凉血通瘀方干预高血压大鼠急性脑出血模型对脑组织中差异miRNA表达的影响＊

目的 研究凉血通瘀方对高血压大鼠急性脑出血模型脑组织miRNA表达的影响，对差异表达的miRNA靶基因进行分析，探索凉血通瘀方可能的药效机制。方法 将自发性高血压大鼠随机分成对照组（B）和实验组（C）。适应性饲养一周后，C组灌胃凉血通瘀方，B组灌胃等体积生理盐水，连续5天，每天1次。构建脑出血模型后收集脑组织，借助全转录组测序技术获得miRNA表达量，与miRBase数据库比对获取已知miRNA，使用miRDeep2预测新miRNA。差异分析软件为DESeq2，筛选阈值为|log₂FC| ≥1 并且P <0.05。对显著差异表达的miRNA进行靶基因预测，对靶基因进行GO功能、KEGG通路富集和PPI网络分析。结果 实验组和对照组对比，共发现21个显著差异表达的miRNA，上调有9个，下调有12个，共预测得到1243个有统计学意义的靶基因。GO富集分析发现，生物过程中突触囊泡分泌的调节、神经递质分泌的调节和神经递质运输的调节占前三位，神经元投射终点、全膜、质膜区域和细胞投射则是主要的细胞成分。分子功能分别为小GTPase绑定、底物特异性跨膜转运蛋白活性和离子跨膜转运体活性。通路分析结果显示，靶基因在癌证通路、pI3K-Akt信号通路、人类乳头瘤病毒感染、神经活性配体-受体相互作用和MAPK通路等分布广泛。采用STRING网站和Cytoscape软件，根据MCC算法筛选出ADRA2C、CASR、CCL28、CCR1、DRD2、GNAT3、GRM2、DYNC1LI1、GABBR1、GNAI1等核心靶基因。结论 凉血通瘀方对脑出血急性期鼠脑组织内miRNA的表达有重要影响；显著差异表达miRNAs可能通过靶向核心基因调控凉血通瘀方干预急性脑出血的病理过程及预后。     

我国弓形虫Chinese 1优势基因型感染对小鼠脑组织铁代谢影响

目的　探讨我国弓形虫Chinese 1优势基因型感染对宿主脑组织铁代谢及脑损伤的影响。方法　将20只C57BL/6（体质量15～17 g）小鼠随机分为对照组和感染组，每组10只。感染组每只小鼠腹腔注射4 000个弓形虫Chinese 1优势基因型TgCtwh3虫株速殖子，对照组小鼠注射等量无菌PBS，饲养6 d后处死小鼠并取其脑组织。采用电感耦合等离子体质谱法（inductively coupled plasma mass spectrometry, ICP⁃MS）检测小鼠脑组织铁元素水平；采用RNA芯片检测两组小鼠脑组织差异基因数目并对功能基因表达情况进行基因本体论（Gene Ontology, GO）功能富集；采用实时荧光定量PCR（fluorescent quantitative real⁃time PCR, qPCR）技术检测小鼠脑组织中弓形虫表面抗原1（Toxoplasma gondii surface antigen 1，TgSAG1）基因及部分锌铁调控蛋白（Zrt⁃ and Irt⁃like protein, ZIP）家族mRNA表达水平；采用光镜和电镜观察小鼠脑组织海马齿轮回（dentate gyrus, DG）超微结构；采用Western blotting检测谷胱甘肽过氧化物酶4（glutathione peroxidase 4, GPx4）蛋白表达水平；采用硫代巴比妥酸（TBA）法检测丙二醛（malondialdehyde, MDA）水平；采用免疫组化检测血管内皮生长因子（vascular endothelial growth factor, VEGF）蛋白表达光密度（optical density, OD）值。结果　光镜下可见感染组小鼠脑组织海马DG区细胞坏死，电镜下见感染组小鼠脑组织海马区出现胞质空泡化、核皱缩坏死、线粒体嵴断裂消融、自噬小体增加等超微结构变化。与对照组相比，感染组小鼠脑组织中铁元素水平上调[（32.92 ± 0.90） µg/g vs.（37.72 ± 1.10） µg/g；t = 3.397, P < 0.01]；RNA芯片检测感染组小鼠脑组织发现721个基因上调、276个基因下调，差异表达基因在金属离子结合能力上有明显富集。与对照组相比，感染组小鼠脑组织金属元素转运体ZIP2 mRNA表达水平上调（t = 8.659，P < 0.05）、GPx4表达下降[（1.046 ± 0.025） vs. （0.720 ± 0.101）；t = 3.129，P < 0.01]）、MDA水平升高[（4.37 ± 0.33） nmol/mgprot vs.（5.93 ± 0.54） nmol/mgprot；t = 2.451，P < 0.05）]、VEGF蛋白平均OD值上调[（0.348 3 ± 0.017 8） vs. （0.490 6 ± 0.010 5）；t = 6.641，P < 0.01]。结论　Chinese 1优势基因型弓形虫感染后，小鼠脑组织中铁元素蓄积、抗氧化能力下调、氧化应激水平升高，提示弓形虫感染可影响宿主脑组织铁代谢而导致脑损伤。     

