原子力显微镜对泌尿系结石超微结构的研究

目的 研究和观察泌尿系结石的超微结构。方法 10枚结石分别做超薄切片(厚度小于3mm),打磨抛光处理,并收集粉末。先行原子力显微镜观察,再做成电镜标本观察分析。结石粉末均行X-线衍射检测。结果 得到肾盂结石和膀胱结石超微结构的AFM图像和电镜扫描图像,X-线衍射检测有7枚是草酸钙结石,3枚是磷酸钙结石。测得草酸钙结石和磷酸钙结石的微小晶粒尺寸。分析AFM的反转片信息,获得结石基质的空间构架图像。结论 应用原子力显微镜技术可以实现对泌尿系结石超微结构直接实时的观察分析。尿结石的超微结构形态与结石所处部位无关,而取决于结石晶体类型。草酸钙结石的晶体和基质的AFM图像与磷酸钙结石不尽相同,晶体尺寸有差异(P<0.05)。结石基质的空间构架为晶体的聚集提供了可能性。     

应用原子力显微镜技术对人膀胱癌细胞株T24活细胞的成像研究

目的实现应用原子力显微镜(AFM)技术对人膀胱癌细胞株T24活细胞的成像,在原子级或纳米级水平上观测T24活细胞的超微结构。方法在生理条件下,使用AFM直接观测培养中的人膀胱癌T24细胞,对其超微结构动态成像。结果获得T24活细胞原子级或纳米级分辨率下的实时扫描图像,细胞胞膜和细胞骨架成像清晰。结论AFM技术可以用于对膀胱癌T24活细胞超微结构的成像研究,所获得的图像可反映其在生活状态下的动态变化。     

原子力显微镜新应用:单细胞诊断肝癌

目的 运用原子力显微镜(AFM)分析正常肝细胞、肝硬化细胞、肝癌细胞膜表面微细结构;绘出细胞膜表面蛋白变化图,并以此图作为诊断肝癌的标准.方法 选取正常肝细胞、肝硬化细胞、肝癌细胞各40组,利用AFM对标本扫描,将细胞膜表面扫描图用分析软件进行分析.结果 正常肝细胞、肝硬化细胞、肝癌细胞的细胞膜表面结构存在明显的差异,从正常肝细胞到肝硬化细胞再到肝癌细胞是一个渐进的过程.每个细胞膜表面的蛋白微细结构分布并不均匀,从膜表面蛋白分析数据中可看到它们存在差异.结论 依据细胞膜表面分析数据做出三组图形,我们可以以此为肝癌的诊断标准,只要获得一个细胞,即可做出肝癌的诊断.根据细胞膜表面蛋白分布的不同,将细胞膜表面分为功能活跃区和功能不活跃区.     

应用原子力显微镜观察脉冲电磁场对大鼠骨质疏松的治疗后骨组织表面超微结构的变化

目的观察实验性大鼠骨质疏松模型股骨颈超微结构的变化特点,并探讨利用原子力显微镜观察的可行性。方法清洁级雌性SD大鼠(鼠龄3个月)60只,体重(200±20)g。随机将其分成对照组(Sham组)、去卵巢组(OVX组)、阿伦膦酸钠治疗组(ALN组)、PEMFs治疗组(PEMFs组),每组大鼠15只。除Sham组外,对其他各组行双侧卵巢切除术,造模后第30天开始分组干预。ALN组给予阿伦膦酸钠灌胃,PEMFs组给予脉冲电磁场治疗。Sham组和OVX组手术后正常喂养,不予任何处理。治疗后30d取大鼠股骨头切片,通过AFM观察大鼠股骨颈的表面超微结构。结果 Sham组原子力显微镜扫描可显示骨组织的骨陷窝、骨小管及其表面的钙磷晶体沉积,骨组织表面粗糙度为(2.59±0.645)μm。OVX组可见骨陷窝的大小变化及其钙磷晶体的排列紊乱,模型组骨组织表面粗糙度明显高于对照组(P0.01),提示造模成功;ALN组表面粗糙度明显低于OVX组(P0.05),PEMFs组的表面粗糙度与OVX组相比,有明显降低,差异具有统计学意义(P0.05);而PEMFs组与ALN组比较,其表面粗糙度无明显差异(P0.05)。结论通过原子力显微镜能清晰观察到脉冲电磁场对大鼠骨质疏松治疗后的骨组织表面超微结构的变化,其疗效与阿仑膦酸钠相当,为脉冲电磁场治疗骨质疏松提供了一定的理论依据。     

