OBJECTIVE To investigate the roles and mechanisms of γ-bungarotoxin(γ-BGT)in inducing respiratory distress in mice.METHODS Six male Kunming mice were selected and anesthe-tized before tracheal intubation and respiratory recording.After stabilizing respiration,the mice were intraperitoneally injected with γ-BGT at a dose of 6 mg·kg-1.Once a decrease in respiratory frequency was observed,the mice were intravenously injected with nikethamide at a dose of 12.5 mg·kg-1.Respi-ratory frequency was monitored using the BL420 signal acquisition and processing system.Male Kunming mice were randomly divided into the normal control group(saline,ip),γ-BGT group(6 mg·kg-1,ip),and γ-BGT+nikethamide group(γ-BGT 6 mg·kg-1,ip,followed by nikethamide 12.5 mg·kg-1,ip,when shal-low breathing and enhanced abdominal respiration were observed).The levels of Glu and GABA in the medulla oblongata were measured using ELISA.The protein expression levels of GAD65 and GAD67 in the medulla oblongata were determined by Western blotting.Primary mouse medullary neurons were cultured in vitro and divided into the following groups:cell control group,γ-BGT group,carbachol group,gallamine group,γ-BGT+H-89 group,and γ-BGT+Y-27632 group.The γ-BGT group,carbachol group,and gallamine group were incubated with γ-BGT(40 mg·L-1),carbachol(100 mmol·L-1),and gallamine(100 mmol·L-1),respectively,for 4 h.The γ-BGT+H-89 and γ-BGT+Y-27632 groups were pretreated with γ-BGT(40 mg·L-1)for 4 h,followed by incubation with the protein kinase A(PKA)inhibitor H-89(50 mmol·L-1)and the Ca2+channel inhibitor Y-27632(50 mmol·L-1)for another 2 h,respectively.ELISA was used to measure the levels of Glu,GABA,cAMP,and calpain in the primary mouse medul-lary neurons.Western blotting was employed to assess the protein expression levels of GAD65 and GAD67,and PKA phosphorylation levels.Fluo-4 fluorescent probe was used to detect the intracellular Ca2+level.RESULTS The respiratory rate of mice significantly decreased after iv administration of γ-BGT(γ-BGT group)(P<0.05).After treatment with nikethamide(nikethamide group),the respiratory rate significantly recovered(P<0.05).Compared with the normal control group,the γ-BGT group exhib-ited a significant decrease in Glu content(P<0.05),a significant increase in GABA content(P<0.05),and a significant decrease in the Glu/GABA ratio.Additionally,the protein expression levels of GAD65 and GAD67 were significantly elevated(P<0.05).Compared with the γ-BGT group,the γ-BGT+niketh-amide group showed a significant increase in Glu content(P<0.05),a significant decrease in GABA content(P<0.05),a significant increase in the Glu/GABA ratio,and a significant reduction in GAD65 and GAD67 protein expression levels(P<0.05).Compared to the cell control group,the γ-BGT group demonstrated a significant decrease in Glu content(P<0.05),a significant increase in GABA content(P<0.05),and a significant reduction in the Glu/GABA ratio.Furthermore,the protein expression levels of GAD65 and GAD67 were significantly elevated(P<0.05).Additionally,cAMP content,PKA phosphor-ylation levels,Ca2+levels,and calpain activity were all significantly increased(all P<0.05).Glu,GABA,Glu/GABA ratio,and GAD expression levels in the γ-BGT group changed in the same way as in the gallamine group;In the γ-BGT+Y-27632 group,calpain activity and expression levels of GAD65 and GAD67 were all significantly decreased(all P<0.05).In the γ-BGT+H-89 group,Ca2+levels and calpain activity were significantly reduced(all P<0.05).CONCLUSION γ-BGT-induced poisoning can lead to respiratory distress in mice,possibly through the antagonism of M2 muscarinic acetylcholine receptors in medullary neurons,activation of the cAMP/PKA signaling pathway,elevation of intracellular Ca2+levels,and increased expression and activity of GAD,resulting in an imbalance of Glu and GABA in the medulla.

