甲状腺乳头状癌患者外周血miR-146a、miR-146b表达水平与预后的关系研究

目的 探讨甲状腺乳头状癌(papillary thyroid cancer,PTC)患者血清miR-146a、miR-146b的表达水平及其与临床病理特征、预后的关系.方法 采用前瞻性研究方法,选取2018年6月至2020年2月本院收治的PTC患者80例为研究对象,50例同期健康人群作为对照组.荧光定量PCR检测外周血...     

miR-146a基因rs2910164 G/C多态性与miR-146a异常表达与胃癌风险相关性的研究

目的:探讨miR-146a基因单核苷酸多态位点rs2910164 G/C是否影响miR-146a的表达并改变其对胃癌的易感性。方法:选取53例胃癌患者和6株胃癌细胞株(AGS、BGC-823、HGC27、MKN-28、MKN-45和SGC-7901),采用Taqman定量PCR检测miR-146a的表达,构建miR-1...     

miR-146a在卵巢癌组织中的表达及其对卵巢癌SKOV3细胞增殖-凋亡的影响

目的探讨miR-146a在卵巢癌组织中的表达及其对卵巢癌SKOV3细胞增殖与凋亡的影响。方法RT-qPCR检测21例卵巢癌组织及对应癌旁正常卵巢组织中miR-146a表达;将miR-146a模拟物和miR-146a阴性对照转染到卵巢癌SKOV3细胞中,RT-qPCR检测转染后SKOV3细胞中miR-146a的表达;CCK8检测转染后miR-146a对SKOV3细胞增殖的影响;流式细胞仪检测转染后miR-146a对细胞凋亡的影响;Western blot检测SKOV3细胞p-Akt和Bcl-2的表达。结果 miR-146a在卵巢癌组织中的表达水平显著低于癌旁组织;转染miR-146a模拟物能显著提高SKOV3细胞miR-146a的表达量,显著降低SKOV3细胞的增殖能力,提高SKOV3细胞的凋亡率,抑制SKOV3细胞p-Akt和Bcl-2的蛋白表达(P0.01)。结论 miR-146a在卵巢癌癌组织低表达,转染miR-146a模拟物能抑制SKOV3细胞增殖,促进SKOV3细胞凋亡。     

Pre-miR-146a基因rs2910164位点单核苷酸基因多态性及miR-146a表达与类风湿关节炎相关性研究

目的 探讨pre-miR-146a基因rs2910164位点单核苷酸基因多态性及miR-146a表达与类风湿关节炎相关性.方法 采用聚合酶链反应-连接酶检测反应检测123例类风湿关节炎(RA)患者和220例健康对照者pre-miR-146a rs2910164位点基因多态性,应用实时荧光定量聚合酶链反应检测68例RA患者、10例骨关节炎(OA)患者及20例健康对照外周血单个核细胞中miR-146a的表达水平,并选取10例RA疾病活动患者行激素加免疫抑制剂正规治疗3个月后miR-146a表达水平的测定.收集并计算RA患者临床参数:发病年龄、性别、类风湿因子(RF)和抗环瓜氨酸肽(抗-CCP)抗体、RA疾病活动(DAS28≥3.2)、骨破坏(X＞Ⅰ期).统计学处理采用X2检验、方差分析、t检验和Pearson相关分析.结果 RA组pre-miR-146a rs2910164位点的基因型频率和等位基因频率与健康对照组比较,差异无统计学意义(P均＞0.05).RA患者pre-miR-146ars2910164位点基因型与发病年龄、性别、RF和抗-CCP抗体阳性率、RA疾病活动、骨破坏阳性率及miR-146a表达量均无相关性(P均＞0.05).RA患者组miR-146a的表达量高于健康对照组和OA组(P均＜0.01),后两组miR-146a的表达量无统计学差异(P＞0.05).RA疾病活动组miR-146a表达高于非活动组和对照组(P均＜0.01),后两组miR-146a的表达量无统计学差异(P＞0.05).RA疾病活动患者治疗后miR-146a表达下降(P＜0.05),DAS28评分降低(P＜0.01).RA患者组miR-146a的表达与红细胞沉降率(ESR,即血沉)、C反应蛋白(CRP)及DAS28评分之间呈正相关(P均＜0.01),与RF、抗-CCP抗体滴度无相关性(P均＞0.05).结论 我国汉族人群中,pre-miR-146a rs2910164位点多态性与RA的易感性、临床参数及miR-146a的表达无相关性,RA患者外周血单个核细胞miR-146a表达上调,其表达水平与RA病情活动有关,miR-146a的检测可能是RA病情活动的一个有用的判断指标.     

