Objective To observe dynamic changes of Kir2. 3 mRNA in the hippocampus of rats with chronic temporal lobe epilepsy, and to discuss the relationship between Kir2. 3 expression and the pathogenesis of chronic temporal lobe epilepsy. Methods We used pilocarpine to induce status epilepticus (SE) in rats,which became chronic temporal lobe epileptic rats in 2 weeks. The expression of Kir2.3 mRNA was measured by reverse transcription polymerase chain reaction (RT-PCR) at the time points of 0, 6, 72 hours and 2 weeks after SE. Results The ratios of Kir2. 3 mRNA to β-actin of normal control and 0, 6, 72 hours, 2 weeks after SE were 0. 080 ± 0. 030, 0. 103 ± 0. 045, 0. 164 ± 0. 026, 0. 132 ± 0. 024, and 0. 011 ± 0. 008, respectively. The ratio was significantly higher 6 and 72 hours after SE and significantly lower 2 weeks after SE than that of the normal control. Conclusions Two weeks after SE, when the rats had spontaneous recurrent seizures, the expression rate of Kir2.3 reached a turning point, which possibly became the basis of epileptogenesis.

Adult ; Diagnosis, Differential ; Female ; Gastrectomy ; methods ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; Humans ; Ki-1 Antigen ; metabolism ; Leukocyte Common Antigens ; metabolism ; Leukosialin ; metabolism ; Lymphoma, Large-Cell, Anaplastic ; metabolism ; pathology ; surgery ; Melanoma ; metabolism ; pathology ; Mucin-1 ; metabolism ; Receptor Protein-Tyrosine Kinases ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; surgery

Antibodies, Monoclonal, Murine-Derived ; metabolism ; Brain Neoplasms ; metabolism ; pathology ; Child ; Choriocarcinoma ; metabolism ; pathology ; Chorionic Gonadotropin, beta Subunit, Human ; metabolism ; Diagnosis, Differential ; Humans ; Male ; Neoplasms, Germ Cell and Embryonal ; metabolism ; pathology ; Pineal Gland ; pathology

Objective To survey gene expression profiles in nonlesional refractory temporal lobe epilepsy(TLE)and to further verify the difference of gene expression.thus to evaluate the possible molecular pathogenesis of this kind of epilepsy that can help to supply a new way for the diagnosis and treatment.Methods The TLE samples and control cases were studied by means of cDNA microarray consisting of 1 8 000 genes.Reverse transcription polymerase chain reaction(RT-PCR)Was performed to measure the expression alterations of SH3GL2.BTNN2A2 and KCNJ4 mRNA in temporal cortex samples from patients who had undergone temporal lobectomy surgery for intractable epilepsy.Tissue from 10 subjects who did not have epilepsy served as controls.Results The known genes differently expressed in those TLE samples involved immunity correlation factor genes,signal conduction genes,ion channel transportation genes;mitochondria function genes and SO on were identified.Among which.the expression of SH3GL2 mRNA Was significantly increased in epileptic brain(1.022±0.547)compared with the controls(0.446±0.171,t=-3.181).In TLE group(0.481±0.196),the expression of BTN2A2 mRNA was also significantly higher than that of control subjects(0.243±0.111,t=3.351).Compared with control group(O.795±0.112),the expression of KCNJ4 mRNA Was significantly decreased in TLE patients(0.438±0.178).Conclusions cDNA microarray is an efficient and high.throughout method to survey gene expression profiles in intractable temporal lobe epilepsy.The variation of those gene expressions might be a potential etiological agent for TLE that may offer a novel target for anticonvulsant therapy.

Objective To establish a multi-drug resistant model of temporal lobe epilepsy,and to observe the changes of γ-aminobutyric acid (GABA) receptor expression in the hippocampal tissues so as to explore its effects in pharmacoresistant epileptogenesis.Methods One hundred rats were selected to prepare the amygdaloid kindled model of epilepsy by chronic stimulation of amygaloid basal lateral nucleus.After the kindled model of epilepsy was prepared successfully(n =52),pharmacoresistant epileptic rats were selected according to their response to the phenobabital and phenytoin.The selected pharmacoresistant epileptic rats (n =8)were sacrificed and the hippocampus was removed to determine the GABA receptor expression,and the same number of pharmacosensitive epileptic rats was used as control.Results The pharmacoresistant epileptic rats displayed degenerative and necrotic hippocampal neurons.The arrangement of hippocampal neurons was disordered,and the structural characteristics of the arrangement of the hippocampal neurons disappeared.The gray values of GABAA-positive neurons in the hippocampal tissues (141.15 ± 14.72) increased significantly compared with the pharmacosensitive epileptic rats (92.56 ± 5.17; t =3.380,P =0.006).Western blot method demonstrated that the band of GABAA became narrowed and thin.The relative quantity of GABAA in the hippocampal tissues (0.38 ± 0.08) decreased significantly as compared with the pharmacosensitive epileptic rats (0.88 ± 0.18).A significant difference was observed (t =5.420,P =0.002).Conclusions GABA receptor expression might be decreased in the hippocampal tissues of pharmacoresistant epileptic rats.It might play a certain role in the formation of pharnmacoresistant epilepsy.

