Neuronal nitric oxide synthase/reactive oxygen species pathway is involved in apoptosis and pyroptosis in epilepsy

Dysfunction of neuronal nitric oxide synthase contributes to neurotoxicity,which triggers cell death in various neuropathological diseases,including epilepsy.Studies have shown that inhibition of neuronal nitric oxide synthase activity increases the epilepsy threshold,that is,has an anticonvulsant effect.However,the exact role and potential mechanism of neuronal nitric oxide synthase in seizures are still unclear.In this study,we performed RNA sequencing,functional enrichment analysis,and weighted gene coexpression network analysis of the hippocampus of tremor rats,a rat model of genetic epilepsy.We found damaged hippocampal mitochondria and abnormal succinate dehydrogenase level and Na+-K+-ATPase activity.In addition,we used a pilocarpine-induced N2a cell model to mimic epileptic injury.After application of neuronal nitric oxide synthase inhibitor 7-nitroindazole,changes in malondialdehyde,lactate dehydrogenase and superoxide dismutase,which are associated with oxidative stress,were reversed,and the increase in reactive oxygen species level was reversed by 7-nitroindazole or reactive oxygen species inhibitor N-acetylcysteine.Application of 7-nitroindazole or N-acetylcysteine downregulated the expression of caspase-3 and cytochrome c and reversed the apoptosis of epileptic cells.Furthermore,7-nitroindazole or N-acetylcysteine downregulated the abnormally high expression of NLRP3,gasdermin-D,interleukin-1βand interleukin-18.This indicated that 7-nitroindazole and N-acetylcysteine each reversed epileptic cell death.Taken together,our findings suggest that the neuronal nitric oxide synthase/reactive oxygen species pathway is involved in pyroptosis of epileptic cells,and inhibiting neuronal nitric oxide synthase activity or its induced oxidative stress may play a neuroprotective role in epilepsy.

Harnessing interventions during the immediate perioperative period to improve the long-term survival of patients following radical gastrectomy

Although the incidence and mortality of gastric cancer(GC)have been decreasing steadily worldwide,especially in East Asia,the disease burden of this malignancy is still very heavy.Except for tremendous progress in the management of GC by multidisciplinary treatment,surgical excision of the primary tumor is still the cornerstone intervention in the curative-intent treatment of GC.During the relatively short perioperative period,patients undergoing radical gastrectomy will suffer from at least part of the following perioperative events:Surgery,anesthesia,pain,intraoperative blood loss,allogeneic blood transfusion,postoperative complications,and their related anxiety,depression and stress response,which have been shown to affect long-term outcomes.Therefore,in recent years,studies have been carried out to find and test interventions during the perioperative period to improve the long-term survival of patients following radical gastrectomy,which will be the aim of this review.

Differential Expression of Alpha-Adrenoceptor Subtypes in Rat Dorsal Root Ganglion after Chronic Constriction Injury

Summary： mRNAs of alpha-adrenoceptor （α-AR） subtypes are found in neurons in dorsal root ganglion （DRG） and change after peripheral nerve injury. In this study, the distribution of α-AR subtype proteins was studied in L5 DRG of normal rats and rats with chronic constriction injury of sciatic nerve （CCI）. Using immunofluorescence technique, it was found that α1A-, α1B-, and α2A-AR proteins were expressed in large, medium, and small size neurons in normal DRG, and significantly increased in all size neurons 14 days after CCI. α1D- and α2C-AR was also expressed in all size neurons in normal DRG. However, α1D-AR was significantly increased and α2C-AR was decreased in small size neurons 14 days post CCI. α2B-AR neurons were not detectable in normal and CCI DRG. Co-expression of α1A- and α2A-AR in the same neuron was observed in normal DRG and increased post CCI. Collectively, these results indicated that there is distinct distribution of α-AR subtypes in DRG neurons, and the distribution and levels of expression of α-AR subtypes change differently after CCI. The up-regulation of α-AR subtypes in DRG neurons may play an important role in the process of generating and transmitting neuropathic pain.

