癫痫患者血浆神经肽Y与免疫细胞的水平变化及其临床意义

目的观察颞叶癫痫患者不同时期外周血T淋巴细胞亚群、NK细胞和神经肽Y(NPY)的水平,探讨癫痫与神经免疫学之间的关系。方法将符合诊断标准36例颞叶癫痫患者入院10 min内和入院24 h的外周血中免疫细胞及NPY的水平与入院时进行比较分析。结果入院10 min内外周血中白细胞总数、中性粒细胞、淋巴细胞、NK细胞及NPY的水平均较入院时明显升高(P<0.05),而CD4细胞则降低13%;NPY与NK细胞水平之间呈正相关(r=0.839,P<0.05);入院24 h外周血中免疫细胞与入院时水平无显著性差异(P>0.05)。结论癫痫患者存在免疫功能低下,癫痫发作后血浆NPY含量明显增加,NPY可能参与免疫功能的调节。     

骨髓增生异常综合征患者红细胞的天然免疫黏附功能及对自然杀伤细胞杀伤活性的影响

目的 探讨骨髓增生异常综合征患者红细胞天然免疫黏附功能及对自然杀伤细胞（NK细胞）杀伤活性的影响.方法 选择我院骨髓增生异常综合征患者14例为试验组,20例健康人作为对照组,将2组外周血红细胞用柠檬酸钠抗凝,检测红细胞在自身血浆条件下对K562细胞的免疫黏附并计算黏附率;并用四甲基偶氮唑盐微量酶反应比色（MTT）法测定红细胞对正常NK细胞杀伤K562细胞活性的影响,并与未加红细胞时进行比较.结果 试验组红细胞使K562细胞形成了＂玫瑰花＂样结合.对照组红细胞对K562细胞的免疫黏附结合率为（15.6±6.7）%,试验组红细胞对K562细胞的免疫黏附结合率为（8.5±7.0）%,2组比较差异有统计学意义（t=3.66,P＜0.01）.不加红细胞条件下,对照组外周静脉血NK细胞杀伤K562细胞活性杀伤率为64%～75%,加入红细胞后为79%～88%;试验组不加入红细胞为45%～56%,加入红细胞后为51%～58%.加入红细胞后,2组NK红细胞对K562细胞的杀伤率均增加（P＜0.01）,且对照组高于试验组（P＜0.01）.结论 红细胞可增加NK细胞对K562细胞的杀伤率;骨髓增生异常综合征患者的红细胞对K562细胞的免疫黏附能力下降,并可减低NK细胞对K562细胞的杀伤率.     

Application of proteomic approaches to assess the effect of anti-epileptic drug on seizure foci

Epilepsy is one of the most common neurological disorders characterized by epileptic seizures. The anti-epileptic drugs (AEDs) are the main form of treatment for people with epilepsy. Classically, people thought that AEDs modify the activities of ion channels to suppress epileptic seizures. However, accumulating evidence suggests that targeting at ion channels cannot completely account for the numerous effects of the AEDs on its broad clinical activity in epileptic patients. In our study, proteomic methods were used to quantify the proteome of hippocampal tissues from patients who were treated or not treated by AEDs (Carbamazepine). Further bioinformatics methods, including Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, were utilized to analyze the differences between two patient groups. We found that more than 400 proteins, including metabolism and immune related proteins, had a higher expression level in the carbamazepine-treated group compared with the controls. These altered proteins were considered to be involved in many different biological pathways. Among these pathways, immune-system related pathway modulated by complement C3 and microglia was highly remarkable, which regulated the synapses elimination in physiological condition. In epilepsy, the carbamazepine induced up-regulation of complement C3 might decrease the abnormal synaptic connections between neurons and thus contribute to the therapeutic role of carbamazepine. The results of our study suggested that apart from ion channels, carbamazepine exerted numerous effects on human epileptic foci, which might be the fundamental mechanisms of AEDs for treatment, adverse-effects and pharmacoresistance of epilepsy.     

