骨髓间充质干细胞在丝素蛋白/骨形态发生蛋白中的生物学行为观察

目的 观察丝素蛋白与牛骨形态发生蛋白的载体系统与骨髓间充质干细胞的体外相容性.方法 将多孔丝素蛋白修剪成20×10×5 mm长方体状物,置入6孔板中;将粉状bBMP用PBS液溶解制成2mg/ml混悬液,0.5ml无菌移液管滴定于多孔丝素蛋白上(0.5毫升/个).骨髓间充质干细胞以107/ml接种到载体系统上,复合4小时后加入生长液继续体外培养,每1、4、8天时进行碱性磷酸酶、矿化结节染色观察以及扫描电镜和激光共聚焦显微镜观察;另外同时设立单纯丝素蛋白与细胞复合培养对照组进行观察.结果 倒置显微镜、扫描电镜和共聚焦显微镜下观察到细胞在两种材料上生长形态、活性正常,体外培养24小时时细胞已贴附材料上,呈匍匐状伸展,以后逐渐融合生长,8天时大量细胞成片状攀附在材料表面,并分泌有大量的网状细胞外基质,实验组可见结节状基质生成;碱性磷酸酶、矿化结节染色观察发现实验组细胞碱性磷酸酶和矿化结节染色阳性反应,对照组阴性.结论 丝素蛋白是一种良好细胞外基质材料,也是一种良好生长因子的缓释载体.     

RGD多肽接枝聚复合导管桥接神经缺损的实验研究

Objective To investigate the effect of RGD peptide conjugated poly[ LA-(Glc-Lys) ]/βTCP/PLA nerve conduit for bridging peripheral nerve regeneration defect. Methods Forty-five male Wister rots were randomly divided into 3 groups, with 15 rats each. A 10 mm defect was created in the right sciatic nerve. In group A the gap was bridged by PLA tube. In group B RGD peptide conjugated poly[ LA-(Glc-Lys) ]/β-TCP/PLA nerve conduit was used to repair the defect. Autologous nerve graft was done in group C which served as control. Twelve weeks postoperatively nerve regeneration was evaluated by gross observation,electrophysiology, muscle weight and muscle morphometry of triceps surae, and ultrastructure of the regenerating nerve. Results Twelve weeks after the operation, nerve conduction velocity and muscle weight recovery of group B were better than those of group A. The differences were statistically significant( P < 0.05). There was no significant difference between group B and group C ( P > 0. 05). The results of histology and ultrastructure showed that nerve regeneration in group B and group C was significantly superior to that in group A.Conclusion RGD peptide conjugated poly[LA-(Glc-Lys)]/β-TCP/PLA conduit can achieve similar results in repairing sciatic nerve defect to that of autogentic nerve graft. It may be an ideal material to repair nerve defect.     

Undersensing of VF in a Patient With Optimal R Wave Sensing During Sinus Rhythm

We describe a case of potentially fatal undersensing of VF by a third generation ICD with predetermined automatic gain control. In this patient, ventricular sensing was optimal, as R wave amplitudes during sinus rhythm were at least 16 mV. Cyclical, high amplitude signals during VF elevated the sensing floor to such an extent that complete undersensing of subsequent lower amplitude local electrograms occurred. This led to bradypacing and complete ICD therapy failure. Therefore, high R wave amplitudes during sinus rhythm do not warrant flawless sensing during VF. (PACE 2004; 27[Pt. I] 833–834)     

