Objective To develop models of epileptic discharge by activating group Ⅰ metabotropic glutamate receptors (mGluR) or by blocking gamma-aminobutyric acid (GABA) receptors on rat hippocampal slices. Methods Rat hippocampal slices were exposed to mGluR group Ⅰ specific agonist dihydroxyphenylglycine (DHPG) or to GABAA receptor antagonists bicuculline methiodide(BMI), and single pyramidal cell in the CA3 region of the slice was recorded by whole cell patch clamp technique. Results Exposure to DHPG or BMI resulted in the induction of spontaneously occurring epileptic discharge in the CA3 region of rat hippocampal slice, and there was no significant difference in the frequency of discharge between them(P>0.05). Conclusion Epileptic discharge can be generated in vitro in response to a loss of balance between excitatory and inhibitory influences.

0.05).However,there was no significant difference in the AMPAR-mediated basal synaptic transmission on hippocampal CA1 between those at the later period(6 weeks) of epilepsy induced by pilocarpine and controls,while LTP was inhibited(P

OBJECTIVETo establish a new culture method to induce the differentiation of embryonic pancreatic cells into mature endocrine cells.

METHODSMouse embryos at day 12.5 were used and embryonic pancreata were isolated. The isolated embryonic pancreata were cultured on the filter for 7 days, which floated in the dish containing medium. During culture, the expression of pancreas duodenum homeobox-1 (PDX-1), a pancreatic stem cell marker, was examined at day 1. The expression of neurogenin 3 (Ngn3), a pancreatic progenitor cell marker, was examined at day 3. The expressions of endocrine and exocrine markers, insulin, glucagon, and carboxypeptidase (CPA) were examined at day 7 by immunohistochemistry. The kinetics of pancreatic marker expression during culture was assayed by real-time PCR.

RESULTSMany pancreatic stem cells still existed in embryonic pancreata cultured for 1 day; meanwhile, these pancreatic stem cells proliferated in high rate. A large amount of pancreatic progenitor cells were found in embryonic pancreata cultured for 3 days.Pancreatic stem/progenitor cells differentiated into mature endocrine and exocrine cells in embryonic pancreata after having been cultured for 7 days. Furthermore, the expression pattern of pancreatic marker is consistent with that in vivo.

CONCLUSIONWe successfully established a new culture method, with which embryonic pancreatic cells can efficiently differentiate into mature endocrine cell.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; Endocrine Cells ; cytology ; Female ; Homeodomain Proteins ; metabolism ; Male ; Mice ; Nerve Tissue Proteins ; metabolism ; Pancreas ; cytology ; Trans-Activators ; metabolism

BACKGROUNDIt was still rare for the quantitative magnetic resonance imaging (MRI) research of regional changes in hippocampus sclerosis (HS) in Chinese patients with epilepsy. This study aimed to study the hippocampal volumes (HVs) with quantitative MRI measurement in Chinese patients with epilepsy.

METHODSForty-six Chinese patients with epilepsy (intractable epilepsy (IE), n = 21; non-intractable epilepsy (NIE), n = 25) and 25 normal controls were collected between July 2007 and March 2008. All of the subjects underwent a 3T high-resolution MRI with oblique coronal thin sections oriented perpendicular to the hippocampal long axis. Hippocampal structures were assessed by visual detection, and HVs were quantitatively studied with a Picture Archiving and Communication System (PACS).

RESULTSOur study suggested that there was no significant difference in gender (P > 0.05) while the right hippocampal head volume (HHV), hippocampal body volume (HBV), and the whole hippocampal volume (HCV) were greater than the left one (P < 0.05), but no significant difference was found in bilateral hippocampal tail volume (HTV) (P > 0.05) in normal controls. That unilateral/diffuse (64%/21%) and bilateral/focal (86%/20%) hippocampal atrophy (HA) were significant in IE and NIE patients, respectively. Anterior hippocampus, especially HHV (26% in IE and 20% in NIE) and HBV (29% in IE and 12% in NIE), had more significant atrophy than the HTV (5% in IE and 0% in NIE) in patients with epilepsy.

CONCLUSIONBy assessing the volumes of the regional hippocampus with 3T MRI, we could better define the range and distribution of HS, since regional or subtle changes in HVs could be detected earlier with 3T MRI.

Adult ; Epilepsy ; diagnosis ; Female ; Hippocampus ; pathology ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Young Adult

BACKGROUNDMuscles present different responses to muscle relaxants, a mechanism of importance in surgeries requiring facial nerve evoked electromyography under general anaesthesia. The non-depolarizing muscle relaxants have multiple reaction formats in the neuromuscular junction, in which pre-synaptic quantal release of acetylcholine was one of the important mechanisms. This study was to compare the pre-synaptic quantal release of acetylcholine from the neuromuscular junctions innervated by normal/damaged facial nerves and somatic nerve under the effect of rocuronium in rats in vitro.