Infantile spasm is an age-related epileptic encephalopathy，most are drug-resistant，as the chirldren grow older，it will transfer into other epileptic syndrome.Most chirldren have a bad prognosis with neurodevelomental disorders.For now，aderenocortitropic hormone，glucocorticoid and vigabatrin are first-line treatment choice.But the control rate of spasm、remission rate of EEG and recurrence rate after first-line treatment are not ideal.It's necessary to find more effective treatment plans desperately.Considering the diversity and complexity of infantile spasm's etiologies，there are a lot of studies about treatment plan based on etiology.Controlling spasm more effectively，improving prognosis，personalized therapy and targeted therapy become the new focus.This article reviews the current treatment status of infantile spasm，also upates some novel methods which is not widely applied in clinical practice.

OBJECTIVE To investigate the roles and mechanisms of γ-bungarotoxin(γ-BGT)in inducing respiratory distress in mice.METHODS Six male Kunming mice were selected and anesthe-tized before tracheal intubation and respiratory recording.After stabilizing respiration,the mice were intraperitoneally injected with γ-BGT at a dose of 6 mg·kg-1.Once a decrease in respiratory frequency was observed,the mice were intravenously injected with nikethamide at a dose of 12.5 mg·kg-1.Respi-ratory frequency was monitored using the BL420 signal acquisition and processing system.Male Kunming mice were randomly divided into the normal control group(saline,ip),γ-BGT group(6 mg·kg-1,ip),and γ-BGT+nikethamide group(γ-BGT 6 mg·kg-1,ip,followed by nikethamide 12.5 mg·kg-1,ip,when shal-low breathing and enhanced abdominal respiration were observed).The levels of Glu and GABA in the medulla oblongata were measured using ELISA.The protein expression levels of GAD65 and GAD67 in the medulla oblongata were determined by Western blotting.Primary mouse medullary neurons were cultured in vitro and divided into the following groups:cell control group,γ-BGT group,carbachol group,gallamine group,γ-BGT+H-89 group,and γ-BGT+Y-27632 group.The γ-BGT group,carbachol group,and gallamine group were incubated with γ-BGT(40 mg·L-1),carbachol(100 mmol·L-1),and gallamine(100 mmol·L-1),respectively,for 4 h.The γ-BGT+H-89 and γ-BGT+Y-27632 groups were pretreated with γ-BGT(40 mg·L-1)for 4 h,followed by incubation with the protein kinase A(PKA)inhibitor H-89(50 mmol·L-1)and the Ca2+channel inhibitor Y-27632(50 mmol·L-1)for another 2 h,respectively.ELISA was used to measure the levels of Glu,GABA,cAMP,and calpain in the primary mouse medul-lary neurons.Western blotting was employed to assess the protein expression levels of GAD65 and GAD67,and PKA phosphorylation levels.Fluo-4 fluorescent probe was used to detect the intracellular Ca2+level.RESULTS The respiratory rate of mice significantly decreased after iv administration of γ-BGT(γ-BGT group)(P<0.05).After treatment with nikethamide(nikethamide group),the respiratory rate significantly recovered(P<0.05).Compared with the normal control group,the γ-BGT group exhib-ited a significant decrease in Glu content(P<0.05),a significant increase in GABA content(P<0.05),and a significant decrease in the Glu/GABA ratio.Additionally,the protein expression levels of GAD65 and GAD67 were significantly elevated(P<0.05).Compared with the γ-BGT group,the γ-BGT+niketh-amide group showed a significant increase in Glu content(P<0.05),a significant decrease in GABA content(P<0.05),a significant increase in the Glu/GABA ratio,and a significant reduction in GAD65 and GAD67 protein expression levels(P<0.05).Compared to the cell control group,the γ-BGT group demonstrated a significant decrease in Glu content(P<0.05),a significant increase in GABA content(P<0.05),and a significant reduction in the Glu/GABA ratio.Furthermore,the protein expression levels of GAD65 and GAD67 were significantly elevated(P<0.05).Additionally,cAMP content,PKA phosphor-ylation levels,Ca2+levels,and calpain activity were all significantly increased(all P<0.05).Glu,GABA,Glu/GABA ratio,and GAD expression levels in the γ-BGT group changed in the same way as in the gallamine group;In the γ-BGT+Y-27632 group,calpain activity and expression levels of GAD65 and GAD67 were all significantly decreased(all P<0.05).In the γ-BGT+H-89 group,Ca2+levels and calpain activity were significantly reduced(all P<0.05).CONCLUSION γ-BGT-induced poisoning can lead to respiratory distress in mice,possibly through the antagonism of M2 muscarinic acetylcholine receptors in medullary neurons,activation of the cAMP/PKA signaling pathway,elevation of intracellular Ca2+levels,and increased expression and activity of GAD,resulting in an imbalance of Glu and GABA in the medulla.

