pSVPoMcat微基因修饰雪旺氏细胞移植对损伤脊髓前角神经元的保护作用研究

目的：探讨pSVPoMcat微基因修饰雪旺氏细胞移植对损伤脊髓前角神经元的保护作用。方法：采用大鼠脊髓半横切损伤模型，将实验动物分为3组：pSVPoMcat微基因修饰雪旺细胞移植组（A组），单纯雪旺氏细胞移植组（B组），损伤对照组（C组）。对脊髓切片行Nissl染色，酸性磷酸酶（ACP）组织化学染色及原位末端标记（TUNEL）法染色，并用联合行为记分（CBS）观察大鼠神经功能恢复情况。结果：A、B、C三组前角神经元存活率呈显著性差异（A＞B＞C，P＜0．01）；A组ACP变化幅度明显降低（A＜B＜C）；细胞凋亡率为C＞B＞A。大鼠神经功能恢复也出现了相同的变化趋势。结论：pSVPoMcat微基因修饰雪旺氏细胞移植对脊髓损伤前角神经元有保护作用并促进大鼠损伤脊髓功能的恢复。     

甲基强的松龙治疗急性脊髓损伤的实验研究及临床疗效观察

目的：通过动物实验和临床疗效观察评价甲基强的松龙（MP）对急性脊髓损伤的治疗作用。方法：动物实验：将90只SD成年大鼠（体重230-250g）随机分为3组（A组：假手术组；13组：尾静脉注射生理盐水组：C组：尾静脉注射甲基强的松龙组），各组分别于术后4h、8h、1d、3d、7d进行运动功能评分、运动诱发电位检查、HE染色观察病理改变。临床研究：对2003-2005年我科收治的85例患者给予大剂量甲基强的松龙治疗（MP组），观察疗效，与前期未用甲基强的松龙治疗的93例（对照组）比较，评价甲基强的松龙的治疗作用。结果：动物实验显示，A、B、C组大鼠在术后8h、1d、3d、7d时相点的BBB评分、运动诱发电位潜伏期、病理改变存在组间差异，A组几乎为正常表现，而C组恢复情况优于B组，更接近于A组。临床观察显示，MP组神经功能恢复速度和程度均优于对照组。结论：MP能够减轻急性脊髓损伤的水肿程度，减少神经细胞凋亡，加快脊髓神经功能恢复速度和程度。     

大剂量甲基强的松龙对脊髓损伤后神经细胞凋亡的影响

目的　探讨大剂量甲基强的松龙对脊髓损伤后神经细胞凋亡的影响。　方法　将60只大鼠分为脊髓损伤后应用大剂量甲基强的松龙组(A组,30只)和单纯脊髓损伤组(B组,30只)。损伤后1,3,7,14,28　d,对脊髓损伤区进行细胞凋亡的检测（原位末端脱氧核糖核苷酸转移酶介导dUTP标记法,TUNEL）以及Bcl-2蛋白表达的测定（免疫组化SP法）。　结果　A、B两组中均发现凋亡细胞及Bcl-2蛋白阳性表达。大鼠脊髓损伤后3d图像分析表明,细胞凋亡率分别为A组（12.43±5.16）,B组（36.27±6.28）;Bcl-2蛋白阳性细胞分别为A组（37.68±5.83）,B组（21.48±7.83）。两组比较,差异有显著性意义(P<0.01和0.05)。　结论　脊髓损伤可导致神经细胞凋亡,大剂量甲基强的松龙能抑制细胞凋亡。     

大鼠脊髓急性损伤后神经细胞的凋亡

目的 研究脊髓急性损伤后神经细胞的凋亡及其分布特点。方法 44只大鼠分为对照组（4只）和损伤组（40只）。损伤组脊髓（T8．9）经中度压迫致伤后,分别在伤后30min、2,4,8,24,48,72h和7,14,21d处死取材（每时相点鼠数＝4只）。应用HE、尼氏染色及凋亡细胞原位末端标记法对脊髓组织进行细胞检测。结果 伤后4h,损伤段及邻近段可见末端标记阳性神经细胞;伤后8h,损伤段灰质中阳性细胞数达高峰;伤后24h,白质中阳性胶质细胞数达高峰;伤后72h,相邻节段阳性细胞数达高峰。阳性细胞以白质中胶质细胞为主,主要分布于相邻手段。结论 脊髓损伤后神经细胞凋亡是继发损伤期的重要病理变化。     

