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Exploring the transition from inter-ictal to ictal epileptiform discharges (IDs) and how GABA receptor-mediated action affects the onset of IDs will enrich our understanding of epileptogenesis and epilepsy treatment. We used Mg-free artificial cerebrospinal fluid (ACSF) to induce epileptiform discharges in juvenile mouse hippocampal slices and used a micro-electrode array to record the discharges. After the slices were exposed to Mg-free ACSF for 10 min-20 min, synchronous recurrent seizure-like events were recorded across the slices, and each event evolved from inter-ictal epileptiform discharges (IIDs) to pre-ictal epileptiform discharges (PIDs), and then to IDs. During the transition from IIDs to PIDs, the duration of discharges increased and the inter-discharge interval decreased. After adding 3 μmol/L of the GABA receptor agonist muscimol, PIDs and IDs disappeared, and IIDs remained. Further, the application of 10 μmol/L muscimol abolished all the epileptiform discharges. When the GABA receptor antagonist bicuculline was applied at 10 μmol/L, IIDs and PIDs disappeared, and IDs remained at decreased intervals. These results indicated that there are dynamic changes in the hippocampal network preceding the onset of IDs, and GABA receptor activity suppresses the transition from IIDs to IDs in juvenile mouse hippocampus.
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Animals , Male , Mice , Animals, Newborn , Bicuculline , Pharmacology , Disease Models, Animal , Epilepsy , Pathology , GABA-A Receptor Agonists , Pharmacology , GABA-A Receptor Antagonists , Therapeutic Uses , Hippocampus , Metabolism , In Vitro Techniques , Magnesium , Metabolism , Pharmacology , Membrane Potentials , Mice, Inbred C57BL , Muscimol , Pharmacology , Nerve Net , Receptors, GABA-A , MetabolismABSTRACT
Objective To explore the inhibitory effect of captopril on acute radiation-induced lung injury in rats and the possible mechanism. Methods Sixty-four female Wistar rats were randomly divided into control group, irradiation group, irradiation+low-dose captopril group, and irradiation+high-dose captopril group. A single dose of 20 Gy was given to the right lung of all rats except those in the control group to establish a rat model of acute radiation-induced lung injury. These rats were sacrificed at 1, 2, 4, and 8 weeks. HE staining was used to observe the pathological changes in lung tissue;RT-PCR and Western blot were used to measure the mRNA and protein expression of CCL-2 in lung tissue;immunohistochemical assay was used to determine the number of monocytes ( CD68 ) in lung tissue. A one-way analysis of variance was performed. Results Captopril significantly reduced lung inflammation in rats with acute radiation-induced lung injury (P<005), inhibited the accumulation of monocytes (CD68) in lung tissue (P<005), and decreased the content of CCL-2 in lung tissue ( P<005 ) . Conclusions For rats with acute radiation-induced lung injury, captopril can reduce the expression of CCL-2 to inhibit the accumulation of monocytes in lung tissue and thus attenuate lung inflammation.
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Objective To explore the inhibitory effect of captopril on acute radiation-induced lung injury in rats and the possible mechanism. Methods Sixty-four female Wistar rats were randomly divided into control group, irradiation group, irradiation+low-dose captopril group, and irradiation+high-dose captopril group. A single dose of 20 Gy was given to the right lung of all rats except those in the control group to establish a rat model of acute radiation-induced lung injury. These rats were sacrificed at 1, 2, 4, and 8 weeks. HE staining was used to observe the pathological changes in lung tissue;RT-PCR and Western blot were used to measure the mRNA and protein expression of CCL-2 in lung tissue;immunohistochemical assay was used to determine the number of monocytes ( CD68 ) in lung tissue. A one-way analysis of variance was performed. Results Captopril significantly reduced lung inflammation in rats with acute radiation-induced lung injury (P<005), inhibited the accumulation of monocytes (CD68) in lung tissue (P<005), and decreased the content of CCL-2 in lung tissue ( P<005 ) . Conclusions For rats with acute radiation-induced lung injury, captopril can reduce the expression of CCL-2 to inhibit the accumulation of monocytes in lung tissue and thus attenuate lung inflammation.