Simple hematological predictors of AF recurrence in patients undergoing atrial fibrillation ablation

Backgound Red cell distribution width(RDW) and neutrophil-to-lymphocyte ratio(NLR) are simple hematologic indices that have been used to predict adverse outcomes in different clinical settings. The aim of our study is to determine whether RDW and NLR can predict atrial fibrillation(AF) recurrence in patients undergoing AF ablation. Methods Consecutive patients, without known hematological disorders, who underwent AF catheter ablation between January 2014 and April 2017 were enrolled into this study. Blood samples were taken one day before and five hours after the ablation procedure. Results A total of 346 patients(224 males(65%), mean age: 59 ± 11 years old) were included. After a mean follow up of 26.2 ± 12.1 months, 80(23.1%) patients experienced late AF recurrence(defined as any recurrence after the blanking period of three months), while 97(28%) patients experienced early AF recurrence during the blanking period. Univariate analysis showed that early arrhythmia recurrence, type of AF and NLR after the procedure were significantly associated with late AF recurrence, while early arrhythmia recurrence and NLR remained significant in multivariate analysis. RDW was not associated with late AF recurrence. None of the parameters above predicted early arrhythmia recurrence. Conclusions Simple and inexpensive hematological indices such as NLR should be evaluated for their ability to predict AF recurrence in patients undergoing catheter ablation in larger prospective studies.

Predictive models for mild cognitive impairment to Alzheimer's disease conversion

Alzheimer's disease(AD) is an irreversible and progressive neurodegenerative disease as well as the most common form of dementia. It usually affects the older population, but early onset AD is still possible(Ritchie et al., 2015). Recent studies propose that AD is a middle-life disease(Ritchie et al.,2015).

Does electrophysiological testing have any role in risk stratification for sudden cardiac death？

Implantation of implantable cardioverter defibrillators （ICD） has widely been accepted for secondary prevention of sudden cardiac death （SCD） in cardiac arrest survivors.1 Currently there are increasing interests in primary preven- tion of SCD in selected high risk patients who have not experienced cardiac arrest.~ Despite extensive investigation for risk stratification, our current ability to accurately iden- tify patients at high risk for SCD remains very poor. The primary reason is probably due to our limited understand- ing of the mechanisms underlying the pathogenesis of ven- trieular tachy-arrhythmias and sudden cardiac death （Fig. 1）.

Arrhythmias,chronic kidney disease and the elderly:a triple jeopardy

Crdiovascular diseases (CVD) incur a heavy burden of morbidity and mortality among patients with chronic kidney disease (CKD),particularly among the elderly.It is estimated that about 22-25% of all adults beyond the age of 65 years have moderate or severe renal dysfunction.1。

Recent concepta regarding“Mahaim”accessory pathway;Implication for catheter ablation.

Mahaim accessory pathway has generally been regarded as an accessorypathway which originates from the atrioventricular node and is inserted at,or adjacent to the right bundle branch or ventricle.Recently.severalreportes have demonstrated that these pathways originate from the atrialfree wall distant from the atrioventriculer node.Electrophysiologicalcharacteristics of Mahaim fiber have also been investigated particularlyconcerning its decremental property by examining electrophysiologicalbehavior of the so-called Mahaim potential.We also tried.to delineate thefanctional and anatomical characteristics along the Mahaim fiber with thedetailed electrophysiological study.In my presentation 1 would like todemonstrate that there is electrophysiological evidence Of functionallongitudinal dissociation or supernormality in the Mahaim fiberextrapolated from the discontinuity or the resumption of the antegradeconduction over the Mahaim fiber.1 also demonstrate the exact procedureto eliminate conduction over the Mahaim pathways using radiofrequanencycatheter ablation targeted at the tricuspid annulus.An inter esting case,which under wenr catheter ablation for its wide QRS complex tachycardiaassciated with Mahaim pathway,will be presented for your reference.

