正中神经显微结构三维可视化研究

目的 将正中神经干全长作连续冰冻组织切片,采集系列切片的神经二维信息,开发3D Nerve可视化系统,实现正中神经内部显微结构的三维可视化.方法 取新鲜成人尸体正中神经标本,作好定位并标记、OCT包埋、采用连续冰冻组织切片、染色、用扫描仪获取切片染色前后的连续的正中神经断面二维信息数据库,开发并应用3D Nerve可视化系统.结果 通过3D Nerve可视化系统观察发现,在不同断面正中神经神经束的数量、位置以及神经束内神经纤维的性质变化较大.在该系统中可在任意断面放大的视野下观察正中神经的显微结构,追踪各神经束在正中神经内的立体行径;重建的正中神经能在空间位置上绕任意轴作任意角度旋转,便于从不同的角度对各神经束的形态、空间位置及相互毗邻关系进行观察,从而动态地展示正中神经内部的复杂结构.结论 正中神经3D Nerve可视化系统真实地再现正中神经干全长及其内部各神经束和束组的三维立体结构,可为临床修复正中神经损伤提供精确的断层解剖图像.     

人体正中神经内部显微结构的三维重建与可视化研究

[目的]探索应用计算机处理人体正中神经全长连续冰冻组织切片的二维图像信息,开发3DNerve神经三维可视化系统,重建人体正中神经内部显微结构并实现三维可视化.[方法]取新鲜人尸正中神经标本1例,以人长发为定位线、OCT包埋剂包埋、采用连续冰冻组织切片、乙酰胆碱脂酶组织化学法染色、高分辨率扫描仪获取二维数码信息后、应用3DNerve对正中神经显微结构进行三维重建.[结果]正中神经在不同断面神经束的数目、位置以及神经束内神经纤维的性质变化较大.连续断面观察显示各神经束均是混合束,未见有纯粹的感觉束或运动束的出现.通过3D Nerve可在任意断面放大的视野下观察正中神经的显微结构,追踪各神经束在正中神经内的立体行径.从而动态地展示正中神经内部神经束的复杂结构.[结论]重建的正中神经三维可视化真实地再现正中神经干全长及其内部各神经束和束组的三维立体结构,可为临床修复正中神经损伤提供精确的断层解剖图像.     

短段腓总神经功能束三维重建的初步研究

目的 基于组织学连续断层切片,利用计算机技术进行短段腓总神经功能束三维重建.方法 取自愿捐献的成人约5 cm长胭窝段腓总神经,连续横断冰冻切片,片厚10 μm,切片间距0.25 mm,共切取200张切片.采用乙酰胆碱酯酶组织化学染色,镜下观察神经束变化规律,通过数码摄像系统将染色切片转化为数字图像,图像拼接获取放大100倍的二维全景图像,人工判断功能束性质,图形处理软件配准分割后,利用Amira 3.1三维重建软件实现腓总神经功能束的三维重建.结果 (月国)窝段腓总神经内部功能束可划分为感觉神经束、运动神经束、混合神经束和以运动神经纤维为主的混合神经束.其中,腓深神经和腓浅神经间无神经束的交叉融合,神经束的交叉融合主要发生在腓深神经和腓浅神经内部的功能束间.三维重建结果能较真实地再现周围神经的三维立体结构及其内部功能束组的三维立体行径,重建结构能单独或搭配显示,还能任意角度显示.结论 基于组织学和计算机技术,可以三维重建短段腓总神经功能束,为长段周围神经功能束的三维重建提供可行性依据.     

臂丛神经显微结构的计算机三维重建

目的：重建臂丛神经的外轮廓及其内部神经束的精细三维行径，同时探索一种臂丛神经显微结构计算机三维重建的实用方法。方法：取健康成年尸体的臂丛神经标本2例(从神经根管出口至正中神经交叉处，平均长20cm)，作好标记，以女性头发作为定位线，采用连续组织切片后胆碱脂酶组织化学染色，高分辨率数码摄像系统获取二维数码信息后对臂丛神经显微结构进行三维重建。结果：三维重建真实地再现臂丛神经的三维立体结构及其内部各神经束的三维立体行径，并可显示臂丛神经中神经束的任意断面及其全长的解剖结构与相互关系，形象地展示臂丛神经内部神经束的复杂重组过程。重建结构均能单独或搭配显示，还能任意角度显示：在臂丛的五个根中，C6-C8内部神经束数目较C5、T1多。在C6-C8中，又以C7神经根内部神经束数目最多。结论：臂丛神经内部神经束结构相当复杂，相互间不断交叉重组，形成独特的神经束网络结构。臂丛神经显微结构三维重建由于采用了较为精确的定位材料和方法，三维图像显示效果较好，是一种较为实用的方法。     

