癫痫患儿应用生酮饮食治疗的效果观察

目的:观察生酮饮食不同时间治疗不同年龄阶段的癫痫患儿的临床效果,为临床治疗提供指导。方法:将接受生酮饮食治疗的49例患儿进行严格筛选后挑选42名患儿进入临床观察,遂饮食逐步调整为(糖+蛋白质)/脂肪比例为1:2或1:3,并进行随访,将其分为治疗时间组及年龄组进行分别观察。时间组观察入院前3个月及生酮饮食治疗3、6、12个月及以上时间患儿癫痫发作的情况。年龄组观察8个月-5岁、5岁-12岁、12岁-15岁患儿治疗相同时间癫痫发作情况其治疗效果按Engel标准分级进行统计。结果:患儿在应用生酮饮食治疗6个月后与治疗3个月后相比,患儿的癫痫发作频率明显降低(X~2=23.011,P0.05),与治疗12个月后相比,12个月后大部分患儿癫痫发作可得到有效控制(X~2=65.798,P0.001).而8个月-5岁的患儿癫痫在治疗3个月时,与5岁-12岁年龄段的患儿相比癫痫发作开始有所减少,但没有明显的差异(X~2=5.782,P=0.123),与12岁-15岁患儿相比有较明显的差异(X~2=10.696,P=0.013),6个月后,8个月-5岁患儿癫痫发作的次数明显降低,与5岁-12岁的患儿癫痫发作次数相比差异更加明显(X~2=12.215,P=0.007).12岁-15岁患儿虽也有改善,但与8个月-5岁相比效果不及低年龄段的患儿(X~2=14.766,P=0.002),治疗12个月后有相同的结果(X~2=11.869、18.290,P=0.007、P0.001)。坚持生酮饮食治疗时间12月的患儿愈后情况明显比治疗时间≥12个月的患儿差,差异具有统计学意义(2=9.548,P=0.023)。年龄越小应用生酮饮食治疗患儿的愈后情况越好。差异具有统计学意义(X~2=14.020,P=0.029)。结论:年龄越小的癫痫患儿接受生酮饮食治疗时间越长,治疗效果越好。生酮饮食对于癫痫患儿癫痫的发作有良好的治疗效果。     

呼吸窘迫综合症患儿外周血单个核细胞中CD24和炎症因子mRNA的表达水平及其预后价值

目的:通过检查呼吸窘迫综合症患儿外周血单个核细胞中CD24和TNF-α、IL-6、和IL-17A炎症因子mRNA的表达,探讨其对呼吸窘迫综合症的诊断和预后价值。方法:选择2015年1月至12月在我院接受治疗的32例非感染型呼吸窘迫综合症患儿为研究组,选择同期的30例健康新生患儿为对照组,采集研究组治疗前后和对照组的外周血,分离单个核细胞,采用RT-PCR检测CD24和TNF-α、IL-6、和IL-17A炎症因子mRNA的表达水平。结果:研究组治疗前CD24mRNA表达水平明显高于对照组,差异有统计学意义(P0.01),而TNF-α、IL-6、和IL-17A mRNA表达水平比较,差异无统计学意义(P0.05)。研究组治疗后CD24mRNA表达水平明显低于治疗前,差异有统计学意义(P0.01),而TNF-α、IL-6、和IL-17A mRNA表达水平比较,差异无统计学意义(P0.05)。结论:呼吸窘迫综合症患儿外周血单个核细胞中CD24mRNA表达水平升高,可能是其诊断和预后的分子标记物。     

滤泡辅助性T 细胞在特发性血小板减少性紫癜患者的变化及其意义*

目的:研究滤泡辅助性T细胞(T follicular helper cells,Tfh)在免疫性血小板减少性紫癜(immune thrombocytopenic purpura,ITP)患者的表达并探讨其临床意义。方法:用流式细胞术检测20例健康人、25例ITP患者外周血CXCR5+CD4+T细胞占CD4+T细胞的比例。结果:与健康对照组相比,ITP患者外周血CXCR5+CD4+T细胞占CD4+T细胞的比例显著增高(P<0.05)。结论:Tfh在ITP患者外周血比例增高,为Tfh能否为ITP的免疫调节和干预提出新的方向提供了证据。     

