BackgroundEpilepsy is a complex neurological disorder characterized by recurrent seizures. Neuroinflammation and excessive neuronal excitation are key pathogenic factors, but current therapies fail to target these mechanisms effectively, highlighting the need for novel therapeutic agents. Zingerone, a bioactive compound derived from ginger (Zingiber officinale Roscoe), exhibits anti-inflammatory, antioxidant, and neuroprotective properties. However, its acute anticonvulsant efficacy and underlying mechanism in temporal lobe epilepsy (TLE) remain unclear. This study aimed to investigate whether zingerone exerts anticonvulsant effects by modulating neuronal excitability.MethodsA lithium chloride-pilocarpine-induced acute TLE rat model was established. Rats were randomly assigned to control, pilocarpine, and zingerone treatment groups (75, 150, and 300 mg/kg, i.p.). Seizure activity was evaluated via behavioral scoring (Racine scale) and electroencephalography (EEG). Immunohistochemistry (IHC), hematoxylin-eosin (HE) staining, and immunofluorescence were used to assess hippocampal microglial/astrocytic activation and neuronal damage. Whole-cell patch-clamp recordings were performed to analyze intrinsic neuronal excitability and synaptic transmission in hippocampal CA1 pyramidal neurons.ResultsAcute administration of zingerone (150 and 300 mg/kg) significantly reduced the number and duration of Racine stage IV/V generalized seizures. Zingerone dose-dependently inhibited microglial (IBA1+) and astrocytic (GFAP+) activation (p < 0.01 for 150 mg/kg; p < 0.001 for 300 mg/kg) and preserved neuronal integrity in the hippocampal CA1 region, as evidenced by reduced neuronal shrinkage, pyknosis, and increased NeuN+ neuron density. Electrophysiological recordings revealed that zingerone (10 μM) decreased the firing frequency of CA1 pyramidal neurons (p < 0.05), prolonged the inter-spike interval (p = 0.0026), reduced the action potential peak (p = 0.041), increased the rheobase current (p = 0.042), and increased afterhyperpolarization amplitude (p = 0.0001). Furthermore, Zingerone also modulated excitatory synaptic transmission onto CA1 neurons.ConclusionZingerone exerts acute anticonvulsant and neuroprotective effects in a TLE rat model by suppressing hippocampal neuroinflammation and reducing the intrinsic excitability of pyramidal neurons. These findings highlight zingerone as a promising natural compound for developing novel adjuvant therapies for drug-resistant TLE.

Circadian rhythms regulate seizure threshold and neuronal excitability in epilepsy. Temporal lobe seizures tend to arise more frequently during the diurnal phase relative to the nocturnal phase, yet dentate gyrus (DG) granule cells display elevated intrinsic excitability and CA1 pyramidal neurons show enhanced synaptic excitability at night. Here, we used male adult ICR mice (8–10 weeks old) as experimental subjects. Systemic pilocarpine injection was applied to pharmacologically trigger acute seizures, while AAV (pAAV2/9-CaMKIIα-hChR2 (H134R) - EYFP) delivery followed by optogenetic stimulation (10 Hz, 20 ms pulse width, 5 mW 473 nm blue light) was implemented to evoke region-specific hippocampal seizures. Combining continuous video-electroencephalography (vEEG) recording, optogenetically induced seizure assays, mRNA sequencing, and whole-cell patch-clamp electrophysiology, we investigated circadian modulation of neuronal excitability and seizure threshold within the DG and CA1 subfields of the hippocampus. Our results revealed divergent circadian regulation of seizure thresholds between the DG and CA1 regions. Notably, the diurnal-nocturnal seizure threshold pattern in CA1 recapitulated that seen in systemic pilocarpien-provoked seizure models. We further confirmed that circadian cycles exert distinct regulatory effects on excitatory/inhibitory synaptic transmission and the intrinsic excitability of hippocampal principal neurons in the DG and CA1. Collectively, this work highlights region-specific circadian control over seizure susceptibility and the brain excitation-inhibition balance. Elucidating circadian influences on hippocampal physiology and seizure threshold will improve the translational value of anti-epileptic therapies and guide time-stratified seizure intervention strategies.

Induced pluripotent stem cells (iPSC)-derived retinal pigment epithelium cells serve as a promising cell source for transplantation therapy in age-related macular degeneration. Here, we present a protocol for purifying the mature retinal pigment epithelium (RPE) cells post-differentiation. We describe steps for sequentially digesting cells with different enzymes and sorting them based on the autofluorescence of mature RPE cells, avoiding fixation and immunostaining. By maintaining high cell viability and purity, this cost-efficient protocol has broad implications for both in vitro research and transplantation applications.