Platelet GABA-aminotransferase in epileptic patients

Platelet GABA-aminotransferase (GABA-T) activity was determined in 12 adults (six healthy volunteers and six long-term treated epileptic patients) and 17 children (six non-epileptic, and 11 long-term treated epileptic patients). Platelet GABA-T activity was about 60% higher in the epileptic patients than in the controls, both in adults (14.7 +/- 8.6 versus 8.8 +/- 3.5 pmol/min/mg of protein,) and children (13.1 +/- 4.8 versus 8.3 +/- 3.3 pmol/min/mg of protein, p less than 0.05). The relationship between this increase and either epilepsy or anti-epileptic treatment should be clarified in further studies.     

癫痫患者血清催乳素的检测及其临床意义

目的 :为了研究癫痫患者下丘脑功能的改变及催乳素 ( PRL)在癫痫诊断中的临床意义。方法 :选择了 2 7例癫痫患者 ,2 5例非癫痫患者及 30例正常人作为对照组 ,分别采集其血样 ,用放射免疫法检测血清 PRL水平的变化。结果 :癫痫患者血清 PRL 水平明显高于非癫痫组及对照组 ,非癫痫组与对照组之间差异无显著性。结论 :癫痫发作对下丘脑 -垂体轴有一定影响 ,血清 PRL 升高的比率可作为癫痫鉴别诊断的一个参考指标     

儿童特发性血小板减少性紫癜患者NK细胞及T淋巴细胞亚群变化及意义

目的检测特发性血小板减少性紫癜(ITP)患者T淋巴细胞亚群及NK细胞的变化,探讨其在ITP发病机制中的作用。方法应用流式细胞术分别检测80例ITP及35例正常对照组T淋巴细胞亚群、NK细胞变化。结果 ITP患者CD3+、CD4+细胞百分比及CD4+/CD8+比值明显低于正常对照组(P<0.01),CD8+细胞百分比高于正常对照组(P<0.01)。NK细胞百分比明显低于正常对照组P<0.01)。结论及T淋巴细胞亚群及NK细胞的变化可较好的反映ITP的病理过程,对提高ITP的诊断水平及指导临床具有一定的使用价值。     

癫痫患者丙戊酸血药浓度监测及个体化给药

目的：为临床癫痫患者丙戊酸的合理用药提供参考。方法：采用荧光偏振免疫法对１２３ 例癫痫患者进行血药浓度监测,并对其结果及疗效进行分析总结。结果：达到丙戊酸有效血药浓度范围未有效控制的有１４ 例,占１１－４ ％ ;不在有效血药浓度范围而控制良好的有１１ 例,占８－９ ％ 。结论：丙戊酸的个体化给药必须综合考虑各方面的因素,不能仅仅以血药浓度为依据,并建议在适当情况下有必要监测游离丙戊酸血药浓度。     

Compared effects of morphine and nickel chloride on NK cell activity in vitro in rats and monkeys.

The measurement of natural killer (NK) cell activity often is recommended as an endpoint for inclusion in the non-clinical immunotoxicity evaluation of environmental chemicals and pharmaceuticals. To date, most data on the impact of immunotoxicants on NK cell activity have been obtained in the rat. Because non-human primates often are used in the safety evaluation of new medicinal products, there is a need to compare chemically induced changes in NK cell activity between rats and primates. In this study, the in vitro effects of nickel chloride and morphine hydrochloride were investigated on NK cell activity in the rat and the cynomolgus monkey. Despite some species-specific differences in the techniques used, rather similar results were obtained in the two species. At the higher concentration, nickel chloride induced a significant decrease in NK cell activity in the ranges of 21.6-24.3% (rat) and 34.4-42.2% (monkey), depending on the effector-to-target cell ratio used, and morphine hydrochloride induced a decrease in the ranges of 23.7-34.7% (rat) and 59.1-68.0% (monkey). These results suggest that NK cell activity can be used as a reliable endpoint for the assessment of immune effects during safety evaluation studies in the cynomolgus monkey.     