玉屏风颗粒联合他克莫司对原发性肾病综合征患儿肾功能、免疫功能以及Th1/Th2细胞平衡的影响

目的:探讨玉屏风颗粒联合他克莫司对原发性肾病综合征(PNS)患儿肾功能、免疫功能以及Th1/Th2细胞平衡的影响。方法:选取2017年1月～2019年6月期间我院收治的PNS患儿97例,根据随机数字表法将患者分为对照组(n=48)和研究组(n=49),对照组患儿给予他克莫司治疗,研究组在对照组的基础上联合玉屏风颗粒治疗,比较两组患儿疗效、肾功能指标[尿素氮(BUN)、血肌酐(Scr)、免疫功能指标[CD3+、CD4+/CD8+、免疫球蛋白G(Ig G)、免疫球蛋白A(Ig A)]以及Th1/Th2细胞平衡因子[Th1细胞分泌的白细胞介素-2(IL-2)、转化生长因子-β1(TGF-β1)及Th2细胞分泌的白介素-6(IL-6)、白介素-10(IL-10)],记录两组治疗期间不良反应发生情况。结果:研究组治疗9个月后的临床总有效率为91.84%(45/49),高于对照组的75.00%(36/48)(P0.05)。两组治疗9个月后Scr、BUN均下降,且研究组低于对照组(P0.05)。两组治疗9个月后CD3+、CD4+/CD8+、Ig G、Ig A均升高,且研究组高于对照组(P0.05)。两组不良反应发生率比较无差异(P0.05)。两组治疗9个月后TGF-β1、IL-2、IL-10、IL-6均下降,且研究组低于对照组(P0.05)。结论:玉屏风颗粒联合他克莫司治疗PNS患儿,疗效显著,可有效改善患儿肾功能、免疫功能以及Th1/Th2细胞平衡,且不增加不良反应发生率,安全可靠。     

益生菌联合常规药物对难治性癫痫患儿的疗效及对脑电图的影响CSCD

目的 探讨益生菌联合常规药物对难治性癫痫患儿的治疗效果及对患儿脑电图的影响,为该类患儿的治疗提供参考。方法 选取2020年1月至2022年5月我院收治的104例难治性癫痫患儿作为研究对象,按照随机数表法分为对照组和联合组。对照组患儿采用常规药物治疗,联合组在对照组的基础上联用益生菌治疗。对比两组患儿治疗前后癫痫发作频率、脑电图情况、炎症因子[白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)]、神经递质[γ-氨基丁酸(GABA)、5-羟色胺(5-HT)]水平及治疗后的临床疗效和用药安全性。结果 治疗前两组患儿癫痫发作频率、脑电图各频段相对功率以及IL-6、TNF-α、GABA和5-HT水平差异均无统计学意义(均P>0.05)。治疗后两组患儿癫痫发作频率、脑电图θ频段相对功率及IL-6、TNF-α水平均降低,且联合组患儿以上指标水平均低于对照组(均P<0.05)。治疗后两组患儿GABA和5-HT水平均升高,且联合组的GABA和5-HT水平高于对照组(均P<0.05)。两组患儿临床疗效分布差异有统计学意义,联合组总有效率高于对照组(94.23%vs 80.77%,P<0.05)。治疗期间两组患儿的不良反应发生率对比差异无统计学意义(P>0.05)。结论 对难治性癫痫患儿采用益生菌联合常规药物治疗能有效降低其癫痫发作频率,改善脑电图相应频段的相对功率,减轻炎症反应,抑制异常兴奋,且疗效显著,安全性高。     

