荧光素双标记和电镜观察神经端侧吻合修复神经缺损的实验研究

目的　观察兔耳大神经端侧吻合后轴突再生情况。方法　采用兔耳大神经端侧吻合模型 ,右侧耳大神经作为供神经 ,在适当部位的外膜上纵行切开 2 .0mm(开窗 ) ,左侧耳大神经在近端切断 ,其远侧段作为受神经 ,随机分为 1月组、2月组、4月组和正常对照组。结果　荧光素组 :右侧C2～ 3背根节内可见 3种不同结构的标记细胞 ,即FB(胞浆 )单标记细胞、NY(胞核 )单标记细胞和FB/NY(胞浆 /胞核 )双标记细胞。每组标记细胞数量不同 ,以 2～ 4月间再生较多。FB标记细胞分布广泛 ,大、中、小型细胞均有 ,NY单标记细胞分布区域相对集中。双标记细胞多分布于背根节的中央区。电镜观察组 :受神经术后 1个月 ,距吻合口 0 .5cm之神经 ,有无髓纤维溃变 ,结构紊乱 ,轴突和髓鞘蹦解 ,在溃变有髓纤维髓鞘外偶见再生的较细的无髓纤维 ,2个月时 ,有大量的无髓纤维再生 ,雪旺氏细胞增殖活跃 ,并可见新生的直径细小的有髓纤维 ,髓鞘较薄 ,内部线粒体结构清晰。术后 4个月 ,新生的有、无髓纤维较 2个月时增多 ,以无髓纤维增多显著。结论　神经端侧吻合后轴突可以再生。     

神经生长因子在位听神经核内的定位分布

 目的 采用免疫细胞化学方法,观察正常豚鼠耳蜗神经核及前庭神经核内神经生长因子(Nerve growth factor,)精确定位,为进一步研究打下基础。方法 采用免疫组化法。结果 NGF-ir免疫阳性产物见于前庭核所属的内侧核,密集,上、外、下核呈散在分布,在4个亚核之间阳性神经纤维分布较多。耳蜗核的腹、背侧核团内阳性细胞相差不多,背略多于腹侧核。结论 提示NGF在豚鼠位听神经核团内,具有营养、保护及维持正常神经功能作用。     

光学方法二维成像前庭核神经元的突触传导

为探索前庭核神经元传导的时空二维特征，用吸光性电压敏感染料RH155染色新生小鼠的脑干切片20min，电刺激前庭神经，用光学记录膜电位技术成像前庭核的神经电活动。结果显示，电刺激前庭神经后光学记录显示前庭核出现神经电兴奋，在部分实验中观察到同侧及对侧前庭核的兴奋传导；前庭核神经兴奋有激发延迟和高峰延迟；所记录的光学信号具有光吸收波长特性，表明光学记录的可靠性；光学信号包括尖峰状快反应信号和持续较长时间的慢反应信号，所有兴奋信号被20μmol/L河豚毒(特异性钠通道阻断剂)不可逆性消除，表明快反应信号来源于钠通道介导的动作电位，慢反应信号被无钙液可逆性阻断，表明慢反应信号可能为兴奋性突触后电位。研究表明，光学记录膜电位方法可以在神经细胞群的水平上直观观察脑干前庭核神经电活动的时空二维方式及其兴奋性突触传递过程。     

神经干细胞超顺磁性氧化铁标记及体内外MRI示踪

目的使用超顺磁性氧化铁(SPIO)纳米粒子对神经干细胞进行体外标记,并在体外及活体移植后对标记细胞进行MRI示踪。方法实验动物包括13只SD大鼠。联合应用SPIO及多聚左旋赖氨酸(PLL)通过受体介导的内吞作用标记神经干细胞,对标记细胞分别进行普鲁士蓝染色、电子显微镜观察、体外MRI示踪及活体移植后体内MRI示踪。体外1.5T及4.7TMR扫描对象分为5组,分别为5×105个标记后培养1d的细胞、5×105个相同时期未标记的细胞、提取标记细胞后的含铁培养基、相应的无铁培养基及蒸馏水。扫描序列包括轴面T1WI、T2WI及T2WI,并在4.7TMR仪上测量标记细胞的弛豫率R2及R2的变化。结果(1)该方法标记神经干细胞的有效率为100%,普鲁士蓝染色证实了每个标记细胞胞质内有多少不等的蓝染铁颗粒。(2)体外1.5T及4.7TMRI显示,标记细胞同未标记细胞相比,T1WI信号强度平均上升分别为20.53%及24.06%,T2WI信号强度平均下降分别为40.78%及50.66%,T2WI信号强度平均下降46.57%及53.70%。1.5TMRI上标记细胞的信号改变在T2WI与T2WI之间差异无统计学意义(F=3.06,P>0.05),而T2WI及T2WI与T1WI之间差异均有统计学意义(F=6.5,P<0.05)。(3)4.7TMRI测量未标记细胞和标记细胞的T2分别为516ms及77ms,其弛豫率R2分别为1.94s-1及12.98s-1;T2分别为109ms及22.9ms,其弛豫率R2分别为9.17s-1及43.67s-1。(4)标记细胞活体移植1周后行1.5TMR检查,见移植部位在T2WI及T2WI上呈显著低信号,而对照侧未见低信号。结论该方法标记神经干细胞简单高效,标记细胞弛豫率R2及R2明显提高,并能采用1.5TMRI对标记细胞进行活体内外示踪研究。     

