CT血管造影对脑血管性疾病的临床诊断价值

目的评价64层螺旋CT血管成像对脑血管病的诊断及临床应用价值。方法64层螺旋CT血管成像检查确诊脑血管疾病患者60例,50例在1周内行DSA检查。采用减影方法进行后处理,分别得到VR,MIP,TSMIP图像,利用这三种方法重建图像综合评价脑血管病变。结果①脑动脉狭窄或闭塞者46例,其中36例有DSA对照,98．8％的狭窄或闭塞的脑血管与DSA诊断一致,CT血管成像的敏感性,特异性,准确性分别为100％、98．5％、98．8％;②烟雾病3例,均有DSA对照,表现为单侧或双侧大脑中动脉重度狭窄,远端分支异常增多;③脑动脉瘤患者12例,均有DSA对照,后交通动脉瘤7例,前交通动脉瘤3例,大脑中动脉瘤1例,大脑后动脉瘤1例,CT血管成像与DSA显示一致。结论64层螺旋CT层血管成像能快速,准确地诊断各种脑血管疾病,结合减影处理方法提高了诊断的效率和准确性,为临床提供了全面的信息,可作为脑血管疾病患者的首选诊断方法。     

早期母子隔离应激对大鼠学习记忆能力的影响

目的探讨早期母子隔离应激对幼鼠学习记忆能力的影响。方法30只SD新生大鼠用于实验,实验随机分成3组,正常对照纽、母子隔离15分钟纽、母子隔离3小时纽。在生后40天,三组大鼠进行Morris水迷宫及记录海马脑片长时程增强电位（LTP）实验。结果Morris水迷宫测试,与正常组比较母子隔离3h组大鼠寻找平台需要的潜伏期延长（P〈0．05）,而母子隔离15分钟组大鼠寻找平台需要的潜伏期与正常组比较则无明显变化（P〉0．05）;海马脑片长时程增强电位提示：正常对照组条件刺激（CS）前后fEPSP斜率变化率为：64．7±22．9％;母子隔离3h组为：35．3±14．2％,低于正常组（P〈0．01）;母子隔离15min组为：66．3±29．7％,与正常组比较无显著性意义（P〉0．05）。结论幼鼠生后早期过度的应激导致大鼠的学习记忆能力减弱。     

胃癌组织中EGFL7的表达及其临床病理特征的关系研究

目的：探讨胃癌组织中表皮生长因子样结构域（EGFL7）的表达及其与胃癌临床病理特征的关系。方法：采用免疫组织化学SP法分别检测45例胃癌组织中EGFL7的表达水平及微血管密度（MVD,CD34标记）,分析EGFL7的表达与MVD及胃癌临床病理特征之间的关系。结果：EGFL7的表达在有淋巴结转移组中的阳性率（84．6％）高于无淋巴结转移组（57．9％）,在浸润浆膜层组中的阳性率（84．4％）高于未浸润浆膜层组（46．2％）。胃癌组织中EGFL7的表达与淋巴结转移和浸润程度呈正相关（P〈0．05）,但与患者的年龄、性别、肿瘤的分化程度无关（P〉0．05）。胃腺癌低分化纽MVD（37．62±10．42）高于高分化组（27．91±9．93）和中分化组（28．40±9．18）;浸润至浆膜层组MVD（34．25±10．43）高于未至浆膜层组（26．69±8．66）;有淋巴结转移组MVD（33．00±9．99）高于无淋巴结转移组（25．94±9．34）。癌组织中MVD值与分化程度、浸润深度及有无淋巴结转移呈正相关（P〈0．05）,但与年龄、性别无关（P〉0．05）。在EGFL7表达阳性的肿瘤纽织中MVD均值为33．80±10．56,高于EGFL7表达阴性的肿瘤组织中MVD均值26．004-7．21（P〈0．05）,采用Pearson积差相关系数进行分析,发现MVD值与EGFL7的表达成正相关（r=0．313,P〈0．05）。结论：EGFL7的高表达可能促进了胃癌的浸润和转移,其机制可能与促进胃癌组织中的血管新生有关。     