Effects of Spider Venom Toxin PWTX-I (6-Hydroxytrypargine) on the Central Nervous System of Rats

The 6-hydroxytrypargine (6-HT) is an alkaloidal toxin of the group of tetrahydro-β-carbolines (THβC) isolated from the venom of the colonial spider Parawixia bistriata. These alkaloids are reversible inhibitors of the monoamine-oxidase enzyme (MAO), with hallucinogenic, tremorigenic and anxiolytic properties. The toxin 6-HT was the first THβC chemically reported in the venom of spiders; however, it was not functionally well characterized up to now. The action of 6-HT was investigated by intracerebroventricular (i.c.v.) and intravenous (i.v.) applications of the toxin in adult male Wistar rats, followed by the monitoring of the expression of fos-protein, combined with the use of double labeling immunehistochemistry protocols for the detection of some nervous receptors and enzymes related to the metabolism of neurotransmitters in the central nervous system (CNS). We also investigated the epileptiform activity in presence of this toxin. The assays were carried out in normal hippocampal neurons and also in a model of chronic epilepsy obtained by the use of neurons incubated in free-magnesium artificial cerebro-spinal fluid (ACSF). Trypargine, a well known THβC toxin, was used as standard compound for comparative purposes. Fos-immunoreactive cells (fos-ir) were observed in hypothalamic and thalamic areas, while the double-labeling identified nervous receptors of the sub-types rGlu2/3 and NMR1, and orexinergic neurons. The 6-HT was administrated by perfusion and ejection in "brain slices" of hippocampus, inducing epileptic activity after its administration; the toxin was not able to block the epileptogenic crisis observed in the chronic model of the epilepsy, suggesting that 6-HT did not block the overactive GluRs responsible for this epileptic activity.     

海马磁共振波谱分析在颞叶癫痫症中的价值

目的探讨海马氢质子磁共振波谱分析（1H—MRS）在颞叶癫痫症中的应用价值。方法采用PhihpsAchieva1．5T双梯度磁共振扫描仪,对38例颞叶癫痫患者和20例健康志愿者行颅脑MRI和海马’H—MRs检查,观察NAA、Cr、Cho浓度和NAA／（Cho＋Cr）、NAA／Cr和NAA／Cho值。分别对癫痫患者病灶侧与正常侧,病灶侧与正常对照组海马的NAA／（Cho＋Cr）、NAA~r和NAA／Cho值进行t检验,分析两者之间差异有无显著性。结果癫痫患者组癫痫侧、正常侧和对照组海马的NAA／（Cho＋Cr）值分别为（O．424-0．06）、（0．67±0．09）、（O．72±0．07）,癫痫组癫痫侧与正常侧、对照组比较差异有显著性（尸〈0．05）,而正常侧与对照组比较差异无显著性（尸〉0．05）。以NAA／fCho＋c0来检出癫痫,灵敏度曲线呈现低一高一低的变化,在0．60时,灵敏度最高。结论-H—MRS不仅能早期诊断海马硬化,而且可进一步提高术前癫痫灶定位的准确性。     

NMDA receptor-mediated long-term alterations in epileptiform activity in experimental chronic epilepsy

When epileptiform activity is acutely induced in vitro, transient partial blockade of N-methyl-d-aspartic acid (NMDA) receptor-mediated calcium influx leads to selective long-term depotentiation of the synapses involved in the epileptic activity as well as a reduction in the probability of further epileptiform activity. If such selective depotentiation occurred within foci of epileptic activity in vivo, the corresponding long-term reduction in seizure probability could form the basis for a novel treatment of epilepsy. Continuous radiotelemetric EEG monitoring demonstrated modest acute anticonvulsant effects but no long-term reductions in the probability of spontaneous seizures after transient partial blockade of NMDA receptors (NMDAR) during ictal and interictal activity in the kainate animal model of chronic epilepsy. In vitro, depotentiation was induced when NMDAR were partially blocked during epileptiform activity in hippocampal slices from control animals, but not in slices from chronically epileptic rats. However in slices from epileptic animals, depotentiation during epileptiform activity was induced by partial block of NMDAR using NR2B- but not NR2A-selective antagonists. These results suggest that chronic epileptic activity is associated with changes in NMDA receptor-mediated signaling that is reflected in the pharmacology of activity- and NMDA receptor-dependent depotentiation.     