BackgroundZingiber officinale Roscoe has been shown to possess analgesic properties. Zingerone (ZO), a bioactive compound derived from Zingiber officinale Roscoe, exhibits a range of pharmacological effects, including anti-inflammatory, anti-cancer, antioxidant, antibacterial, and anti-apoptotic activities. However, the analgesic properties of zingerone remain unclear.MethodsComplete Freund’s adjuvant (CFA) was administered to the left hind paw of C57BL/6 mice to induce a model of inflammatory pain. The analgesic effects of zingerone were assessed using the Von Frey and Hargreaves tests. In vivo fiber photometry and whole-cell patch clamp techniques were employed to investigate the potential mechanisms.ResultsBoth acute and long-term treatment with zingerone resulted in a significant increase in mechanical and thermal pain thresholds in mice experiencing CFA-induced inflammatory pain. Mechanical stimulation led to a pronounced increase in calcium levels within the anterior cingulate cortex (ACC) neurons of the inflammatory pain model, which was alleviated by zingerone administration. Furthermore, zingerone was found to modify synaptic transmission to ACC neurons and decrease their intrinsic excitability by prolonging the refractory period of these neurons.ConclusionZingerone demonstrates potential for alleviating CFA-induced inflammatory pain by reducing the intrinsic excitability of ACC neurons in a mouse model.

The regulation of protein degradation through the ubiquitin-proteasome system is essential for normal brain development, axon growth, synaptic growth and plasticity. The E3 ubiquitin ligase RFWD2 plays a key role in the onset and development of neurological diseases, including the pathogenesis of Alzheimer's disease (AD), but the mechanisms controlling the homeostasis of neuronal synaptic proteins are still poorly understood. Here, we showed that the expression level of RFWD2 gradually decreased with the age of the rats and was negatively correlated with the development of cerebral cortical neurons and dendrites in vivo. RFWD2 was shown to localize to presynaptic terminals and some postsynaptic sides of both excitatory synapses and inhibitory synapses via colocalization with neuronal synaptic proteins (SYN, PSD95, Vglut1 and GAD67). Overexpression of RFWD2 promoted dendrite development and dendritic spine formation and markedly decreased the expression of synaptophysin and PSD95 by reducing the expression of ETV1, ETV4, ETV5 and c-JUN in vitro. Furthermore, the whole-cell membrane slice clamp results showed that RFWD2 overexpression resulted in greater membrane capacitance in neuronal cells, inadequate cell repolarization, and a longer time course for neurons to emit action potentials with decreased excitability. RFWD2 regulates dendritic development and plasticity, dendritic spine formation and synaptic function in rat cerebral cortex neurons by activating the ERK/PEA3/c-Jun pathway via a posttranslational regulatory mechanism and can be used as an efficient treatment target for neurological diseases.

Objective To investigate the anticonvulsant effect of retigabine（RTG） on acute convulsion and explore its protective effect on convulsive hippocampal injury.Methods Forty ICR male mice were randomly divided into control group（n=10）, convulsion group（n=10）, RTG group（n=10）, and phenobarbital sodium group（n=10）. The above symptomatic treatment were given in the course of acute seizures in mice, respectively. The seizure grades of mice were analyzed to observe the anticonvulsant effect of RTG. HE staining was applied to detect pathological morphology of mouse hippocampi. Whole cell patch current clamp technique was performed to record the discharge of granule cells in dentate gyri of the hippocampi in the brains collected from anesthetized mice in convulsion group. RTG was perfused to the brain slice during recording to observe its effects on granule cell’s discharge.Results RTG reduced the grade and frequency of convulsion. HE staining results showed that RTG significantly improved neuronal damage in the CA1 region of mouse hippocampus after convulsion（P＜0.05）. RTG decreased the excitability of granule cells in the dentate gyri of convulsive mice（P＜0.05）. Conclusion RTG can inhibit mouse convulsion, alleviate neuronal damage in mouse hippocampus convulsion, and reduce the discharge of overexcited granular cells after mouse convulsion.

