| Epilepsy is caused by a variety of groups of brain cells caused by the onset of abnormal discharge and temporary brain dysfunction syndrome. Epilepsy prevalence rate is about 7‰in China, an annual increase of over 55 million patients with epilepsy as much to society and the family has created a significant financial burden and psychological stress. Most patients with epilepsy can be controlled by the rational use of drugs seizures, but there are still about 20% to 30% of drug refractory epilepsy patients had surgical treatment aimed at destruction of abnormal discharge through the conduction path or the removal of epileptic foci, so that seizures to reduce or stop seizures. It is reported that about 72%-90% of patients had seizures after surgery can be eased. However, the traditional epilepsy surgery to remove a range of normal brain tissue, devastating, and will only be used for non-functional area epilepsy. Functional area of epilepsy surgery, it is necessary to effective control of epilepsy, but also to maximize the retention of the functional cortex of normal function, or after resection of epileptic foci often cause severe neurological dysfunction. At present, the epileptogenic focus resection, multiple subpial transection and cortical thermocoagulation for the functional areas common method of surgical treatment of epilepsy, but three methods are craniotomy, there is a larger wound, treatment under the scope of open-skull window restrictions, the relatively high incidence of complications such as inadequate. With the development of minimally invasive neurosurgery, gamma knife radiosurgery in the treatment of intracranial tumors, cerebral arteriovenous malformations, functional disorders, etc. shows the advantages of minimally invasive, have become a craniotomy an important means of treatment of intracranial lesions can be whether the use of minimally invasive gamma knife treatment for epilepsy, has attracted widespread attention at home and abroad scholars. At present, the application of gamma knife treatment of refractory epilepsy increasing number of reports, and achieved good results, but the important functional areas of brain gamma knife treatment of epilepsy is rare relevant reports. The exact mechanism of gamma knife treatment of epilepsy is not yet very clear, may be related to nerve conduction block induced by epilepsy, epileptic neurons highly sensitive to radiation, reduced excitability and the epileptogenic zone related to radiation necrosis. Therefore, the functional areas of GKS the effectiveness of epilepsy treatment and its mechanism, is an important issue to be addressed. To this end, this study was through the production of functional area epilepsy animal model, low-dose application of Gamma Knife and high radiation dose group compared to the functional area epilepsy treatment, and to detect relevant biological indicators, in order to explore the Gamma Knife treatment on the functional area epilepsy the feasibility and therapeutic mechanism of action.Now that the Gamma Knife treatment of functional areas related to epilepsy and a variety of factors, such as apoptosis, excitatory amino acids and inhibitory amino acid imbalance, oxygen free radicals, ion channels and other relevant, By studying the functional area in this experiment the cat model of epilepsy is established, compared to Gamma Knife irradiation before and after the performance of optical microscopy, apoptosis index, SOD, MDA,γ-aminobutyric acid (GABA), glutamate (Glu), NMDA receptor subunits R1, R2A, R2B and other changes, to explore the biological mechanism of functional areas of epilepsy and the intervention of the role of gamma knife irradiation.Part 1 The motor areas in patients with epilepsy preoperative evaluation of gamma knife treatment methods and optimal dosage analysisObjective: To establish a sound system for preoperative evaluation of gamma knife treatment and the development of optimal dose plan.Methods: functional neurosurgery, Hebei Province People's Hospital from October 2002 to December 2008 preoperative evaluation (clinical features, VEEG, MRI, MEG, SPECT / PET, consider the areas of motor function epilepsy who do exercise functional areas plus location) at least four matches, diagnosed as primary areas of motor function in patients with epilepsy a total of 11 cases to determine the epileptic foci area, Line Gamma Knife treatment, concurrent mental state before and after surgery and intelligence assessment. Peripheral dose 12-14GY/50%.Results:In accordance with international efficacy of epilepsy surgery Classification Society:â… grade 5 cases,â…¡grade 5 cases,â…¢grade 1 case.