小鼠皮肤局部缺血模型的建立

目的研究建立稳定的昆明小鼠皮肤局部缺血模型。方法取40只雌性昆明小鼠，随机分为A／B两组，每组20只。分别在小鼠背部建立深达皮下筋膜层的u型皮瓣。A组为3．3×1．5cm皮瓣组．B组为2．5×1．5CITI皮瓣组。皮瓣游离缘垫以医用橡胶，缝合切口，造成皮肤缺血模型，术后观察皮瓣发绀水肿及坏死情况，并用散斑全景实时血流成像仪（MoorFLH）检测皮瓣血流。结果肉眼观察，3．3×1．5cm皮瓣组在术后第2天，50％小鼠皮瓣末端坏死，坏死面积达皮瓣面积的（37．20±4．83）％。术后第3天100％小鼠皮瓣末端坏死。并随时间的延长，坏死面积逐渐扩大。2．5×1．5皮瓣组在术后第1天皮瓣出现发绀，水肿，发绀水肿面积占（77．46±4．51）％，术后第3，第5天发绀水肿区面积逐渐减小，至术后第7天，发绀、水肿完全消退。散斑全景实时血流成像仪（MoorFLPI）检测皮肤血流显示，术后第1天皮瓣显著性缺血（P〈0．05），术后第7天血流基本恢复到术前水平。结论成功建立了昆明小鼠皮肤局部缺血模型，为进一步研究组织微血管再生的机理和策略提供简单而稳定的动物模型。     

darbufelone对胃癌SGC-7901细胞裸鼠移植瘤血管生成的影响

目的观察环氧合酶-2／5-脂氧合酶双重抑制剂darbufelone对人胃癌皮下移植瘤血管生成的影响,并初步探讨其机制。方法建立裸鼠实体瘤模型,随机分为darbufelone组和对照组,darbufelone及生理盐水分别连续灌服4周。测量肿瘤质量、体积,计算抑瘤率;免疫组化检测CD34并计算微血管密度;RT—PCR法及Western blot法分析移植瘤组织中MMP-9、VEGF的表达。结果darbufelone可明显抑制裸鼠移植瘤的生长,质量抑瘤率为58．42％,体积抑瘤率为67．13％。darbufelone组的微血管密度（MVD）（15．36±0．30）明显低于对照组（29．47±0．63）（P〈0．05）;darbufelone组肿瘤组织中VEGF及MMP-9在基因水平及蛋白水平的表达（P〈0．05）。结论darbufelone能有效抑制裸鼠移植瘤的生长,减少移植瘤组织中VEGF及MMP-9的表达,抑制肿瘤的微血管生成,具有抗血管生成的作用。     

难治性癫痫患者脑组织中SCG10的表达及其意义

目的了解难治性癫痫患者脑组织中SCG10和MAPK的表达,探讨其在难治性癫痫发生、发展中的作用。方法按随机化原则,在我们建立的难治性癫痫患者术后脑组织库中抽取36例患者的脑组织,用免疫组织化学分别检测SCG10和MAPK的表迭,与16例对照组进行比较。并采用免疫荧光双标的方法,检测SCG10和MAPK在病例组中的表达情况。结果免疫组化检测到SCG10在实验组表达（0．4674±0．0258）,高于对照（0．3405±0．0207）,两组比较有显著性差异（P〈0．05）;MAPK在实验组表达（0．4217±0．0141）,同样也高于对照组（0．3189±0．1422,P〈0．05）。免疫荧光双标法,显示SCG10和MAPK表达在同一细胞上。讨论SCG10与MAPK在难治性癫痫患者的脑组织中表达增加,以及它们的共同表达,提示两者的相互作用可能是难治性癫痫发生发展中的一个重要因素。     