托吡酯影响成年癫痫患者甲状腺激素的前瞻性研究

目的：探讨托吡酯（TPM）对成年癫痫患者甲状腺激素水平的影响。方法：入选患者分为试验组63例和健康对照组40例，试验组用TPM进行单药治疗，测定对照组和试验组治疗前及治疗后3，6及12个月时的血清甲状腺激素水平，并进行比较。结果：未经治疗的癫痫患者的甲状腺激素水平与对照组比较无统计学差异（P〉0．05）；TPM治疗后3，6及12个月时的甲状腺激素水平与治疗前及对照组比较无统计学差异（均P〉0．05）。结论：癫痫本身并不引起甲状腺激素的改变；TPM对成年癫痫患者的甲状腺激素没有影响，安全性好。     

Malformation rates in children of women with untreated epilepsy: a meta-analysis.

BACKGROUND: It is widely quoted that women with epilepsy have a higher than baseline risk for giving birth to a child with malformations, independent of the effects of antiepileptic drugs. OBJECTIVE: To determine, based on available evidence, if epilepsy per se represents a teratogenic risk. To systematically review all studies investigating the occurrence of major malformation rates among children of treated or untreated women with epilepsy and non-exposed controls who do not have epilepsy. METHODS: A meta-analysis, using a random effects model, was conducted of all cohort and case-control studies reporting malformation rates in children of women with epilepsy exposed or unexposed to antiepileptic drugs compared with that of children of nonepileptic women. Medline (1966-2001), EMBASE, the Cochrane database as well as REPROTOX (an information system on environmental hazards to human reproduction and development) databases were accessed. RESULTS: We found ten studies reporting results of untreated epilepsy (n = 400) and their non-epileptic healthy controls (n = 2492). Nine out of ten studies also reported results on 1443 patients exposed to antiepileptic drugs and their 2526 unexposed healthy controls. The risk for congenital malformations in the offspring of women with untreated epilepsy was not higher than among nonepileptic controls (odds ratio [OR] = 1.92; 95% CI 0.92-4.00). There was evidence of publication bias, thus with bias removed the OR was 0.99 (95% CI 0.49-2.01). In contrast, the offspring of epileptic women who received antiepileptic drugs had higher incidences of malformation than controls (OR 3.26; 95% CI 2.15-4.93). CONCLUSION: Our study does not support the commonly held view that epilepsy per se represents a teratogenic risk. Our study suggests that this view is the result of a publication bias, with several small (< 100 participants) positive studies leading to a premature conclusion.     

Effect of cianidanol on natural killer cell activity in patients with chronic B virus hepatitis

The hepatoprotective antioxidant bioflavonoid cianidanol has beneficial therapeutic and immunomodulatory effects in chronic hepatitis. Its action on natural killer (NK) cell activity has not yet been studied in hepatitis B virus (HBV) infection. In the present study, the in vitro and in vivo effects of the drug on NK cell activity have been determined in six patients with chronic HBV hepatitis and in ten healthy control subjects. Two methods were used: an enzyme release assay and a cytotoxicity test based on the assessment of endogenous alkaline phosphatase activity of the target cells. The in vitro effect of the drug was assessed using cianidanol at 10(-6), 10(-5) and 10(-4) M concentrations. For in vivo studies, HBV hepatitis patients were treated with cianidanol at a daily dose of 3.0 g cianidanol for seven days and were investigated before and after the treatment. Chronic HBV hepatitis patients showed a moderate decrease in NK cell activity compared to the controls, but after the cianidanol therapy their NK cell activity significantly rose to 68.0% +/- 9.5% (p less than 0.01). Cianidanol in vitro inhibited the NK cell activity both in hepatitis and healthy groups when using K-562 target cells and the lactic acid dehydrogenase enzyme release assay, but did not influence or even slightly enhance the NK activity when human embryonic fibroblast cells and alkaline phosphatase assay were used for the test. After the 7-day in vivo treatment, the in vitro inhibitory action of the drug was diminished or absent.(ABSTRACT TRUNCATED AT 250 WORDS)     