面中份缝牵引成骨过程中颅面骨缝的组织与形态学变化

目的：观察犬颅面骨缝的组织结构的改变,对经缝牵引技术作用下的颅面骨缝的生长改建进行初步探讨。方法：18只12周龄雌性反牙合京叭犬随机分为牵引成骨组12只和正常对照组6只,在牵引第5、10、15天,固定10、20天,去固定2月分别处死实验犬2只和对照犬1只,光镜下观察各骨缝的组织学和形态学变化。结果：在牵引与固定过程中,骨缝宽度逐渐增大,骨缝细胞发生分化、增殖,骨缝中出现大量新生血管,骨缝重新进行改建,形成新的骨缝。结论：受张力影响的上颌骨周围骨缝出现适应性重建。     

原子力显微镜下观察急性压力损伤后脊髓背根神经节神经元的结构变化

目的:利用脊髓背根神经节神经元细胞体外机械压力损伤模型,探计单纯机械压力因素对背根神经节神经元细胞的损伤效应,尤其是细胞膜骨架蛋白的变化情况。方法:新生SD大鼠脊髓背根神经节神经元体外原代培养,利用多功能全自动细胞压力仪(国家实用新型专利ZL 200820030390.7),将培养细胞分为A、B、C、D四组,其中A组为未施加机械压力损伤的对照组,B、C、D组分别给予0.3mPa、0.5mPa.、0.7mPa的压力损伤10min,每组观察细胞爬片6个,于加压后培养8h取细胞爬片固定,送原子力显微镜检测,观察神经元细胞的细胞膜骨架蛋白纳米级变化情况,获取并分析胞膜表面颗粒高度分布数据及原子力显微镜图片。结果:对照组细胞膜表面平滑,无明显粗糙颗粒;0.3mPa组细胞膜表面颗粒突起较对照组增多,膜颗粒总体高度增加,局部变粗糙,胞膜上蛋白高度分布直方图峰值右移;0.5mPa组较0.3mPa组胞膜粗糙度更加明显;0.7mPa组膜表面蛋白颗粒总体高度较0.3mPa及0.5mPa组降低,粗糙颗粒减少,但出现胞膜骨架蛋白解聚与构像变化,胞膜出现孔道样改变。胞膜颗粒高度分布峰值、分布宽度及最高颗粒值的单因素方差分析显示,压力损伤各组间及损伤组与对照组均有统计学差异(P<0.01)。结论:机械压力损伤因素可引起细胞膜骨架蛋白变化和表面颗粒高度变化,为压力损伤的细胞保护研究提供了实验数据。     

兔椎体软骨终板血管芽增龄性变化

目的 探讨不同年龄段兔椎体软骨终板内血管芽的形态变化与椎间盘退变的关系.方法 15只不同年龄段新西兰兔,2月龄5只,1年龄5只,3年龄5只,雌雄不限.X线观察终板有无钙化及椎体周围有无骨赘形成,组织学观察软骨终板和椎间盘的形态学变化,血管铸型后扫描电镜观察终板各部位血管的形态变化.按Miyamoto等分级标准将椎间盘分为1～5级,分别规定为1～5分.采用t检验,分别对2月龄组和1年龄组髓核区与内层纤维环区,2月龄组及1年龄组兔髓核区、内层纤维环区椎体终板内血管直径进行比较.结果 2月龄组终板无明显钙化、软骨终板结构良好、潮标清晰,髓核及纤维环结构饱满清晰;1年龄组终板轻度钙化,髓核皱缩,纤维环出现裂隙;3年龄组终板钙化明显,密度增高,并可见骨赘形成,髓核消失.随着年龄增加,终板内微血管逐渐减少,最终消失,软骨终板中心部位较周围部血管减少更为明显.组织学显示软骨终板出现钙化,软骨终板退变重于椎间盘;钙化的程度与椎间盘退变程度呈正相关.2月龄组髓核区与内层纤维环区椎体终板内血管直径比较,差异有统计学意义;2月龄组与1年龄组髓核区椎体终板内血管直径比较,差异有统计学意义.结论 随着兔年龄增加终板内血管逐渐减少,终板发生退变,其程度重于椎间盘,提示椎体终板内微血管的变化可能是椎间盘退变的重要促进因素.