多排螺旋CT扫描对胃癌的诊断价值

目的评价多排螺旋CT扫描对胃癌的诊断价值。方法对62例胃癌病例行气体对比薄层动态增强CT扫描三维重建技术,层厚0．5mm,在此基础上进行CT仿真胃镜和VRT技术,对获取的图像进行分析。全部病例均经手术和病理证实。结果检出痛变位于胃窦部31例（50％）.胃底贲门部10例（16％）,体部9例（15．7％）,弥漫型12例（19．3％）,病变检出率为100％。多薄层动态增强cT扫描三维重建技术表现为局限性胃壁增厚50例,广泛性胃壁增厚12例;其中早期胃癌10例,进展期胃癌52倒;局部和远处淋巴结转移24例;远处转移17例;本组CT定性诊断准确率为96％。结论薄层动态增强CT扫描三维重建技术能清晰显示胃壁及腔内、外病变,并直观反映胃癌大体形态及肿瘤的范围,在胃癌的定位、定性和定量诊断中发挥其独特的优势,是一种最具潜力的检查方法。     

人结肠癌CW-2干细胞获取方法研究

目的运用3种不同的方法分离纯化人结肠癌CW-2干细胞,并对其分离纯化效率进行比较,探讨获得癌干细胞的有效方法。方法采用单纯无血清悬浮培养、无血清悬浮培养联合化疗药物、流式细胞分选技术分别富集人结肠癌细胞株CW-2干细胞;然后运用流式细胞术、NOD—SCID小鼠致瘤实验和Transwell侵袭实验分析比较3种方法的富集效率。结果无血清悬浮培养细胞．无血清悬浮培养联合化疗药物处理细胞和流式细胞仪分选技术分选后细胞中具有结肠癌干细胞特性的CD44＋EPCAM_细胞分别为（59．39±4．55）％、（74.36±6．78）％、（86．43±8．43）％;3群细胞的成瘤能力和侵袭能力都存在显著统计学差异（P值〈0．05）：流式细胞分选技术分选后细胞〉无血清悬浮培养联合化疗药物处理细胞〉单纯无血清悬浮培养细胞。结论流式细胞分选技术富集癌干细胞的能力强于单纯无血清悬浮培养和无血清悬浮培养联合化疗药物,无血清悬浮培养联合化疗药物又强于单纯无血清悬浮培养。     

刚地弓形虫RH株感染对BALB／c小鼠情绪行为的影响

目的研究刚地弓形虫RH株感染对BALB／c小鼠学习记忆行为的影响及可能机制。方法将72只周龄、大小相近的雄性BALB／c小鼠采用随机数字表分为生理盐水对照组与不同数量（3×10^3／ml、3×10^4／ml、3×10^5／m1）弓形虫RH株感染组,每组18只。于感染第5周从每组各随机抽取6只分别进行高架十字迷宫实验、旷场实验及强迫游泳实验,观察各组小鼠在实验中情绪行为变化。并记录各项指标进行统计分析。结果在高架迷宫试验与旷场实验中,3×10^3／ml弓形虫感染组小鼠与生理盐水对照组小鼠比较,各项行为学指标无明显差异。在3×10^4／ml、3×10^5／ml弓形虫感染组小鼠与对照组小鼠比较出现明显降低（P〈0．05）．以3×10^5／ml感染组最为明显,其小鼠运动活力（0E＋cE）、进入开放臂次数比例（OE％）、开放臂停留时间比例（OT％）小鼠爬行总格数、中央格在总格数中比例分别为11．08±2．12、28．73±0．59％、25．62±2．33％、32．30±17．26、2．42±0．65％。在强迫游泳试验中,各弓形虫感染组小鼠游泳的静止时间均高于对照组小鼠（P〈0．05）,3×10^5／ml感染组静止时间最长,达226．6±1．9S。结论刚地弓形虫RH株感染可引起小鼠情绪行为改变,具有焦虑抑郁倾向。     