Reliability and validity of the gait classification system in children with cerebral palsy (GCS-CP)

BackgroundGait classification systems (GCS) may enable clinicians to differentiate gait patterns into clinically significant categories that assist in clinical decision-making and assessment of outcomes. Davids and Bagley in 2014 [1] described a GCS for children with cerebral palsy (GCS-CP). The purpose of our study was to use the GCS-CP for the first time on a sample of patients with CP and to evaluate the reliability and utility of the classification system.MethodsThe gait of 131 children with CP was retrospectively reviewed and classified according to Davids and Bagley’s classification using two-dimensional (2D) video and three-dimensional (3D) lower limb kinematics and kinetics. Gross Motor Function Classification System (GMFCS) levels were determined, and the Gait Profile Scores (GPS) calculated to characterize the sample concerning gait classification. The comparison between the groups was performed using the Kruskal-Wallis test with respect to the non-normal distribution of the data. The intrarater and interrater reliability was determined using the Kappa index (k) statistics with 95% CI.ResultsAll GCS-CP groups were represented within the evaluated sample. Of the 131 cases evaluated, 127 (96.95%) were able to be classified with respect to sagittal plane stance phase gait deviations. All patients in the sample were able to be classified with respect to sagittal plane swing phase and transverse plane gait deviations. The interrater reliability was 0.596 and 0.485 for the first and second levels of the classification, respectively, according to the Fleiss’s Kappa statistics. Intrarater reliability was 0.776 and 0.714 for the raters one and two, respectively, according to the Cohen’s Kappa statistics.SignificanceThe GCS-CP exhibited clinical utility, successfully classifying almost all subjects with CP in two planes, based upon kinematic and kinetic data. The classification is valid and has moderate interrater and moderate to substantial intrarater reliability.     

Potentilla anserine L. polysaccharide protects against cadmium-induced neurotoxicity

Cadmium is a toxic metal that can damage the brain and other organs. This study aimed to explore the protective effects of Potentilla anserine L. polysaccharide (PAP) against CdCl₂-induced neurotoxicity in N2a and SH-SY5Y cells and in the cerebral cortex of BALB/c mice. In addition, we aimed to identify the potential mechanisms underlying these protective effects. Relative to CdCl₂ treatment alone, pretreatment with PAP prevented the reduction in cell viability evoked by CdCl₂, decreased rates of apoptosis, promoted calcium homeostasis, decreased ROS accumulation, increased mitochondrial membrane potential, inhibited cytochrome C and AIF release, and prevented the cleavage of caspase-3 and PARP. In addition, PAP significantly decreased the CdCl₂-induced phosphorylation of CaMKII, Akt, and mTOR. In conclusion, PAP represents a potential therapeutic agent for the treatment of Cd-induced neurotoxicity, functioning in part via attenuating the activation of the mitochondrial apoptosis pathway and the Ca²⁺-CaMKII-dependent Akt/mTOR pathway.     

Reliability and validity of the Turkish version of Fullerton Advanced Balance Scale in cerebral palsy

BackgroundThe Fullerton Advanced Balance Scale (FAB) is a multi-item balance assessment test designed to measure balance in relatively higher functioning individuals. The aim of this study was to examine the reliability and validity of the Turkish version of the FAB (FAB-T) in children with cerebral palsy (CP).Research questionIs the Turkish version of the Fullerton Advance Balance Scale valid and reliable in determining balance problems in children with cerebral palsy and determining the underlying cause of this condition?MethodsForty-six children with CP participated in this study. Rasch analysis was used to investigate item adherence. Internal consistency of the FAB-T was established using Cronbach's alpha coefficient. Test-retest reliability was also evaluated. In addition, to assess concurrent validity, FAB-T scores were compared with the Pediatric Balance Scale (PBS) using the Spearman correlation coefficient.ResultsThe FAB-T showed satisfactory internal consistency (Cronbach's alpha value=0.94) and excellent test-retest reliability (ICC=0.99). The FAB and the PBS exhibited concurrent positive validity (r = 0.913; p < 0.001). All items of the FAB-T were found to fit the Rasch Model (Chi-square 16.01(df=20), p = 0.716).SignificanceThe FAB-T is a reliable and valid tool that can be used to measure balance skills and to identify the source of the problem in children with CP.