METHODSAcute right-sided facial nerve injury was induced by nerve crush axotomies. Both sided facial nerve connected orbicularis oris strips and tibial nerve connected gastrocnemius strips were isolated to measure endplate potentials (EPP) and miniature endplate potentials (MEPP) using an intracellular microelectrode gauge under different rocuronium concentrations. Then, the pre-synaptic quantal releases of acetylcholine were calculated by the ratios of the EPPs and the MEPPs, and compared among the damaged or normal facial nerve innervated orbicularis oris and tibial nerve innervated gastrocnemius.

RESULTSThe EPP/MEPP ratios of the three neuromuscular junctions decreased in a dose dependent manner with the increase of the rocuronium concentration. With the concentrations of rocuronium being 5 µg/ml, 7.5 µg/ml and 10 µg/ml, the decrease of the EPP/MEPP ratio in the damaged facial nerve group was greater than that in the normal facial nerve group. The decrease in the somatic nerve group was the biggest, with significant differences.

CONCLUSIONSRocuronium presented different levels of inhibition on the pre-synaptic quantal release of acetylcholine in the three groups of neuromuscular junctions. The levels of the inhibition showed the following sequence: somatic nerve > damaged facial nerve > normal facial nerve. The difference may be one of the reasons causing the different sensitivities to rocuronium among the muscles innervated by the normal/injured facial nerves and the somatic nerve. The results may provide some information for the proper usage of muscle relaxants in surgeries requiring electromyographic monitoring for the pre-surgically impaired facial nerves.

Acetylcholine ; metabolism ; Androstanols ; pharmacology ; Animals ; Facial Nerve ; drug effects ; metabolism ; Male ; Neuromuscular Junction ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley

BACKGROUNDKnowledge about factors influencing the prognosis of resective epilepsy surgery can be used to identify which patients are most suitable for surgical treatment. The aim of this study was to identify preoperative prognostic factors associated with the chance of achieving long-term seizure freedom.

METHODSWe retrospectively reviewed seizure outcomes and clinical, electroencephalography (EEG), magnetic resonance imaging (MRI), histopathology, and surgical variables from 99 epilepsy surgery patients with at least one year of postoperative follow-up. Seizure outcomes were categorized based on the modified classification by the International League Against Epilepsy.

RESULTSWe found that the seizure-free rate was 27.9% after one year, and that it stabilized at about 20.0% between two and six years after surgery. Univariate analysis showed that medial temporal lobe epilepsy with hippocampal sclerosis, MRI with visible focal lesions concordant with EEG, and regional ictal EEG and electrocorticography patterns were associated with a favorable surgical outcome. On the other hand, seizure recurrence within six months, incomplete focus resection, and surgical complications were associated with a poor outcome. Multivariate analysis showed that medial temporal lobe epilepsy with hippocampal sclerosis and MRI with visible focal lesions were independent presurgical predictors of a favorable outcome (P < 0.01). Seizure recurrence within six months was the only significant independent predictor associated with a poor outcome (P < 0.01).

CONCLUSIONHippocampal sclerosis and abnormal MRI findings are strongly associated with a favorable surgical outcome, whereas seizure recurrence within six months is associated with a poor outcome.

Adolescent ; Adult ; Child ; Epilepsy ; surgery ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult

Objective To provide scientific evidences for improving children early integrated development,through analyzing the mental developmental status and characteristics of 322 cases of 2-year-old children in Nanjing.Methods Intelligence and motor development condition in 322 cases of 2-year-old children were assessed by using Bayley Scales of Infant Development test,and the assessed results were analyzed.Results 1.The incidences of the children whose mental development index(MDI)or psychomotor development index(PDI)were under 69 were 3.1% and 5.6%,respectively;2.The MDI mean score(114.34?19.65)was significantly higher than that of PDI(101.73?21.53)(t=9.71,P0.05).Conclusions The incidences of mental retardation in this study were consistent with the result reported by World Health Organization.There were differences between motor and intelligence development in children,as well as the intelligence development between male and female.Therefore,it should be implemented early childhood developmental screening in child health care.Parents should be given scientific guides about intelligence and motor development of children.