人类免疫缺陷病毒1型慢性感染者B细胞表型分析及抗反转录病毒治疗的修复作用

人类免疫缺陷病毒1型（HIV-1）感染会造成严重的免疫功能损伤,除引起CD4＋ T细胞不断耗损和功能损伤外,体液免疫应答也受到损伤。本研究通过检测HIV-1慢性感染者和慢性感染治疗者外周血B细胞数目和亚群比例,以及活化、凋亡和共刺激分子的表达,探讨 HIV-1感染者中B细胞损伤及抗反转录病毒治疗（ART）对B细胞损伤的修复作用。结果显示,HIV-1慢性感染者外周血B细胞数目显著低于健康对照组,其中未成熟B细胞、初始B细胞、静息记忆B细胞和浆母细胞显著降低,而组织样记忆B细胞显著增加, ART可恢复初始B细胞和组织样记忆B细胞比例,但不能恢复静息记忆B细胞比例。与健康对照组相比, HIV-1感染者未成熟B细胞、初始B细胞、静息记忆B细胞和组织样记忆B细胞中CD38的表达上调;CD95的表达在所有B细胞亚群中均上调;而Bcl-2在初始B细胞、组织样记忆B细胞和浆母细胞中的表达显著降低;静息记忆B细胞和浆母细胞中PD-1的表达上调;共刺激分子CD40在所有B细胞亚群中的表达降低,而CD70的表达在未成熟B细胞以外的亚群中均显著下调。ART仅能部分修复以上分子的表达。结果表明, HIV-1感染引起B细胞及其亚群比例异常,B细胞表现为过度活化、易凋亡及与T细胞作用受损,ART不能完全修复B细胞损伤,有效的免疫干预策略亟待开发。     

滤泡辅助性T细胞在特发性血小板减少性紫癜患者的变化及其意义

目的：研究滤泡辅助性T细胞（T follicular helper cells,Tfh）在免疫性血小板减少性紫癜（immune thrombocytopenic purpura,ITP）患者的表达并探讨其临床意义。方法：用流式细胞术检测20例健康人、25例ITP患者外周血CXCR5＋CD4＋T细胞占CD4＋T细胞的比例。结果：与健康对照组相比,ITP患者外周血CXCR5＋CD4＋T细胞占CD4＋T细胞的比例显著增高（P〈0.05）。结论：Tfh在ITP患者外周血比例增高,为Tfh能否为ITP的免疫调节和干预提出新的方向提供了证据。     

雌激素受体阳性乳腺癌中mTOR 蛋白的表达及与内分泌辅助治疗预后的关系

目的:研究雌激素受体阳性乳腺癌中m TOR蛋白的表达以及与内分泌辅助治疗预后的关系。方法:选取2010年6月到2011年8月我院收治的雌激素受体阳性乳腺癌且接受内分泌辅助治疗的患者110例,应用免疫组化法检测患者乳腺癌组织中的m TOR蛋白的表达,观察其与临床特征和预后的关系。结果:m TOR蛋白表达阳性者68例,m TOR蛋白表达阴性者42例,m TOR蛋白表达阳性者存在较多的组织学特征;m TOR蛋白表达阴性者无病中位生存期为(56.8±1.1)个月,显著优于m TOR蛋白表达阳性者的(32.8±2.1)个月,比较差异具有统计学意义(P<0.05);多因素分析显示,m TOR蛋白表达阳性者出现肿瘤复发转移的风险是m TOR蛋白表达阴性者的3.21倍,且是影响预后的独立性影响因子。结论:m TOR蛋白的表达阳性是雌激素受体阳性乳腺癌复发转移的独立因素,在临床上可以联合m TOR抑制剂来治疗。     

生酮饮食在神经系统疾病中的临床应用

生酮饮食（ketogenic diet，KD）是一种由高脂肪、低碳水化合物和适量蛋白质及营养素组成的配方饮食。传统上KD主要用于癫痫的治疗，近年来越来越多的研究表明，KD对神经系统也具有一定的保护作用，可用于多种神经系统疾病的临床治疗。KD不仅在Dravet综合征、结节性硬化症和葡萄糖载体蛋白Ⅰ型缺陷综合征等有较好的疗效，对其他神经系统疾病如创伤性颅脑损伤及脑胶质瘤等也具有明显的改善症状的作用。KD治疗神经系统疾病的作用机制尚不明确，可能涉及的神经保护机制包括抗氧化应激、抗炎、抗细胞凋亡、维持能量供应、调节去乙酰化酶活性等。本文综述了KD在神经系统疾病中的作用机制及其应用。     

