脊柱损伤的法医临床学鉴定研究进展

随着社会和现代交通、建筑事业的发展,脊柱损伤的病人发病率呈上升趋势,尤其道路交通事故为其损伤的首要原因。脊柱损伤的现场急救、临床诊断与治疗为人们所关注。然而,伴随着事件的进一步处理,对病人的损伤进行法医学鉴定显得尤为重要。目前,有关脊柱损伤的鉴定标准仅仅是确定的标准,没有规定标准的适用条件。同时,也易导致鉴定人对标准的理解与应用存在差异,由此导致了鉴定结论的争议。本文对脊柱损伤的法医临床学应用研究进展作一综述。     

法医DNA鉴定中常染色体STR三等位基因型的研究进展

在法医DNA鉴定中,常染色体STR基因座是法医学个体识别和亲子鉴定中最常用的遗传标记。常染色体STR三等位基因型在DNA图谱上可表现为三个等位基因峰、不平衡双等位基因峰以及异常单等位基因峰,通常被认为是一种异常分型。因在法医DNA分析中比较罕见,故而需要对其进行谨慎判定。文章对三等位基因型的特征、发生频率、形成机制、遗传模式等进行了综述,着重探讨了法医学个体识别和亲子鉴定中三等位基因型存在的问题和应对策略,并展望了三等位基因型在法医DNA鉴定及临床实践中的应用。     

脊柱脊髓损伤病人的康复护理

脊柱脊髓损伤是由于各种外力作用于脊柱造成的脊髓压迫或断裂致患者四肢感觉、运动功能发生不同程度的障碍以及大小便功能障碍,严重影响患者生活质量。因此恢复期做好患者的康复护理能有效地预防并发症,改善患者的生活质量,最大程度地让患者重返社会。现将我科2005年1月-2010年12月收治的i00例脊柱脊髓损伤患者的康复护理体会介绍如下。     

串联质谱技术在法医毒物鉴定中的优势

串联质谱技术可同时分析原体及代谢物,不仅适用于分析强极性、热不稳定性、非挥发性及大分子化合物,还可直接分析结合型代谢物,具有样品前处理简便、快速、不需要衍生化等优点。     

真菌的分子生物学鉴定方法研究进展

综述了真菌的分子生物学鉴定方法,包括DNA碱基组成分析,核酸分析技术,真菌核型的脉冲电泳分析。     

30例病毒性心肌炎法医病理学分析

目的:回顾性分析30例病毒性心肌炎的法医病理学特点,为病毒性心肌炎的法医学鉴定提供依据。方法:采用昆明医科大学司法鉴定中心2003年至2014年的30例病毒性心肌炎法医病理鉴定案例进行回顾性分析。结果:30例死亡原因均为病毒性心肌炎致急性心功能障碍死亡,大多为重型病毒性心肌炎,均累及心肌损害,心肌细胞水肿,基质溶解坏死;心肌肌间多量淋巴细胞浸润;局部心肌束排列紊乱,间质纤维组织增生,同时不同程度伴有肺、脑、扁桃体等器官组织的损害。结论:重型病毒性心肌炎起病急,临床症状不典型,进程快,易造成误诊误治,死亡率高,产生医疗纠纷,需要法医学鉴定。病毒性心肌炎法医学鉴定时,以心脏大体所见和组织学观察为主要依据,同时还要特别注意其他机械性损伤,辨别每个死亡原因之间的关系,必要时结合特殊检查,例如对趋化因子MCP-1的检测。     

脑血管畸形致猝死的法医病理研究

目的:分析脑血管畸形致猝死的法医病理理论。方法:选择法医学工作中31例脑血管畸形致猝死病例作为研究对象,分析病例的猝死原因以及病理类型等情况。结果:31例中有25例有明显的外伤史,其中大部分为斗殴造成的外伤,脑血管破裂出血之后导致死亡。脑动脉或静脉畸形是最为常见的脑血管畸形病理类型,28例患者为此类病理类型,畸形的主要发生部位有脑底部、脑室、蛛网膜下腔和脑干;最为常见的出血类型是蛛网膜下腔出血,占14例。结论:就法医病理学研究而言,脑血管畸形致猝死的鉴定具有一定的难度,再结合法医病理学特点的同时可以联合脑血管造影、磁共振检查等措施,确保得到科学、准确的结论。     