致心律失常性右室发育不良患者室性心动过速的基质标测和消融:近期和长期成功率研究

Background-Multiple morphologies,hemodynamic instability, or noninducibility may limit ventricular tachycardia(VT) ablation in patients with arrhythmogenic right ventricular dysplasia(ARVD). Substrate-based mapping and ablation may overcome these limitations. We report the results and success of substrate based VT ablation in ARVD. Methods and Results-Twenty-two patients with ARVD were studied. Traditional mapping for VT was limited because of multiple/changing VT morphologies(n=14), nonsustained VT(n=10),or hemodynamic intolerance(n=5). Sinus rhythm CARTO mapping was performed to define areas of “scar”(< 0.5 mV) and “abnormal”myocardium(0.5 to 1.5 mV). Ablation was performed in “abnormal”regions, targeting sites with good pace maps compared with the induced VT(s). Linear lesions were created in these areas to(1) connect the scar/abnormal region to a valve continuity or other scar or(2) encircle the scar/abnormal region. Eighteen patients had implanted cardioverter defibrillators, 15 had implanted cardioverter defibrillator therapies, and 7 had sustained VT(6 with syncope). VTs(3±2 per patient) were induced(cycle length, 339±94 ms), and scar was identified in all patients. Scar areas were related to the tricuspid annulus, proximal right ventricular outflow tract, and anterior/inferior-apical walls. Lesions connected abnormal regions to the annulus(n=12) or other scars(n=4) and/or encircled abnormal regions(n=13). Per patient, a mean of 38±22 radiofrequency lesions was applied. Shortterm success was achieved in 18 patients(82%). VT recurred in 23%, 27%, and 47%of patients after 1, 2, and 3 years’follow-up, respectively. Conclusions-Substrate-based ablation of VT in ARVD can achieve a good short-term success rate. However, recurrences become increasingly common during long-term follow-up.

铝抑制拟南芥根尖PIN2循环和囊泡运输

铝对植物毒害作用最明显的症状是迅速抑制根尖生长.然而,铝抑制根尖生长的机制并不清楚.本文研究了铝对生长素和生长素运输载体(PIN2)囊泡运输的影响.结果表明,铝抑制拟南芥根尖生长素运输,其中过渡区生长素抑制率最高,达66%.布雷菲尔德菌素(Brefeldin A,BFA,一种囊泡运输抑制剂)明显诱导PIN2囊泡在细胞内形成点状结构,铝处理降低点状结构的大小,表明铝抑制PIN2囊泡在细胞内的运输.实时定量PCR和蛋白印迹反应发现,铝增加PIN2基因的转录表达,促进PIN2蛋白在细胞膜水平方向累积.细胞骨架解聚药物处理表明,铝抑制PIN2囊泡的运输,主要通过破坏肌球蛋白微丝来完成.铝处理下,拟南芥根尖伸长区细胞比过渡区具有较少的铝吸收和较低的囊泡运输频率.上述结果表明,通过调节生长素运输载体(PIN2)在质膜与胞内移动,阻碍生长素的运输,铝抑制了拟南芥根尖的生长.

Heat induces adenosine triphosphate release from mast cells in vitro:a putative mechanism for moxibustion

OBJECTIVE:To investigate the role of adenosine triphosphate(ATP) purinergic signaling in mast cells(MCs) modulated by heat to further understand the molecular mechanisms of moxibustion.METHODS:Skin temperatures induced by monkshood cake moxibustion were evaluated by measuring the Neiguan acupoint(PC 6) from 31 participants with a digital thermocouple thermometer.Temperatures of 43 ℃ and 52 ℃ were applied to cultured human leukemia mast cell line HMC-1 in vitro.Calcium fluorescence was applied to detect intracellular Ca^(2+)([Ca^(2+)]_i).Extracellular ATP contents were measured by luciferin-luciferase assay.RESULTS:Maximum skin temperatures mostly ranged from 40-45 ℃,but some reached up to50 ℃.Both 43 ℃,and 52 ℃ induced MC degranulation,which was accompanied by an increase in[Ca^(2+)]_i and ATP release.Complexing extracellular Ca^(2+) with 5 mM ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid(EGTA) inhibited the noxious heat-induced elevation of[Ca^(2+)]_i and prevented the enhanced ATP secretion by those cells at 52℃ but not 43 ℃.CONCLUSION:Monkshood cake moxibustion can generate heat sufficient to trigger cellular events of MCs,including degranulation,[Ca^(2+)]_i elevation,and ATP release,suggesting that purinergic signals originating from MCs are possibly the initiating response of acupoints to moxibustion.

Aluminium toxicity targets PIN2 in Arabidopsis root apices: Effects on PIN2 endocytosis, vesicular recycling, and polar auxin transport