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Objective To investigate the role of autophagy in radiation-induced death process of human esophageal squamous carcinoma Eca-109 cells.Methods Esophageal carcinoma cell line Eca-109 was divided into 6 groups of control,5 mmol/L 3-Methyladenine treatment,10 mmol/L treatment,6 Gy irradiation,irradiation + 5 mmol/L drug,and irradiation + 10 mmol/L drug.Some cells were transferred with GFP-LC3 plasmid and the changes of autophagosome were obserred.After each treatment,the expression of autophagy marker LC3B was measured by Western Blot,cell viability was detected by MTT,morphological characteristics of apoptosis cells were stained with a fluorescein of Hoechst 33342 and the percentage of apoptotic cells and cell cycle distribution were measured by flow cytometry.Clonogenic survival were used to evaluate the cell radiosensitivity.Results Autophagy level was increased after radiation,and the LC3B Ⅱ expression and LC3B Ⅱ/LC3B Ⅰ ratio were significantly decreased by autophagy inhibitor 3-Methyladenine (F =25.64,P ＜ 0.05).The number of autophagosome fluorescent foci were significantly increased in the GFP-LC3 transfected cells after radiation,but reduced by 3-Methyladenine (F =127.36,P ＜ 0.05).Compared with radiation alone group,autophagy inhibition combined with radiation significantly decreased cell viability (F =129.54,P ＜ 0.05) and colony formation,increased apoptosis and the percentage of G2/M-phase cells.Conclusions 3-Methyladenine enhances the radiosensitivity of esophageal squamous carcinoma Eca-109 cells,suggesting that inhibition of autophagy could be used as an adjuvant treatment of radiotherapy in esophageal squamous carcinoma.
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Objective To investigate the radioprotective function of lianhuaqingwen (LHQW) in rat acute radiation-induced lung injury.Methods Totally 36 female Wistar rats were randomized into 3 groups as administered group (treated by LHQW plus radiation),radiation group irradiated with a single of 20 Gy in 6 MV X-ray by Elekta Synergy VMAT,and blank control group without radiation.Performance status (PS) was estimated during 31 d of LHQW instragastric administration.After rats being sacrificed at 1,14,28 d of LHQW adminstration,the pathomorphological changes were observed in trauma lung tissue,the cell number in BALF (Bronchoalveolar lavage fluid) was counted,the levels of TNF-α and IL-6 in serum were measured by ELISA,and TNF-α and IL-6 mRNA expressions in lung tissue were assayed by RT-PCR.Results After LHQW treatment,the PS of rat was significantly elevated with less inflammation in morphous,and the cell number in BALF was markedly decreased in compare with radiation alone group.Furthermore,the serum levels of TNF-α and IL-6 were obviously reduced (tTNF-α =7.372,2.891,tIL-6 =6.335,3.257,P ＜ 0.05) and the TNF-α and IL-6 mRNA levels in lung tissue were also decreased (tTNF-αmRNA =3.714,2.144,tIL-6mRNA =3.589,2.883,P＜0.05).Conclusions LHQW plays a protective role against acute radiation-induced lung injury in rats and the down-expressions of TNF-α and IL-6 may be involved.
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Objective To explore the inhibitory effect and possible mechanisms of lianhuaqingwen capsules on radiation-induced acute lung injury in rats. Methods Rats were randomly divided into control group, radiation group and radiation plus lianhuaqingwen group, the control group and the radiation group rats were given 0. 9% sodium chloride solution, the radiation plus lianhuaqingwen group rats were given lianhuaqingwen 0. 9% chlorine sodium solution. HE staining was applied to test the lung tissue inflammation; quantitative RT-PCR and ELISA were used to measure the content of IL-6, TNF-α and MCP-1 in rats;immunohistochemical assay was taken to detect the infiltration of macrophage in lung tissues. Results The relative mRNA expression of IL-6, TNF-α and MCP-1 in the control, radiation model control and radiation plus Lianhuaqingwen groups were (0. 002 1±0. 000 20),(0. 006 6±0. 000 32),(0. 003 9±0. 000 22); (0. 003 7±0. 000 16),(0. 007 4±0. 000 33),(0. 005 5± 0.000 24);(0.001 4±0.000 15),(0.005 4±0.000 72),(0.003 2±0.000 17),respectively; the concentration (pg·mL-1) of IL-6,TNF-αand MCP-1 in the serum were (35. 2±10. 9),(111. 8±26. 1),(68. 2±15. 2); (229. 3±28. 5),(837. 5±57. 6), (566. 9±39. 8);(96. 85±8. 20),(314. 53±12. 76),(191. 32±10. 97),respectively; and the macrophages at high magnification field in each group were (59. 5±4. 3),(503. 9±25. 8)and (106. 2±12. 6), respectively. Lianhuaqingwen capsules significantly alleviated the lung inflammation in rats with radiation-induced acute lung injury,inhibited the accumulation of macrophage in lung tissue,reduced the expression of IL-6 and TNF-α,and decreased the content of MCP-1 in lung tissues and sera(P<0. 05). Conclusion Lianhuaqingwen capsules attenuated the lung inflammation developed in rats with radiation-induced acute lung injury through inhibiting the expression of MCP-1 and reducing the accumulation of macrophage in lung tissues.
