Noninvasive 3D vital imaging and characterization of notochordal cells of the intervertebral disc by femtosecond near-infrared two-photon laser scanning microscopy and spatial-volume rendering

The central region of the intervertebral disc (IVD) in infant humans is made and maintained by notochordal cells (NCs). These cells disappear during maturation to be replaced by mature chondrocyte-like cells. NCs are completely different morphologically from the mature chondrocyte-like IVD cells and have complex and essential functions but little is known about them. Recently, two-photon laser scanning microscopy (TPLSM) using near-infrared (NIR) femtosecond pulsed lasers has emerged as a promising noninvasive optical technique for observing unfixed living 3D biological specimens in situ and in vitro. Several lines of evidence suggest that compared with conventional laser scanning confocal microscopy (LSCM), femtosecond NIR laser-based TPLSM has any number of advantages including 3D resolution without a spatial filter (confocal pinhole), minimal photobleaching, and photodamage above and below the focal plane, and importantly, greater depth penetration. We have thus taken advantage of these unique features of femtosecond laser-based TPLSM for vital 3D imaging in conjunction with advanced spatial-volume rendering modalities to compare morphologies of NCs/clusters from pig caudal discs with chondrocyte-like IVD cells from bovine caudal discs, both in ex vivo tissue and when isolated and grown in vitro within 3D alginate scaffolds. Our results provide evidence that (a) ex vivo notochordal tissue consists of areas with NC clusters, and those dominated by tubular structures of low cell density (b) within 3D in vitro scaffolds the morphology of NC is heterogeneous and the cells contain distinct cytoplasmic vacuole-like structures occasionally including acidic subinclusions (c) a quantitative determination based on 3D spatial and volumetric-rendering reveals an average NC diameter of 22.05 microm (range 11.96-46.63 microm) and NC volume of 9701 microm(3) (2041-36427 microm(3)) whereas chondrocyte-like cells have a mean volume of 3279 microm(3) and diameter of 12.20 microm. Taken together, this study demonstrates that femtosecond TPLSM has unique advantages over other conventional histological and in particular LSCM for high resolution noninvasive vital characterization of notochordal and chondrocyte-like cells of IVD over extended depths beyond 300-500 microm.     

Molecular Mechanisms and Physiological Changes behind Benign Tracheal and Subglottic Stenosis in Adults

Laryngotracheal stenosis (LTS) is a complex and heterogeneous disease whose pathogenesis remains unclear. LTS is considered to be the result of aberrant wound-healing process that leads to fibrotic scarring, originating from different aetiology. Although iatrogenic aetiology is the main cause of subglottic or tracheal stenosis, also autoimmune and infectious diseases may be involved in causing LTS. Furthermore, fibrotic obstruction in the anatomic region under the glottis can also be diagnosed without apparent aetiology after a comprehensive workup; in this case, the pathological process is called idiopathic subglottic stenosis (iSGS). So far, the laryngotracheal scar resulting from airway injury due to different diseases was considered as inert tissue requiring surgical removal to restore airway patency. However, this assumption has recently been revised by regarding the tracheal scarring process as a fibroinflammatory event due to immunological alteration, similar to other fibrotic diseases. Recent acquisitions suggest that different factors, such as growth factors, cytokines, altered fibroblast function and genetic susceptibility, can all interact in a complex way leading to aberrant and fibrotic wound healing after an insult that acts as a trigger. However, also physiological derangement due to LTS could play a role in promoting dysregulated response to laryngo-tracheal mucosal injury, through biomechanical stress and mechanotransduction activation. The aim of this narrative review is to present the state-of-the-art knowledge regarding molecular mechanisms, as well as mechanical and physio-pathological features behind LTS.     

Direct Reprogramming and Induction of Human Dermal Fibroblasts to Differentiate into iPS-Derived Nucleus Pulposus-like Cells in 3D Culture

Intervertebral disc (IVD) diseases are common spinal disorders that cause neck or back pain in the presence or absence of an underlying neurological disorder. IVD diseases develop on the basis of degeneration, and there are no established treatments for degeneration. IVD diseases may therefore represent a candidate for the application of regenerative medicine, potentially employing normal human dermal fibroblasts (NHDFs) induced to differentiate into nucleus pulposus (NP) cells. Here, we used a three-dimensional culture system to demonstrate that ectopic expression of MYC, KLF4, NOTO, SOX5, SOX6, and SOX9 in NHDFs generated NP-like cells, detected using Safranin-O staining. Quantitative PCR, microarray analysis, and fluorescence-activated cell sorting revealed that the induced NP cells exhibited a fully differentiated phenotype. These findings may significantly contribute to the development of effective strategies for treating IVD diseases.     

峡部裂性与退变性腰椎滑脱 MSCT扫描解剖学因素的对照研究

目的：利用多层螺旋 CT（multi-slice spiral CT,MSCT）对峡部裂性与退变性腰椎滑脱解剖学因素进行对比分析。方法选取108例腰椎滑脱患者,滑脱椎体均为 L4,其中峡部裂性腰椎滑脱（峡部裂组）54例,退变性腰椎滑脱（退变组）54例,均经 MSCT 检查确诊。分别测量2组病例的骶骨倾斜角、腰椎前凸角、椎间关节角度、椎弓峡部宽度、滑脱距离、滑脱指数。结果峡部裂组年龄、腰椎前凸角、椎弓峡部宽度小于退变组,关节突关节面与矢状面夹角、滑脱指数大于退变组（P ＜0．05或 P ＜0．01）;2组骶骨倾斜角差异无统计学意义（P ＞0．05）。结论峡部裂性腰椎滑脱患者发病年龄早于、滑脱指数大于退变性腰椎滑脱患者;椎弓峡部发育不良或形态改变（纤细）为峡部裂性腰椎滑脱发病的重要因素;椎间关节面与矢状面夹角变小以及腰椎前凸角增大在退变性腰椎滑脱形成中起一定作用。     

Sustained attention in children with two etiologies of early hydrocephalus.