基于二次成像技术标志点获取和自动配准的长段周围神经功能束三维可视化关键技术研究

目的探讨四肢周围神经功能束三维可视化重建过程中的关键技术问题及解决方法,并验证其可行性。方法取自愿捐献新鲜成人尸体右上臂20cm尺神经标本。以4根成年女性头发作为标志线,连续横断面冰冻切片,厚15μm,层距0.5mm,共400张切片,行乙酰胆碱酯酶(acetylcholinesterase,AchE)染色;采用二次成像技术,以配置MSHTO MD90显微数码成像装置的Olympus Z61型体视显微镜于AchE染色前、后分别获取同一切片的2张全景图像,通过Photoshop图像处理软件图层叠加进行图像处理,获取含4个完整定位标志点的神经断面二维全景图像;以优化最小二乘支持向量机方法作为分类器自动识别处理图像,并利用空间变换方法实现自动配准,结合人工辅助获取功能束轮廓,通过Amira 4.1医学三维重建软件进行长段尺神经功能束三维重建,评估重建逆向还原效果。结果利用二次成像Photoshop图像处理软件处理同一张切片染色前、后的图像,图像轮廓吻合度良好,合成图像获得的标志点定位精确,操作简便快捷。优化最小二乘支持向量机方法识别精度高,误差率为8.250%;三维模型可直观观察神经内部不同性质功能束交叉融合的变化,其任意水平切割逆向还原基本吻合。结论基于二次成像技术和Photoshop图像处理软件图层处理技术的计算机自动配准可应用于长段周围神经的三维可视化重建,重建快捷,效果较好。     

尺神经功能束组走行模式的三维重建

目的 在组织学切片基础上，应用计算机三维成像技术，重建尺神经功能束组走行的三维图象，以期指导临床臂丛与尺神经损伤后的显微修复，为臂丛神经根性撕脱伤后手内肌功能的恢复提供必要的形态学基础。方法 制作尺神经连续断面组织切片，AchE组织化学染色，将切片转化为数字图象，人工判断功能束组性质，图形处理软件配准后，采用体数据场轮廓线表面重建方法，应用VC语言编制三维图象存储构建程序，在程序中重建尺神经功能束组走行的三维图象。结果 在程序中实现了尺神经功能束组走行的三维重建。对重建的各神经功能束的外表面进行三维显示，实现人机交互的任意三维旋转、任意断面切割。通过对感兴趣的显示对象(神经干和各束组)的选择，可以在同一三维空间同时显示任意神经功能束组的立体结构，结果反映出它们的在空问位置上的结构、毗邻关系和走行。通过与显微解剖结果对照，发现其显示图象与解剖结果基本吻合。结论 构建的尺神经功能束组的三维图象，能较真实的反映束组的走行分布情况，对临床医生理解神经内部结构，设计手术方案和在术中对照束组的分布方位调整手术方法有积极的作用。     

面神经的角神经三维可视化研究

目的 通过三维可视化技术,验证前期解剖学研究结果,并为失神经支配手术提供一定的理论依据.方法 取自愿捐献的甲醛固定的成人尸头,切取内眦角内侧全层软组织长3 cm、宽1 cm,以皮肤表面2平行刀痕为定位线,连续横断石蜡切片,片厚10 μm,切片间距0.25 mm,共切取120张切片.采用HE组织化学染色,高分辨率扫描仪获取二维数码图像后进行三维重建.结果 ①真实再现了角神经的三维立体结构及角神经与内眦动静脉的三维立体行径,且重建结构不但能单独或搭配显示,并可从任意角度显示.②证实了角神经显微解剖学研究的正确性.③角神经毗邻关系的三维重建技术可以应用到临床.结论 基于组织学和计算机技术,可以三维重建角神经的毗邻关系,为角神经的失神经支配手术提供更加准确的可行性依据.     

周围神经功能束三维可视化中完整信息的二维全景图像获取

目的 探讨用于周围神经功能束三维可视化重建的带完整信息的神经断面二维全景图像获取的关键技术.方法 新鲜成人截肢腓总神经及尺神经标本各一段,以4根成年女性头发作为标志线,连续冰冻切片,乙酰胆碱酯酶组织化学染色:分别以100倍光学显微镜下分区摄影后拼接及光学扫描仪和体视全景数码成像仪一次性获取全景图像,观察染色前后人发标志点贴片情况,比较不同全景图像效果;以数码全景成像仪获取染色前、后同一切片的全景图像,通过Photoshop CS2处理获取含4个完整定位标志点的神经断面二维全景图像.结果 体视全景数码成像仪一次全景成像精确,功能束易于鉴别,基本具备高倍光学显微镜下拼接图像的特点,且获取简单快捷;二次成像Photoshop合成图像功能束轮廓吻合良好,标志点定位精确,操作简单快捷.结论 结合高像素体视全景数码成像装置一次性全景数码成像、二次成像技术和Photoshop图层处理技术,可用于海量获取带完整信息的神经断面二维全景图像进行三维重建,提高重建效率及精度,可为临床上周围神经损伤修复方式的选择以及组织工程神经的构建提供直观的解剖依据.     