The most obvious symptom of Al toxicity is the inhibition of root growth. However, the mechanism of Al-inhibiting root growth remains to be elucidated. In this study, auxin transport and vesicle movement of an auxin-efflux carrier (PIN2) were investigated in Arabidopsis roots in response to Al stress. Results indicated that Al inhibited the apical transport of auxin in root tips of Arabidopsis significantly. The severe inhibition was localized in the cells of transition zone, where the concentration of auxin was only 34% that of the control. Brefeldin A (BFA), an inhibitor of vesicle transport, induced the dot-like structure of PIN2 vesicle significantly. Al decreased the size of dot-like structure of PIN2 vesicles. Re- sults of real-time RT-PCR and Western-blotting analysis showed that Al increased the transcript level of PIN2 and the accumulation of PIN2 protein in horizontal direction of plasma membrane, but decreased its distribution in endosomes, suggesting that Al inhibited the transport of PIN2 vesicles from plasma membrane to endosomes. Results of cytoskeleton-depolymering drugs indicated that it was via the pathway of disruption of actin microfilaments that Al inhibited the transport of PIN2 vesicles. Exposed to Al stress, the cells of elongation zone had less Al uptake and less transport frequency of vesicles than cells of transition zone. Taken together, our results suggested that Al inhibited root growth mainly by modulating the transport of PIN2 vesicles between plasma membrane and endosomes, thus block- ing auxin transport and root growth.

Potential functions of esophageal cancer-related gene-4 in the cardiovascular system

Esophageal cancer-related gene-4(Ecrg4)is cloned from the normal epithelium of the esophagus.It is constitutively expressed in quiescent epithelial cells and downregulated during tumorigenesis,and Ecrg4 expression levels are inversely correlated with the malignant phenotype of tumor cells,validating that Ecrg4 is a real tumor suppressor gene.Unlike other tumor suppressor genes that usually encode membrane or intracellular proteins,Ecrg4 encodes a 148-amino acid pre-pro-peptide that is tethered on the cell surface in epithelial cells,specialized epithelial cells,and human leukocytes,where it can be processed tissue dependently into several small peptides upon cell activation.Ecrg4 is expressed in a wide variety of other cells/tissues,including cardiomyocytes and conduction system of the heart,,the glomus cells of the carotid body,adrenal glands,choroid plexus,and leukocytes among others,where it exerts distinct functions,such as promoting/suppressing inflammation,inducing neuron senescence,stimulating the hypothalamus--pituitary--adrenal axis,maintaining the stemness of stem cells,participating in the rhythm and rate control of the heart,and possibly gauging the responsiveness of the cardiovascular system(CVS)to hypoxia,in addition to tumor suppression.Here,we briefly review the latest discoveries on Ecrg4 and its underlying molecular mechanisms as a tumor suppressor and focus on the emerging roles of Ecrg4 in the CVS.

脂布福吉宁和乙酰毒毛花甙诱发离体浦肯野氏纤维延时性后去极化及触发性心律失常的比较(英文)

用细胞外电图信号叠加技术微电极,于羊浦肯野纤维上,比较脂布福吉宁(RBG)(?)乙酰毒毛花甙元(AS)对诱发延时性后去极化(DAD)和触发性心律失常(TA)的作用,结果:1)低中毒剂量于刺激为990和690 ms时诱发出胞内和胞外的DAD;2)高中毒剂量诱发出DAD及TA,包含过早动作电位及触发性心动过速等;3)均引起动作电位振幅、静息电位及最大舒张期电位的降低、50％APD及Q-T间期的缩短,故RBG属于洋地黄类药物。

Single-vesicle imaging quantifies calcium’s regulation of nanoscale vesicle clustering mediated by α-synuclein

Although numerous studies have shown that the proteinα-synuclein(α-Syn)plays a central role in Parkinson’s disease,dementia with Lewy bodies,and other neurodegenerative diseases,the protein’s physiological function remains poorly understood.Furthermore,despite recent reports suggesting that,under the influence of Ca^(2+),α-Syn can interact with synaptic vesicles,the mechanisms underlying that interaction are far from clear.Thus,we used single-vesicle imaging to quantify the extent to which Ca^(2+)regulates nanoscale vesicle clustering mediated by α-Syn.Our results revealed not only that vesicle clustering requiredα-Syn to bind to anionic lipid vesicles,but also that different concentrations of Ca^(2+)exerted different effects on howα-Syn induced vesicle clustering.In particular,low concentrations of Ca^(2+)inhibited vesicle clustering by blocking the electrostatic interaction between the lipid membrane and the N terminus of α-Syn,whereas high concentrations promoted vesicle clustering,possibly due to the electrostatic interaction between Ca^(2+)and the negatively charged lipids that is independent of α-Syn.Taken together,our results provide critical insights intoα-Syn’s physiological function,and how Ca^(2+) regulates vesicle clustering mediated by α-Syn.