â… ~â…¡grade for the cure and effective, with a total effective rate 91% (Fig.1, Fig.2); 12Gy groupâ… level the highest rate of 75%, 14Gy the highest rate of complications, 66.7%.Gamma Knife treatment in patients with pre-show norm compared with the normal state of mind that there are adverse (P<0.01), after 3 months after Gamma Knife treatment, psychological state of varying degrees of ease, in interpersonal relationships, depression, anxiety , hostility and terrorism, etc. have improved significantly (P<0.05), intelligence has also been significantly improved (P<0.05).All patients were reviewed head CT or MRI, no abnormal reactions occurred. Antiepileptic drug plasma concentration controlled within the normal range.Conclusion: Gamma knife treatment is more satisfactory control of motor function areas seizures, security, damage small; can improve the psychological status in patients with epilepsy and intellectual levels; treatment is the key to preoperative evaluation and and seize the radiation dose, low-dose Gamma Knife treatment better-performing, 12Gy dose better.PART 2 Ribbon model of epilepsy cats production, to determine, and Gamma Knife functional areas before and after irradiation epilepsy cats EEG, optical mirror changes inObjective: Successful functional area to establish a cat model of epilepsy, compared different doses of gamma knife irradiation at different time points before and after the cat frontal motor areas of epileptic foci and EEG changes in morphology.Methods: 36 healthy cats, male, weighing 2500 to 3500 grams, were divided into normal control group (n=6), epilepsy model, non-irradiated group (n=6), Gamma Knife 12Gy irradiation group (12 rats) and Gamma Knife 24Gy irradiation group (12 rats); Due to the subkect have been carried out 15Gy and 50Gy, 30 and 60Gy-control study showed that relatively low doses of radiation are better, so this study to further set 12Gy and 24Gy compared to explore the optimal dose) of the animals with 3% amyl barbital sodium anesthesia craniotomy in the left frontal motor area of the brain into the 8% aluminum hydroxide emulsion 100ul or saline 100ul. After a day with video surveillance system records changes in cat behavior, namely, living habits, eating, with or without convulsive seizures and seizure duration; epileptic 8 weeks after the success of Gamma Knife irradiation, observation time point after the two epileptogenic weeks, 4 weeks, 8 weeks, 16 weeks (ie, 8 weeks after irradiation, gamma knife), and 32 weeks (ie 16 weeks after irradiation, gamma knife) a total of five points in time. Each group selected a point in time every 1 or 2 healthy cats, the normal control group and the epilepsy model, non-irradiated animals without gamma knife irradiation, and the other two groups of animals were given different doses of gamma knife radiation, at different times of the above points with MS4000U electrophysiological monitor trace recorded epileptic cortex of laboratory animals were observed and recorded electroencephalogram EEG changes, executed after the specimens obtained by optical microscopy and transmission electron microscopy observation of morphological changes.Results:The experimental animals on the day after eating less apathetic, two to three days after the return to normal eating, the spirit of improvement in epileptic model group and Gamma Knife irradiation group of 30 cats have on the cortical inkection of aluminum hydroxide Suspension appear 6-8 weeks after the right rear lower limbs kitter, clonic and tonic, and some followed the limbs twitching and body clonus, to Racine classification gradeâ…¤. Control group, 6 cats after cortical inkection of normal saline were not apparent abnormal behavior change. Gamma Knife irradiation did not re-appear after irradiation seizures.Induced epileptic model group, and Gamma Knife irradiation group of 30 cats 4 weeks, began to spike and spike and slow wave discharge, low volatility, for accidental discharge, through the statistical treatment of results of 3.5±1.8 beats/min; at 8 weeks the highest frequency of abnormal discharge, the maximum amplitude, showing clustering of spikes or spike and slow wave abnormal discharge, after statistical processing results of 