Wallerian degeneration in a patient with Schilder disease: MR imaging demonstration

Wallerian degeneration in the corticospinal tract was demonstrated by magnetic resonance (MR) imaging in a patient with Schilder disease. The histochemical stages of myelin breakdown that allow its demonstration by MR imaging are reviewed.     

用细口径反相柱HPLC分离多种前列腺素

HPLC与放射性同位素联用,可同时分离鉴定多种花生四烯酸代谢产物。目前国外一般是使用普通口径(4.6 mm)的反相柱,以30%左右乙腈为流动相,流速为1～3 ml/min。这种HPLC方法乙腈用量较大,费用太贵,不适合国内实验室常规操作。据报道,细口径     

Childhood scoliosis: MR imaging

The spinal cords of 28 scoliosis patients between the ages of 1 month and 17 years were examined with magnetic resonance (MR) imaging. Complete visualization was obtained in all cases. In 15 patients (53%) neuropathologic abnormalities demonstrated by MR imaging significantly affected their clinical course, including tethered cords (n = 7), syringomyelia (n = 5), Arnold-Chiari I malformation (n = 4), spinal cord tumors (n = 2), Arnold-Chiari II malformation (n = 3), and diastematomyelia (n = 1). The advantages of MR imaging in the evaluation of the scoliotic spine in children include a high sensitivity for the occult conditions associated with scoliosis, good anatomic demonstration of the cord, and absence of bone artifacts. MR imaging is recommended as a primary imaging modality in scoliosis, following conventional radiography.     

Monoclonal antibody to human high-molecular-weight kininogen recognizes its prekallikrein binding site and inhibits its coagulant activity

We developed a mouse monoclonal antibody (MoAb 115-21) to human high- molecular-weight kininogen (HK) that recognizes its prekallikrein binding site (residues 565 through 595 of HK). The corresponding synthesized 31-amino acid peptide (peptide IV) was recently shown to retain native HK's prekallikrein binding property. The same peptide bound factor XI also, although less avidly. Our MoAb recognizes purified HK, peptide IV, and the light chain moiety of HK (where the peptide IV resides), as shown by enzyme-linked immunosorbent assay (ELISA) and Western blotting experiments. The apparent dissociation constant for the HK and MoAb 115-21 interaction was 2.2 nmol/L. It does not recognize low-molecular-weight kininogen (LK) with which HK shares its heavy chain moiety or any antigens in human plasma congenitally deficient in kininogens. The binding of MoAb 115-21 to purified light chain of HK was competitively inhibited by peptide IV. In addition, the antibody inhibits HK-dependent clotting activity of normal human plasma and dextran sulfate-mediated activation of prekallikrein in plasma and retards cleavage of HK in normal plasma after contact activation with dextran sulfate. Also, purified Fab fragments of MoAb 115-21 inhibited the HK-dependent coagulant activity and dextran sulfate-mediated prekallikrein activation in normal plasma. Since the kd for HK-MoAb 115- 21 interaction is ten times lower than that of HK-prekallikrein, our data suggest that binding of MoAb 115-21 to HK's peptide IV site increases the free prekallikrein concentration in plasma and thus results in the decreased efficiency of factor XIIa-mediated activation of prekallikrein. Decreased levels of kallikrein thus formed may be responsible for the inhibition of HK-dependent clotting activity and the decrease in rate and extent of HK cleavage in normal plasma on contact activation with dextran sulfate. MoAb 115-21 may thus prove very useful, especially with its high affinity for HK, in further delineation of the role of HK and prekallikrein in contact activation and kinin-related human pathology.     

