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目的探讨神经元特异性烯醇化酶(NSE)、S-100β对肠道病毒71型(EV71)脑炎的早期诊断及脑损伤程度的评估价值。方法选取2008年5月至2010年12月山东大学齐鲁儿童医院重症监护室(PICU)收治的EV71脑炎患儿(EV71脑炎组)100例,同期住院的其他病毒性脑炎患儿89例为其他病毒性脑炎组,非神经系统感染患儿22例为阴性对照组。采用ELISA法检测所有EV71脑炎组及其他病毒性脑炎组患儿急性期及恢复期脑脊液与血清NSE及S-100β蛋白含量,检测阴性对照组患儿急性期及恢复期血清NSE及S-100β蛋白含量及急性期脑脊液NSE及S-100β蛋白含量,并分别进行比较。结果 EV71脑炎组急性期脑脊液、血清及恢复期血清中NSE与S-100β高于阴性对照组,差异均有统计学意义(P均<0.01)。EV71脑炎组急性期脑脊液中NSE低于其他病毒性脑炎组,差异有统计学意义(P<0.05)。EV71脑炎组急性期血清与恢复期脑脊液、血清中S-100β蛋白低于其他病毒性脑炎组,差异有统计学意义(P<0.05)。EV71脑干脑炎组急性期与恢复期脑脊液、血清中NSE显著高于普通脑炎组,差异有统计学意义(P<0.05或P<0.01);EV71脑干脑炎组急性期与恢复期脑脊液、血清中S-100β蛋白显著高于EV71普通脑炎组,差异有统计学意义(P<0.01)。血清与脑脊液中NSE与S-100β有明显相关性(r=0.782、0.734,P均<0.01)。结论监测手足口病患儿脑脊液与血清NSE、S-100β可早期诊断是否合并中枢神经感染,并可评估脑损伤程度及预后。     

基于STM32芯片的肿瘤细胞荧光检测系统设计

为了实现肿瘤细胞早期筛查以达到早期防治,本研究开发了一种基于微流控芯片的肿瘤细胞荧光检测系统。光路采用反射式共焦距光路结构;硬件电路基于光电探测器模块、STM32F103C8T6、A/D转换芯片与串口芯片对激发荧光信号实时采集,并向上位机传输。同时设计了一套基于荧光检测的软件系统,实现检测动态可视化与报告生成及打印功能。采用FITC荧光素标记的乳腺癌细胞MDA-MB-453进行试验。结果表明,该系统在6 mL/h最佳进样速度下,对肿瘤细胞的平均检测率达到85.36%,为临床的应用提供可靠的诊断方法。     

缺氧对新生大鼠海马神经干细胞增殖和分化的影响

目的探讨单纯性缺氧对新生大鼠海马神经干细胞(NSCs)增殖和分化的影响。方法将36只新生1d大鼠随机均分为N组(正常对照组)、Q2组(缺氧2h组)、Q5组(缺氧5h组)。将海马组织在无血清培养液(均含有碱性成纤维生长因子bFGF、表皮生长因子EGF和白细胞抗原B27)中,分别进行原代培养、单克隆培养和传代培养,单克隆培养细胞进行巢蛋白(Nestin)和Hoechst33342免疫荧光双标染色;传代培养的细胞传三代诱导分化,诱导分化3d的细胞分别行神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)和Hoechst3342免疫荧光双标染色,计数NSE阳性细胞比例,进行统计学分析。结果单克隆培养的细胞均呈Nestin阳性表达,诱导分化的细胞部分呈NSE或GFAP阳性表达。Q2组、N组、Q5组细胞原代培养克隆球直径达2μm的时间分别为7d、10d、13d。传3代后,对各组细胞进行诱导分化,Q2组细胞诱导分化为神经元的比例明显高于N组,Q5组的细胞诱导分化为神经元的比例明显低于N组。结论短时间缺氧可促进在体海马神经干细胞增殖和诱导其向神经元分化,长时间缺氧则抑制在体神经干细胞增殖和向神经元分化。     

Generic effectiveness measures: sensitivity to symptom change in anxiety disorders