Abstract：

Objective To observe the morphologic changes of of vascular buds in vertebral cartilage endplate in age-specific rabbits and also to investigate the correlation between the changes of vascular buds and interverbral disc degeneration. Methods There were 15 New Zealand white rabbits in our study,which include three groups, 2-week-old rabbits, 1-year-old rabbits and 3-year-old rabbits, and each groups had five rabbits. The X-ray radiograph, histology and scanning electron microscope were used to observe the changes of vertebral cartilage endplate. According to Miyamoto standard, the interverbral disc was graded 1-5, and scored 1-5 respectively. Results The changes of micro-vascular structure of vertebral cartilage endplate were observed during aging. Under the scanning electron microscope, the vascular structure degenerated gradually, and disappeared in the end. The blood vessels in the central region of the vertebral cartilage endplate reduced more obviously than those in periphery region. The severe degeneration was found in vertebral endplate, compared with intervertebral disc. The changes of vascular buds in rabbits vertebral cartilage endplate had positive correlation with the vertebral endplate calcification and the interbertebral disc degeneration. Conclusion Changes of vascular buds in vertebral endplate may accelerate intervertebral disc degeneration.     

药物对兔模拟唇裂术后瘢痕和颅面发育影响的初步研究

用兔经手术形成单侧完全性模拟唇裂，于广泛组织潜行分析后直接缝合。术后伤口周围皮下和粘膜下定期注射具有改善伤口愈合，减少瘢痕形成的药物盐酸罂粟碱和透明质酸酶。与对照组比较和不用药组比较术后唇压低、颅颌骨全长和上颌骨全长的长度大。     

兔未成熟心肌增龄性变化及成熟时限的研究

目的 研究兔未成熟心肌的成熟时限及其特性。方法 分别以１、３、５、７、１６、２４周龄正常兔心肌为对象,每组１０只,从形态结构、钙代谢、能量代谢、氧自由基代谢、内分泌功能及其基因表达等方面进行研究。结果 正常未成熟心肌结构、功能有其特点,随周龄增长主要变化多发生在３～７周龄,至７周龄时绝大多数研究指标趋于成熟,各指标成熟时间先后不一,心肌基本成熟时间早于性成熟期。结论 正常未成熟心肌结构、功能 在７     

原子力显微镜对角质形成细胞与黑素细胞共培养模型的观察研究

目的：用原子力显微镜观察人表皮黑素细胞（MC）、角质形成细胞（KC）单独培养和共培养的表面形态以及α-促黑素（α-MSH）对细胞表面形态的影响。方法：分别纯化培养来自人包皮的表皮MC和KC,MC以自配的添加MC生长物质的MCDB153培养基培养,KC以KC无血清培养基（K-SFM）常规培养。传第2代后以1：10的比例将两细胞接种到3Cm×3Cm的小培养皿中,以K-SFM培养基继续培养,单独或混合培养的细胞经添加含或不含100nMα-MSH的培养基干预3天后,0．5％戊二醛固定10min,原子力显微镜常温常压下,触摸式扫描。结果：正常人表皮MC有3个树突,每个树突有明显的二级分枝,除主干和分支见到膨出的颗粒物质,我们在树突的侧缘底侧和顶端还发现有丝状伪足结构,经α-MSH刺激后树突明显变长、变细,主干和分支表面膨出颗粒物质更为密集,许多已脱离枝干,丝状伪足则未有明显变化。表皮KC表面可见许多片状或钩状突起。共培养后,KC与MC接触部位可见明显的丝状伪足样结构,未连接部位则未见到丝状伪足样结构,添加α-MSH后,两细胞连接处的丝状伪足样结构明显增多。结论：通过胞吐和丝状伪足输送可能是黑素小体从MC向KC传递的两种主要方式,α-MSH可能通过促进这两种结构的发生而发挥促黑素传递的作用。     