GIRK1、2在慢性颞叶癫痫大鼠海马内的表达变化

目的:探讨G蛋白偶联内向整流钾通道(GIRK)在海人酸诱导慢性颞叶癫痫大鼠海马内的表达变化及其在癫痫中的作用。方法:采用海人酸颞叶癫痫模型,运用原位杂交、免疫细胞化学检测海马齿状回、CA1、CA3区GIRK1、2mRNA和蛋白表达;Westernblotting检测海马整体GIRK1、2蛋白变化。结果:GIRK1、2mRNA和蛋白在大鼠海马内分布广泛;慢性颞叶癫痫海马DG区GIRK2mRNA和蛋白表达增高有显著差异(P＜0.01);GIRK1mRNA在海马DG区表达有差异(P＜0.05);Westernblotting检测海马GIRK1、2未有明显变化。结论:GIRK1、2在慢性癫痫大鼠海马内的表达增高,特别在齿状回,提示是机体对神经元网络过度兴奋的代偿或适应性反应;其合成增加将降低神经元的兴奋性,阻止海马内过度兴奋的扩散(DG-CA3-CA1)。     

miR-146a对小鼠巨噬细胞IL-18表达的影响

目的:探讨miR-146a对小鼠单核-巨噬细胞株RAW264.7以及小鼠原代腹腔巨噬细胞Thl/Th2类细胞因子表达的影响。方法:体外培养的小鼠单核-巨噬细胞系RAW264.7细胞和腹腔新鲜分离的巨噬细胞分别瞬时转染miR-146a mimics、阴性对照mimics(NC mimics)、抑制性miR-146a(miR-146a inhibitor)和抑制性miR-146a阴性对照(NC in-hibitor)。转染后,利用real time PCR定量检测IL-18、IL-5和IL-10的表达情况。结果:RAW264.7细胞和原代腹腔巨噬细胞在转染miR-146a mimics后IL-18的表达水平明显降低(P<0.05),转染miR-146a inhibitor后IL-18的表达水平明显升高(P<0.05),但对IL-5和IL-10的表达,两种转染形式均没有影响。结论:巨噬细胞RAW264.7及原代巨噬细胞表达的miR-146a能够负向调控Thl类细胞因子IL-18的表达,但是不能调控Th2类细胞因子IL-5及IL-10的表达。     

IL-1β和NF-κB在慢性内侧颞叶癫痫模型中的相互作用

目的:观察白介素1β(IL-1β)与核因子-κB(NF-κB)在大鼠内侧颞叶癫痫(mesial temporal lobe epilepsy,MTLE)模型中的表达变化;体外培养的星形胶质细胞经IL-1β和吡咯烷二硫氨基甲酸(PDTC)预处理后,观察其增殖和NF-κB表达变化。方法:利用匹罗卡品诱导SD大鼠发作癫痫制成MTLE模型,根据自发发作出现和稳定时间分为急性期对照组(AC,制模后2 h)、急性期癫痫组(AS)、潜伏期对照组(LC,制模后3周)、潜伏期癫痫组(LS)、慢性期对照组(CC,制模后8周)和慢性期癫痫组(CS)。体外培养的星形胶质细胞分为对照组、IL组和PDTC+IL组,利用凝胶迁移电泳(EMSA)、免疫印迹(WB)、酶联免疫吸附(ELISA)和免疫组织化学(IHC)的方法观察IL-1β和NF-κB的表达变化;MTT检测星形胶质细胞的增殖活化程度。结果:匹罗卡品诱导的MTLE模型大鼠海马内IL-1β和NF-κB在急性期、潜伏期和慢性期表达均增加,但以急性期和慢性期明显;IL-1β可以促进体外培养的星形胶质细胞增殖、上调星形胶质细胞内NF-κB的表达,而PDTC可以明显抑制IL-1β的上述作用。结论:IL-1β通过NF-κB促进大鼠星形胶质细胞的活化增殖,这一改变可能与MTLE发病密切相关,探讨其机制可能为MTLE的治疗提供新的靶点。     