富血小板血浆在脊柱脊髓损伤治疗中的应用进展

富血小板血浆是近些年来比较热门的一种血液制品,其来源于自体,且制备方法简单,又富含大量血小板及多种生长因子,能够加速骨愈合,增强骨再生,促进软组织及神经损伤恢复,因此得到了广泛的关注。国内外的研究人员根据富血小板血浆所具有的特点,针对各个方面对其进行了大量的研究实验,并且在临床骨科疾病的治疗中也已经开始了实验性应用,如骨缺损、骨再生,肌腱、韧带及软组织损伤,脊柱脊髓损伤等。尤其是在脊柱脊髓损伤的治疗方面,无论是单独应用富血小板血浆治疗,还是联合应用富血小板血浆与脊髓神经前体细胞、骨髓间充质干细胞等有利于脊髓神经损伤恢复的细胞因子复合物共同治疗,均取得了突破性的进展,为研究脊柱脊髓损伤的治疗提供了新的方向。     

植物无融合生殖鉴定方法的研究进展

由于利用无融合生殖途径可以固定杂种优势,从而改良现有植物的育种策略,因此对无融合生殖的研究已成为生物学科的新生长点,为了利用植物的无融合生殖,首先必须建立,发展和完善一套简单,准确的鉴定体系,目前有关植物无融合生殖的鉴定的方法,主要可以分为5大类型：（1）形态学观察法;（2）显微观察法;（3）生化鉴定法;（4）分子生物学的方法;（5）其它方法,主要对以上5类方法的各各鉴定方法进行了介绍,对几种已被证实有效方法的优缺点进行了比较,并就今后的改进方向进行了探讨。     

《法医人类学经典》评介

法医人类学是体质人类学以及现代法医学的分支学科,它应用体质人类学及其它相关学科的理论和方法,研究、解决法律或司法实践(法律制订、侦查、审判等)涉及的个人特征识别、鉴定(如种族、性别、年龄、身高、面貌特征等)等问题,为案件侦破及审判提供证据.     

间充质干细胞-透明质酸-多聚赖氨酸治疗脊髓损伤的实验研究

目的研究间充质干细胞—透明质酸—多聚赖氨酸复合物治疗脊髓损伤的可行性,评价其治疗效果并探讨其可能机制。方法从人骨髓中分离、培养人骨髓间充质干细胞（human bone marrow mesenchymal stem cell,hBMSC）;制作大鼠脊髓半横断模型,按照实验分组分别将hBMSC、透明质酸-多聚赖氨酸（hyaluronic acid-poly-L-lysine,HA-PLL）、hBMSC-HA-PLL复合物注入损伤区域,单纯损伤组作为对照。术后按照不同时间点评价损伤和移植后的大鼠运动功能。8周后杀死大鼠,观察不同移植组体内轴突和血管生长的情况,对不同细胞、材料及复合物移植对大鼠脊髓损伤修复效果进行评估。结果 hBMSC移植组和hBMSC-HA-PLL移植组的大鼠运动功能的改善显著好于单纯损伤及HA-PLL移植组。电镜结果证实复合物移植组可显著促进轴突和血管生长,新生的轴突和血管结构较为完整。结论 hBMSC具有促进神经功能恢复的作用,将其与HA-PLL相结合,可以促进大鼠脊髓损伤修复,其机制可能包括材料框架作用和hBMSC在体内对大鼠神经细胞的营养作用以及促进微血管的生成。     

Spatiotemporal Patterns of SSeCKS Expression After Rat Spinal Cord Injury

Src suppressed C kinase substrate (SSeCKS) was identified as a PKC substrate/PKC-binding protein, which plays a role in mitogenic regulatory activity and has a function in the control of cell signaling and cytoskeletal arrangement. However its distribution and function in the central nervous system (CNS) lesion remain unclear. In this study, we mainly investigated the mRNA and protein expression and cellular localization of SSeCKS during spinal cord injury (SCI). Real-time PCR and Western blot analysis revealed that SSeCKS was present in normal whole spinal cord. It gradually increased, reached a peak at 3 days for its mRNA level and 5 days for its protein level after SCI, and then declined during the following days. In ventral horn, the expression of SSeCKS underwent a temporal pattern that was similar with the whole spinal cord in both mRNA and protein level. However, in dorsal horn, the mRNA and protein for SSeCKS expression were significantly increased at 1 day for its mRNA level and 3 days for its protein level, and then gradually declined to the baseline level, ultimately up-regulated again from 7 to 14 days. The protein expression of SSeCKS was further analysed by immunohistochemistry. The positively stained areas for SSeCKS changed with the similar pattern to that of protein expression detected by immunoblotting analysis. Double immunofluorescence staining showed that SSeCKS immunoreactivity (IR) was found in neurons, astrocytes, oligodendrocytes of spinal cord tissues within 5 mm from the lesion site. Importantly, injury-induced expression of SSeCKS was co-labeled by active caspase-3 (apoptotic marker), Tau-1 (the marker for pathological oligodendrocyte) and β-1,4-galactosyltransferase 1 (GalT). All the results suggested that SSeCKS might play important roles in spinal cord pathophysiology and further research is needed to have a good understanding of its function and mechanism. Feng Xiao and Min Fei contributed equally to this work.     