Study of inhibition mechanisms of glycosaminoglycans on calcium oxalate monohydrate crystals by atomic force microscopy

Atomic force microscopy (AFM) was applied to the (-101) faces of calcium oxalate monohydrate (COM) crystals grown from calcium oxalate (CaOx) solutions. Microstructures of many spiral hillocks with step height of 1 nm were observed on the faces. Then using AFM in situ, we analysed the re-growth process of the spiral steps on the face of COM seed-crystals in CaOx growth solutions that contained growth inhibitors of glycosaminoglycans and studied their inhibition mechanisms on COM crystals. The total morphology of the faces of COM seed crystals re-grown in the CaOx growth solutions was assessed by scanning electron microscopy (SEM). In the growth solution without glycosaminoglycans (control experiment) or with chon-droitin sulphate (ChS), AFM images and SEM micrographs of the faces of the re-grown seed crystals showed two-dimensional (2D) nucleation although 2D nucleation was delayed in the presence of ChS. However, the addition of dermatan sulphate (DS) to the growth solution resulted in isotropic growth by a step flow mode and spiral mechanism. With regard to the main inhibition mechanisms of two glycosaminoglycans (ChS and DS) on COM crystals, it can be concluded from these results that ChS delays 2D nucleation and DS inhibits 2D nucleation. Received: 23 November 1998 / Accepted: 25 June 1999     

The use of interosseous dental wires and sutures for internal fixation in a patient with multiple comminuted middle facial fractures and facial nerve injury: A demonstrative case report

IntroductionThe use of plates and screws for facial fractures is considered the gold standard; providing accurate reduction and rigid fixation.Case reportWe report on a case with multiple comminuted middle facial fractures and concurrent facial nerve injury. The fractures were fixed with a combination of interosseous dental wires and polypropylene sutures with a satisfactory outcome.DiscussionWe aim to demonstrate two main advantages of wire/suture fixation in such cases when compared to plates and screws. Wire/suture fixation does not require periosteal dissection for fixation; and hence there is more preservation of the blood supply of the bony fragments. Furthermore, in the setting of concurrent facial nerve repair, the use of plates may risk re-injury of the repaired nerve during the late removal of the hardware. Interosseous wires/sutures do not require late removal and this is another advantage in these cases.ConclusionInterosseous dental wires and polypropylene sutures may be considered for fixation of multiple comminuted middle facial fractures and concurrent facial nerve injury.     

黄精赞育胶囊优选方处理前后精子的超微结构研究

目的 观察黄精赞育胶囊优选方作用前后精子的超微结构变化.方法 运用原子力显微镜(AFM)技术对比观察正常精子和病理性精子在黄精赞育胶囊优选方作用前后超微结构的动态变化.结果 获得精子头体、颈部和鞭毛等部位的实时超微结构图像,发现药物T预后活动力低精子超微结构的缺陷有逐步被修复的现象.结论 黄精赞育胶囊通过修复活动力低精子超微结构的病理形态学缺陷可能足优选方改善弱精子质量的机制之一.     

应用去神经足(足母)短、趾短伸肌游离移植结合埋没导引缝合技术治疗晚期面瘫

目的探讨治疗晚期面瘫更为有效的手术方法。方法采用足短伸肌、趾短伸肌游离移植和带蒂颞肌筋膜瓣转移结合埋没导引缝合技术综合治疗晚期面瘫432例。结果354例患者获得面部表情静态对称和动态基本对称的良好效果;52例患者面部表情静态对称较理想而动态对称不足;26例患者面部表情具有一定的动态恢复,而静态对称不足,经再次手术悬吊获得比较满意的效果。有4例患者出现术后感染,2例在足部供区,2例在面部,经引流和抗炎治疗,均于3周内治愈,对手术效果没有明显影响。我们对46%的患者进行了复诊或电话随访,随访时间最长12年,最短1年,平均28个月。93%的患者对治疗效果满意,7%的患者认为治疗虽然使面部畸形明显改善,但表情对称性尚不充分。结论足短伸肌、趾短伸肌游离移植结合埋没导引缝合技术可以有效地治疗晚期面瘫,手术方法简单,组织损伤小,术后并发症少,值得推广。     