目的:调查分析安徽省蚌埠市4 所二甲以上医院弥漫性血管内凝血(DIC)的患病情况、病因、死亡率及相关影响因素,为临床对DIC的预防和治疗提供依据.方法:采用2001 年ISTH SSC DIC专业委员会及《弥散性血管内凝血诊断中国专家共识(2017 年版)》修订的DIC诊断标准,回顾性调查分析2018 年1 月—2022 年10 月4 所二甲以上医院住院人数及死亡率、DIC的患病情况及病因、病死率及相关影响因素等.结果:共收治住院患者1 135 906 例,死亡8 035 例,病死率为0.71％.其中DIC患者319 例,占同期住院患者的0.03％;DIC患者死亡占58.31％.DIC患者其致病主要因素是感染,占67.71％,其次是创伤、肿瘤、产科意外等.单纯DIC的患者死亡率为 17.39％,DIC患者累及器官发生功能障碍数目越多,死亡率越高.结论:DIC因感染出现较高的患病率,并易诱发多器官功能障碍导致临床高死亡率.因此针对相关影响因素制定防治策略,以期早期诊断和早期干预治疗.

目的:分析颞叶癫痫患者海马样本中星形胶质细胞和小胶质细胞标志基因的表达情况,探索其功能改变及影响.方法:利用现有的公共数据库,定量分析两类胶质细胞标志基因在不同病理条件下的表达量,评估两类胶质细胞的激活情况,并通过相关性分析探索二者之间可能的相互作用.结果:与对照样本相比,部分颞叶癫痫患者海马样本中这两类胶质细胞标志基因表达水平均增高,二者存在相关性(P＜0.05).其中GFAP、S100B、TMEM119和AIF1的升高有统计学意义(均P＜0.05).进一步分析显示,出现海马硬化后,星形胶质细胞标志基因的表达进一步上调,但与小胶质细胞标志基因的表达没有显著相关性(P＞0.05).结论:标志基因表达水平的显著升高提示了这两类胶质细胞在颞叶癫痫发病过程中都被激活.海马硬化时,星形胶质细胞可能通过不同的调控通路发生进一步激活.

Breast cancer is characterized by some types of heterogeneity, high aggressive behaviour, and low immunotherapeutic efficiency. Detailed immune stratification is a prerequisite for interpreting resistance to treatment and escape from immune control. Hence, the immune landscape of breast cancer needs further understanding. We systematically clustered breast cancer into six immune subtypes based on the mRNA expression patterns of immune signatures and comprehensively depicted their characteristics. The immunotherapeutic benefit score (ITBscore) was validated to be a superior predictor of the response to immunotherapy in cohorts from various datasets. Six distinct immune subtypes related to divergences in biological functions, signatures of immune or stromal cells, extent of the adaptive immune response, genomic events, and clinical prognostication were identified. These six subtypes were characterized as immunologically quiet, chemokine dominant, lymphocyte depleted, wounding dominant, innate immune dominant, and IFN-γ dominant and exhibited features of the tumor microenvironment (TME). The high ITBscore subgroup, characterized by a high proportion of M1 macrophages:M2 macrophages, an activated inflammatory response, and increased mutational burden (such as mutations in TP53, CDH1 and CENPE), indicated better immunotherapeutic benefits. A low proportion of tumor-infiltrating lymphocytes (TILs) and an inadequate response to immune treatment were associated with the low ITBscore subgroup, which was also associated with poor survival. Analyses of four cohorts treated with immune checkpoint inhibitors (ICIs) suggested that patients with a high ITBscore received significant therapeutic advantages and clinical benefits. Our work may facilitate the understanding of immune phenotypes in shaping different TME landscapes and guide precision immuno-oncology and immunotherapy strategies.

Numerous epilepsy-related genes have been identified in recent decades by unbiased genome-wide screens. However, the available druggable targets for temporal lobe epilepsy (TLE) remain limited. Furthermore, a substantial pool of candidate genes potentially applicable to TLE therapy awaits further validation. In this study, we reveal the significant role of KCNQ2 and KCNQ3, two M-type potassium channel genes, in the onset of seizures in TLE. Our investigation began with a quantitative analysis of two publicly available TLE patient databases to establish a correlation between seizure onset and the downregulated expression of KCNQ2/3. We then replicated these pathological changes in a pilocarpine seizure mouse model and observed a decrease in spike frequency adaptation due to the affected M-currents in dentate gyrus granule neurons. In addition, we performed a small-scale simulation of the dentate gyrus network and confirmed that the impaired spike frequency adaptation of granule cells facilitated epileptiform activity throughout the network. This, in turn, resulted in prolonged seizure duration and reduced interictal intervals. Our findings shed light on an underlying mechanism contributing to ictogenesis in the TLE hippocampus and suggest a promising target for the development of antiepileptic drugs.