To investigate the spatiotemporal properties of epileptiform activity in vitro, 400 microm-thick transverse hippocampal slices were prepared from juvenile rat and planar multi-electrode array (MEA) containing 60 electrodes was used to record the electrical activity induced by bath application of high potassium artificial cerebrospinal fluid (ACSF) on slices. Following successful induction of epileptiform bursts, phenobarbital sodium was applied to test for its inhibitory effects on bursting activity in different regions of slice. Region-specific characteristics of epileptiform activity and anticonvulsant actions of phenobarbital sodium in the hippocampal network were determined by comparing the population activity obtained from MEA. The results showed that: (1) 15 min after high-K+ ACSF application, rhythmic and synchronous epileptiform bursts could be detected from all CA sub-regions. Quantitative analysis indicates that the firing patterns of different CA sub-regions were not statistically different (P>0.05). However, no bursting activity was recorded from granular cells in dentate gyrus, only sparse spikes were observed, with frequency significantly lower than that in CA regions (P<0.05). (2) The high-K+-induced bursting activity could last for more than 40 min with stable bursting activities. (3) Bath application of 60 micromol/L phenobarbital sodium inhibited the bursting activities on hippocampal slice. Bursting activities in CA3c and CA1 were firstly suppressed. 10 min after the phenobarbital sodium application, strong bursting activities persisted only in some of pyramidal cells in CA3a and CA3b. These results show that MEA could be applied for studying the spatial and temporal properties of epileptiform activity in vitro, as well as the region-specific effects of anti-epileptic drugs.
Action Potentials ; physiology ; Animals ; Electrodes ; Electroencephalography ; Electrophysiological Phenomena ; physiology ; Epilepsy ; physiopathology ; Hippocampus ; physiopathology ; In Vitro Techniques ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Processing, Computer-Assisted

Hematopoietic stem cells (HSC) shift from fetal liver and spleen to bone marrow at neonatal stages and this movement may be due to inductive signals from different microenvironments. Mesenchymal stem cells (MSC) are the precursors of stromal cells in bone marrow microenvironments such as osteoblasts and endothelial cells. Some researchers speculated that fetal bone marrow before birth might be not perfectly suit HSC growth. However, it is still lack of direct evidence to prove this hypothesis. This study was aimed to compare the hematopoietic supportive capacity between human fetal and adult bone marrow MSC in vitro. Adult bone marrow MSC (ABM-MSC) were isolated from three healthy donors and fetal bone marrow MSC (FBM-MSC) were isolated from three fetuses between gestations of 19 to 20 weeks. After irradiation, MSC were co-cultured with CD34(+) cells isolated from umbilical cord blood in long-term culture-initiating cell (LTC-IC) assay. The colony number of colony forming cells (CFC) was counted and the phenotypic changes of co-cultured CD34(+) cells were analyzed by flow cytometry. Cytokine expressions in both kinds of MSC were detected by reverse transcription polymerase chain reaction (RT-PCR). The results showed that ABM-MSC had a stronger hematopoietic supportive capacity than FBM-MSC. Both of them enhanced the differentiation of CD34(+) cells into myeloid lineages. Cytokines were expressed differently in ABM-MSC and FBM-MSC. It is concluded that ABM-MSC possess more potential application in some treatments than FBM-MSC, especially in hematopoietic reconstitution.
Adult Stem Cells ; cytology ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Fetus ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology

This study was undertaken to observe the effect of acute stress on seizure occurrence in chronic period of epileptic model rats. Lithium-pilocarpine (LiCl-PILO)-induced epileptic rat model was constructed. At the spontaneous recurrent seizure period, acute stress stimulations such as cat's urine and foot electrical shock were applied to observe the behavioral changes and seizure occurrence. The results showed that after the cat's urine stimulation, the self-directed behaviors of the epileptic model rats decreased significantly, while the risk assessment behaviors increased significantly. The seizure occurrence, however, was not observed during the 45 min after the stimulation. Applying electrical foot shocks also did not evoke seizures in epileptic model rats. On the contrast, intra-peritoneal injection of low dose of pentylenetetrazole (PTZ, 30 mg/kg) evoked seizure more efficiently, and the duration of seizure activity was extensively prolonged in epileptic model rats than that of control rats. Taken together, these results indicate that although applying stress stimulations such as cat's urine and electrical foot shock cause several behavioral changes, they are not severe enough to evoke seizure in epileptic model rats.
Animals ; Behavior, Animal ; Disease Models, Animal ; Epilepsy ; chemically induced ; physiopathology ; Lithium Chloride ; adverse effects ; Pentylenetetrazole ; adverse effects ; Pilocarpine ; adverse effects ; Rats ; Seizures ; physiopathology ; Stress, Physiological