 60. These results suggest that Slit2 may play an important role in the pathogenesis of TLE.

Proteomic analysis of cerebrospinal fluid (CSF) from patients with temporal lobe epilepsy (TLE) and controls was carried out using two-dimensional gel electrophoresis followed by liquid chromatography electrospray ionization tandem mass spectrometry. Five protein spots showed significant differential expression (p<0.05): vitamin D-binding protein (DBP) was elevated in the CSF of TLE patients whereas cathepsin D, apolipoprotein J, Fam3c, and superoxide dismutase 1 (SOD1) were decreased in the CSF of TLE patients. Additional six protein spots presented only in the CSF of epilepsy patients were identified as tetranectin (TN), talin-2, apolipoprotein E, immunoglobulin lambda light chain (IGL@), immunoglobulin kappa variable light chain 1-5 (IGKV1-5), and procollagen C-endopeptidase enhancer 1 (PCOLCE). Expression of DBP, SOD1 and talin-2 was validated by western blot. Our results may provide better understanding of the pathophysiologic mechanisms underlying epileptogenesis and possible epilepsy biomarkers.