大鼠放射性脊髓损伤脊髓血流量变化规律

目的:放射性脊髓损伤(Radiation spinal cord injury,RSCI)是头颈部、胸部及上腹部肿瘤放射治疗和射线意外照射时的常见并发症,一般认为,白质坏死、脱髓鞘为其主要的病理学变化.然而,越来越多的证据表明血-脊髓屏障破裂和血管通透性增加等血管损伤远早于白质坏死和脱髓鞘改变.所以本文阐明大鼠放射性脊髓损伤病理生理过程中脊髓血流量变化规律.方法:将60只Sprague-Dawley (SD)大鼠随机分为12组,1组为对照,其余11组采用60Co放射治疗机行30 Gy大鼠颈髓C2-T2单次照射,剂量率为153 cGy/min,源皮距为80 cm,照射时长为1153 s,照射范围为2.0× 1.0 cm,对照组大鼠于麻醉后置于60Co放射治疗机下,佯照,照射前及照射后分别采用激光多普勒法测量脊髓血流量,11组大鼠于照射前以及照射后1、3、7、14、21、30、60、90、120、150、180天进行测量,以照射前测量值为基数,各时间点以基数的百分比表示该时间点脊髓血流量.结果:大鼠放射性脊髓损伤后,脊髓血流量在照射早期即有降低,照射后90天达到最低,随后脊髓血流量进入平台期.结论:阐明了大鼠放射性脊髓损伤后脊髓血流量的变化规律.大鼠放射性脊髓损伤可影响脊髓血流量,导致脊髓长期处于持续低灌流、缺血缺氧状态,最终导致脊髓不可逆性损伤.临床上放射性脊髓损伤的病人感到疲乏无力,出现神经系统的症状体征,通常死于脑疝.本文为临床上疲乏无力,出现神经系统的症状体征,死于脑疝放射性脊髓损伤的病人的早期防治提供病理生理基础.     

Combination of Dexamethasone and Aminoguanidine Reduces Secondary Damage in Compression Spinal Cord Injury

The study was performed to investigate the effect of combination therapy with aminoguanidine (AG) and dexamethasone (DEX) on the compression spinal cord injury (SCI) in rat. Compared to the control group, the combination therapy group with AG (75 mg/kg) and DEX (0.025 mg/kg) significantly reduced the degree of (1) spinal cord edema, (2) the permeability of blood spinal cord barrier (measured by ^99mTc-Albumin), (3) infiltration of neutrophils (MPO evaluation), (4) cytokines expression (tumor necrosis factor-α and interleukin-1β), and (5) apoptosis (measured by Bax and Bcl-2 expression). In addition, we have also clearly demonstrated that the combination therapy significantly ameliorated the recovery of limb function (evaluated by motor recovery score). Taken together, our results clearly indicated for the first time that strategies targeting multiple proinflammatory pathways may be more effective than a single effector molecule for the treatment of SCI.     

A Surgery Protocol for Adult Zebrafish Spinal Cord Injury

Adult zebrafish has a remarkable capability to recover from spinal cord injury,providing an excellent model for studying neuro-regeneration. Here we list equipment and reagents,and give a detailed protocol for complete transection of the adult zebrafish spinal cord. In this protocol,potential problems and their solutions are described so that the zebrafish spinal cord injury model can be more easily and reproducibly performed.In addition,two assessments are introduced to monitor the success of the surgery and functional recovery:one test to assess free swimming capability and the other test to assess extent of neuroregeneration by in vivo anterograde axonal tracing.In the swimming behavior test,successful complete spinal cord transection is monitored by the inability of zebrafish to swim freely for 1 week after spinal cord injury,followed by the gradual reacquisition of full locomotor ability within 6 weeks after injury.As a morphometric correlate,anterograde axonal tracing allows the investigator to monitor the ability of regenerated axons to cross the lesion site and increasingly extend into the gray and white matter with time after injury,confirming functional recovery.This zebrafish model provides a paradigm for recovery from spinal cord injury,enabling the identification of pathways and components of neuroregeneration.     