OBJECTIVE: The purpose of this study was to compare a newly developed screening measure of disability, the WHODAS II, to three established generic effectiveness measures in terms of its sensitivity to symptom change in people with anxiety disorders. METHOD: Patients who had undergone treatment for social phobia or panic disorder/agoraphobia at an anxiety disorders clinic were administered generic effectiveness measures and symptom measures before and after treatment. The design was naturalistic and observational. Data analysis included correlations between generic effectiveness and symptom measures; and effect size calculations regarding the ability of each generic effectiveness measure to discriminate between patients whose symptoms improved and patients whose symptoms did not improve over the course of treatment. RESULTS: The WHODAS II was consistently the most sensitive generic effectiveness measure in its capacity to detect symptom changes in patients with social phobia. The SF-12 and K-10 also showed moderate sensitivity to symptom change. In the sample of patients with panic disorder/agoraphobia, the SF-12 was the most sensitive measure overall, closely followed by the K-10 and WHODAS II. The NCS Disability Days were the least sensitive to symptom change in both samples. CONCLUSION: The WHODAS II is at least as sensitive as other generic effectiveness measures to anxiety symptom changes, and is particularly sensitive to changes in social anxiety symptoms. It may prove to be a valuable measurement tool for informing public health policy in relation to anxiety disorders.     

枕下极外侧入路应用解剖学探讨及手术体会

目的探讨枕下极外侧入路切除延颈交界区腹侧病变的效果,并对一些手术技巧加以讨论。方法在10例成人尸头上模拟枕下极外侧入路进行延颈交界区腹侧局部显微解剖探讨,观测延颈交界区有关结构。采用枕下极外侧入路对26例延颈交界区腹侧及腹外侧病变进行手术治疗。结果本组26例患者,9例痊愈,17例好转。14例肿瘤全切除,12例次全切除。全组无手术死亡。术后并发症包括面瘫2例,后组脑神经轻度麻痹1例,脑脊液漏1例。结论枕下极外侧入路可以满足延颈交界区腹侧及腹外侧病变手术野的显露,是切除该部位病变的一种有效的手术方法。     

UHRF1 regulation of the Keap1–Nrf2 pathway in pancreatic cancer contributes to oncogenesis

The cellular defence protein Nrf2 is a mediator of oncogenesis in pancreatic ductal adenocarcinoma (PDAC) and other cancers. However, the control of Nrf2 expression and activity in cancer is not fully understood. We previously reported the absence of Keap1, a pivotal regulator of Nrf2, in ～70% of PDAC cases. Here we describe a novel mechanism whereby the epigenetic regulator UHRF1 suppresses Keap1 protein levels. UHRF1 expression was observed in 20% (5 of 25) of benign pancreatic ducts compared to 86% (114 of 132) of pancreatic tumours, and an inverse relationship between UHRF1 and Keap1 levels in PDAC tumours (n = 124) was apparent (p = 0.002). We also provide evidence that UHRF1‐mediated regulation of the Nrf2 pathway contributes to the aggressive behaviour of PDAC. Depletion of UHRF1 from PDAC cells decreased growth and enhanced apoptosis and cell cycle arrest. UHRF1 depletion also led to reduced levels of Nrf2‐regulated downstream proteins and was accompanied by heightened oxidative stress, in the form of lower glutathione levels and increased reactive oxygen species. Concomitant depletion of Keap1 and UHRF1 restored Nrf2 levels and reversed cell cycle arrest and the increase in reactive oxygen species. Mechanistically, depletion of UHRF1 reduced global and tumour suppressor promoter methylation in pancreatic cancer cell lines, and KEAP1 gene promoter methylation was reduced in one of three cell lines examined. Thus, methylation of the KEAP1 gene promoter may contribute to the suppression of Keap1 protein levels by UHRF1, although our data suggest that additional mechanisms need to be explored. Finally, we demonstrate that K‐Ras drives UHRF1 expression, establishing a novel link between this oncogene and Nrf2‐mediated cellular protection. Since UHRF1 over‐expression occurs in other cancers, its ability to regulate the Keap1–Nrf2 pathway may be critically important to the malignant behaviour of these cancers. © 2015 The Authors. Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.     

母亲行为与儿童行为问题的探讨

目的 :探讨母亲行为与儿童行为问题之间的关系。方法 :以山东省 765名 6-11岁儿童的母亲为被试 ,采用Achenbach’s儿童行为量表 (CBCL)和母亲行为问卷分别对儿童及其母亲进行评定。结果 :儿童行为问题的检出率为 12 0 % ,有行为问题儿童母亲的支持行为显著低于正常儿童 ,有行为问题儿童母亲的不支持行为显著高于正常儿童。结论 :母亲的不支持行为增加儿童的行为问题 ,母亲支持行为减少儿童的行为问题。母亲行为对儿童的行为问题有显著影响     