脊髓减压病大鼠神经细胞凋亡及神经生长因子的保护作用

目的探讨脊髓减压病对大鼠脊髓神经细胞凋亡的影响及神经生长因子的保护作用。方法SD大鼠随机分为正常对照组、安全减压组、生理盐水组和神经生长因子治疗组，建立大鼠脊髓减压病模型．于出舱后0，10，30，60，120min经蛛网膜下腔各注入20μl生理盐水或神经生长因子，在6，24，48，72h用TUNEL法标记脊髓神经细胞凋亡，免疫组织化学方法检测TNF-α蛋白表达。结果脊髓减压病大鼠出舱后6h组出现少量脊髓神经细胞凋亡及TNF—α蛋白表达，24h组阳性染色明显增强，48h组达到高峰，神经生长因子治疗后神经细胞凋亡及TNF—α蛋白表达明显减少，与生理盐水组比较差异有统计学意义（P〈0．05）。TNF—α蛋白表达与神经细胞凋亡存在正相关（P〈0．05）。结论神经细胞凋亡是脊髓减压病大鼠的重要病理变化，TNF—α可能参与了脊髓减压病继发损伤，神经生长因子可以减轻脊髓减压病的神经细胞损伤。     

高压氧治疗脊髓损伤后脊髓组织中增殖细胞核抗原的表达变化

为探讨高压氧(HBO)对脊髓损伤(SCI)大鼠脊髓组织中增殖细胞核抗原(PCNA)表达的影响，笔者观察了HBO治疗后脊髓的组织学改变及PCNA的表达变化。结果显示，损伤后，脊髓组织出血、水肿、空泡变性、神经纤维排列紊乱、断裂；PCNA强阳性表达于神经元细胞中。治疗组中，0．1MPa HBO组的HE及免疫组织化学染色结果与损伤组相似；而0．25MPa HBO组则组织水肿消失，出血、空泡变性减轻，神经细胞形态恢复，结构排列相对完整；PCNA的表达强度较损伤组下降。说明PCNA能明确提示细胞的增殖指数；HBO治疗可阻止或减轻脊髓损伤的病理变化，有效修复脊髓组织的结构和功能。     

脑源性神经营养因子转基因细胞移植和甲基强的松龙对损伤脊髓caspase-3表达的影响

目的 探讨大鼠脊髓损伤后腺病毒介导的脑源性神经营养因子（AxCA-BDNF）体外转基因成肌细胞移植和静脉内注射大剂量甲基强的松龙（MP）对大鼠脊髓细胞caspase-3表达的影响。方法 将120只大鼠分为脊髓挫伤组（A组）、脊髓挫伤后AxCA-BDNF基因转染的成肌细胞移植组（B组）、脊髓挫伤后静脉内注射大剂量甲基强的松龙治疗组（C组）、AxCA-BDNF基因转染的成肌细胞移植和静脉内注射大剂量，甲基强的松龙治疗组（D组）。伤后1，3，7，14，28d，应用免疫组化法对脊髓损伤区进行细胞凋亡caspase-3蛋白表达的测定，并采用计算机图像分析技术进行定量分析。应用行为学和电生理检查方法观察大鼠运动功能恢复情况。结果A、B、C、D四组中均发现凋亡caspase-3蛋白阳性表达细胞；各组caspase-3免疫反应阳性细胞数依次为A〉B〉C〉D组（P〈0．05），与大鼠后肢功能恢复有平行的变化趋势。结论 体外转基因成肌细胞移植和大剂量甲基强的松龙能抑制脊髓损伤后的细胞凋亡。     

神经节苷脂对大鼠脊髓损伤后神经元的保护作用

目的 通过观察单唾液酸四己糖神经节苷脂( monosialotetrahexosyl gangliosides,GM-1)对大鼠脊髓损伤后微管相关蛋白-2（microtubule - associated protein 2,MAP -2）和胆碱乙酰转移酶( choline acetyhransterase,ChAT)的表达及运动功能的影响,探讨GM -1对大鼠脊髓损伤后神经元的保护作用及其机制。方法 Wistar雌性大鼠66只（体重260～ 300 g）,随机取6只作为正常对照组,余60只采用改良Allen打击法于Tt0节段制作大鼠急性脊髓损伤模型,并按随机数字表法分为GM -1组（A组）和等渗盐水对照组（B组）,每组30只大鼠。分别于术后1,3,7,14和28 d采用改良Tarlov评分法评价大鼠脊髓损伤后后肢运动功能恢复程度后处死取材,每组各时相点6只大鼠。以免疫荧光染色观察大鼠脊髓损伤后MAP -2和ChAT的表达情况。结果 术后7d起,Tarlov评分在A、B组间比较差异有统计学意义(P＜0.05),即A组大鼠运动功能恢复优于B组。免疫荧光染色发现,A组术后各时相点MAP -2及ChAT呈阳性表达的细胞数量较B组同时相点明显增多（P＜0.05）。结论 脊髓损伤后大鼠神经功能的恢复与MAP -2及ChAT的表达呈一定的相关性;GM -1可通过增强脊髓损伤后MAP -2及ChAT的表达来保护神经元。     