循环雌二醇及代谢物与绝经后乳腺癌的相关性

目的探讨循环雌二醇（E2）、4-羟雌二醇（4-OHE2）、2-甲氧雌二醇（2-MOE2）水平及4-羟雌二醇：2-甲氧雌二醇（4-OHE2：2-MOE2）比值与绝经后乳腺癌危险性的相关性。方法采用ELISA法检测绝经后34例乳腺癌、40例乳腺良性（乳腺纤维腺瘤与乳腺囊性增生病）疾病患者和20例正常对照循环雌二醇、4-羟雌二醇、2-甲氧雌二醇水平并计算4-羟雌二醇：2-甲氧雌二醇比值。结果乳腺癌组及乳腺良性疾病组循环2-甲氧雌二醇水平差异无统计学意义（P〉0．05）,但均较正常对照组低（P〈0．05）;乳腺癌组及乳腺良性疾病组4-羟雌二醇：2、甲氧雌二醇比值差异无统计学意义（P〉0．05）,但均较正常对照组升高（P〈0．05）。乳腺癌Ⅰ期～Ⅲ期各期循环雌二醇、4．羟雌二醇、2-甲氧雌二醇、4-羟雌二醇：2-甲氧雌二醇比值差异无统计学意义（P〉0．05）。4-羟雌二醇、2．甲氧雌二醇、4．羟雌二醇：2-甲氧雌二醇比值在乳腺癌ER（-）组与ER（＋）组间差异无统计学意义（P〉005）,但ER（＋）组雌二醇水平较ER（-）组明显升高（P〈0．05）。结论2．甲氧雌二醇与绝经后乳腺疾病负相关;4-羟雌二醇：2-甲氧雌二醇比值可能预示绝经后乳腺疾病危险。     

鸟苷酸交换因子DOCK1对心肌细胞存活的影响

目的 探讨鸟苷酸交换因子DOCK1对心肌细胞存活的影响.方法 用重组真核质粒pCXN2-Flag-hDOCK1(人DOCK1过表达质粒),pCXN2-Flag(空质粒)及空白试剂分别转染大鼠源H9C2心肌细胞,并给予缺氧/复氧(H/R)干预.将心肌细胞分为空白组,空质粒组,DOCK1过表达组,空白+-H/R组,空质粒+H/R组,DOCK1过表达+H/R组.用RT-PCR测定DOCK1mRNA水平.流式细胞术、MTT分别测定细胞的凋亡率和增殖率.结果 DOCK1过表达组(1.51 ±0.169)％,(2.49±0.442)％,(38.94±0.580)％,(P＜0.05)较空白组(-),(3.61±0.334)％,(20.64±0.720)％,(P＜0.05)和空质粒组(-),(3.66±0.373)％,(22.29±0.838)％,(P＜0.05),DOCK1过表达+H/R组1.03±0.171,(5.38±0.431)％,(17.33±0.343)％,(P＜0.05)较空白+H/R组[(-),(2.49±0.442)％,(7.95±0.322)％,(P＜0.05)和空质粒+H/R组(-),(10.32±0.388)％,(7.92±0.351％,(P＜0.05),人的DOCK1RNA分别显著增加、心肌细胞凋亡率分别显著降低及增殖率分别显著增加.较DOCK1过表达组.DOCK1过表达+H/R组人的DOCK1 mRNA显著降低.较空白组(0.64±0.145),(P＜0.05)、空质粒组(0.60±0.182),(P＜0.05)及DOCK1过表达组(0.60±0.182),(P＜0.05),空白+H/R组(0.30±0.115),(P＜0.05)、空质粒+H/R组(0.36±0.101),(P＜0.05)及DOCK1过表达+H/R组(1.03±0.171),(P＜0.05)大鼠的DOCK1 mRNA分别显著降低、心肌细胞凋亡率分别显著增加及增殖率分别显著降低.结论 DOCK1可抑制H9C2心肌细胞的凋亡,促进H9C2心肌细胞的增殖、存活；且DOCK1可抑制H/R诱导的H9C2心肌细胞凋亡增加及增殖、存活的降低.     