应用micro-CT实现兔坐骨神经显微三维结构可视化研究

目的探讨通过micro-CT扫描新西兰大白兔坐骨神经标本,利用三维可视化软件Mimics17.0重建兔坐骨神经内部显微三维结构。方法取6只成年新西兰大白兔坐骨神经组织标本分成A、B组(n=3),分别用1%、5%Lugol液对两组标本染色,于染色0.5、1.0、1.5、2.0、2.5、3.0、3.5 h时,行光镜及micro-CT观察两组标本的显像变化,将显像良好的micro-CT图像序列导入Mimics软件,采用三维重建工具重建兔坐骨神经神经显微三维结构。结果 A组标本在染色2.5 h、B组标本在染色1.5 h时,经光镜及micro-CT观察可获得较为清晰的显微三维结构图像。图像显示新西兰大白兔的坐骨神经主要分3组神经束,且各神经束立体行径相对固定,Mimics软件测量各神经束横截面积分别为(0.425±0.013)、(0.038±0.007)、(0.242±0.026)mm~2,生成的数字化三维模型可在任意横断面观察坐骨神经内部显微结构。结论应用micro-CT可清晰真实显示兔坐骨神经显微三维结构,为建立大样本量周围神经显微解剖学数据库提供了可靠方法。     

胰腺及其周围结构的三维重建研究

目的 建立胰腺及其周围结构三维可视化模型,为胰腺疾病的影像学诊断及外科手术方案选择提供形态学依据.方法 应用中国数字化可视人体数据集,选取从脾上缘至十二指肠下段连续断面图像,对断面图像轮廓进行数据分割,并运用体数据绘制及表面绘制的重建方法,在计算机上对胰、脾外形及其内部管道系统与其毗邻结构进行三维重建和立体显示.结果 重建出了胰腺及其周围结构的三维可视化模型,该模型具有多种显示功能,可显示任意切面的结构,同时也可以任意放大、缩小和任意角度旋转观察.结论 该模型直观揭示了胰、脾及其毗邻区域的三维解剖结构,有助于临床解剖教学以及该区域病变的诊断及手术方案的制定.     

Three‐dimensional reconstruction and visualization of the median nerve from serial tissue sections

The purpose of this study was to definitively implement the three‐dimensional visualization of sensory and motor fascicles in the human median nerve by means of acetylcholinesterase (AChe) histochemical staining and under the assistance of the computer technology. One fresh human median nerve was harvested from a male adult cadaver. The median nerve was fixed at a special holder. Then, the whole holder was embedded and rapidly frozen in the liquid nitrogen. The processed median nerve was then cut coronally every 100 μm at a 20 μm thickness along its long axis in a sliding freezing microtome. The total number of sections was 4,650 slices. All sections were stained with the AChe histochemical method. The stained sections were scanned and saved as Joint Photographic Experts Group files. These images with positively and negatively stained sections were acquired to an Intel dual Pentium computer. The Adobe Photoshop CS2 software was used to compare the reference points of images before and after staining. The two‐dimensional intraneural microstructure database of median nerve was then acquired. A software of 3D nerve visualization system was developed. With the 3D nerve visualization system, the 3D visualization result of intraneural microstructure of median nerve was created. The findings may provide more accurate and detailed anatomic information for nerve repairs, specifically for the fascicular nerve repairs. The 3D nerve visualization technique may have potential for future studies of topography of peripheral nerve. © 2009 Wiley‐Liss, Inc. Microsurgery, 2009.     

Study of sensory and motor fascicles in brachial plexus and establishment of a digital three-dimensional graphic model

To investigate a 3-dimensional (3D) model of human brachial plexus including its topography of sensory and motor fascicles with the assistance of the computer technology, 2 brachial plexus were serially horizontally sliced. Each slice was stained by Karnovsky-Roots acetylcholinesterase histochemical method. The stained sections were scanned, and the image was put into the computer serially. At last, the 3D diagram of brachial plexus was made. The internal structure of the brachial plexus was found to be very complicated. The fascicles bifurcated and recombined with one another with no fixed rules. All fascicles were mixed sensory and motor fibers. Acetylcholinesterase histochemical staining from a serial tissue section is a useful technique to distinguish sensory fibers from motor ones. The 3D visualization of the brachial plexus may help to develop a computerized database of the fascicle topography to provide an anatomical reference in fascicular repair of brachial plexus.     