细胞外三磷酸腺苷对大鼠脊髓损伤后微管相关蛋白-2表达及运动功能恢复的影响

目的 研究细胞外三磷酸腺苷（ATP）对大鼠脊髓损伤后微管相关蛋白-2（MAP-2）表达运动功能恢复的影响。方法 健康成年Wistar大鼠66只，随机取6只作为正常对照组，余60只制作成脊髓打击伤动物模型，随机分为两组：A组（ATP组）和B组（对照组），每组30只大鼠。伤后1，3，7，14，28d取材，应用免疫组织化学方法观察MAP-2的表达，采用计算机图像分析系统，进行定量分析；并用改良的Tarlov评分观察大鼠脊髓损伤后运动功能的恢复情况。结果脊髓损伤后14d和28d，A组大鼠MAP-2的表达明显强于B组（P＜0．05）；损伤后14d和28d，A组大鼠改良Tarlov评分明显大于B组（P＜0．05）．结论细胞外ATP能促进大鼠损伤脊髓表达MAP-2，并能促进大鼠脊髓损伤后运动功能的恢复。     

雌激素影响急性大鼠脊髓损伤后细胞凋亡的相关性实验研究

目的在建立大鼠脊髓损伤模型基础上,观察不同时间点雌激素对脊髓胶质细胞,神经元凋亡的影响,以期为临床急性脊髓损伤的治疗提供理论依据。方法健康成年大鼠72只,随机分为两组,其中单纯损伤组（单纯打击损伤）36只,雌激素组（手术加雌激素）36只。脊髓损伤动物模型的制备：用自制Allen打击器25gcm致伤力撞击脊髓,制成脊髓损伤动物模型。雌激素组每日肌注雌激素（100μg／kg）直至处死,单纯损伤组仍每日肌注生理盐水0．5ml直至处死。组织切片的制备：脊髓损伤后1d、3d、5d、8d、14d、21d共6个时间段分批处死动物。4％多聚甲醛心肌灌注固定24h,切取每只大鼠T5～T13节段脊髓组织,常规制成切片,给于Bcl－2检测,TUNEL原位末端标记,检测大鼠脊髓损伤后细胞凋亡的情况。脊髓损伤后2周给予Gale评分及斜板维持试验。结果Bcl－2蛋白检测：在脊髓损伤后的第1天,脊髓组织即开始较高表达Bel－2蛋白。单纯损伤组Bcl－2高峰发生在损伤后3天,而雌激素组伤后8天达到高峰,此时Bcl－2蛋白不仅表达在神经细胞中,更多的在胶质细胞中大量表达,且此状态一直维持到伤后14天才开始下降,伤后21天仅少量表达（P〈0．05）。TUNEL原位标记检测：单纯损伤组24小时已出现不少阳性细胞,以胶质细胞为主．3～8天达到高峰,此后逐渐回落,但21天时仍有阳性细胞,应用雌激素治疗后,凋亡细胞明显减少（P〈0．05）。脊髓损伤后2周Gale评分及斜板维持率雌激素组优于单纯损伤组（P〈0．01）。结论雌激素在脊髓损伤中通过改善微循环,促进Bcl－2蛋白的表达,清除氧自由基,抗氧化作用能够抑制脊髓损伤早期神经细胞和胶质细胞的凋亡,从而减轻脊髓继发性损伤．促进脊髓神经功能的恢复。所以脊髓损伤后早期应用雌激素配合其它药物阻止神经细胞和胶质细胞的凋亡可能有助于减轻脊髓损伤的损害程度,促进脊髓神经功能的恢复,为SCI的治疗提供一个新的途径。     

Knee injury and Osteoarthritis Outcome Score (KOOS) - validation of a Swedish version

The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living.     