Coriolis加速度刺激适应豚鼠前庭传出系统神经元活性的变化

目的 探索动物前庭适应产生的神经生理基础。方法 采用胃电图波幅值作为观察植神经反应的指标,利用小剂量、多次重复的Ｃｏｒｉｏｌｉｓ加速度刺激,建立前庭适应动物模型。采用免疫组化染色和显微相对灰度分析方法,观察动物在产生前庭适应前后,以及前庭适应消退后,脑干前庭传出系统神经元细胞活性的变化,及其与胃电图波幅值变化的联系。结果 建模成功豚鼠受到运动刺激时,刺激前后的胃电图波幅相近。在前庭适应的豚鼠身上,其脑干前庭传出系统神经元内胆碱乙酰化酶的含量明显增加;但在前庭适应消退后,这种含量的差别消失。这表明在豚鼠出现适应性时此神经元对于前庭终器感觉细胞的抑制性作用明显增强。结论 脑干前庭传出系统神经元可能通过降低前庭终器感觉细胞向中枢神经系统的信号发放的程度,参与了适应性的产生过程。     

用光学成像研究前庭核神经元谷氨酸受体的兴奋性传导

目的　在神经细胞群的水平上二维动态观测前庭核 (VN)神经元兴奋性谷氨酸递质受体的传导机制。方法　自新生小鼠制备脑干切片 ,用吸光性电压敏感染料RH 15 5染色 ,并成像于 16× 16elementsPhotodiodeArrays光学记录系统 ,电刺激前庭神经断端。为了观察谷氨酸是否为前庭核神经元的兴奋性递质 ,用受体拮抗剂 (NMDA受体拮抗剂APV ,non NMDA受体拮抗剂CNQX)灌流脑片。结果　电刺激前庭神经断端后光学记录显示兴奋传导至VN区域 ;神经兴奋有激发延迟和高峰延迟 ;光学记录可以同步观察氨基酸拮抗剂对多个神经元核团兴奋性传递的作用 ,NMDA受体拮抗剂APV和non NMDA受体拮抗剂CNQX对兴奋性突触后电位 (EPSP)的抑制作用。APV和CNQX灌流后神经元的EPSP被抑制 ,减低比率分别为 6 4 9%± 9 0 6 %和 35 1%± 9 0 6 % (n =18)。结论　光学记录膜电位方法可以在神经细胞群的水平上直观观察VN神经电活动的时空二维方式及其兴奋性突触传递过程 ;谷氨酸是VN的神经元EP SP传导的兴奋性递质 ;谷氨酸NMDA和non NMDA受体均参与介导VN神经元EPSP,在新生期以NMDA受体为主     

尾吊大鼠脑内和核团iNOS和NPY变化研究（摘要）

尽管大量研究表明微重力环境能够导致神经系统一些功能改变,但是其基础生理改变等还不是很清楚。为此,我们采用尾吊SD大鼠制作模拟微重力动物模型,用IMC的方法观察脑内孤束核、网状核、前庭神经核、基底核等核团的iNOS、     

胎鼠神经干细胞超顺磁性氧化铁颗粒标记移植后MRI研究

目的:探讨超顺磁性氧化铁颗粒(SPIO)标记神经干细胞的方法,以及标记细胞正常大鼠脑内移植后MR成像的方法学研究。方法:多聚左旋赖氨酸介导的SPIO标记胎鼠神经干细胞,进行台盼兰染色和普鲁士兰染色分别检测标记细胞的存活率和标记率。选取SD大鼠15只,简单随机法分为3组:第1组于大鼠右侧尾状核移植未标记的NSCs,第2组于大鼠右侧尾状核移植标记的NSCs,第3组右侧尾状核移植游离的SPIO颗粒,移植后第1、4、8周进行MRI。8周后处死大鼠,行组织切片普鲁士兰染色。结果:体外标记的神经干细胞普鲁士兰染色发现铁颗粒聚集于细胞浆内,标记率为100%;标记细胞与未标记细胞的台盼兰染色结果无显著差异。移植后MRI,第1组注射点未见低信号影;第2组注射点T2WI及GRE序列均可见类圆形低信号影;第3组大鼠注射后1周注射点可见低信号影,4周后低信号影变淡且边缘变模糊,8周后低信号影T2WI已不明显。与T2WI序列比较,GRE序列显示标记细胞更清晰,但显示范围较扩散。脑组织切片的普鲁士兰染色显示,第1组大鼠脑组织切片未见异常蓝染细胞,第2组注射点可见蓝染细胞,第3组注射点可见稍许散在蓝色颗粒状物质。结论:多聚左旋赖氨酸介导下SPIO可用于标记神经干细胞,标记细胞移植后MRI可以无创性观察移植神经干细胞的位置及分布情况。     

