核糖核酸的酶消化方法直接顺序分析

应用硷基特异性核酸内切酶进行RNA的顺序分析。以~(32)P标记RNA分子的5′或3′末端的四种硷基能被酶试剂部分裂解。放射性的水解产物经胶电泳按其大小而分离。放射自显影后它的核苷酸顺序从带谱上可以直接读出。核糖核酸酶(RNasc)T_1(G特异性)、U_2(A特异性)、Phy M(U+A特异性)和B.cereus(C+U特异性)是最常用于顺序RNA的四种酶。     

Synergistic effects of Nell-1 and BMP-2 on the osteogenic differentiation of myoblasts.

Osteogenesis is synergistically enhanced by the combined effect of complimentary factors. This study showed that Nell-1 and BMP-2 synergistically enhanced osteogenic differentiation of myoblasts and phosphorylated the JNK MAPK pathway. The findings are important because of the osteochondral specificity of Nell-1 signaling and the potential therapeutic effects of coordinated BMP-2 and Nell-1 delivery. INTRODUCTION: BMPs play an important role in the migration and proliferation of mesenchymal cells and have a unique ability to alter the differentiation of mesenchymal cells toward chondrogenic and osteogenic lineages. Signaling upstream of Cbfa1/Runx2, BMPs effects are not limited to cells of the osteoblast lineage. Thus, additional osteoblast-specific factors that could synergize with BMP-2 would be advantageous for bone regeneration procedures. NELL-1 (NEL-like molecule-1; NEL [a protein strongly expressed in neural tissue encoding epidermal growth factor like domain]) is a novel growth factor believed to preferentially target cells committed to the osteochondral lineage. MATERIALS AND METHODS: C2C12 myoblasts were transduced with AdLacZ, AdNell-1, AdBMP-2, or AdNell-1+AdBMP-2 overexpression viruses. Effects were studied by cell morphology, alkaline phosphatase activity, osteopontin production, and MAPK signaling. Additionally, in a nude mouse model, viruses were injected into leg muscles, and new bone formation was examined after 2 and 8 wk. RESULTS: C2C12 myoblasts co-transduced with AdNell-1+AdBMP-2 showed a synergistic effect on osteogenic differentiation as detected by alkaline phosphatase activity and osteopontin production. Nell-1 stimulation on AdNell-1 + AdBMP-2 preconditioned C2C12 cells revealed significant activation of the non-BMP-2 associated c-Jun N-terminal kinase (JNK) MAPK signaling pathway, but not the p38 or extracellular signal-regulated kinase (ERK1/2) MAPK pathways. Importantly Nell-1 alone did not induce osteogenic differentiation of myoblasts. In a nude mouse model, injection of AdNell-1 alone stimulated no bone formation within muscle; however, injection of AdNell-1+AdBMP-2 stimulated a synergistic increase in bone formation compared with AdBMP-2 alone. CONCLUSIONS: These findings are important because of the confirmed osteochondral specificity of Nell-1 signaling and the potential therapeutic effects of enhanced BMP-2 action with coordinated Nell-1 delivery.     

股骨头骨坏死样本空间结构的Micro-CT评估

目的以股骨头骨坏死样本的Micro-CT断层图像为基础，对其进行空间结构评估。方法2003年11月～2005年6月，收集股骨头骨坏死患者行全髋关节置换术时取出的股骨头样本6个，对样本进行Micro-CT断层扫描，获取股骨头骨坏死样本的计算机三维图像，图像空间分辨率为36μm×36μm×36μm。手工选取兴趣区，随后采用骨体积分数（BV／TV）、骨小梁间隙（Tb．Sp）、骨小梁厚度（Tb，Th）、骨小梁数目（Tb．N）、骨表面积体积比（BS／BV）、结构模型指数（SMI）、骨小梁模型因子（Tb．Pf）等三维空间参数分别对股骨头骨坏死标本的正常区域、硬化带和塌陷区进行评价。结果晚期股骨头骨坏死硬化区和塌陷区的骨小梁空间结构明显改变：硬化区Bv／Tv明显增加，Tb．Th明显增厚，Tb，Sp变窄，SMI与正常区域的骨小梁无差别；而塌陷区Bv／Tv明显减少，BS／BV增大，Tb．N和Tb．Pf增加，Tb，Th改变不明显。结论Micro-CT作为一种新的检测手段，能够在不损伤样本的条件下快速获取股骨头骨坏死样本断层图像，具有精度高、检测快、可进行三维重建分析的优点。晚期股骨头骨坏死不同区域的骨三维结构呈现出不同的空间结构特征。     

应用ROC曲线评价DR后处理技术对胸部影像的影响

目的:运用ROC曲线,探讨DR胸部摄影最佳的影像后处理技术。方法:使用DR系统设置的正常胸部摄影条件,对胸部体模与专用于X线摄影的ROC曲线评价体模(TRG)进行组合并曝光。分析未使用后处理技术及使用不同后处理技术,对DR胸部摄影胸片影像的影响。结果:使用单纯噪声消除技术、噪声消除加图像反转及噪声消除加全屏放大三种后处理技术所得的ROC曲线下面积Az值依次为0.85、0.825及0.925,均明显大于未使用后处理技术的面积Az值0.6;单纯噪声消除技术获得的ROC曲线下面积略大于噪声消除加图像反转技术的面积;噪声消除加全屏放大技术获得的ROC曲线下面积Az值最大。结论:应对胸部原始影像实施后处理调节。单纯噪声消除技术可明显增加影像的信息量,若在噪声消除基础上实施全屏放大技术,可获得DR胸部摄影的最大影像信息量。     

Effects of cerulenin on the endogenous fatty acid synthetic activity in squamous cell carcinoma of the oral cavity.