55.1±6.2 beats/min; 16 weeks compared with 8 weeks to reduce the number of discharges, there are still more spikes discharge the discharge frequency of 15.5±1.6 beats/min; 32 weeks the frequency of discharge 12.8±2.5 beats/min, discharge compared with 16 weeks no significant difference. Gamma Knife irradiation group at 16 weeks (ie 8 weeks after irradiation, Gamma Knife), only a small amount of low-amplitude discharge, after statistical processing results of 4.7±2.5 beats/min; in 32 weeks (ie 16 weeks after irradiation, Gamma Knife), only sporadic low-amplitude discharge, after statistical processing result is 1.6±0.9 beats/min. The high and low doses of gamma knife irradiation no significant difference between the two groups. And normal control group had no abnormal spike-wave discharges, after statistical processing, at the same time point, induced epilepsy model group and normal control group, the number of spike-wave discharges are significant differences, gamma knife irradiation-induced epilepsy model group and there are significant differences between the two groups .HE staining pathological results: light microscopy: light microscope, normal control cats no obvious abnormalities in brain tissue. Modeling of epileptic model group, after 4 weeks, 8 weeks with epilepsy can see that the red cat cortical neurons, macrophages neurological phenomena and satellite phenomena, while aluminum hydroxide can be seen epileptogenic zone and the surrounding crystal clear proliferation of glial cells (mainly astrocytes cells and microglia cells); 8 weeks can also be found in varying degrees of degeneration and necrosis of neurons, while low magnification microscope, cortical thinning, gray matter and white matter cells decreased conduction fiber replacement. Gamma Knife irradiation 8 weeks before the performance with the previous 8 weeks later showed cell degeneration and necrosis compared with light-induced epilepsy model group, most of cells were apoptotic status, nuclear osteoporosis, nuclear pyknosis, but the high-dose irradiation necrosis a lower dose of weight, and the cellular edema heavier.4 Ultra-structural pathological results: normal control group of cats no obvious abnormalities in brain tissue. Modeling of epileptic model group, after 4 weeks, 8 weeks epilepsy cat cortex are mainly manifested degeneration: the dissolution of Nissl bodies, water-based degeneration. After 8 weeks, cells showed widespread necrosis of the state: cell body swelling, and some of a nuclear fragmentation and nuclear dissolution of Nissl bodies, mitochondria, the organelles and other makor neurofilament dissolved; glial cell hypertrophy and hyperplasia; nerve fibers: cell body and dendrites symmetric on the reduction of synaptic vesicles, some axonal swelling, and some myelinated nerve fibers myelin disintegration, disorder, axonal atrophy. Gamma Knife irradiation 8 weeks before the performance with the previous 8 weeks later showed cell degeneration and necrosis compared with light-induced epilepsy model group, most of cells were apoptotic status, nuclear osteoporosis, nuclear pyknosis, but the high-dose irradiation necrosis A lower dose of weight.Conclusion:1 Induced epilepsy model group, the Gamma Knife irradiation group model emerged before radiation are complex partial seizures, cortical and scalp EEG spikes continued to discharge and abnormal morphological changes, indicating that functional area of aluminum hydroxide induced epilepsy cats modeling success. Gamma Knife irradiation did not re-appear after irradiation seizures. Description Gamma Knife treatment of epilepsy and effective.2 Iinduced epileptic model group, the Gamma Knife irradiation model of spike-wave discharge started after the first 4 weeks or so, the model group of epileptic peak at 8 weeks or so, continuous spike-wave discharges are still 32 weeks. 