CD8~+CD122~+T细胞在脑缺血过程中作用的研究

目的:探讨CD8+CD122+T细胞在脑缺血过程中的作用及其机制。方法:线栓法建立小鼠大脑中动脉栓塞模型(MCAO);激光共聚焦显微镜检测小鼠脑缺血组织中CD8+CD122+T细胞浸润情况;流式细胞仪检测脑缺血组织中CD8+CD122+T细胞/CD3+T细胞的比例及脾和胸腺中CD8+CD122+T细胞数量变化;RT-PCR方法检测CD8+CD122+T细胞对氧糖剥夺(Oxygen-glucose deprivation,OGD)条件下星形胶质细胞表达TNF-α,IL-1β,IFN-γ的影响。结果:各时间点脑缺血组织中均有CD8+CD122+T细胞浸润,且随脑缺血时间延长,缺血侧脑组织中CD8+CD122+T细胞/CD3+T细胞比例逐渐增加,5 d和7 d组差异显著,与非缺血侧相比,P5d0.05,P7d0.05;MCAO小鼠脾及胸腺中CD8+CD122+T细胞呈现先增高后降低的趋势。星形胶质细胞经OGD处理后,与对照组相比,IFN-γ、TNF-α、IL-1β表达显著增高,PIFN-γ0.01、PTNF-α0.001、PIL-1β0.01;CD122-blocked组与CD8+组相比,IFN-γ、TNF-α、IL-1β表达明显增高,PIFN-γ0.05、PTNF-α0.05、PIL-1β0.01;CD8+组与HBSS组相比,IFN-γ表达降低,P0.05;IL-1β表达有降低的趋势。结论:CD8+CD122+T细胞在脑缺血过程中发挥保护性作用,其保护作用通过CD122抑制星形胶质细胞TNF-α,IL-1β,IFN-γ炎症因子表达实现的。     

The ketogenic diet has no effect on the expression of spike-and-wave discharges and nutrient transporters in genetic absence epilepsy rats from Strasbourg

The genetic absence epilepsy rat from Strasbourg is considered an isomorphic, predictive, and homologous model of typical childhood absence epilepsy. It is characterized by the expression of spike-and-wave discharges (SWDs) in the thalamus and cortex. The ketogenic diet (KD) is successfully used in humans and animals with various types of seizures, but was not effective in children with intractable atypical absence epilepsy. Here, we studied its potential impact on the occurrence of SWDs in genetic absence epilepsy rat from Strasbourg. Rats were fed the KD for 3 weeks during which they were regularly subjected to the electroencephalographic recording of SWDs. The KD did not influence the number and duration of SWDs despite a 15–22% decrease in plasma glucose levels and a large increase in β-hydroxybutyrate levels. Likewise, the KD did not affect the level of expression of the blood–brain barrier glucose transporter GLUT1 or of the monocarboxylate transporters, MCT1 and MCT2. This report extends the observation in humans that the KD does not appear to show effectiveness in intractable atypical absence epilepsy to this model of typical childhood absence epilepsy which responds to specific antiepileptic drugs.     

Inhibition of Increased Circulating Tfh Cell by Anti-CD20 Monoclonal Antibody in Patients with Type 1 Diabetes

Objectives

Follicular helper T (Tfh) cells exert an important role in autoimmune diseases. Whether it might be involved in type 1 diabetes (T1D) is unknown. Our aim was to investigate the role of Tfh cells in patients with T1D and the effect of anti-CD20 monoclonal antibody (rituximab) on Tfh cells from T1D patients.

Patients and Methods

Fifty-four patients with T1D and 37 healthy controls were enrolled in the current study. 20 of those patients were treated with rituximab. The frequencies of circulating CD4⁺CXCR5⁺ICOS⁺T cells were analyzed by flow cytometry. The serum autoantibodies were detected by radioligand assay. The levels of IL-21, IL-6 and BCL-6 were assessed using ELISA and/or real-time PCR.