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<p><b>BACKGROUND</b>It 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.</p><p><b>METHODS</b>Forty-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).</p><p><b>RESULTS</b>Our 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.</p><p><b>CONCLUSION</b>By 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.</p>
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Adult , Female , Humans , Male , Middle Aged , Young Adult , Epilepsy , Diagnosis , Hippocampus , Pathology , Magnetic Resonance Imaging , MethodsABSTRACT
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.
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Animals , Rats , Behavior, Animal , Disease Models, Animal , Epilepsy , Lithium Chloride , Pentylenetetrazole , Pilocarpine , Seizures , Stress, PhysiologicalABSTRACT
Chicken anemia virus (CAV) is an important viral pathogen that causes anemia and severe immunodeficiency syndrome in chickens worldwide. In this study, a potential diagnostic monoclonal antibody against the CAV VP1 protein was developed which can precisely recognize the CAV antigen for diagnostic and virus recovery purposes. The VP1 gene of CAV encoding the N-terminus-deleted VP1 protein, VP1Nd129, was cloned into an Escherichia (E.) coli expression vector. After isopropyl-beta-D-thiogalactopyronoside induction, VP1Nd129 protein was shown to be successfully expressed in the E. coli. By performing an enzyme-linked immunoabsorbent assay using two coating antigens, purified VP1Nd129 and CAV-infected liver tissue lysate, E3 monoclonal antibody (mAb) was found to have higher reactivity against VP1 protein than the other positive clones according to the result of limiting dilution method from 64 clones. Using immunohistochemistry, the presence of the VP1-specific mAb, E3, was confirmed using CAV-infected liver and thymus tissues as positive-infected samples. Additionally, CAV particle purification was also performed using an immunoaffinity column containing E3 mAb. The monoclonal E3 mAb developed in this study will not only be very useful for detecting CAV infection and performing histopathology studies of infected chickens, but may also be used to purify CAV particles in the future.
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Animals , Mice , Antibodies, Monoclonal/biosynthesis , Antigens, Viral/analysis , Capsid Proteins/genetics , Chicken anemia virus/genetics , Chickens , Circoviridae Infections/blood , Escherichia coli/genetics , Immunohistochemistry/veterinary , Liver/virology , Mice, Inbred BALB C , Microscopy, Fluorescence/veterinary , Poultry Diseases/blood , Specific Pathogen-Free Organisms , Thymus Gland/virologyABSTRACT
<p><b>BACKGROUND</b>Knowledge 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.</p><p><b>METHODS</b>We 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.</p><p><b>RESULTS</b>We 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).</p><p><b>CONCLUSION</b>Hippocampal 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.</p>
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Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Young Adult , Epilepsy , General Surgery , Retrospective Studies , Treatment OutcomeABSTRACT
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.
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Animals , Male , Rats , Action Potentials , Physiology , Electrodes , Electroencephalography , Electrophysiological Phenomena , Physiology , Epilepsy , Hippocampus , In Vitro Techniques , Rats, Sprague-Dawley , Signal Processing, Computer-AssistedABSTRACT
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.
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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
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Objective To study the effects of a rotating magnetic field(RMF)on Li pilocarpine-in- duced seizure activity and the expression of mGluR1 and mGluR5 in the hippocampus.Methods Thirty rats were divided into 5 groups.Each rat in the model(M),short treatment(ST)and long treatment(LT)groups was treated with intra-peritoneal injections of lithium chloride(60 mg/kg),followed by an intra-peritoneal injec- tion of pilocarpine(35 mg/kg)24 h later.The rats in the ST group were exposed to 20 mT RMF for 20 min ev- ery day for 3 d before seizure induction,while the rats in LT group were exposed to the same RMF for 8 d.The latency,severity and duration of seizure,as well as accompanying symptoms and electroencephalogram data, were recorded,and the expression of mGluR1 and mGluR5 was calculated using an electrophoretic imaging anal- ysis system.Results The duration,times and accompanying symptoms of seizure were significantly decreased in the LT group.The mGluR1 mRNA level and mGluR1/mGluR5 ratio in the M group were markedly increased, but the mGluR5 mRNA level was obviously decreased,while the expression of mGluR1 in the ST and LT groups was decreased,and mGluR5 was increased.Conclusions Seizure activity in rats can be inhibited by 20 mT RMF,and the expression of mGluRl and mGluR5 in the hippocampus of rats suffering seizures can be markedly influenced by longer-term RMF.