计算机辅助导航骨科手术及医用机器人技术在创伤骨科的应用

本文介绍了计算机辅助导航骨科手术(CAOS)及医用机器人技术在创伤骨科应用中的主要进展、当前在临床应用中存在的主要问题和相关对策,并对其未来的发展趋势进行了预测,同时简要介绍了北京积水潭医院创伤骨科在计算机辅助导航骨科手术及医用机器人技术方面的研究进展。当前骨科手术导航定位所应用的医学图象导引系统已经由使用单一的C型臂、CT等传统影像设备向应用三维C型臂、多模态图像处理系统等新型影像设备转变,基于多模态图像的导航系统将有可能成为导航手术的主流。医用机器人已经在自动化程度和人机交互模式方面,有了长足进展,摆脱了原有工业机器人的结构模式。医学图像后处理技术及其它相关信息技术极大地丰富了导航和机器人外科,只有在不断完善光学定位技术的同时,加大对其它定位方法的研究,才能够提高定位精度;要对相关设备进行开放式结构设计,使不同导航系统的注册软件能够互相兼容,手术器械能够通用,降低设备成本。骨科医生要正确认识计算机辅助导航骨科手术及医用机器人技术,在充分了解CAOS的技术特点、基本原理、操作程序的基础上,对要实施的手术具有深刻的理解,才能开展CAOS手术。目前,迫切需要建立CAOS技术标准、临床适应证和手术操作规范,进行CAOS产品之间的技术比较和评估,便于医生选择合适的CAOS产品。伴随快速发展的信息技术,数字化手术室、智能化微创导航手术系统、医用机器人辅助的远程医疗将有可能成为未来CAOS技术的主要组成部分。     

多聚酶链反应检测细菌DNA对大鼠空、回肠吻合口瘘的早期诊断价值

目的:探讨PCR检测大鼠外周血及腹水中细菌DNA对空肠-空肠、回肠-回肠吻合口瘘的早期诊断价值。方法:健康Wistar雌性大鼠50只，随机分成5组，每组10只：A组为假手术组;B组为空肠-空肠吻合组;C组为空肠吻合口瘘组;D组为回肠-回肠吻合组;E组为回肠吻合口瘘组。采集手术前后外周血及术后腹水，抽提DNA, 比较lacZ基因和16SrRNA基因的PCR阳性率，并观察各组的病理学情况。结果:（1）C，E组术后外周血lacZ基因PCR阳性率与B，D组无显著性差异（P>0.05）;C，E组术后外周血16SrRNA基因PCR阳性率显著高于B，D组（P<0.05）。（2）C，E组腹水lacZ基因和16SrRNA基因PCR阳性率均显著高于B，D组（P<0.05）。（3）C，E组腹水lacZ基因阳性率显著高于外周血（P<0.05）;C，E组腹水16SrRNA基因阳性率与外周血无显著性差异（P>0.05）。结论:（1）PCR检测术后外周血16SrRNA基因对空、回肠吻合口瘘的早期诊断有一定意义;（2）检测术后腹水lacZ基因和16SrRNA基因对空肠-空肠、回肠-回肠吻合口瘘的早期诊断也有一定意义。     

Variation of multifocal electroretinogram with axial length

INTRODUCTION: The first-order kernel response of multifocal electroretinogram (mfERG) decreases in myopia. A recent study indicates that the flash ERG is also reduced with increased axial length. The aim of this study was to investigate the variations in the first-order response (K1) and the first slice of second-order response (K2.1) across the retina for different axial lengths. METHODS: Thirty healthy subjects with axial length from 23.72 to 28.13 mm (spherical equivalent refractive errors from plano to -10.50 D) were recruited for mfERG measurement using VERIS 4.0. All subjects were fully corrected after cycloplegic refraction and pupils were dilated prior to mfERG recording. There is one trough, n1, and one peak, p1, in the K1 response and three troughs, n1, n2, n3, and three peaks, p1, p2, p3, in the K2.1 response. The amplitudes and implicit times of K1 and K2.1 responses were analysed to determine the characteristic of the responses across retina and the correlation to axial length. RESULTS: The amplitudes of p1 (in the first-order kernel-K1) decreased in the central region and the paracentral region (ring 3) as the axial length increased. The central retinal region showed high rates of reduction in both n1 and p1 (in K1). The amplitudes of n1p1 and n2p2 (in the first slice of the second-order kernel-K2.1) were reduced in the paracentral region (from ring 2 to ring 5) as axial length increased. The average n1 and p1 in K1, and n1p1 and n2p2 in K2.1 mfERG responses are decreased in amplitude by 6-10% per millimetre elongation of axial length. CONCLUSION: Eyes with longer axial lengths, usually with high myopia, have a weaker mfERG response and this attenuation is across the measured retina (from central to paracentral regions) but different kernel responses show a different pattern of attenuation at different retinal eccentricities. The weaker mfERG responses may be related to the morphological changes associated with increased axial length.     