miR-146a在哮喘患儿痰液的表达及对哮喘小鼠气道炎症的影响

目的探讨miR-146a在哮喘患儿痰液中的表达及其对气道炎症的作用。方法应用real-time PCR方法检测哮喘患儿和健康儿童痰液,以及卵清蛋白诱导的哮喘小鼠和正常小鼠肺组织中miR-146a的表达;化学合成miR-146a模拟物与阴性对照,并分别滴鼻至哮喘小鼠,HE染色验证哮喘小鼠模型的成功建立,免疫荧光方法和Western blot方法检测各组小鼠肺组织白介素-1β(IL-1β)与白介素-4(IL-4)的蛋白表达。结果与对照组相比,哮喘患儿痰液及哮喘小鼠肺组织中miR-146a表达显著升高(P0.01);HE染色显示哮喘模型组小鼠炎症细胞浸润明显高于正常对照组,哮喘小鼠模型成功建立。miR-146a模拟物滴入后,哮喘小鼠肺组织IL-1β与IL-4的蛋白表达显著降低(P0.01)。结论 miR-146a在支气管哮喘患儿及支气管哮喘小鼠模型中高表达,miR-146a模拟物能够抑制哮喘小鼠的气道炎症。     

MiR-146a在免疫反应中的调节作用

microRNA是一类非编码的小分子RNA,通过在转录后水平抑制蛋白的表达,在许多生理和病理进程中发挥重要作用。MiR-146a在固有免疫调节、炎症反应以及抗病毒中发挥重要作用。本文就最近对miR-146a在固有免疫、炎症应答、病毒感染和人类疾病等方面的研究进行了综述。研究显示,可能可以通过对miR-146a表达的调控来治疗一些人类疾病。     

Hamartia in hippocampal sclerosis-associated mesial temporal lobe epilepsy

Hamartia are small collections of rounded glioneuronal cells that are thought to be due to aberrant cell migration. Their presence has been recognized in association with mesial temporal lobe epilepsy; their prevalence among cases of hippocampal sclerosis (HS) and any potential association with patient demographics and outcomes is unknown. This study examines hamartia in a series of 292 patients with pathologically confirmed HS. Medical records were reviewed for pertinent patient clinical information (follow-up mean 5 years). Hamartia were identified in 96 cases (33%) and were seen primarily in the amygdala (n = 88) and less commonly in the hippocampus (n = 10) and temporal lobe (n = 4). A statistically significant relationship was found between the presence of hamartia and male gender, younger age of seizure onset, and history of childhood febrile seizures and developmental delay. It is unclear if these associations represent a real association or are a result of the underlying pathologies related to chronic epilepsy. At follow-up, there were no significant differences between patients who had hamartia and those who lacked this finding. Hamartia were observed in all subtypes of HS and there was a significant difference found in subtype distribution as well as proportion of cases between subtypes, but no association with any specific subtype overall. The presence of hamartia was not associated with the coexistence of focal cortical dysplasia or any specific histologic pattern of dysplasia. Hamartia are a common concomitant finding in HS and indicates evidence of aberrant cell migration in the hippocampal and parahippocampal regions in these patients.     

Defining clinico-neuropathological subtypes of mesial temporal lobe epilepsy with hippocampal sclerosis

Hippocampal sclerosis (HS) is the most frequent cause of drug-resistant focal epilepsies (ie, mesial temporal lobe epilepsy with hippocampal sclerosis; mTLE-HS), and presents a broad spectrum of electroclinical, structural and molecular pathology patterns. Many patients become drug resistant during the course of the disease, and surgical treatment was proven helpful to achieve seizure control. Hence, up to 40% of patients suffer from early or late surgical failures. Different patterns of hippocampal cell loss, involvement of other mesial temporal structures, as well as temporal neocortex including focal cortical dysplasia, may contribute to the extent of the epileptogenic network and will be discussed. An international consensus is mandatory to clarify terminology use and to reliably distinguish mTLE-HS subtypes. High-resolution imaging with confirmed histopathologic diagnosis, as well as advanced neurophysiologic and molecular genetic measures, will be a powerful tool in the future to address these issues and help to predict each patient's probability to control their epilepsy in mTLE-HS conditions.     

Evaluation of corpus callosum morphometry in patients with mesial temporal lobe epilepsy with hippocampal sclerosis

Purpose

The purpose of the study was to evaluate the effects of mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS) on corpus callosum (CC) morphometry in large and pathologically proven MTLE-HS patients.

Methods

We measured certain CC dimensions in 103 patients and 33 healthy controls using in vivo magnetic resonance imaging. In particular, we compared the two groups in relation to the clinical (localization of the HS, duration of epilepsy, frequency of seizures and length of seizures) and demographical (age, gender, handedness) features. Students’ t test, two-way ANOVA and Spearman test were used for statistical analysis.