Analysis of Rat Hindlimb Muscle Proteins by Two-Dimensional Gels Following Spinal Cord Injury

Proteins from extensor digitorum longus (EDL), plantaris (PLN), and soleus (SOL) muscles of adult, female rats were examined by high resolution two-dimensional gel electrophoresis up to 4 weeks following spinal cord transection. The electrophoretograms were analyzed by computer imaging and densitometry. Reproducible and significant changes in the relative concentrations of several proteins in each muscle type were detected. Whereas changes involving the largest number of proteins were observed in SOL, changes in EDL and PLN were also detected. In SOL, approximately 50% of the altered proteins increased in concentration and the remaining decreased: Actin and myosin light chains LCF-1 and LCF-2 were among those proteins whose concentrations increased, whereas myosin light chains LCS-1 and LCS-2 were among those proteins whose concentrations decreased. The present findings regarding the reversal in myosin light chain composition provide biochemical support for previously published data on changes in contractile properties of muscles following spinalization. In EDL, the relative concentration of only one protein was elevated in a time-dependent manner. The concentrations of two protein species in PLN were increased following cord transection. These findings provide new biochemical markers on the effects of spinal cord on gene expression in specific hindlimb skeletal muscles.     

Possible Involvement of Interleukin-1 in Cyclooxygenase-2Induction After Spinal Cord Injury in Rats

Abstract : A standardized compression injury of rat spinal cord brought about a time-dependent biphasic production of thromboxane A₂ (detected as thromboxane B₂) and prostaglandin I₂ (detected as 6-ketoprostaglandin F_1α. Thromboxane B₂ was predominant during the first 1 h, whereas the 6-ketoprostaglandin F_1α level exceeded that of thromboxane B₂ at 8 h postinjury. As examined by inhibitor experiments and northern blotting, cyclooxygenase-1 was responsible for the first phase, and cyclooxygenase-2 was involved in the second phase. On compression injury the levels of interleukin-1α and -1β detected as mRNA and protein increased and peaked at 2-4 h. Injection of exogenous interleukin-1 α into the spinal cord resulted in an increase of cyclooxygenase-2 mRNA content and a predominant production of 6-ketoprostaglandin F_1α resembling the second phase of eicosanoid production. Concomitantly, extravascular migration of polymorphonuclear leukocytes was enhanced after the interleukin-1α injection. These cells together with vascular endothelial cells and glial cells were stained positively with an anti-cyclooxygenase-2 antibody. The results suggest that the immediate eicosanoid synthesis after spinal cord injury was due to the constitutive cyclooxygenase-1 and the delayed synthesis of eicosanoids was attributable to the induction of cyclooxygenase-2 mediated by interleukin-1 α.     

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.     

G—CSF对大鼠脊髓损伤后神经细胞凋亡及caspase-3表达的影响

目的：通过观察粒细胞集落刺激因子（G—CSF）对大鼠急性脊髓损伤后神经细胞凋亡及Caspase-3的表达的影响,探讨其对脊髓保护的作用机制。方法：32只Vistar大鼠随机分成2组：对照组和治疗组,每组16只,采用改良的Allen’s装置制成大鼠急性脊髓损伤模型。在术前及术后对大鼠进行BBB功能评分观察大鼠的神经功能变化;用免疫荧光法检测脊髓损伤后个时间点Caspase-3表达;原位脱氧核糖核苷酸末端转移酶介导的缺口末端标记法（Tunel法）检测凋亡细胞。结果：大鼠急性脊髓损后Caspase-3表达与细胞凋亡均呈现先升高后下降的趋势,损伤后3d可见大量的Caspase-3和TUNEL阳性细胞,7d时达到高峰,此后表达逐渐减少,21d时仍可见少量阳性细胞。与对照组比较,G—CSF治疗组各时间点Caspase-3表达和细胞凋亡显著降低,功能恢复显著优于对照组,差异具有统计学意义。结论：G-CSF可以减轻大鼠脊髓损伤后的神经元凋亡,从而发挥神经保护作用,其作用可能是通过抑制Caspase-3的表达使脊髓损伤周围神经细胞凋亡显著下降而实现的。