力竭运动大鼠心室肌蛋白质组表达特征

目的:采用蛋白质组学技术,建立安静和递增运动负荷训练后力竭大鼠心室肌蛋白质组的差异性表达谱,初步筛选出心室肌对力竭运动产生反应的目标蛋白质。方法:实验于2007-03在湖南师范大学生命科学学院蛋白质化学与蛋白质组学国家教育部重点实验室和省级运动人体科学实验室完成。①实验分组:10只SD雄性大鼠随机分为对照组和运动组,每组5只。②实验方法:运动组经过7周的大强度递增运动负荷训练后(最后一次力竭),对两组心室肌组织的全蛋白进行双向凝胶电泳分离。结果:经图像分析,在运动组的电泳图谱上共展现蛋白质点(338±17)个,对照组展现蛋白质点(352±17)个。运动后差异表达的蛋白质点共有99个。对其中差异表达的9个蛋白质点进行质谱鉴定,共鉴定出7个蛋白质,Stress-70protein,NADH-ubiquinone oxidoreductase Mr75000subnunit,Long-chain specific acyl-CoA dehydrogenase,Tropomyosin-1alphachain在运动后"缺失",Nitrilase family,member2在运动后表达上调在5倍以上,一个相对分子质量为21000的未知蛋白在运动后表达下调在5倍以上,另外有两个点经鉴定均为Myosin-6,在运动后表达量相反。这些蛋白质属于收缩蛋白、能量代谢酶、分子伴侣等。结论:递增运动负荷训练后力竭时,大鼠心室肌蛋白质组明显地发生了反应。运动后"缺失"和下调的蛋白质点与心肌收缩的调控和能量代谢的方式转变以及细胞的应激反应有关,其中,成功筛选出6种在运动医学领域尚未涉足的、具有运动应激特点的目标蛋白质。     

急性心肌梗死后外周血单个核细胞表面CD147与基质金属蛋白酶-9 mRNA表达的相关性

目的：基质金属蛋白酶在急性心肌梗死后的心室重构中起着重要作用，但其调节机制目前尚未明确。实验拟通过动物模型的建立及体外细胞培养，观察急性心肌梗死后单个核细胞表面CD147与心肌成纤维细胞基质金属蛋白酶-9 mRNA表达的关系。 方法：实验于2006—08／2007-06在河北省人民医院临床实验中心完成。实验材料：SD大鼠及SD仔鼠（出生1～3d）购自河北医科大学试验动物中心。实验过程中对动物处置符合动物伦理学标准。实验方法：①将30只大鼠随机分为急性心肌梗死组（n=15）和假手术组（n=15），假手术组只过线不结扎。流式细胞分析法检测大鼠术后24h外周血单个核细胞表面CD147表达。②选择SD仔鼠制备心肌成纤维细胞。将单个核细胞与心肌成纤维细胞以细胞数0．5：1，1：1，2：1混合培养24h后，半定量反转录一聚合酶联反应法检测基质金属蛋白酶-9 mRNA表达。当单核细胞与心肌成纤维细胞2：1混合时，加入CD147单克隆抗体1，2，4μL/L，培养24h后检测基质金属蛋白酶-9 mRNA表达。 结果：①急性心肌梗死后外周血单个核细胞表面CD147表达明显增加。②单个核细胞与心肌成纤维细胞混合培养，随着单个核细胞比例的增加，心肌成纤维细胞基质金属蛋白酶-9 mRNA表达增加。③在单个核细胞与心肌成纤维细胞2：1混合培养体系中，随着加入CD147单克隆抗体浓度的增加，基质金属蛋白酶-9 mRNA生成减少。 结论：急性心肌梗死后单个核细胞表面CD147表达明显增加，对心肌成纤维细胞基质金属蛋白酶-9生成起上游调节作用。     

多普勒超声诊断移植肾排斥反应与病理检查及肾功能指标的对比分析

目的：以病理活检结果为金标准，评估多普勒超声检查对移植肾排斥反应的诊断价值。方法：选择2003—01/2006—12在中国医科大学附属第一医院器官移植科行肾移植并在术后行超声检查的患者176例，均知情同意。①实验分组：根据术后移植肾功能分为2组，移植肾功能不良组78例，其中30例次行病理活检；移植肾功能正常组98例。②实验方法及评估：对患者移植肾行多普勒超声检查，参数选择峰收缩期流速、平均舒张期流速、阻力指数及血管显示率。血管显示率的评估标准（0～5级）：0级为肾动脉及其远侧血管未显示；5级为肾各级血管均显示良好。以病理活检结果为金标准，分别选取阻力指数=0．7，0．75，0．8，0．85为诊断界值进行诊断试验。结果：169例患者进入结果分析，脱落7例。①峰收缩期流速、平均舒张期流速不呈正态分布，无法作为肾功能评价指标。30例次病理活检中共有28例次被确诊为排斥反应，急性排斥反应15例次，慢性排斥反应13例次。②肾功能正常组患者中血管显示率5级者占63．30％，4级者占36．73％。肾功能不良组患者中血管显示率4级者占41．03％，3级者占46．15％，2级者占10．30％，1级者占2．60％。③移植肾功能不良组患者阻力指数显著高于移植肾功能正常组（P〈0．01）。移植肾功能不良组患者移植肾功能恢复后阻力指数显著低于移植肾尚未恢复时（P〈0．01），其中99％以上的患者△（阻力指数）≥0.20。④界值阻力指数=0．75的诊断试验的敏感性、特异性和准确性最高，均达到100％。结论：当移植肾血管阻力指数升高至0．75以上，特别是同一患者自身对照升高超过0.2以上和或血管显示率低于4级，结合临床表现和生化结果，提示可能出现移植肾排斥反应。     