Dual effect of cannabinoid CB<Subscript>1</Subscript> receptor stimulation on a vanilloid VR1 receptor-mediated response

Cannabinoid CB1 receptors and vanilloid VR1 receptors are co-localized to some extent in sensory neurons of the spinal cord and dorsal root ganglia. In this study, we over-expressed both receptor types in human embryonic kidney (HEK)-293 cells and investigated the effect of the CB1 agonist HU-210 on the VR1-mediated increase in intracellular Ca2+ ([Ca2+]i), a well-known response of the prototypical VR1 agonist capsaicin. After a 5-min pre-treatment, HU-210 (0.1 microM) significantly enhanced the effect of several concentrations of capsaicin on [Ca2+]i in HEK-293 cells over-expressing both rat CB1 and human VR1 (CB1-VR1-HEK cells), but not in cells over-expressing only human VR1 (VR1-HEK cells). This effect was blocked by the CB1 receptor antagonist SR141716A (0.5 microM), and by phosphoinositide-3-kinase and phospholipase C inhibitors. The endogenous agonist of CB1 and VR1 receptors, anandamide, was more efficacious in inducing a VR1-mediated stimulation of [Ca2+]i in CB1-VR1-HEK cells than in VR1-HEK cells, and part of its effect on the former cells was blocked by SR141716A (0.5 microM). Pre-treatment of CB1-VR1-HEK cells with forskolin, an adenylate cyclase activator, enhanced the capsaicin effect on [Ca2+]i. HU-210, which in the same cells inhibits forskolin-induced enhancement of cAMP levels, blocked the stimulatory effect of forskolin on capsaicin. Our data suggest that in cells co-expressing both CB1 and VR1 receptors, pre-treatment with CB1 agonists inhibits or stimulates VR1 gating by capsaicin depending on whether or not cAMP-mediated signalling has been concomitantly activated.     

Distinct features of recombinant rat vanilloid receptor-1 expressed in various expression systems

In this study, we expressed rat vanilloid receptor 1 (VR1) in various heterologous expression systems using different VR1-encoding vectors, and examined how the VR1 agonists capsaicin and resiniferatoxin affected intracellular calcium. Our results clearly show that the magnitude and kinetics of response as well as the extent of tachyphylaxis differ markedly between systems. Using green fluorescent protein-tagged VR1, we show that much of the VR1 is localized to intracellular membranes. Consistent with this localization, VR1 agonists are able to liberate calcium from intracellular stores in the absence of extracellular calcium. As with other parameters of response, the three expression systems differ in the degree to which, in the absence of extracellular calcium, capsaicin and resiniferatoxin can liberate calcium from the intracellular stores. Our findings emphasize the influence of the expression system on characteristics of the response of VR1 to its ligands and the need for caution in extrapolating such results to other settings.Received 16 June 2003; received after revision 26 July 2003; accepted 31 July 2003     

Anandamide induces cell death independently of cannabinoid receptors or vanilloid receptor 1: possible involvement of lipid rafts

Anandamide triggers various cellular activities by binding to cannabinoid (CB1/CB2) receptors or vanilloid receptor 1 (VR1). However, the role of these receptors in anandamide-induced apoptosis remains largely unknown. Here, we show that SR141716A, a specific inhibitor of cannabinoid receptor (CB1-R), did not block anandamide-induced cell death in endogenously CB1-R expressing cells. In addition, CB1-R-lacking Chinese hamster ovary (CHO) cells underwent cell death after anandamide treatment. SR144528, a specific inhibitor of CB2-R also failed to block anandamide-induced cell death in HL-60 cells. Capsazepine, a specific antagonist of VR1 could not prevent anandamide-induced cell death in constitutively and endogenously VR1 expressing PC12 cells. Moreover, anandamide noticeably triggered cell death in VR1-lacking human embryonic kidney (HEK) cells. In contrast, methyl-beta cyclodextrin (MCD), a membrane cholesterol depletor, completely blocked anandamide-induced cell death in a variety of cells, including PC12, C6, Neuro-2a, CHO, HEK, SMC, Jurkat and HL-60 cells. MCD also blocked anandamide-induced superoxide generation, phosphatidyl serine exposure and p38 MAPK/JNK activation. Thus, our data imply a novel role for of membrane lipid rafts in anandamide-induced cell death.     