PURPOSES: Inhibition of cerulenin on the endogenous fatty acid synthetic activities of oral squamous cell carcinoma (OSCC) and normal oral mucosa was assayed. METHODS: Squamous cell carcinoma and normal oral mucosa were collected fresh from surgical specimens. The collected tissues were minced in RPMI 1640 and divided into 3 groups: cerulenin treated, dimethylsulfoxide treated, and control. The tissues were incubated in [1(2)-(14)C]acetic acid, sodium salt for the last 2.5 hours of the treatment at 37 degrees C in 5% CO(2). After labeling, total lipids were extracted and counted for (14)C by scintillation counting. RESULTS: Endogenous fatty acid synthetic activities of oral squamous cell caranoma in the cerulenin-treated group decreased by 19% at 1 hour, 64% at 2 hours, and 87% at 4 hours; remained nearly unchanged in the dimethylsulfoxide-treated group; and increased slightly in the control group. The oral mucosa tissues were only mildly affected by cerulenin in fatty acid synthesis. CONCLUSIONS: Cerulenin significantly inhibits fatty acid synthetic activity in squamous cell carcinoma and only mildly affected the oral mucosa, indicating that the fatty acid synthetic pathway may be exploited as a target for developing anticancer drugs.     

神经干细胞超顺磁性氧化铁纳米粒子标记和体内MRI示踪

神经干细胞（neural stem cell，NSC）是当今医学的前沿课题，困扰其临床应用的一个难题是如何对移植进入体内的神经干细胞进行监测，观察其存活和迁徙的情况。在中枢神经系统的不同病理、生理条件下，移植的神经干细胞有着不同的迁徙和分化能力，对干细胞这一特性的研究目前多为在移植后的一定时间内处死实验动物，对神经组织进行切片。而这些侵袭性的方法，不能对神经干细胞在同一动物体内的迁徙、增殖情况进行动态的观察，故需要非侵袭的手段来对移植入体内的神经干细胞进行监测。     

细胞内游离Ca2+在发育中大鼠皮层神经元无镁诱导惊厥性损伤中的作用

目的: 观察细胞内游离Ca2+([Ca2+]i)在培养的不同发育阶段皮层神经元无镁诱导惊厥性损伤中的作用,探讨惊厥性脑损伤年龄依赖性的可能机制.方法:体外培养6 d、17 d的胚胎大鼠皮层神经元用无镁细胞外液处理3 h,或于无镁处理前用NMDA(N-甲基-D-门冬氨酸)受体拮抗剂或Ca2+通道阻滞剂预处理,用MTT代谢率测定的方法检测神经元损伤,以Fluo-3作标记用激光共聚焦显微镜扫描的方法检测[Ca2+]i.结果:体外培养6 d、17 d的神经元单纯无镁组MTT代谢率较同期对照组降低.应用MK-801 10 μmol*L-1、AP-5 50 μmol*L-1、尼莫地平10 μmol*L-1预处理后再给无镁处理,培养6 d、17 d的神经元MTT代谢率均不同程度高于同期单纯无镁组.培养6 d、17 d的神经元相对荧光强度之间差异有显著性,两者与基线荧光强度比较差异亦有显著性.应用上述各种拮抗剂后,[Ca2+]i改变的峰值均明显低于同期单纯无镁组.结论: 在体外不同发育阶段的神经元,短暂无镁处理诱导惊厥样放电所引起的神经元线粒体功能损伤以及[Ca2+]i改变程度不同.这种[Ca2+]i改变的年龄依赖性可能是惊厥导致神经元损伤的年龄依赖性的机制之一.NMDA受体-Ca2+通道激活是导致这种[Ca2+]i改变及神经元损伤的关键环节.     

定量组织速度成像对右室起搏患者左室收缩运动的研究

目的　应用定量组织速度成像 (QTVI)评价右室心尖起搏 (RVAP)VVI型对左心收缩功能的影响。方法　应用GEVivid 7彩色多普勒超声显像仪对 2 0例RVAP患者和 2 0例正常人的心尖四腔切面的室间隔和左室外侧壁速度和位移曲线进行观察 ,测量心电图Q波分别至室间隔和左室外侧壁收缩期峰速度的时间 ,并除以R R间期进行校正。结果　QTVI显示右室起搏器置入者的室间隔与左室外侧壁速度曲线的收缩期S波非同步出现。Q波至室间隔收缩期峰速度的时间短于Q波至左室外侧壁收缩期峰速度的时间 ,两者分别为 ( 0 .12± 0 .0 2 )s和 ( 0 .14± 0 .0 2 )s,P <0 .0 5。结论　右室起搏后早期的左室整体收缩功能虽未见明显下降 ,但QTVI可以发现室间隔与左室壁收缩明显的不协调 ,可作为早期分析左室收缩运动的定量方法。     

闭合性肾损伤的诊断和治疗

目的探讨闭合性肾损伤的诊断与治疗方法。方法对205例闭合性肾损伤的临床资料进行回顾性分析。结果205例闭合性肾损伤中,肾挫伤113例,肾裂伤67例,肾碎裂伤31例,肾蒂血管伤12例,合并伤81例(39.5%)。血尿18例(91.7%),血尿是主要的临床症状。B 超检查199例,其中165例(82.9%)有异常表现;CT 检查80例,均显示肾损伤(100%)。保守治疗153例(74.6%);手术治疗52例,死亡5例(3.5%);超选择性肾动脉栓塞术29例。结论 B 超和 CT 检查是诊断肾损伤的有效方法。严格掌握手术时机是治疗闭合性肾损伤的关键。