1.A Study In Screening Mutation Of The Nicotinic Acetylcholine Receptor Gene In Sporadic Nocturnal Frontal Lobe Epilepsy 2.A Study In Assessing Hemispheric Language Dominance In Drug-resistant Temporal Lobe Epilepsy Before Surgery By Using FMRI

Background and ObjectivesThe prevalence of epilepsy is estimated at 0.5-1.5%in the world and there are at least 7-8 million epileptics in china.Forty percent of them are idiopathic epilepsy, in which the genetic etiology play an important role to the pathogenesis.It is very important to find out the genetic etiology to diagnose and treat the epilepsy.ADNFLE is sometimes associated with mutations in the transmemberane domain(TM) 1-3 in the genes encoding theα4(CHRNA4),β2(CHRNB2)andα2(CHRNA2) subunits of the neuronal nicotinic acetylcholine receptors(nAChRs).Clinically,sporadic cases of nocturnal frontal lobe epilepsy(NFLE) with no demonstrable structural brain lesion are more common than that of ADNFLE and there are no phenotypic difference between sporadic and ADNFLE.In a study,a de novo mutation(CHRNA4-C755T) which had been reported in ADNFLE was found in a lebanese women with sporadic NFLE and was also found in a son with NFLE of the women.Because there is few patient with ADNFLE and a few patients with NFLE in china,we performed a clinical and molecular genetic study in 33 chinese patients of sporadic NFLE to assess the role of the mutations of CHRNA4, CHRNB2 and CHRNA2 in the pathogenesis of sporadic NFLE.Materials and MethodsWe obtained clinical information from 33 unrelated outpatients(19 male) with sporadic NFLE and the diagnosis of these patients was based on the clinical history, EEG/ictal VEEG and cranial magnetic resonance imaging(MRI).DNA was extracted from peripheral blood of these patients and 100 healthy control persons by using a standard proteinase K digestion and phenol-chloroform method,we performed a mutation screening in 33 unrelated patients with sporadic NFLE by amplifying and sequencing bidirectionally the part of CHRNA4,CHRNB2 and CHRNA2 which containing the TM 1-3 and the mutaions reported in ADNFLE.We used PCR-RFLP analysis with restriction enzyme Cac8Ⅰand MscⅠto confirm the novel two substitutions presented in sequencing in CHRNA4 and screen them in 100 healthy controls.We investigated the the role of the mutations of CHRNA4,CHRNB2 and CHRNA2 in the pathogenesis of sporadic NFLE by analysising the mutation property,the transferring of the mutation in family and the conservative extent of the amino acid residue resulted from the mutation among species and other nAChR subunits.ResultsIn 33 chinese patients with sporadic NFLE,the segment containing the TM 1-3 of exon 5 in CHRNA4 was sequenced bidirectionally for mutations.It identified a heterozygous mutation in one patient,G923A,while it was a missense mutation and resulted in a change from arginine to histidine(R308H) located in the second intracellular loop between the TM 3 and TM 4 and adjacent to TM 3.We also used the same method to show that the mutation was absent in 100 healthy control individuals.We found a potentially novel polymorphism(A1054G) and the polymorphism T1125G known previously.No mutation was found by sequencing bidirectionally a 468-bp segment containing the TM 1-3 ofexon 5 in CHRNB2 in 33 patients with sporadic NFLE. Nor was in CHRNA2.Conclusion1.The missense mutation R308H in CHRNA4 in chinese patients with sporadic NFLE in our study may be a genetic etiology and take a important role in the pathogenesis of sporadic NFLE.2.Mutations external to TM of exon 5 in CHRNA4 may be associated with NFLE/ADNFLE.3.Our study seems to confirm that exon 5 of CHRNA4 is an hot spot mutation site for NFLE/ADNFLE,while the mutations in CHRNB2 and CHRNA2 are an extremely rare cause of the diseases. Background and ObjectivesThe prevalence of epilepsy is estimated at 0.5-1.5%in the world and there are at least 7-8 million epileptics in china,of which 30-40%are medically refractory. Operation to resetting the epilepsy genesis is an effective therapy for medically refactory epilepsy,while localizating the functional areas around the epilepsy genesis before surgery is important to minimizing the impairment of the surgery and became a hotspot in presurgical assessment in patients with epilepsy.Many studies showed that fMRI has become a routine clinical technique for language lateralization and mapping before epilepsy surgery,because of the noninvasive and reliability.It was demonstrated that using the laterality index and the semantic task in fMRI were the best way to lateralizing the hemispheric language dominance(HLD),while the LI in inferior frontal gyms was more consistent with LI in Wada test than LI in temporopariental region and LI was associated with many factors such as age at onset,epilepsy duration and so on.Several studies suggested that the lateralization were different in reading Chinese characters and English words in same patients in fMRI.Recent studies demonstrated that fMRI-LI in presurgery assessment could predict the naming function decline after left anterior temporal lobectomy(L-ATL),in which the LI in temporopariental had the more predictive effect.However,few studies were performed to investigate the effect of LI aboved in language fMRI in chinese patients with temporal lobe epilepsy.In this study,we used fMRI to evaluated presurgical lateralisation of language in chinese patients with drug-resistant temporal lobe epilepsy,investigating the relationship between the total LI and the focal LI(frontal LI and temporoparietal LI),the predictive effedct of LI in the naming decline after anterior temporal lobectomy and the factories associated with LI.MethodsA prospective cohort study was performed in PUMCH,including 10 patients with drug-resisitant temporal lobe epilepsy,while the diagnosis of these patients was based on the clinical history,EEG/ictal VEEG and cranial magnetic resonance imaging (MRI),and the temporal lobe origin of epilepsy was determined by natural history, clinical history,ictal and interictal VEEG.interictal PET,interictal and/or ictal SPECT, MRI and MRS.The clinical data of them were reviewed.All subjects were examinated with language examination by Aphasia Battery of Chinese(ABC) before the surgery and 6-9 months after surgery.In assessing HLD by using fMRI with chinese semantic task, We took the left inferior frontal region,the left temporoparietal region and the homologous region in right hemisphere as the distinct region of interest(ROI) to calculate each laterality index.Statistical analysis was performed by Cohen's kappa coefficient of agreement comparing the HLD results of the LI in inferior frontal region (LI-IF) or temporoparietal region(LI-TP) with the LI in total.In addition,the correlation between the LI in total and the demographic factors and the clinical factors was analysised by a spearman's correlation analysis or X~2 test.Base on the score of ABC before and after surgery,we investigated the predictive effect of fMRI-LI for the naming changes in patients suffered the ATL. ResultsThe HLD indicated by LI-total showed more a baddish agreement with the HLD indicated by LI-IF(kappa=0.375),whereas the HLD indicated by LI-total showed more a good agreement with the HLD indicated by LI-TP(kappa=1).Among the demographic and clinical factors(actal age,age at onset,epilepsy duration,and the side of epilepsy origin),age at onset(r=0.778,p＜0.01) and epilepsy duration were significantly correlation with LI(r=-0.744,p＜0.01).The naming changes predicted by the LI was agreement with the naming changes determined by the score changes of naming in L-ATL group(kappa=0.5),while the LI were significantly negative correlation with the score changes of naming in R-ATL group(r=-0.924,p＜0.01).ConclusionsThe results showed that the LI-TP was more consistent with LI-total by using fMRI to indicating the HLD in non-lesional temporal epilepsy with senmantic chinese task in presurgical estimate,while the age at onset and duration of epilepsy could influence the generation of HLD.The fMRI-LI could predict naming function changes after ATL.