Results

There was no significant difference between CC morphometry with respect to age and handedness among patients. The differences between the genders, however, were significant favouring longer diameters in males. We found significant decrease in the dimensions of the genu, body, isthmus and splenium of the CC in the MTLE-HS group, but there was no reduction in the size of the rostrum.

Conclusions

This general reduction in the size of the CC except for the rostrum was thought to be the result of cortical atrophy secondary to the disease. Concerning the preserved rostral part of the CC, it was thought that the fibers of the frontal lobe pass through different pathways than the tracts in the rostrum.     

视频脑电图对颞叶内侧癫癎致癎灶定侧价值探讨

目的：探讨视频脑电图（V—EEG）对颞叶内侧癫癎（MTLE）致痼灶定侧的价值。方法：回顾性分析34例MTLE患者的发作间期及发作期V—EEG提供的致癎灶的侧别信息,并与术中深部电极检测到颞叶内侧部位的痫样放电的侧别进行比较。结果：发作间期、发作期V-EEG能定侧的分别是23例（68％）,29例（85％）。与术中深部电极检查定侧符合的分别是20例（87％）,26例（90％）,两者的定侧符合率比较差异无统计学意义（P〉0．05）。结论：MTLE患者发作间期痫样放电呈单侧分布或分布有绝对的侧别优势时,定侧意义较大,与发作期EEG对致癎灶定侧意义相当。     

Expression of the group I metabotropic glutamate receptor in the hippocampus of patients with mesial temporal lobe epilepsy

Immunocytochemical studies at light and electron microscopic levels in hippocampi of patients with mesial temporal lobe epilepsy showed mGluR1 and mGluR5 immunoreactivity in the molecular layer of the dentate gyrus and CA1 area, especially at the border between stratum oriens and alveus. By electron microscopy, degenerating neuronal elements were found in all areas studied. There were glial filamentous tangles which appeared similar to intranuclear inclusions in astrocytes in all areas studied. Reactive product for mGluR1 was localised only in post-synaptic elements. However, mGluR5 immunoreactivity was demonstrated in both post- and pre-synaptic elements in the molecular layer of the dentate gyrus and CA1 area. Reactive product for mGluR5 was also demonstrated in astrocytes and in the periphery of fibrillary tangles. We postulate that in patients with mesial temporal lobe epilepsy, mGluR1 may increase hippocampal excitability through postsynaptic activation, and mGluR5 may do so through both pre- and post-synaptic mechanisms.     

Expression of the group I metabotropic glutamate receptor in the hippocampus of patients with mesial temporal lobe epilepsy

Immunocytochemical studies at light and electron microscopic levels in hippocampi of patients with mesial temporal lobe epilepsy showed mGluR1 and mGluR5 immunoreactivity in the molecular layer of the dentate gyrus and CA1 area, especially at the border between stratum oriens and alveus. By electron microscopy, degenerating neuronal elements were found in all areas studied. There were glial filamentous tangles which appeared similar to intranuclear inclusions in astrocytes in all areas studied. Reactive product for mGluR1 was localised only in post-synaptic elements. However, mGluR5 immunoreactivity was demonstrated in both post- and pre-synaptic elements in the molecular layer of the dentate gyrus and CA1 area. Reactive product for mGluR5 was also demonstrated in astrocytes and in the periphery of fibrillary tangles. We postulate that in patients with mesial temporal lobe epilepsy, mGluR1 may increase hippocampal excitability through postsynaptic activation, and mGluR5 may do so through both pre- and post-synaptic mechanisms.     

AMPA receptor alterations precede mossy fiber sprouting in young children with temporal lobe epilepsy

Following neurological injury early in life numerous events, including excitotoxicity, neural degeneration, gliosis, neosynaptogenesis, and circuitry reorganization, may alone or in concert contribute to hyperexcitability and recurrent seizures in temporal lobe epilepsy. Our studies provide new evidence regarding the temporal sequence of key elements of hippocampal reorganization, mossy fiber sprouting and glutamate receptor subunit up-regulation, in a subset of young temporal lobe epileptic patients. Without evidence of mossy fiber sprouting, the youngest age group (3-10 years old) of mesial temporal lobe epileptic patients demonstrated enhanced glutamate receptor subunit profiles, suggesting that the dendritic change precedes axonal sprouting. However, sclerotic hippocampal specimens from epileptic patients ages 12-15 years old had the characteristic features of glutamate receptor up-regulation and mossy fiber sprouting first identified in the adult, indicating that reconstructed circuits appear early in the course of the disease. Non-sclerotic hippocampal specimens from lesion associated temporal lobe epileptic patients of all age groups showed minimal cell loss, sparse staining of glutamate receptor subunits in the dentate gyrus, and little or no mossy fiber sprouting. These compelling findings suggest a progressive sequence of events in the reorganization of the dentate gyrus of sclerotic hippocampal specimens. We suggest that cell loss and up-regulation of glutamate receptor subunits appear early in temporal lobe epilepsy and contribute to the synaptic plasticity that may facilitate the subsequent sprouting of mossy fiber collaterals which compound an already precipitous state of decline. The combination of pre-synaptic and post-synaptic changes serves as a potential substrate for hyperexcitability.     