Endovascular management of carotid-cavernous fistulas

Objective To investigate endovascular treatment of traumatic direct carotid-cavernous fistulas （CCF） and their complications such as pseudoaneurysms. Methods： Over a five-year period, 22 patients with traumatic direct CCFs were treated endovascularly in our institution. Thirteen patients were treated once with the result of CCF occluded, 8 twice and 1 three times. Treatment modalities included balloon occlusion of the CCF, sacrifice of the ipsilateral internal carotid artery with detachable balloon, coll embolization of the cavernous sinus and secondary pseudoaneurysms, and covered-stem management of the pseudoaneurysms. Results All the direct CCFs were successfully managed endovascularly. Four patients developed a pseudoaneurysm after the occlusion of the CCF with an incidence of pseudoaneurysm formation of 18.2% （4/22）. A total number of 8 patients experienced permanent occlusion of the ICA with a rate of ICA occlusion reaching 36.4% （8/22）. Followed up through telephone consultation from 6 months to 5 years, all did well with no recurrence of CCF symptoms and signs. Conclusion Traumatic direct CCFs can be successfully managed with endovascular means. The pseudoaneurysms secondary to the occlusion of the CCFs can be occluded with stent-assisted coiling and implantation of covered stents.     

The acutely limping child

Acute limping may be the result of multiple pathologies in children. The differential diagnosis varies based on the age of the child. Irrespective of age, the initial imaging work-up includes AP and frog leg radiographs of the pelvis and ultrasound; MRI may sometimes be helpful. In children less than 3 years, infections and trauma are most frequent. MRI is the imaging modality of choice when osteomyelitis is clinically suspected. Between the ages of 3 and 10 years, transient synovitis of the hip and Legg-Calvé-Perthes disease are main considerations but infection, inflammation and focal bony lesions are also considered. In children over 10 years, slipped capital femoral epiphysis also is considered.     

Do Balance Board Training Programs Reduce the Risk of Ankle Sprains in Athletes?

Introduction Ankle sprains are the most common musculo-skeletal injury that occurs in athletes,particularly in sports that require jumping and landing on one foot such as soccer,and basketball(1-4).These injuries often result in significant time loss from participation,long-term disability,and have a major impact on health care costs and resources(5-8).     

High Intensity Interval Training:New Insights

KEY POINTS ·High-intensity interval training(HIT)is characterized by repeated sessions of relatively brief，intermittent exercise.often performed with an“a11 out”effort or at an intensity close to that which elicits peak oxygen uptake(i.e.，≥90%of VO2 peak).     

Developmental validation of the PowerPlex(®) ESI 16 and PowerPlex(®) ESI 17 Systems: STR multiplexes for the new European standard

In response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega^® developed a suite of four new DNA profiling kits. This paper describes the developmental validation study performed on the PowerPlex^® ESI 16 (European Standard Investigator 16) and the PowerPlex^® ESI 17 Systems. The PowerPlex^® ESI 16 System combines the 11 loci compatible with the UK National DNA Database^®, contained within the AmpFlSTR^® SGM Plus^® PCR Amplification Kit, with five additional loci: D2S441, D10S1248, D22S1045, D1S1656 and D12S391. The multiplex was designed to reduce the amplicon size of the loci found in the AmpFlSTR^® SGM Plus^® kit. This design facilitates increased robustness and amplification success for the loci used in the national DNA databases created in many countries, when analyzing degraded DNA samples. The PowerPlex^® ESI 17 System amplifies the same loci as the PowerPlex^® ESI 16 System, but with the addition of a primer pair for the SE33 locus. Tests were designed to address the developmental validation guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), and those of the DNA Advisory Board (DAB). Samples processed include DNA mixtures, PCR reactions spiked with inhibitors, a sensitivity series, and 306 United Kingdom donor samples to determine concordance with data generated with the AmpFlSTR^® SGM Plus^® kit. Allele frequencies from 242 white Caucasian samples collected in the United Kingdom are also presented. The PowerPlex^® ESI 16 and ESI 17 Systems are robust and sensitive tools, suitable for the analysis of forensic DNA samples. Full profiles were routinely observed with 62.5 pg of a fully heterozygous single source DNA template. This high level of sensitivity was found to impact on mixture analyses, where 54–86% of unique minor contributor alleles were routinely observed in a 1:19 mixture ratio. Improved sensitivity combined with the robustness afforded by smaller amplicons has substantially improved the quantity of data obtained from degraded samples, and the improved chemistry confers exceptional tolerance to high levels of laboratory prepared inhibitors.