Results

Increased frequencies of circulating Tfh cells together with enhanced expression of IL-21 were detected in patients. The correlation between the frequencies of circulating Tfh cells and the serum autoantibodies or C-peptide level was comfirmed. After rituximab therapy, follow-up analysis demonstrated that the frequencies of circulating Tfh cell and serum IA2A were decreased. The levels of IL-21, IL-6 and Bcl-6 mRNA were decreased after treatment. Furthermore, beta cell function in 10 of 20 patients was improved.

Conclusions

These data indicate Tfh cells may participate in the T1D-relatede immune responses and B cells might play a role in the development of Tfh responses in the disease progression.     

Effects of the ketogenic diet in the glucose transporter 1 deficiency syndrome

The ketogenic diet (KD), established to treat intractable childhood epilepsy, has emerged as the principal treatment of GLUT1 deficiency syndrome (OMIM 606777). This defect of glucose transport into the brain results in hypoglycorrhachia causing epilepsy, developmental delay, and a complex motor disorder in early childhood. Ketones provided by a high-fat, low-carbohydrate diet serve as an alternative fuel to the brain. Glucose, lactate, lipids, and ketones in blood and cerebrospinal fluid were investigated in five GLUT1-deficient patients before and on the KD. Hypoglycorrhachia was detected in the non-ketotic and ketotic state. In ketosis, lactate concentrations in the cerebrospinal fluid increased moderately. The CSF/blood ratio for acetoacetate was higher compared to beta-hydroxybutyrate. Free fatty acids did not enter the brain in significant amounts. Blood concentrations of essential fatty acids determined in 18 GLUT1-deficient patients on the KD were sufficient in all age groups. The effects of the KD in GLUT1 deficiency syndrome, particularly the course of blood lipids, are discussed in an illustrative case. In this syndrome, the KD effectively restores brain energy metabolism. Ketosis does not influence impaired GLUT1-mediated glucose transport into brain: hypoglycorrhachia, the biochemical hallmark of the disease, can be identified in GLUT1-deficient patients on a KD. The effects of ketosis on the concentrations of glucose, lactate, ketones, and fatty acids in blood and cerebrospinal fluid in this entity are discussed in view of previous data on ketosis in man.     

The effect of aging on the frequency,phenotype and cytokine production of human blood CD4 + CXCR5 + T follicular helper cells: comparison of aged and young subjects

Background

T cell-dependent B-cell responses decline with age, indicating declined cognate helper activity of aged CD4?+?T cells for B cells. However, the mechanisms remain unclear. T follicular helper (Tfh) cells, a novel T helper subset, play an essential role in helping B cells differentiation into long-lived plasma cells in germinal center (GC) or short-lived plasma cells. In the present study, we proposed that there might existe changes of proportion, phenotype or cytokine production of blood Tfh cells in healthy elderly individuals compared with healthy young individuals.

Results

The results showed that frequencies of aged blood CXCR5?+?CD4?+?Tfh cells increased compared with young subjects. Both aged and young blood CXCR5?+?CD4?+?Tfh cells constitutively expressed CD45RO, CCR7 and CD28, and few of these cells expressed CD69 or HLA-DR, which indicated that they were resting memory cells. There was no significant difference of IL-21 frequency production by aged blood CXCR5?+?CD4?+?Tfh determined by FACS compared with young individuals, however, aged PBMCs produced significantly higher levels of IL-21 evaluated by ELISA. Furthermore, there were no significant differences of percentages of IFN-γ, IL-4, IL-17 or IL-22 production by aged Tfh cells compared with their counterparts of young individuals respectively. However, frequencies of IL-17+ cells within aged CD4?+?CXCR5-T cells were markedly lower than in the young individuals. Furthermore we observed different frequencies of IFN-γ, IL-17, IL-4 or IL-22 production by Tfh or by CD4?+?CXCR5- cells in aged and young subjects respectively.

Conclusions

Our data demonstrated that the frequencies of blood memory CXCR5?+?CD4?+?Tfh cells increased in the elderly population. There were similar frequencies of Th characterized cytokine production such as IL-21, IFN-γ, IL-4, IL-17 or IL-22 in aged and young Tfh cells. However, aged PBMCs produced a significantly higher amount of IL-21 compare to young subjects.