Impaired memory following repeated pentylenetetrazol treatments in kindled mice

Epilepsy is characterized by recurrent seizures, which are caused by excessive discharges from cerebral neurons. Currently, antiepileptic drugs that possess sodium channel blocking activities and also mediate GABA-ergic systems are primarily used to prevent epileptic seizure. However, approximately 40% of patients with epilepsy suffer from interictal psychiatric comorbidities in clinical practice. Furthermore, it is unclear whether epilepsy is associated with psychic function. The aim of the present study was to clarify the effects of kindling-induced epileptic seizures on psychic functioning using behavioral pharmacological tests. Pentylenetetrazol (PTZ)-kindled mice demonstrated no significant differences in locomotor activity and muscle relaxation compared with na?ve mice. PTZ-kindled mice also demonstrated cognitive impairment in the objective location test, but no significant effects of PTZ-kindling were observed in the Y-maze test. These findings suggested that PTZ-kindling impairs reference memory, but not working memory. These results suggest that, with respect to their psychic functioning, PTZ-kindled mice have specific characteristics.     

Neuronal MDR-1 gene expression and persistent low levels of anticonvulsants in a child with refractory epilepsy

SUMMARY: It is estimated that 20-25% of epileptic patients fail to achieve good control with antiepileptic drug (AED) treatment; thus, refractory epilepsy (RE) has been described in patients who have adequate therapeutic levels of AEDs without control of seizures. Multidrug resistance genes have been reported to be highly expressed in brain of patients with RE. Persistent low plasma levels of AEDs and high brain expression of the multidrug resistance product P-glycoprotein (P-gp) have been previously communicated in a case report of RE secondary to tuberous sclerosis. Here, the authors report a case of an 8-year-old boy diagnosed with partial RE with focal seizures who was admitted to hospital for a severe episode of subintrant crisis. The patient received polytherapy with carbamazepine (CBZ), phenytoin (PHT), and valproic acid (VA); however, habitual doses of these AEDs failed to control the patient's symptoms. AED blood levels were monitored for 25 consecutive days and showed low values in 8/25 (33%) for CBZ, 10/25 (40%) for PHT, and 25/25 (100%) for VA of samples studied. Because the patient developed focal status epilepticus, surgical treatment by callosotomy was done, resulting in a significant improvement in epileptic symptoms. The immunostaining of brain specimens showed significantly increased expression of P-gp not only in vascular endothelial cells and related astrocytes but also in neurons. Overexpression of P-gp in the brain does not explain the low blood levels of AEDs described in these cases. Different mechanisms such as drug-drug interactions and drug transporters can be involved in the results observed. The P-gp overexpression and/or its pharmacologic induction should be considered as a potential mechanism responsible for drug resistance to epilepsy treatment and highly suspected in patients with persistent subtherapeutic AEDs plasma levels.     

急性髓性白血病患者外周血淋巴细胞亚群的检测及其临床意义

目的探讨急性髓系白血病(AML)患者外周血T淋巴细胞亚群、DNT细胞、NK细胞的表达水平及其临床意义。方法采用流式细胞术对40例初发AML患者、32例AML完全缓解(CR)患者及20例正常人外周血T淋巴细胞亚群、DNT细胞及NK细胞水平进行检测。结果初发AML组与正常对照组比较,CD4+细胞、CD4+/CD8+比值、DNT细胞及NK细胞明显降低(P<0.05);CD3+细胞、CD8+细胞变化不显著(P>0.05)。复治CR组与对照组比较,NK细胞明显降低(P<0.05),其他各值差异无统计学意义。结论初发AML患者的T淋巴细胞亚群、DNT细胞、NK细胞变化明显,而治疗缓解后淋巴亚群基本恢复正常,说明T淋巴细胞亚群、DNT细胞、NK细胞水平检测在评价AML疗效及判断预后方面具有一定的临床价值。     