Objective:To investigate the clinical outcome of endovascular treatment vs. drug treatment in patients with symptomatic non-acute long-segment occlusion of the internal carotid artery. Methods:Based on prospective cohort registration research data, patients with symptomatic non-acute long-segment occlusion of internal carotid artery were retrospectively included. They were divided into a drug treatment group and an endovascular treatment group according to the actual treatment received. The latter was further divided into a successful recanalization group and an unsuccessful recanalization group. The endpoint events included ipsilateral ischemic stroke, any stroke, and all-cause death. Multivariate logistic regression analysis was used to compare the endpoint events between groups during the perioprocedural period (within 30 days), and multivariate Cox proportional hazards model was use to compare the endpoint events between the groups during the long-term follow-up. Results:A total of 684 patients were included, of which 570 (83.33%) were male, median aged 63 years (interquartile range, 56-70 years). Three hundred and fifty-three patients (51.6%) received drug treatment; 331 (48.4%) received endovascular treatment, of which 161 (48.6%) had successful recanalization. The median follow-up time was 1 223 days (interquartile range, 646.5-2 082 days), with 109 patients (15.9%) experiencing stroke recurrence events (including 87 ipsilateral ischemic stroke) and 78 (11.4%) experiencing all-cause mortality. The risk of any stroke during the perioprocedural period in the successful recanalization group was significantly higher than that in the drug treatment group (odds ratio 3.679, 95% confidence interval 1.038-13.036; P=0.044), but the risk of ipsilateral ischemic stroke recurrence (risk ratio 0.347, 95% confidence interval 0.152-0.791; P=0.012) and all-cause mortality (risk ratio 0.239, 95% confidence interval 0.093-0.618; P=0.003) during the long-term follow-up were significantly lower than those in the drug treatment group. Conclusions:In patients with symptomatic non-acute long-segment occlusion of the internal carotid artery, endovascular treatment can increase the risk of stroke recurrence within 30 days, but successful recanalization can reduce the risks of long-term ipsilateral ischemic stroke recurrence and all-cause mortality.

Objective:To investigate predictive factors for successful endovascular recanalization in patients with non-acute symptomatic internal carotid artery occlusion (SICAO), to develop a decision tree model using the Classification and Regression Tree (CART) algorithm, and to evaluate the predictive performance of the model.Methods:Patients with non-acute SICAO received endovascular therapy at 8 comprehensive stroke centers in China were included retrospectively. They were randomly assigned to a training set and a validation set. In the training set, the least absolute shrinkage and selection operator (LASSO) algorithm was used to screen important variables, and a decision tree prediction model was constructed based on CART algorithm. The model was evaluated using the receiver operating characteristic (ROC) curve, Hosmer-Lemeshow goodness-of-fit test and confusion matrix in the validation set.Results:A total of 511 patients with non-acute SICAO were included. They were randomly divided into a training set ( n=357) and a validation set ( n=154) in a 7:3 ratio. The successful recanalization rates after endovascular therapy were 58.8% and 58.4%, respectively. There was no statistically significant difference ( χ2=0.007, P=0.936). A CART decision tree model consisting of 5 variables, 5 layers and 9 classification rules was constructed using the six non-zero-coefficient variables selected by LASSO regression. The predictive factors for successful recanalization included fewer occluded segments, proximal tapered stump, ASITN/SIR collateral grading of 1-2, ischemic stroke, and a recent event to endovascular therapy time of 1-30 d. ROC analysis showed that the area under curve of the decision tree model in the training set was 0.810 (95% confidence interval 0.764-0.857), and the optimal cut-off value for predicting successful recanalization was 0.71. The area under curve in the validation set was 0.763 (95% confidence interval 0.687-0.839). The accuracy was 70.1%, precision was 81.4%, sensitivity was 63.3%, and specificity was 79.7%. The Hosmer-Lemeshow test in both groups showed P>0.05. Conclusion:Based on the type of ischemic event, the time from the latest event to endovascular therapy, proximal stump morphology, the number of occluded segments, and the ASITN/SIR collateral grading constructed the decision tree model can effectively predict successful recanalization after non-acute SICAO endovascular therapy.