Objective To establish a multi-drug resistant model of temporal lobe epilepsy and observe the effect of hippocampal stimulation on pharmacoresistant epileptic rats and its possible mechanism with the use of indexes including after discharges (AD) of the amygdalae,the stimulus-induced seizures and the extracellular levels of γ-aminobutyrate (GABA).Methods Totally,120 Wistar rats were used for the amygdaloid kindled model of epilepsy by chronic stimulation of amygaloid basal lateral nucleus.Based on the successful kindled model of epilepsy,we selected the pharmacoresistant and pharmacosensitive epileptic rats according to their response to phenobabital and phenytoin.We then divided the pharmacoresistant epileptic rats into the hippocampal stimulation group and control group,with 8 rats in each group.Low-frequency stimulus was conducted for two weeks in the hippocampal stimulation group.The hippocampus extracellular fluid collected by microdialysis was then used to determine the levels of GABA by a high performance liquid chromatography method after hippocampal stimulation.Results The stimulus-induced seizures were inhibited significantly,and the frequency of the AD was decreased,as well as the amplitude,compared with the control group.The extracellular level of GABA in the 8:00-9:00 am and the 8:00-9:00 pm was (32.69 ± 7.80) and (35.76 ± 6.27) μg/ml,respectively,both significantly increased as compared with the control ((26.58 ± 6.87) μg/ml,t =-21.45,P =0.000 ; (31.50 ± 4.87) μg/ml,t =-15.74,P =0.000).Conclusions The low-frequency hippocampal stimulation inhibits the seizures of the kindled model of epilepsy and decreases the frequency,the amplitude,as well as the duration of the amygdale AD in the pharmacoresistant epileptic rats,which may be contributed to the increased GABA content in extracellular fluid of the brain after stimulation.

Objective To observe the change of expression of inwardly rectifying K+(Kir)2.3 mRNA and protein of Kir in the hippocampus of rats with chronic temporal lobe epilepsy(TLE)in different time points and the effect of Tenidap a Kir2.3 channel opener on its expression,investigate the relationship between Kir2.3 and the pathogenesis of TLE and to explore the potential of Kir agonists as anti-epileptic drugs.Methods The pilocarpine TLE rat model was used.Animals were randomly assigned to the control or the status epilepticus(SE)groups,which were further divided into four time point subgroups consisting of 0.6,72 hours,and 2 weeks post-SE termination.Another subgroup was given Tenidap,a Kir2.3 channel opener,and tested 2 weeks post-SE.Hippoeampi were removed and the expression of Kir2.3 mRNA and protein at different time points was measured by reverse transcription polymerase chain reaction(RT-PCR)and western blotting.Results The ratios of Kir2.3 mRNA and β-actin in normal control and 0,6,72hours and 2 weeks after SE termination were 0.080±0.030,0.103±0.045,0.164±0.026,0.132±0.024.0.011±0.008,respectively(F=23.684,P<0.01).The ratios of Kir2.3 protein and GAPDH in propotional groups were0.305±0.030,0.263±0.028,0.767±0.167,0.498±0.077,0.176±0.026(F=44.183.P<0.05).The expression of Kir2.3 channel in the epileptic rats was bimodal,increasing immediately after SE,relative to controls,and declining in the chronically epileptic period.Tenidap administration upregulated both the mRNA(0.021±0.006)and protein expression(0.636±0.140) of Kir2.3(F=25.216 and 47.355,P<0.05 and 0.01).Condusion These findings suggest that the pathogenesis of TLE is accompanied by a decrease in Kit2.3 expression,which may be ameliorated by the administration of tenidap.