TP53 is the most frequently mutated gene in lung adenocarcinoma (LUAD). The tumor immune microenvironment (TIM) is considered a vital factor that influences tumor progression and survival rate. The influence of TP53 mutation on TIM in LUAD has not been fully studied. Here we systematically investigated the relationship and potential mechanisms between TP53 mutation status and immune response in LUAD. We constructed an immune prognostic model (IPM) using immune associated genes, which were expressed differentially between the TP53 mutant and wild type LUAD patients. We discovered that TP53 mutations were significantly associated with 5 immune related biological processes. Thirty-six immune genes were expressed differentially between TP53 mutant and wild type LUAD patients. An IPM was constructed using 3 immune genes to differentiate the prognostic survival in LUAD. The high-risk LUAD group displayed significantly higher proportions of dendritic cell resting, T cell CD4 memory resting and mast cell resting, and significantly low proportions of dendritic cell activated, T cell CD4 memory activated, and mast cell activated. Moreover, IPM was found to be an independent clinical feature and can be used to predict immunotherapy responses. In summary, we constructed and validated an IPM using 3 immune related genes, which provides a better understanding of the mechanism from an immunological perspectives.

探讨极域电子教室软件在医学机能学实验虚拟仿真教学课堂上的应用.以本学期2019级临床医学两个带教班级为研究对象,在虚拟仿真教学项目中使用极域电子教室软件,以采用班级为实验组,未采用班级为对照组,比较两个班学生测试成绩并对实验组学生发放调查问卷.测试成绩结果显示实验组成绩(98.000±4.549)与对照组成绩(88.807±9.478)比较差异具有统计学意义(P＜0.001).问卷调查结果显示100％的学生表示采用极域软件教学能更有助于学习,75.86％的学生更倾向使用该软件.极域电子教室软件在医学机能学实验虚拟仿真教学课堂中具有积极的促进作用.

目的 探索形成性评价在病理生理学教学中的应用及效果.方法 于2017年9月—2020年1月在本校部分班级开展形成性评价教学,围绕课堂教学设计前中后三个测评环节,平时测评成绩和期末考试成绩按比例组成本课程综合成绩.选择本校两个年级卓越医师班进行教学对比研究,并在2017级临床本科和2018级护理本科开展教学效果调查.结果 授课教师根据测评结果可以及时发现教学问题并进行调整;实施形成性评价的实验班级综合分显著提高(P<0.05);调查结果反馈学生非常认可形成性评价教学模式.结论 开展形成性评价对提高病理生理学的教学效果具有积极意义.

目的:对微课嵌入式教学模式在病理生理学教学中的应用及效果进行评价.方法:选择蚌埠医学院2016级临床医学专业两个班作为研究对象,其中一个班为实验班,采用微课嵌入式教学模式进行病理生理学教学;另一班为对照班,采用传统教学模式教学.课程结束后对两个班学生进行阶段性理论测试、临床病例分析能力测试及学习满意度问卷调查,评价教学效果.结果:与对照班比较,实验班理论考试成绩和病例分析测试成绩均得到显著提升(P<0.01),课程满意度调查结果显示实验班学生对微课嵌入式教学模式各项指标的满意度均达90％以上.结论:微课嵌入式教学模式对病理生理学的教学效果有较为显著的促进作用.

目的:探讨2型糖尿病(T2DM)病人血清甲状腺激素(TH)和促甲状腺激素(TSH)水平变化及其影响因素.方法:选取T2DM病人75例(T2DM组)及同期健康体检者75名(对照组),比较2组血清TH和TSH水平;另将T2DM组按性别、年龄、病程、糖化血红蛋白(HbA1c)、空腹血糖(FBG)进行划分,比较血清TH和TSH水平.结果:T2DM组病人血清游离三碘甲状腺原氨酸(FT3)水平明显低于对照组(P<0.01),2组游离甲状腺素(FT4)和TSH水平差异无统计学意义(P>0.05).不同性别、是否服用二甲双胍T2DM病人的血清FT3、FT4和TSH水平差异均无统计学意义(P>0.05);不同年龄段和病程病人间的FT3水平差异均有统计学意义,随年龄和病程增加,FT3水平降低(P<0.05～P<0.01);不同HbA1c病人间的FT3、FT4水平差异均有统计学意义,随HbA1c升高,FT3、FT4水平均降低(P<0.05～P<0.01);FBG≥7.0 mmol/L病人的FT3、FT4水平均明显低于FBG<7.0 mmol/L病人(P<0.01).Spearman相关分析显示,T2DM病人FT3水平与年龄、病程、HbA1c、FBG均呈负相关关系(P<0.05);FT4与HbA1c、FBG呈负相关关系(P<0.05).结论:与健康成人比较,T2DM病人存在甲状腺功能异常情况,主要表现为FT3降低;T2DM病人的FT3水平变化与年龄、病程、HbA1c和FBG密切相关.