8 weeks or so may be the peak of spike-wave discharges. Gamma Knife irradiation significantly reduced at 16 weeks of discharge, 32 weeks sporadic discharge. Between high and low dose groups no significant difference.3 Functional areas of aluminum hydroxide-induced epileptic seizures in the cat model of the pathological basis of the epileptogenic neuronal degeneration, necrosis results in loss of neurons; metal deposition peri-glial cell hypertrophy and hyperplasia; neural circuit reconstruction; cell body and dendrites on the symmetric reduction of synaptic vesicles.4 Gamma Knife irradiation of epileptic neurons degeneration and necrosis reduced, but high-dose group heavy cell damage, while the high and low dose in reducing the abnormal discharge in neurons does not differ greatly, so 12Gy exposure is the best.Part 3 Gamma knife irradiation on the functional area changes and apoptosis of epilepsy cats SOD, MDA ofObjective: To compare different doses of gamma knife irradiation at different time points before and after the cat frontal motor areas of epileptic foci of apoptosis, bcl-2 protein expression and SOD, MDA investigate the changes of apoptosis, bcl-2 protein and the expression of SOD, MDA and the Gamma Knife radiation treatment of functional area epilepsy mechanism.Methods: The observation at various time points, that is, epileptic after 2 weeks, 4 weeks, 8 weeks, 16 weeks (ie, 8 weeks after irradiation, gamma knife), and 32 weeks (ie 16 weeks after irradiation, gamma knife) six time points Each cats to end the first part of treatment, take samples made of paraffin sections, and then used immunohistochemistry to detect bcl-2 protein, using in situ nick end labeling (TUNEL method) to detect brain cell apoptosis, At the same time taking epileptogenic zone by spectrophotometer measurement of brain tissue SOD, MDA changes. All data are mean±standard deviation using (x±s), using SAS statistical software for analysis and comparison to P<0.05 for the difference was significant.Result: Epileptic SOD content in model group 153.45±21.52 U/mgprot, low-dose Gamma Knife irradiation SOD content was 172.45±17.62 U/mgprot, high-dose Gamma Knife irradiation SOD content was 170.62±15.37 U/mgprot, the normal control group, SOD content of 182.01±27.49 U/mgprot; aluminum hydroxide MDA content model group 0.5791±0.1366 umol/gprot, low-dose Gamma Knife irradiation MDA content was 0.3106±0.0546 umol/gprot, high-dose Gamma Knife irradiation MDA content was 0.3076±0.0342 umol/gprot, the normal control group, MDA content was 0.2596±0.0628 umol/gprot.Changes of apoptosis: (1) at each point in time, high-dose Gamma Knife irradiation apoptotic cells positive expression rate of "low-dose Gamma Knife irradiation" epileptic model group and control group by statistical analysis, high-dose Gamma Knife Gamma Knife irradiation with low-dose irradiation group, Gamma Knife irradiation group and non-irradiated group difference was significant (P<0.01). (2) Every one Gamma irradiation group at different time points after irradiation, the dynamic expression of apoptotic cells does not differ greatly.Cells, bcl-2 protein expression level changes: (1) at each point in time, high-dose Gamma Knife irradiation bcl-2 positive expression rate of "low-dose Gamma Knife irradiation" epileptic model group and control group, statistically analysis of Gamma Knife irradiation group and non-irradiated group difference was significant (P<0.01). (2) Every one Gamma irradiation group at different time points after irradiation, bcl-2 positive expression rate does not differ greatly.Conclusion:1 Epileptogenic epileptogenic zone in model group than the control group in brain tissue MDA content was increased, while the SOD were significantly reduced; Gamma Knife irradiation group compared with MDA content of epileptic model group decreased, but still higher, while the SOD content in the more epileptogenic the model group increased, but still lower than the control group. Description of epileptic seizures induced by model group, region of brain tissue oxidation and antioxidant system imbalance, resulting in an excessive amount of oxygen free radicals into a damage brain tissue, while the scavenging oxygen free radicals due to excessive consumption reduced SOD and increased cell degeneration, necrosis, leading to loss of neurons and neuronal abnormal discharge, promote seizures. The Gamma Knife irradiation re-establishment of brain tissue balance between oxidation and antioxidant system to reduce the oxygen free radicals, protect nerve cells, inhibit seizures.2 Gamma Knife apoptotic cells after irradiation significantly increased the rate of positive expression, inhibition of apoptosis protein bcl-2 decreased significantly. Note the promotion of epileptogenic zone neuronal apoptosis in abnormal discharge is a Gamma Knife treatment of epilepsy mechanisms. At the same time high-dose gamma knife radiation caused apoptosis than low-dose gamma knife radiation, may cause neuronal loss and reduce the therapeutic