运用磁珠分选法建立免疫细胞共培养体系

目的以相互作用的几种免疫细胞上共同表达的抗原物质为标志,运用磁珠分选技术,从慢性乙型肝炎患者外周血单个核细胞中同时获得并建立体外多细胞共培养体系,体外观察细胞之间的相互作用.方法采用梯度离心法分离30例慢性乙肝病毒感染者外周血单个核细胞(PBMC),采用免疫磁珠分选法分离获得 CXCR5+细胞,通过流式细胞仪检测 CXCR5+细胞纯度和组成,并将 CXCR5+细胞进行体外培养.在分离得到的 CXCR5+细胞的基础上,将 CXCR5+细胞分为3组,分别为对照组:不加任何刺激;实验组一:加入 IL 21刺激培养,实验组二:与人肝癌细胞系(HepG2)2215细胞混合培养.体外培养一周后,流式细胞仪检测实验组与对照组细胞 B细胞数量变化,ELISA测定培养体系中上清液 HBe抗体的含量.结果1)用流式细胞仪鉴定 CXCR5+细胞的纯度和构成:CXCR5+细胞纯度为85.5±5.8%,其中 Tfh细胞占23.8±7.4%,B细胞占35.6±7.6%;2)培养7天后,IL 21刺激组 B细胞百分率为47.2±1.8%,与(HepG2)2215混合培养组 B细胞百分率为40.2±3.5%,空白对照组 B细胞百分率为36.6±7.5%,IL 21刺激组与对照组,(HepG2)2215混合培养组与对照组 B细胞百分率均有显著差异(p<0.05);3)ELISA检测培养液上清 HBeAb定量,IL 21刺激组为0.668±0.094pg/ml,空白对照组为0.378±0.088pg/ml,两组有显著差异(p<0.05).结论使用间接免疫磁珠分离法可成功建立 CXCR5+B淋巴细胞和 Tfh细胞的共培养体系,为进一步体外研究细胞之间相互关系奠定基础     

Micromechanical analysis of interaction energy for SMA reinforced composite

The energy of the interaction between the matrix and the inclusions in shape memory alloy (SMA) reinforced composite is one of the most important and complicated parts in thermodynamic constitutive theory.In this paper,the interaction energy is derived based on the classical theory of micromechanics and the thermodynamic theory.The SMA composite is treated as three phases,namely the austenitic phase,the martensite phase and the matrix phase.The interaction among the three phases is analyzed in a way close t...     

CBP-mediated SMN acetylation modulates Cajal body biogenesis and the cytoplasmic targeting of SMN

The survival of motor neuron (SMN) protein plays an essential role in the biogenesis of spliceosomal snRNPs and the molecular assembly of Cajal bodies (CBs). Deletion of or mutations in the SMN1 gene cause spinal muscular atrophy (SMA) with degeneration and loss of motor neurons. Reduced SMN levels in SMA lead to deficient snRNP biogenesis with consequent splicing pathology. Here, we demonstrate that SMN is a novel and specific target of the acetyltransferase CBP (CREB-binding protein). Furthermore, we identify lysine (K) 119 as the main acetylation site in SMN. Importantly, SMN acetylation enhances its cytoplasmic localization, causes depletion of CBs, and reduces the accumulation of snRNPs in nuclear speckles. In contrast, the acetylation-deficient SMNK119R mutant promotes formation of CBs and a novel category of promyelocytic leukemia (PML) bodies enriched in this protein. Acetylation increases the half-life of SMN protein, reduces its cytoplasmic diffusion rate and modifies its interactome. Hence, SMN acetylation leads to its dysfunction, which explains the ineffectiveness of HDAC (histone deacetylases) inhibitors in SMA therapy despite their potential to increase SMN levels.