Akt Inhibitor Perifosine Prevents Epileptogenesis in a Rat Model of Temporal Lobe Epilepsy / 神经科学通报·英文版

Accumulating data have revealed that abnormal activity of the mTOR (mammalian target of rapamycin) pathway plays an important role in epileptogenesis triggered by various factors. We previously reported that pretreatment with perifosine, an inhibitor of Akt (also called protein kinase B), abolishes the rapamycin-induced paradoxical increase of S6 phosphorylation in a rat model induced by kainic acid (KA). Since Akt is an upstream target in the mTOR signaling pathway, we set out to determine whether perifosine has a preventive effect on epileptogenesis. Here, we explored the effect of perifosine on the model of temporal epilepsy induced by KA in rats and found that pretreatment with perifosine had no effect on the severity or duration of the KA-induced status epilepticus. However, perifosine almost completely inhibited the activation of p-Akt and p-S6 both acutely and chronically following the KA-induced status epilepticus. Perifosine pretreatment suppressed the KA-induced neuronal death and mossy fiber sprouting. The frequency of spontaneous seizures was markedly decreased in rats pretreated with perifosine. Accordingly, rats pretreated with perifosine showed mild impairment in cognitive functions. Collectively, this study provides novel evidence in a KA seizure model that perifosine may be a potential drug for use in anti-epileptogenic therapy.

Inhibiting mammalian target of rapamycin signaling pathway improves cognitive function in mice with chronic cerebral ischemia / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of mammalian target of rapamycin(mTOR) inhibitor-rapamycin on cognitive function after chronic cerebral ischemia in mice and its molecular mechanism.</p><p><b>METHODS</b>The chronic cerebral ischemia model was induced by ligation of right common carotid artery (rUCCAO) in 6-week-old ICR mice. The expressions of mTOR, S6K, S6 and corresponding phosphorylated proteins were detected by Western blotting at different time interval (1 h, 3 h, 6 h, 24 h, 3 d, 7 d, 2 w, 4 w, 6 w) after rUCCAO to determine the changes of mTOR signaling pathway. Rapamycin was administrated i.p. at the dose of 3.0 mg/kg 24 h after rUCCAO. Fluoro Jade B staining was used to detect the apoptotic cells. The expressions of Beclin and LC3-Ⅱ were detected by Western blotting to determine the status of autophagy. Morris water maze test and Y maze test were performed to evaluate cognitive functions.</p><p><b>RESULTS</b>The mTOR signaling pathway was abnormally activated from 6 h to 6 w after rUCCAO in mouse cortex. The activation of mTOR signaling pathway induced by rUCCAO was reversed by administration of rapamycin, and the apoptotic cell number was significantly decreased (146.1±16.3 vs 84.5±9.6,<0.05). Meanwhile, the elevation of Beclin and LC3-Ⅱ protein induced by rUCCAO was reversed by rapamycin administration. Furthermore, compared with vehicle-treated mice, the latent period[(11.1±2.3) s vs (8.1±1.8) s,<0.05] and swimming distance[(672.8±128.5) cm vs (558.2±124.9) cm,<0.05] were significantly decreased and the number of crossing the platform quadrant in Morris water maze increased(2.8±0.9 vs 5.2±0.8,<0.05) in rapamycin-treated mice. Correct response rate in the Y maze was also increased significantly in rapamycin-treated mice[(38.5±9.2)% vs (64.9±7.9)%,<0.05].</p><p><b>CONCLUSIONS</b>Inhibiting mTOR pathway by rapamycin reverses the rUCCAO-induced cognitive impairment partly through the suppression of apoptosis and autophagy.</p>

Effective dosage of sirolimus for seizure treatment of immature C57BL/6 mice induced by kainic acid / 中国药理学与毒理学杂志

OBJECTIVE To explore the safe and effective dose of sirolimus (Rapamycin,Sir) and its effect on seizure comorbidities. METHODS Immature C57BL/6 mice at postnatal 10 d of age were administered with kainic acid(KA) 12.0 mg · kg-1 intraperitoneally by a single injection to induce acute seizure. Sir 0.3, 1.0 and 3.0 mg · kg-1 was injected 24 h after seizure every other day until 3 d, 1 week, 3 weeks, 5 weeks and 6 weeks. Western blotting analysis was used to detect the expression and phos?phorylation level of S6 protein and to determine the minimum effective dose of Sir. Effect of the mini?mum effective dose of Sir on cognitive function and body growth was observed by several evaluations. Immunofluorescent intensity of Doublecortin (DCX) immunofluorescent staining was conducted to evaluate the development of neurons in the hippocampus. Morris water maze was used to assess the cognitive function. Tail suspension test, O maze and new object recognition test were used to study the anxiety-like behaviors of mice. RESULTS The result of Western blotting showed that Sir 0.3 mg · kg-1 had no significant effect on the phosphorylation of S6 protein in normal mice or KA mice, whereas 1.0 and 3.0 mg · kg- 1 could significantly inhibit the phosphorylation of S6 protein in KA mice (P<0.05). Sir 1.0 mg·kg-1 had no obvious effect on DCX-positive cells or body wass. Morris water maze showed that KA-induced seizure resulted in prolonged escape latency and swimming length (P<0.05), and a decreased crossing number of target quadrant (P<0.05). Sir 1.0 mg·kg-1 significantly reversed the deficit of cognitive function of KA-induced seizure mice (P<0.05), whereas no significant difference was found between Sir group and normal control group. Compared with normal control group, model group showed increased freezing time in tail suspension test (P<0.05), decreased migration length and reten?tion time in open arms in O maze (P<0.05), decreased retention time and touch frequency with new objects, migration length and average speed in new object recognition test (P<0.05). Sir 1.0 mg · kg-1 significantly reversed the above anxiety and depression status, whereas no significant difference was found between sirolimus group and normal control group. CONCLUSION Sir 1.0 mg · kg-1 inhibits the abnormal activation of mTOR pathway and the formation of epilepsy comorbidity in immature mice. Along with its mild side effect in development, Sir 1.0 mg · kg-1 will be an ideal dose to be used in the treatment of seizure in immature mice.