The study of 3-dimensional structures of IgG with atomic force microscopy

Tsihmteimp eouxrlctaaittnianttog rr oyclae nti eoiunnr-o cbtorrnaanidnsumc itinitnjtuegrr y igo lnaunt cadhma fanutnneec lptsiloa nwyssh iabchnyhave distinct biophysical and pharmacologicalproperties and differ in their binding sites for selectiveagonists and antagonists.1,2NMDA(anti-N-methyl-D-aspartate)receptor widely distributes in the centralnervous system.It is essential not only for its normaldevelopment and plasticity but also for such specializedfunctions as memory or learning.3Espec…     

Ultrastructure of Dental Enamel afflicted with Hypoplasia: An Atomic Force Microscopic Study

The ultrastructure of the human tooth enamel from a patient diagnosed with hypoplasia (HYP) was investigated using atomic force microscopy (AFM) and compared with the surface of normal human tooth enamel. Hypoplasia is a hereditary defect of dental enamel in which the enamel is deficient in either quality or quantity. AFM results presented for the HYP tooth enamel clearly demonstrate that the apatite crystal morphology in hypoplasia tooth enamel is perturbed in the diseased state which could result from a defective synthesis of the extracellular matrix proteins, e.g., amelogenin, by the ameloblasts. HYP enamel consisting of loosely packed, very small grains does not present a tendency for association, as in the case of the normal healthy tooth. Indeed, the enamel surface affected by HYP is porous and is made of much smaller grains. In some samples, the HYP part of enamel surface appeared in the form of a point-defect, which we believe may be associated with the early stages of the HYP deformation.     

Chronic changes in cerebrospinal fluid pathways produced by subarachnoid kaolin injection and experimental spinal cord trauma in the rabbit: their relationship with the development of spinal deformity

Post-traumatic cystic changes in cerebrospinal fluid (CSF) pathways such as ventriculomegaly and/or hydrosyringomyelia are not uncommon, but their characteristics have not yet been fully clarified. This study was designed to investigate the alterations affecting the CSF pathways in rabbits at a late stage, and to clarify the relationship between these changes and the development of spinal deformity. In this study, a total of 60 New Zealand white rabbits were used and they were segregated into four different groups of 15 animals each: sham-operation group, kaolin group, and kaolin plus mild trauma group and kaolin plus severe trauma group. The animals were subjected to radiological investigation using direct X-ray study and magnetic resonance imaging (MRI) after 4 months. The thoracic spinal cords of the animals were dissected after intracardiac perfusion–fixation with 10% formalin for light microscopy and 2.5% glutaraldehyde for transmission electron microscopic study. Following the sectioning and staining procedures, the histological characteristics of the spinal cords were evaluated with light microscopy and transmission electron microscopy. A spinal deformity developed in 90% in rabbits in both kaolin injection group and spinal trauma groups. MRI revealed generalized dilatation of the ventricular system and the central canal of the spinal cord after the kaolin injection with/without trauma in this study. Gross morphologic examination showed some enlargement of entire CSF pathways in these groups. All animals with central canal dilatation had mild or severe scoliotic and kyphotic deformities. In a light microscopic study, a denuded ependymal line and multicyst formations in periependymal areas were found in both kaolin injection and spinal trauma groups. Ultrastructurally, an apical flattening of the ependyma, microcysts in the ependymal cells, axonal degeneration, demyelination, and loss of ependymal cells adjacent mild spongy were found in the spinal cords of animals in these groups. To the best of our knowledge, this is the first study to investigate the chronic effects of spinal cord injury (SCI) on the CSF pathways and their relationship with the development of spinal deformity in an experimental model of kaolin injection and trauma, using MRI as well as light and transmission electron microscopy. In the light of this study, the severity of spinal cord injury on the development of some degenerative findings in the spinal cord was clear, but further clinical and experimental studies using dynamic imaging techniques will be valuable.