Lateralization of epileptic foci by neuropsychological testing in mesial temporal lobe epilepsy

This study investigated the lateralizing value of neuropsychological testing in presurgical evaluation of mesial temporal lobe epilepsy (MTLE). This study differed from previous ones in that the cutoff values were determined to yield high positive predictive values (PPVs), multiple neuropsychological predictors were considered in combination, and patients with atypical language dominance or low intelligence were not excluded from the sample. The participants were 92 patients with MTLE (left, n = 47; right, n = 45) who showed good postoperative seizure control. With a stringent cutoff criterion, the multiple neuropsychological predictors considered in combination yielded a sensitivity of 15% and a PPV of 93%, and with a less stringent cutoff criterion, a sensitivity of 37% and a PPV of 83%.     

Evolution of hippocampal epileptic activity during the development of hippocampal sclerosis in a mouse model of temporal lobe epilepsy

Unilateral intrahippocampal injection of kainic acid in adult mice reproduces most of the morphological characteristics of hippocampal sclerosis (neuronal loss, gliosis, reorganization of neurotransmitter receptors, mossy fiber sprouting, granule cell dispersion) observed in patients with temporal lobe epilepsy. Whereas some neuronal loss is observed immediately after the initial status epilepticus induced by kainate treatment, most reorganization processes develop progressively over a period of several weeks. The aim of this study was to characterize the evolution of seizure activity in this model and to assess its pharmacological reactivity to classical antiepileptic drugs.Intrahippocampal electroencephalographic recordings showed three distinct phases of paroxystic activity following unilateral injection of kainic acid (1 nmol in 50 nl) into the dorsal hippocampus of adult mice: (i) a non-convulsive status epilepticus, (ii) a latent phase lasting approximately 2 weeks, during which no organized activity was recorded, and (iii) a phase of chronic seizure activity with recurrent hippocampal paroxysmal discharges characterized by high amplitude sharp wave onset. These recurrent seizures were first seen about 2 weeks post-injection. They were limited to the injected area and were not observed in the cerebral cortex, contralateral hippocampus or ipsilateral amygdala. Secondary propagation to the contralateral hippocampus and to the cerebral cortex was rare. In addition hippocampal paroxysmal discharges were not responsive to acute carbamazepine, phenytoin, or valproate treatment, but could be suppressed by diazepam.Our data further validate intrahippocampal injection of kainate in mice as a model of temporal lobe epilepsy and suggest that synaptic reorganization in the lesioned hippocampus is necessary for the development of organized recurrent seizures.     

Expression of the group II and III metabotropic glutamate receptors in the hippocampus of patients with mesial temporal lobe epilepsy

By immunocytochemical study by both light and electron microscopy of the hippocampus of patients with mesial temporal lobe epilepsy, we have shown that mGluR2/3 and mGluR4 immunoreaction product was mainly localised in the molecular layer of the dentate gyrus and CA2 area. Electron microscopy showed that most of the immunoreaction product due to mGluR2/3, 4a and 8 was deposited in the postsynaptic elements of the CA2 pyramidal layer and the inner molecular layer of the dentate gyrus. Only mGluR8 immunoreaction product in the CA2 area and mGluR2/3 in the inner molecular layer of the dentate gyrus were demonstrated in presynaptic elements, suggesting that mGluR2/3 and 8 may be involved in presynaptic inhibition of glutamate release in these areas. The demonstration of some degenerating axon terminals in the inner molecular layer of the dentate gyrus suggests that degeneration of interneurons caused by repeated seizures was still in progress. The finding of mGluR2/3, 4 and 8 immunoreactive astrocytes in patient hippocampus suggests that mGluR2/3, 4 and 8 receptors may be involved in gliosis.