42例缺血性卒中后癫痫发作危险因素探讨

目的:研究缺血性卒中患者继发早期癫痫发作和晚期癫痫发作的发病率、发作类型及缺血性卒中继发癫痫发作的临床危险因素。方法:分析山西省人民医院近3年来首发缺血性卒中的408例患者的年龄、性别、入院时意识、糖尿病、高血压、心源性疾病和卒中部位等的临床特征,随访6~12个月,其中缺血性卒中后继发癫痫发作患者42例,未继发癫痫发作患者366例。缺血性卒中后癫痫发作的患者依据癫痫发作的发生时间分为早期癫痫发作组(18例)和晚期癫痫发作组(24例)。分析缺血性卒中后早期癫痫发作和晚期癫痫发作的发病率、发作类型以及缺血性卒中后发生继发癫痫发作的临床危险因素。结果:缺血性卒中后癫痫发作的发病率为10.3%(42/408),其中早期癫痫发作的发病率为4.4%(18/408);晚期癫痫发作的发病率为5.9%(24/408)。早期癫痫发作的50.0%(9/18)为全面性强直-阵挛发作,晚期癫痫发作的58.3%(14/24)为单纯部分性发作。缺血性卒中后癫痫发作组与非癫痫发作组的临床特征经统计学比较,两组间心源性脑栓塞病例的发生率和累及皮质梗死的发生率比较差异有显著性(P<0.05)。结论:缺血性卒中后早期癫痫发作以全面性发作为主,晚期癫痫发作以单纯部分性发作为主;心源性脑栓塞和累及皮质梗死是缺血性卒中后癫痫发生的危险因素。     

The source of Mycobacterium tuberculosis-specific IFN-γ production in peripheral blood mononuclear cells of TB patients

Mycobacterium tuberculosis (Mtb)-specific IFN-γ secretion plays important roles in anti-tuberculosis (TB) immunity. Mtb-specific IFN-γ response can be induced in HIV/TB co-infected patients with a low CD4 lymphocyte count; this suggests that the source of Mtb-specific IFN-γ production is not limited in CD4⁺ T lymphocytes. Currently, the major sources of Mtb-specific IFN-γ production and the function and phenotype of Mtb-specific IFN-γ-producing cells still remain unclear. Thirty-nine participants (24 active TB patients, 10 HIV/TB co-infected patients, and 5 healthy volunteers) were recruited according to conventional tests and Mtb-specific IFN-γ ELISPOT assay. Multicolor flow cytometry was used to investigate the production of intracellular IFN-γ in peripheral blood mononuclear cells (PBMCs) after Mtb-specific antigen stimulation. Our results showed that CD4⁺, CD8⁺ T cells and NK cells are all major sources of Mtb-specific IFN-γ production in PBMCs of TB patients. Moreover, CD8⁺ T cells are the highest number of Mtb-specific IFN-γ-producing cells in HIV/TB co-infected patients. Although the activity of NK cells is significantly reduced in TB patients when compared with healthy controls, Mtb-specific antigen stimulation induces a significant increase in NK cell activity. We also showed that CD45RO is the characteristic marker of Mtb-specific IFN-γ-producing T cells but not that of Mtb-specific IFN-γ-producing NK cells in peripheral blood. High expression of CD11a may be the characteristic feature of Mtb-specific IFN-γ-producing NK cells. This study put forward a new insight on the source of antigen-specific IFN-γ-production in PBMCs of TB patients.     

Decreased percentage of NKG2D+NK cells in patients with incident onset of Type 1 Diabetes

Type 1 diabetes mellitus (T1DM) is characterized by absolute insulin deficiency owing to autoimmune destruction of the pancreatic β cells. A significant decrease in natural killer (NK) cells in peripheral blood has been observed in patients with untreated T1DM. In the present study, we aimed to explore the role of NK cells and their subsets in young T1DM patients. A total of 30 children and adolescents with untreated T1DM and 27 healthy controls (HC) were recruited in this study. Flow cytometry analysis indicated that the percentage of peripheral blood CD3‐CD56+ NK cells and NK cells subsets (CD56bright, CD56dim and CD56neg), were significantly decreased in the T1DM patients compared to healthy controls. In addition, the percentage of inducible CD107a+ and IFN‐γ‐secreting NK cells was significantly decreased compared to HC. Interestingly, the percentage of NKG2D+ NK cells negatively correlated with the level of serum TCHOL and TG in T1DM patients. Our data indicate that decreased number and impaired function of NK cells may have a role in the pathogenesis of T1DM.