用噬菌体肽库筛选重组日本血吸虫线粒体相关蛋白的表位

目的：筛选和鉴定重组的日本血吸虫（中国大陆株）线粒体相关蛋白rSj38的表位。方法：用纯化的rSj338/26GST免疫家兔获得抗rSj338/26GST的多克隆抗体IgG，将抗体进一步纯化，获得抗rSj338单特异多克隆抗体IgG。用纯化抗rSj338抗体对噬菌体12肽库进行5轮免疫学筛选，挑取克隆。采用Western blot免疫识别，核苷酸序列测定分析其获得的表位并与rSj338/26GST进行同源性比较。将获得的不同表位的阳性克隆分别免疫小鼠，并采用Western blot及dot-ELISA方法筛选能刺激小鼠产生较高滴度抗rSj338抗体的阳性克隆，并将阳性噬菌体免疫的小鼠血清对纯化的rSj338/26GST,26GST，日本血吸虫成虫及虫卵抗原进行Western blot识别。结果：经5轮免疫学筛选后挑取的32个克隆，用Western blot方法30个克隆能被抗rSj338抗体识别，核苷酸序列分析发现共有11种表位，与rSj338无一级结构的同源性。经动物免疫初步实验筛选。共获得4个免疫原性较强的阳性克隆，其免疫鼠血清均可识别rSj338/26GST，日本血吸虫成虫及虫卵抗原。结论：获得了4种日本血吸虫中国大陆株线粒体相关蛋白rSj338的表位，均为模拟表位，这将为日本血吸虫病的疫苗研究开辟新的途径。     

一氧化氮合酶与缺氧复氧所致神经细胞凋亡及银杏叶提取物的保护作用

目的　探讨一氧化氮 (NO)在缺氧复氧诱导神经细胞凋亡中的作用及中药银杏叶提取物的保护机制。方法　实验使用胎龄 16～ 17日Wistar大鼠的大脑皮层神经细胞进行原代分离培养 ;采用Wright Giemsa染色 ,光镜、透射电子显微镜观察 ;原位末端标记法确立缺氧复氧神经细胞凋亡病理模型 ;应用NADPH d组织化学方法检测神经细胞一氧化氮合酶 (NOS)的表达并用计算机图像分析系统进行定量检测。　结果　缺氧复氧可以使大鼠大脑皮层神经细胞发生凋亡 ,随缺氧时间的延长 ,凋亡细胞数渐多 ,至缺氧 8h复氧 18h达高峰 ;在缺氧 2h(H2 R0 组 )和缺氧 8h复氧 18h(R8R1 8)组中神经细胞NOS表达均显著增高 ,与正常对照组比有显著性差异 (P <0 0 1;P <0 0 5 )。EGB能显著抑制此双时相NOS活性的增强 ,并明显降低神经细胞凋亡率。　结论　缺氧复氧损伤可诱导培养的大鼠大脑皮层神经细胞发生凋亡。NOS表达增强从而使NO产生增加可能是缺氧复氧诱导神经细胞凋亡的机制之一。银杏叶提取物 (EGB)经下调NOS表达活性 ,抑制NO的产生保护培养的大鼠大脑皮层神经细胞免于凋亡。     

甲状腺癌中肿瘤坏死因子相关凋亡诱导配体及其受体的表达

目的　研究肿瘤坏死因子相关凋亡诱导配体 (TRAIL)和肿瘤坏死因子相关凋亡诱导配体受体(TRAILR)在甲状腺癌中的表达及意义。　方法　采用免疫组织化学方法 ,检测 5例正常甲状腺、13例乳头状甲状腺癌、3例滤泡状甲状腺癌和 12例甲状腺癌旁组织中TRAIL和TRAILR的表达和分布。　结果　乳头状、滤泡状甲状腺癌组织和正常甲状腺组织中的甲状腺滤泡细胞均表达TRAIL和全部的TRAILR ,其中诱捕受体TRAILR4在正常甲状腺组织和甲状腺癌旁组织表达较弱。　结论　甲状腺癌组织中的甲状腺滤泡细胞表达TRAIL和全部的TRAILR ,提示癌变的甲状腺滤泡细胞通过自身表达TRAIL ,以自分泌或旁分泌的形式和其死亡受体TRAILR1、TRAILR2结合 ,诱导癌变的甲状腺滤泡细胞发生凋亡 ,诱捕受体TRAILR3、TRAILR4的存在也不能影响其对TRAIL诱导的细胞凋亡的敏感性