正常和异常内耳道的MR表现

作者用1.5TMR机检查了20例(40耳)正常人和48例异常患者的内耳道。正常内耳道直径最小2～3mm,最大12mm,平均5 mm;血管-神经束直径2～3 mm。面神经位于内耳道前上部,蜗神经在前下部,前庭神经的上、下支则位于后部。约30～50%可见小脑前下动脉在内耳道内形成一血管袢。听神经外侧部由髓鞘包裹而内侧部则由胶质纤维包裹;大多数听神经瘤起自内耳道内二者结合     

移植神经干细胞促进脊髓全横断大鼠结构与功能修复的研究

目的　探讨移植神经干细胞对脊髓全横断性大鼠部分结构与功能修复的影响。方法　在脊髓全横断处移植神经干细胞 6 0d后 ,在横断处下方 3mm注射荧光金逆行标记轴突再生的上运动神经元 ,用免疫组织化学检测神经干细胞在宿主内的分化 ,同时用体视学方法观测脑干细核和大脑皮质感觉运动区内锥体细胞层的神经元密度变化。用BBB评分法和爬网格法观测大鼠后肢运动功能的恢复等。结果　移植的神经干细胞能在宿主内存活并向前后方向迁移到脊髓内 ,部分神经干细胞分化为GFAP、NF - 2 0 0和GAP -43阳性细胞。移植神经干细胞后在红核和感觉运动区内锥体细胞层可见有被荧光金荧光金标记的神经元胞体 ,脊髓横断处附近脊髓组织的溃变程度减轻 ,红核及躯体感觉运动区内神经元密度高于未移植组 ,大鼠后肢的自主运动功能明显好于未移植组。结论　神经干细胞移植入损伤脊髓后能分化为神经元及神经胶质细胞 ,能减轻脊髓的继发性损伤 ,保护受损伤的神经元 ,促进运动功能的恢复     

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.     

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).     

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.     

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.     

Prevalence and severity of hip and groin pain in sub‐elite male football: a cross‐sectional cohort study of 695 players

The purpose of this study was twofold: (a) to investigate the prevalence of hip and groin pain in sub‐elite male adult football in Denmark and (b) to explore the association between prevalence and duration of hip and groin pain in the previous season with the Copenhagen Hip and Groin Outcome Score (HAGOS) in the beginning of the new season. In total 695 respondents from 40 teams (Division 1–4) were included. Players completed in the beginning of the new season (July–Sept 2011) a self‐reported paper questionnaire on hip and/or groin pain during the previous season and HAGOS. In total 49% (95% CI: 45–52%) reported hip and/or groin pain during the previous season. Of these, 31% (95% CI: 26–36%) reported pain for >6 weeks. Players with the longest duration of pain during the previous season had the lowest HAGOS scores, when assessed at the beginning of the new season, P < 0.001. This study documents that half of sub‐elite male adult football players report pain in the hip and/or groin during a football season. The football players with the longest duration of pain in previous season displayed the lowest HAGOS scores in the beginning of the new season.     

Bronchial stump fistula :treatment with covered retrievable hinged metallic stents-preliminary clinical experience

Objective To evaluate the preliminaily clinical efficacy and retrievability of a retrievable hinged covered metallic stent in the treatment of the bronchial stump fistula （BSF）. Methods Between April 2003 and March 2005, 8 patients with bronchial stump fistula after pneumonectomy or lobectomy were treated with two types （A and B） of retrievable hinged covered metallic stents. Type A stent was placed in 6 patients and type B in 2 under fluoroscopic guidance. The stent was removed with a retrieval set when BSF was healed or complications occurred. Results Stent placement in the bronchial tree was technically successful in all patients, without procedure-related complications. Immediate closure of the BSF was achieved in all patients after the procedure. Stents were removed from all patients but one. Removal of the stents was difficult in two patients due to tissue hyperplasia. Patients were followed up for 6 - 21 months. Placement of the stents remained stable in all patients except one due to severe cough. Permanent closure of BSF was achieved in 7 （87.5%） of 8 patients. Conclusion Use of a retrievable hinged covered expandable metallic stent is a simple, safe, and effective procedure for closure of the BSF. Retrieval of the stent seems to be feasible. （J Intervent Radiol, 2007, 16： 253-257）