Thrombophilic risk factors and unusual clinical features in three Italian CADASIL patients

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a genetically transmitted cerebrovascular disease. Typically, the first clinical manifestation is migraine and the full clinical spectrum of the disease with recurrent strokes of the subcortical type, cognitive, and mood disorders is seen during the fourth and fifth decades of life. Vascular risk factors are usually absent in CADASIL patients and the diagnosis of the disease is particularly suspected in young adults with cerebrovascular events of unknown cause, diffuse leukoencephalopathy on computed tomography or magnetic resonance imaging, and a history of cerebrovascular diseases or dementia in many family members. We describe three Italian CADASIL patients who presented to medical attention for cerebrovascular events occurred after the age of 55 and had, in addition to hypertension and hyperlipidemia, thrombophilic risk factors such as hyperhomocysteinemia, elevated levels of lipoprotein(a), and antiphospholipid antibodies. Symptoms possibly related to cortical involvement, such as dysphasia and visual field deficits, were reported by two of these patients. We conclude that a diagnosis of CADASIL should not be disregarded in patients with vascular risk factors and presenting with symptoms not immediately referable to subcortical damage at ages more advanced than commonly reported.     

等距和非等距前交叉韧带重建的对比实验

目的 观察等距和非等距前交叉韧带(anterior cruciate ligament，ACL)重建对膝关节功能的影响。方法 采用新鲜尸体观察等距ACL的解剖结构，在尸体和模型上重建等距和非等距ACL，分别观察重建后ACL长度和胫骨平台表面压强的变化。结果 等距ACL的重建在膝关节的全范围活动中长度的变化值最小，胫骨平台所受的压强也最小。结论 只有等距的ACI。的重建才能恢复膝关节的正常生理功能，而非等距重建的ACL会造成膝关节的不稳定(或活动受限)，或者使膝关节表面的压强增加。     

慢性粒细胞白血病骨髓细胞体外培养净化作用实验研究

目的 :采用 FISH方法直接在干细胞水平对慢性粒细胞白血病 (CML )患者自体骨髓体外培养前后的间期细胞进行 bcr/abl融合基因检测 ,从而探讨 CML 骨髓细胞体外培养对自体骨髓移植物的净化作用。方法 :分离初治的慢性期 Ph+ CML 7例骨髓单个核细胞 (MNCs) ,体外培养 10 d,用免疫磁珠 (MACS)富集培养前后的 CD34+ 细胞 ,通过流式细胞仪检测培养前后 MNCs中 CD34+ 干细胞比例 ,然后用 FISH方法检测其中 bcr/abl融合基因 ,同时分别用正常人细胞及 K5 6 2细胞株作阴性及阳性对照。结果 :(1) 7例患者骨髓细胞培养前后 CD34+ 细胞中 bcr/abl融合基因平均检出率分别为 85 .3%± 4 .9%和 78%± 5 .1% ,有统计学意义 (P<0 .0 5 ) ,(2 )培养前培养体系中 CD34+ 细胞数量为 (6 .0 6 0± 1.5 6 4 )× 10 5个 ,培养后体系中的 CD34+细胞数量为 (5 .974± 1.4 2 4 )× 10 5个 ,两者无明显差异 (P>0 .0 5 )。 (3)在对照组中假阳性率为 2 .5 % ,假阴性率为 2 %。结论 :自体骨髓细胞体外培养对 CML 病人的骨髓肿瘤细胞有一定程度的净化作用 ;FISH技术直接在干细胞水平检测 bcr/abl融合基因 ,且能定量分析 ,较传统方法更适合对骨髓净化进行评价 ,为骨髓净化提供了一种更方便、可靠的评定方法