Recent evidence has demonstrated that microRNA-19b (miR-19b) is elevated and functions as a prognosis predictor in hepatocellular carcinoma and melanoma. However, its expression and clinical significance in colorectal cancer (CRC) remain unclear. The study aimed to identify the correlation between miR-19b expression and the clinicopathological features and prognosis of patients with CRC. In this study, we found that the levels of miR-19b were significantly up-regulated in CRC tissues and cell lines compared with matched adjacent non-cancerous tissues and human colon mucosal epithelial cell lines, and its expression was also increased in patients with lymph node metastasis compared with those patients with no lymph node metastasis. Meanwhile, the patients with distal metastasis have a higher miR-19b expression than those patients with no distal metastasis. The high expression of miR-19b in patients with CRC was associated with lymph node metastasis and distant metastasis. miR-19b expression was an independent prognostic indicator for overall survival of CRC patients. Moreover, patients with a high miR-19b expression have shorter overall survival times than those patients with a low miR-19b expression. In addition, an in vitro functional assay showed that miR-19b knockdown restrained the migration and invasion of HCT116 and SW480 cells. In summary, the study provides the first convincing statistical and experimental evidence that the up-regulation of miR-19b is associated with metastasis and predicts unfavorable prognosis in patients with CRC, suggesting that miR-19b may serve as a novel and promising prognostic biomarker in CRC.

Cerebral ischemia leads to neuronal death for stroke, in which the imbalance between glutamatergic neurons and GABAergic neurons toward neural excitotoxicity is presumably involved. GABAergic neurons are vulnerable to pathological factors and impaired in an early stage of ischemia. The rescue of GABAergic neurons is expected to be the strategy to reserve ischemic neuronal impairment. As protein kinase C (PKC) and calmodulin-dependent protein kinase II (CaMK-II) are activated during ischemia, we have investigated whether the inhibitions of these kinases rescue the ischemic impairment of cortical GABAergic neurons. The functions of GABAergic neurons were analyzed by whole-cell recording in the cortical slices during ischemia and in presence of 1-[N,O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (CaMK-II inhibitor) and chelerythrine chloride (PKC inhibitor). Our results indicate that PKC inhibitor or CaMK-II inhibitor partially prevents ischemia-induced functional deficits of cortical GABAergic neurons. Moreover, the combination of PKC and CaMK-II inhibitors synergistically reverses this ischemia-induced deficit of GABAergic neurons. One of potential therapeutic strategies for ischemic stroke may be to rescue the ischemia-induced deficit of cortical GABAergic neurons by inhibiting PKC and CaMK-II.

以岗位胜任力为核心的新一代医学教育理念更加注重培养医学生的综合素质.为适应新的形式需求,本研究以全科医学专业为试点,对病理生理学的教学过程进行改革,以理论为基础、围绕临床、注重实践.多元化的教学方式充分调动了学生的学习主动性,达到了较好的改革效果.

Conditional associative memory is an important form of human and animal learning and memory.When the conditioned and unconditioned stimuli are correlated in time and space,the central nervous system processes the original information and stores it in the brain with the relevant mechanism.The extinction of conditional associative memory is the suppression of the formed conditional reflex.The study of its mechanism is of great value in neurobiology and clinical medicine.This article reviewed recent research at home and abroad,and analyzed the main mechanisms involved in the memory extinction,provided theoretical basis and experimental experience for the future study of learning and memory.

Neural plasticity occurs in learning and memory. Coordinated plasticity at glutamatergic and GABAergic neurons during memory formation remains elusive, which we investigate in a mouse model of associative learning by cellular imaging and electrophysiology. Paired odor and whisker stimulations lead to whisker- induced olfaction response. In mice that express this cross-modal memory, the neurons in the piriform cortex are recruited to encode newly acquired whisker signal alongside innate odor signal, and their response patterns to these associated signals are different. There are emerged synaptic innervations from barrel cortical neurons to piriform cortical neurons from these mice. These results indicate the recruitment of associative memory cells in the piriform cortex after associative memory. In terms of the structural and functional plasticity at these associative memory cells in the piriform cortex, glutamatergic neurons and synapses are upregulated, GABAergic neurons and synapses are downregulated as well as their mutual innervations are refined in the coordinated manner. Therefore, the associated activations of sensory cortices triggered by their input signals induce the formation of their mutual synapse innervations, the recruitment of associative memory cells and the coordinated plasticity between the GABAergic and glutamatergic neurons, which work for associative memory cells to encode cross-modal associated signals in their integration, associative storage and distinguishable retrieval.