Risk Factors Analysis for Prevalence of Acute Myocardial Infarction in Young and Middle-aged Population / 中国循环杂志

Objective: To study the risk factors for prevalence of acute myocardial infarction (AMI) in young and middle-aged population. Methods: A prospective cohort study was conducted in 110100 subjects at the age of (18-98) years who received physical examination in Kailuan Group from 2012-06 to 2014-10. Based on the limitations of male≤53 years and female≤63 years, a total of 62367 subjects were enrolled in our study. The subjects were followed-up for 2 years by the end point event of AMI to analyze the risk factors ofAMI occurrence. Results: According to AMI occurrence at the follow-up period, the subjects were divided into 2 groups: AMI group, n=56 and Control group, n=62152. Compared with Control group, AMI group had increased BMI, SBP, DBP and elevated blood levels of LDL-C, TG; AMI group also showed the higher ratios of subjects with the history of diabetes and taking anti-hypertension medication. Cox proportional hazard regression analysis indicated that age (RR=1.37), male (RR=60.54), LDL-C (RR=1.12), and TG (RR=5.93) were the risk factors forAMI occurrence in young and middle-aged population, allP<0.05. Conclusion: Age, male gender, blood levels of LDL-C, and TG were the risk factors for AMI occurrence in young and middle-aged population.

Entity of acupoint: kinetic changes of acupoints in histocytochemistry / 中国针灸

The present study aims to investigate the kinetic histocytochemical changes of acupoints in different condition. The expression of tryptase (+) mast cells, histamine (HA) , serotonin (5-HT) and nociceptive neuropeptides including calcitonin gene-related peptide (CGRP) and substance P (SP) were observed by immunohistochemistry combined with confocal technology. Mast cells were labeled with anti-mast cell tryptase antibody and simultaneously with HA or 5-HT primary antibodies to observe their co-expression. The results showed that: (1) SP and CGRP were expressed more highly on the cutaneous nerve fibers of "Hegu" (LI 4) after acupuncture stimulation than that of the control. Mast cells aggregated in close proximity to the blood vessels in intra-epidermis and dermis, and some of them with degranulation in the lower dermis and subcutaneous tissue of "Hegu" (LI 4). Both mast cells and their granules appeared with HA (+) and 5-HT (+) expression at stimulated LI 4 sites, while a few intact mast cells with a little expression of 5-HT and HA were distributed in areas of non-stimulated Ll 4. (2) The acupoints in different locations such as Baihui (GV 20), Weishu (BL 21), Zhongwan (CV 12) and LI 4 had the same constituent but the contents were different. (3) The histocytochemical responses of acupoints sensitized by the Gastric mucosa injury (GMI) were also investigated. GMI resulted in neurogenic plasma extravasation by Evans Blue (EB) in the skin of the acupoints over the back and abdomen, which mostly occurred in the T9-T11 dermatomere. The EB extravasation dots just like acupoints sensitization appeared after GMI and disappeared gradually during the natural self-recovery of the gastric mucosa. More SP and CGRP positive nerve fibers were distributed in EB dots than in regions beside EB dots and in the control, mostly distributed in the nerve fibers around both the vessels and root of hair follicle. Mast cells also aggregated and degranulated to release algogenic substances of 5-HT and HA around the vessels in areas of the EB dots. Collectively the acupoints displayed the same histocytochemical responses due to either acupuncture stimulation or GMI. This may potentially be the histocytochemical basis in the local acupoints and acupoints displayed kinetic changes in different condition.

Myocardial systolic function of local left ventricle in patients with hypertrophic cardiomyopathy evaluated bystrain and strain rate imaging / 中国基层医药

ObjectiveTo evaluate regional myocardial systolic function in patients with hypertrophic cardiomyopathy(HCM) by strain and strain rate imaging.Methods38 patients with HCM(observe group) and 36 healthy volunteers(control group) were involved in this study.All subjects were received conventional 2D-Color Doppler echocardiography and Color Doppler myocardial image(CDMI) ,then analyzed the regional myocardial function by strain and strain rate imaging.ResultsComparwed with the control group, left atrial diameters (LAD), left ventricular diameters (LVD), left ventricular end-systolic dimension (LVDs), interventricular ventricular septal (IVS) and left ventricular posterior wall(LVPW) were significantly increased in patients of observe group (all P < 0.05), but left ventricular ejection fraction(LVEF) was decreased(P <0.05).Patients with HCM showed regional longitudinal peak systolic myocardial deformation properties lower than those of counterparts at inferior, lateral, posterior, frontal and septum waLl of left ventricular(all P < 0.05).ConclusionHypertrophic cardiomyopathy was associated with significant reduction in systolic function of left ventricle and strain and strain rote imaging was useful in evaluating the change.