经冠状动脉移植自体骨髓单个核细胞和间充质干细胞对急性心肌梗死模型心功能的改善

目的:为保护濒死心肌提供机会窗口,对比观察经冠脉移植自体骨髓单个核细胞或间充质干细胞后,实验性急性心肌梗死动物心功能变化及心肌组织核转录因子кB、心肌细胞凋亡情况。方法:实验于2005-03/2006-11在河北省人民医院实验中心完成。选用24只雄性冀中白猪,随机数字表法分为4组:正常对照组、模型组、单个核细胞组、间充质干细胞组,6只/组。①24只猪均以盐酸氯胺酮200mg臀部肌肉注射麻醉后,分别于各自右侧股骨抽取骨髓20mL,采用Fercoll法分离获得骨髓单个核细胞,加入胶体金溶液,培养12～16h待用。分离过程中取出含有骨髓单个核细胞成分的细胞层,常规培养传代,每3d换液1次,贴壁生长细胞即为骨髓间充质干细胞,加入胶体金溶液,培养24h待用。②除正常对照组外,其余各组均经导管球囊封闭第一对角支以远的前降支,复制猪急性心肌梗死模型。单个核细胞组、间充质干细胞组均于造模后立即开通前降支,分别经球囊注入预先分离的骨髓单个核细胞6×108个、间充质干细胞6×108个。模型组造模后于梗死1h开通前降支,经球囊注入磷酸盐缓冲液10mL。③各组分别于术前及术后4周经心脏超声检测心功能,取材行病理学检查、心肌组织核转录因子кB的免疫组织化学检测及心肌细胞凋亡检测。结果:24只雄性白猪均进入结果分析。①心功能变化:术前各组左心室收缩末内径、左心室舒张末内径、左心室射血分数、短轴缩短率基本相似。移植术后4周,正常对照组、单个核细胞组、间充质干细胞组左心室舒张末内径均明显低于模型组(F=4.68,P=0.01),左心室射血分数及短轴缩短率均明显高于模型组(F=5.14,P=0.01;F=3.32,P=0.04),各组左心室收缩末内径差异无显著性意义(F=1.64,P=0.21)。②心肌组织病理学改变:电镜下单个核细胞组、间充质干细胞组在梗死边缘区可见有胶体金颗粒的不成熟的心肌细胞,胞质中散在肌丝结构,肌丝排列紊乱不规则。③心肌组织核转录因子кB阳性率表达:与模型组比较,单个核细胞组、间充质干细胞组的梗死边缘区核转录因子кB阳性率明显降低(F=25.59,P=0.0001);正常心肌区核转录因子кB阳性率亦明显降低(F=18.20,P=0.0001)。④心肌细胞凋亡检测结果:与模型组比较,单个核细胞组、间充质干细胞组在心肌梗死区细胞凋亡率均明显降低(F=6.63,P=0.0027),梗死边缘区细胞凋亡率亦明显降低(F=36.07,P=0.0001)。正常心肌区单个核细胞组细胞凋亡率与模型组基本相似(F=9.69,P=0.004),但间充质干细胞组有所降低。⑤心功能与心肌细胞凋亡及心肌组织NF-кB的相关性:急性心肌梗死4周时,左心室射血分数与心肌细胞凋亡、心肌组织核转录因子кB均呈负相关(r=0.613,P=0.001;r=-0.437,P=0.033)。心肌细胞凋亡与心肌组织核转录因子кB呈正相关(r=0.672,P=0.002)。结论:经冠脉移植骨髓单个核细胞和间充质干细胞均可改善实验性急性心肌梗死动物的心功能,与梗死边缘区核转录因子кB表达降低及心肌细胞凋亡减少有关。骨髓单个核细胞移植的促血管增生作用优于间充质干细胞移植。     