Histologic structure of the ulnar nerve in the hand

Serial histologic sections of 20 ulnar nerves revealed broad individual variations in the number and diameter of the fascicles at all levels. The number of fascicles increased from the proximal portion to the distal portion, and interfascicular and perifascicular connective tissue occupied about 50% of the entire nerve section. These configurations explain the difficulties and setbacks encountered in the repair of injuries to the ulnar nerve. The counting of myelinated nerve fibers also revealed a great number of variations.     

Nerve fibre composition of the palmar cutaneous branch of the median nerve and clinical implications.

Fifteen fresh human cadaver hands were dissected, using x2.8 loupe magnification, to study the subcutaneous innervation at the site of the incision (in the line with the radial border of the ring finger) for standard open carpal tunnel decompression. Subcutaneous nerve branches were detected and traced proximally to determine their origin. Morphometric analysis of nerve cross sections from the site of the incision and from the main nerve trunk proximal to cutaneous arborisation was performed using light and transmission electron microscopy and a computer-based image analysis system. At the site of the incision, the ulnar sub-branch (US) of the palmar cutaneous branch of the median nerve (PCBMN), which innervates the skin over the hypothenar eminence, was found in 10 of 15 cases. Branches from the ulnar side were not detected. The main trunk of PCBMN consisted on average of 1000 (SD 229) myelinated axons arranged in 1-4 fascicles. In the US of the PCBMN there were on average 620 (SD 220) myelinated axons, 80% of them smaller than 40 microm(2) i.e. thin myelinated axons, and on average 2037 (SD 1106) unmyelinated axons, arranged in 1-3 fascicles. The ratio of the number of myelinated axons in the US and the main trunk of the PCBMN was on average 63% (SD 19%). Frequency distribution of cross-sectional areas of myelinated axons shows no significant difference between the US and the main nerve trunk of the PCBMN. The importance of incision trauma to subcutaneous innervation of palmar triangle is emphasised and possible mechanisms of scar discomfort are discussed.     

Topography of the vestibulocochlear nerve

OBJECTIVE: The terms superior vestibular nerve and inferior vestibular nerve have been used in the field of neurosurgery to indicate anatomically the two respective vestibular components of the vestibulocochlear nerve. To reappraise the aptness of this terminology, fascicular patterns and the anatomic relationship of the vestibular and cochlear components were examined. METHODS: Twenty vestibulocochlear nerve specimens were obtained from cadavers. The nerves were excised, with care taken to sustain their spatial relationships, then embedded in paraffin blocks and cross sectioned in 10-microm-thick slices. Serial cross sections were stained and examined with a light microscope. RESULTS: The vestibular component was separated into two parts only at the lateral fundus of the internal auditory canal, lateral to the vestibular ganglion. In the internal auditory canal of all specimens, the vestibular component was represented by numerous fascicles. Around the porus acusticus, the fascicular pattern among the specimens was diverse: 13 of the 20 specimens were still polyfascicular, 4 specimens consisted of two large, distinct fascicles, and, in the remaining 3 specimens, a portion of the vestibular fascicles had fused with the cochlear component. Near the root entry zone, all vestibular fascicles fused and merged with the cochlear nerve to form a single trunk. CONCLUSION: There was no evidence to support the anatomic correctness of specifying the superior and inferior vestibular nerves, except in the lateral fundus of the internal auditory canal.     

比较两种带血供长段尺神经移植术神经再生远期结果的实验研究

目的 对带尺上副血管的尺移植和带尺侧上副血管加尺血管的尺神经移植中,神经再生的远期结果进行比较以供临床参考。方法 新西兰大白兔9只,将双侧上肢的尺神经游离后,于尺侧上副动脉起始水平及腕部切断后作原位移植。右侧：尺神经带尺侧上副血管。左侧：尺神经肘上带尺侧上副血管,肘下段则带尺血管。两侧尺神经原位移植后均置于屈肌群表面。术后7个月进行尺神经肌电、再箐髓纤维数与截面积、髓鞘（mt）和轴突／纤维的直径比（d／D）及肌肉组织学的检测。结果 术后7个月,9只大白兔双侧前足的溃疡均愈合。右侧尺神经移植段神经干的波幅、运动神经传导速度均低于左侧（P＞0．01）;而在腕部缝合口近端3cm水平,两侧再生有髓纤维数及截面积的差异均无显著性意义（P＞0．05）;而在腕部缝合口近端1cm、远端0．5cm水平,左侧优于右侧（P＜0．05,P＜0．01)。双侧小鱼际肌肌湿重及肌纤维截面积无明显差别。髓鞘厚度与d／D比值的差异无显著性意义。结论 两种带血供的尺神经移植术效果均好。再生神经纤维数量上的差异主要发生在移植神经远端部分。临床作带尺侧上副动脉的尺神经转位或移植时,应增加尺神经肘上段的长度,缩短肘下段的长度。