Several studies have shown that children with spina bifida meningomyelocele (SBM) and hydrocephalus have attention problems on parent ratings and difficulties in stimulus orienting associated with a posterior brain attention system. Less is known about response control and inhibition associated with an anterior brain attention system. Using the Gordon Vigilance Task (Gordon, 1983), we studied error rate, reaction time, and performance over time for sustained attention, a key anterior attention function, in 101 children with SBM, 17 with aqueductal stenosis (AS; another condition involving congenital hydrocephalus), and 40 typically developing controls (NC). In SBM, we investigated the relation between cognitive attention and parent ratings of inattention and hyperactivity and explored the impact of medical variables. Children with SBM did not differ from AS or NC groups on measures of sustained attention, but they committed more errors and responded more slowly. Approximately one-third of the SBM group had attention symptoms, although parent attention ratings were not associated with task performance. Hydrocephalus does not account for the attention profile of children with SBM, which also reflects the distinctive brain dysmorphologies associated with this condition. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Heat transport in magnetohydrodynamic physiological blood flow through a constricted artery

In this paper, we study how the magnetohydrodynamic (MHD) pulsatile flow of blood and heat transfer works through a constricted artery with a flexible wall. The human circulatory network consists of veins and arteries that sometimes contain constrictions, allowing the impact of the applied magnetic field on flow fields to be observed. The walls of the flowing medium are considered to be a function of time. The flowing blood is hypothesized as shear-thinning fluid, emulating Yeleswarapu's viscosity replica. Additionally, we consider the energy equation to understand the impact of a magnetic field on heat transfer rates for such flows. The vorticity transport equation along with the stream function equation is obtained using the vorticity–stream function technique. Numerical solutions of the governing nonlinear MHD equations and energy equation in addition to physically pertinent flow conditions were achieved by adapting a finite difference scheme. Considerable attention has been paid to ensure an accurate comparison between the current and previous results. The two sets of numbers appear to match closely. For an even deeper understanding of the flow and heat transport process, the effects of height of stenosis and diverse physiological parameters on time-averaged wall shear stress (TAWSS), rate of heat transport, and so on are explored in depth through their graphical depiction. In the vicinity of the constriction, it is observed that the separation becomes longer with increasing constriction height. Higher magnetic force strength leads to a reduction in separation length. Newtonian fluids transfer heat more rapidly in their narrowing regions and downstream than fluids with non-Newtonian behavior.     

A proteomics analysis for certain signature proteins of rabbit lacrimal passages after 125I seeds brachytherapy

To search for certain signature proteins and the expression profiles in lacrimal passage stenosis,rabbit models of lacrimal passage stenosis were treated by 125I seed brachytherapy.All the signature proteins were separated by two-dimensional electrophoresis,and identified by mass spectrometry.The results show that the up-regulated proteins are peptidyl-prolyl cis-trans isomerase A(PPIase A),and epidermal fatty acid-binding protein(E-FABP),while the down-regulated proteins are myosin light chain 1(isomer of skeletal muscle),myosin light polypeptide 6(isomer 1 of smooth muscle and non-muscle),myosin light chain 1(isomer of slow-twitch muscle A),isomer 2 of ERC protein 2,and α-crystallin family protein.The proteins may play a role in healing the wound and regulating synaptic active zone of neurons due to correlation to cell apoptosis,proliferation and migration of smooth muscle cell.These provide molecular mechanism for preventing stenosis and restenosis of lacrimal passage.     

一个椎间盘黏弹生物力学模型

椎间盘生物力学模型的研究,由于在临床医学中的重要应用价值而受到了广泛的重视,也是生物力学研究领域中最具挑战性的课题之一.2009年,根据椎间盘的解剖结构及生理特征,曾建立了一个椎间盘横观各向同性复合材料弹性模型,由于椎间盘髓核是流固二相体,因此该模型存在一定的缺陷.为了弥补这种缺陷,也为了使模型更加接近真实,应用弹性力学和渗流力学理论,在以前模型的基础上,建立一个人体椎间盘黏弹模型.结果表明新建的黏弹模型与以前的弹性模型以及相关文献等均具有良好的吻合度,从而证明了新建模型的有效性.新建模型可以合理计算椎间盘在不同体位下的受力和位移等,为临床医学提供最直接的指导和服务.     

狭窄血管多普勒血流信号的频谱仿真分析

通过数值仿真的方法分析狭窄血管内定常流多普勒血流信号的频谱特性,可为利用超声多普勒技术临床诊断血管狭窄提供依据.利用涡度－流函数方法计算狭窄管内的流场分布,估计出血管内不同位置的多普勒信号的功率谱密度函数,从中计算最大频率、平均频率和频谱宽度等参数,并分析它们在不同流速和狭窄程度下所表现出来的特征.