自体骨髓间充质干细胞移植后心肌缝隙连接蛋白43表达与室性心动过速发生的关系

目的:缝隙连接蛋白43对维持心肌细胞的连接通讯功能、电信号传导和正常的节律性收缩起重要作用,其表达和分布的异常是多种室性心律失常的解剖学基础,建立小型猪急性心肌梗死模型.观察自体骨髓间充质干细胞移植后室性心动过速的发生及心肌缝隙连接蛋白43的表达方法:实验于2006-01/2007-01在河北省人民医院导管室完成。①材料:选取8～12月龄小型猪22头,由河北医科大学实验动物中心提供,体质量20～30 kg,随机数字表法分为细胞移植组12头、模型对照组10头.实验过程中对动物的处置符合动物伦理学际准②实验方法:无菌条件下抽取猪双侧股骨骨髓20 mL,percoll法 贴壁法分离培养骨髓间充质干细胞,待细胞生长达75%融合时用胰酶消化传代。将传至第2代细胞加入终浓度为10μmol/L的5-氮胞苷进行诱导,用胶体金标记12 h后继续培养20 d用于移植:两组小型猪均采用球囊堵闭法建立急性心肌梗死模型.心电图监测示相关至少2个导联ST段抬高大于0.2 mV、术后血肌钙蛋白和肌酸磷酸激酶同工酶升高超过正常的两倍为建模成功标准:细胞移植组于造模成功后经OTW球囊于第一对角支远端1 cm处再次阻断血流,注入经胶体金标记的10×10~7个骨髓间充质干细胞。③实验评估:于细胞移植后2 h及4周行电生理程序刺激.观察室性心动过速的发生情况。末次电生理检查后、采用免疫组化染色法检测心肌缝隙连接蛋白43的表达.计算其积分吸光度值。结果:①模型建立指标检测:与术前比较.造模后所有小型猪血肌钙蛋白含量和肌酸磷酸激酶同工酶活性均增高,峰值浓度分别为(21.3±3.6)μg/L和(178.3×41.4)IU/L,术中心电图ST段平均抬高(10.67±1.43)mm.证明急性心肌梗死模型成功建立。②骨髓间充质干细胞移植后室性心动过速的发生情况:与模型对照组诱发出室性心动过速的动物数量比较,术后2 h细胞移植组无明显变化(X~2=0.201,P=0.650),术后4周细胞移植组明显降低(X~2=4.455.P=0.035)。②骨髓间充质干细胞移植后梗死心肌缝隙连接蛋白43的表达:术后4周移植到梗死心肌的骨髓间充质干细胞与宿主心肌生长为一体,移植部位颜色变黑,苏木精-伊红染色示移植细胞的胞浆呈紫红色。细胞移植组心肌梗死区缝隙连接蛋白43积分吸光度值明显高于模型对照组(t=16,82.P=0.00),细胞移植组中未发生室性心动过速小型猪的梗死心肌缝隙连接蛋白43的表达明显高于发生室性心动过速小型猪(t=5.06,P=0.00)。结论:自体骨髓间充质干细胞移植可促进急性心肌梗死猪心肌缝隙连接蛋白43的表达,其表达程度可能与急性心肌梗死室性心动过速的发生有关。     

PGE1 accelerates thrombolysis by tissue plasminogen activator

Platelets are an active element in the generation of thrombus and may influence rates of thrombolysis during the administration of plasminogen activators. To assess the potential importance of platelet aggregation in the thrombolytic response to plasminogen activators, we measured rates of thrombolysis induced by tissue plasminogen activator in the presence and absence of a coinfusion of prostaglandin E1 in a rabbit jugular vein model of thrombosis. Rates of lysis were quantified by measuring the half-time for lysis of the thrombus. At all concentrations of tissue plasminogen activator used, prostaglandin E1 markedly reduced the half-time for clot lysis and enhanced somewhat the overall extent of thrombolysis, without affecting significantly either the degree of fibrinogen depletion or the animals' mean arterial pressures. These effects on thrombolytic efficacy were accompanied by ex vivo evidence of platelet inhibition. These data suggest that the antiplatelet prostaglandin E1 may be a very useful adjunctive agent in thrombolytic therapy primarily by virtue of the significant improvement in the rate of thrombolysis that its use affords.