effect. Therefore, 12Gy exposure is the best.PART 4 Gamma knife irradiation on the functional area epilepsy catsγ-aminobutyric acid (GABA) and glutamate (Glu) ofObjective: To compare different doses of gamma knife irradiation at different time points before and after the cat frontal motor areas of epileptic foci ofγ-aminobutyric acid (GABA) and glutamate (Glu) investigate the changes ofγ-aminobutyric acid (GABA) and glutamic acid (Glu) and gamma knife radiation treatment of epilepsy mechanisms of the relationship between functional areas.Methods: The observation at various time points, that is, epileptic after 2 weeks, 4 weeks, 8 weeks, 16 weeks (ie, 8 weeks after irradiation, gamma knife), and 32 weeks (ie 16 weeks after irradiation, gamma knife) five time points Each cats to end the first two parts of the treatment, take samples centrifuged homogenate supernatant was placed in a 80℃after cryopreservation under test until the sample collection is complete, you use high-performance liquid chromatography detection of brainγ- aminobutyric acid (GABA) and glutamate (Glu) was observed. All data are mean±standard deviation using (x±s), using SAS statistical software for analysis and comparison to P <0.01 for the difference was significant.Results:At the same time point, induced epilepsy model group than in the normal control group, Glu increased, GABA reduced; Gamma Knife irradiation group as compared Glu-induced epilepsy model group lower, GABA significantly higher difference between low and high dose group is not; but Gamma Knife irradiation group than in the normal control group, Glu is still slightly higher, GABA was significantly higher than the normal control group. The statistical analysis of epileptic model group and normal control group, Gamma Knife irradiation group and the difference between epileptic model group was significantly (P<0.01).Gamma irradiation of each group at different time points after irradiation, the dynamic expression of amino acids does not differ greatly.Conclusion: The excitatory amino acid Glu increased inhibitory amino acid GABA reduced due to inhibition of excitatory amino acids and amino acid imbalance is one of the mechanisms seizures; Gamma Knife irradiation and a decrease of excitatory amino acids Glu, increased inhibitory amino acid GABA, which the establishment of excitatory amino acids and inhibitory amino acids a new balance, thereby inhibiting seizures.PART 5 Gamma knife irradiation on the functional area epilepsy cats NMDA receptor subunits R1, R2A, R2B ofObjective: To compare different doses of gamma knife irradiation at different time points before and after the cat frontal motor areas of epileptic foci of the NMDA receptor subunits R1, R2A, R2B investigate the changes of NMDA receptor subunits R1, R2A, R2B and gamma knife radiation therapy functional area epilepsy mechanismMethods: The observation at various time points, that is, epileptic after 2 weeks, 4 weeks, 8 weeks, 16 weeks (ie, 8 weeks after irradiation, gamma knife), and 32 weeks (ie 16 weeks after irradiation, gamma knife) five time points Each cats to end the first three parts of the treatment, take samples made of paraffin sections, and then used immunohistochemistry to detect NMDA receptor subunits R1, R2A, R2B changes, all data are used both the number of±standard deviation x±s, using SAS statistical software for analysis and comparison to P <0.01 for the difference was significant.Results:At the same time point, induced epilepsy model group than in the normal control group, increased expression of NMDAR; Gamma Knife irradiation group than in the expression of NMDAR-induced epilepsy model group decreased low and high dose groups does not differ greatly; but Gamma Knife irradiation group than in the normal NMDAR expression in the control group was still higher. The statistical analysis of epileptic model group and normal control group, Gamma Knife irradiation group and the difference between epileptic model group was significantly (P<0.01).Gamma irradiation of each group at different time points after irradiation, the dynamic expression of amino acids does not differ greatly.Conclusions: NMDA receptor subunits R1, R2A, R2B The increase is one of the mechanisms seizures; Gamma Knife irradiation a decrease in NMDA receptor subunits R1, R2A, R2B content, reduce the excitability of nerve cells, thereby inhibiting seizures.
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