Objective To summarize the clinical characteristics of small intestinal stromal tumors(SIST), and evaluate the diagnostic values of various imaging or endoscope examinations for SIST. Methods From July 2004 to June 2009, 74 patients whose operation or endoscopy biopsy tissues pathologically confirmed SIST were collected. The clinical data, imageology including enteroclysis, abdominal ultrasound, spiral computered tomography (CT) and the double-balloon enteroscopy report of the patients were analyzed retrospectively. According to biological behavior, the SIST was divided into four risk degree such as extremely low risk, low risk, moderate risk and high risk. The correlation between pathologic characters and spiral CT feature was analyzed. Results The most predilection site of SIST was jejunum in 43 patients (58.1 %); secondarily duodenum in 17 cases (23%); and 10 cases (13.5%) in ileum. About 94.6 percent of patients (70/74) showed clinical signs, the most common symptom was gastrointestinal bleeding in 46 cases (67. 2 % ), abdominal pain in 23 cases (31.1%). Of various photogrammetry examinations and endoscopy, spiral CT has the highest diagnosis rate and diagnosis coincidence rate, which was 100% and 72. 1% respectively. Among the 74 SIST lesions, 14 cases were extremely low biological risk (18. 9 % ), 21 at low risk (28.4 % ), 15 at moderate risk (20. 3%) and 24 at high risk (32.4%). Spiral CT is helpful for the SIST risk diagnosis. Conclusions The onset of SIST was concealed and early diagnosis was very difficult. Spinal CT which could help to predict the tumor's risk degree and prognosis was noninvasive, convenient and reliable. Therefore, it could be the first choice for SIST examination at present.

Objective To establish a multi-drug resistant model of temporal lobe epilepsy,and then the sodium current of pyramidal neurons in CA1 areas of the hippocampus was used as as index to observe the effect of hippocampal stimulation on pharmacoresistant epileptic rats.Methods Eighty Wistar rats were selected to prepare an amygdaloid kindled model of epilepsy by chronic stimulation of amygaloid basal lateral nucleus.When the kindled model of epilepsy was prepared successfully,the pharmacoresistant epileptic rats were selected according their response to phenobabital and phenytoin.The selected pharmacoresistant epileptic rats were divided into a hippocampal stimulation group (HS group) and a pharmacoresistant control group (PR group).A low-frequency hippocampal stimulation was performed in the HS group,while the PR group received sham stimulation.The whole-cell recording technique by patch-clamp was used to observe the changes of sodium current of hippocampal pyramidal neurons after the hippocampal stimulation.Results Compared with the PR group,the pharmacoresistant epileptic rats in HS group underwent low-frequency stimulation for 2 weeks showed that the amygdale stimulus-induced seizures were decreased (2.32 ± 0.38 in HS group and 4.45 ± 0.42 in PR group,t =84.600,P =0.000) and the parameters of the after-discharges were improved significantly.In HS group,the peak current shifted towards depolarization,the sodium channels were difficult to activate,and were more susceptible to inactivation.Moreover,the recovery time after the sodium channel inactivation was slower in HS group ((17.9 ±0.6) s) than in PR group((16.3 +0.3) s,t =-25.420,P =0.000).Conclusions Hippocampal stimulation may inhibit the sodium channel current of pyramidal neurons in CA1 areas of hippocampus.The mechanism of hippocampal stimulation in the treatment of pharmacoresistant epilepsy might be achieved partly by inhibiting the sodium channel current so as to decrease the excitability of hippocampal neurons.

OBJECTIVE:To develop an HPLC method for the determination of 5-ISMNconcerntrations in human plasma and study the pharmacokinetics of 5-ISMN orally disintegrating tablets and the reference tablets.METHODS:A single oral dose test capsule(orally disintegrating tablets)or reference capsule of 5-ISMN(60 mg)were administered by randomized crossover way in 20 healthy male volunteers with plasma 5-ISMN concentrations determined by HPLC.The pharmacokinetic parameters were calculated with 3p87 pharmacokinetic program and the bioavailability of the two preparations was evaluated.RESULTS:The main pharmacokinetic parameters of the test preparation vs.the reference preparation were as follows:Cmax:(613.42?73.83)vs.(368.64?38.66)?g?mL-1;tmax:(26.52?2.00)vs.(95.73?4.16)min;AUC0～∞:(73 872.24?543.89)vs.(77 978.47?646.37)ng?min?mL-1,respectively.CONCLUSION:The assay was proved to be sensitive,accurate and suitable for pharmacokinetic study of 5-ISMN.The results of pharmacokinetic study after oral administration of test and reference preparations of 5-ISMN showed that the two preparations were bioequivalent while the test preparation had a higher peak concentration and could reach peak level in less time,and the test preparation showed a rapid drug releasing behavior.