神经元损伤修复机制研究的离体模型--体外培养原代神经元损伤模型的建立及损伤后c-Fos蛋白表达

目的：建立一个模拟脊髓全横断损伤后脊髓组分——原代培养的脊髓神经元损伤的模型。方法：取Wistar大鼠(孕14d)脊髓，培养10～12d后，采用所设计的标准模板，直视下进行损伤。观察损伤前后不同时间神经元的形态及即刻早期基因c-fos的表达变化。结果：在损伤8～12h后，损伤两侧存活的神经元突起便开始朝着损伤部位生长，随着时间的延长，神经元突起甚至可跨越损伤处而延伸至对侧。c-fos基因在损伤后10min即有表达，2h达到最高峰，随后逐渐下降。结论：此模型可用于模拟体外培养的脊髓神经元受到机械损伤以及观察神经元损伤后进行修复的一些分子事件，简便而可靠，实用性较强。     

Rho-A mRNA 在大鼠脊髓损伤中的表达及与损伤程度的相关性

目的观察大鼠不同时间和不同损伤程度下，损伤段脊髓内Rho—A mRNA的表达及与损伤程度的相关性。方法按WD法制模，将SD大鼠135只随机分成3组，A组为全瘫组，B组为不全瘫，C组为空白手术对照组，每组分别为45只。致伤后8h、24h、3d、7d、28d，提取损伤段脊髓内的总RNA，荧光定量PCR和凝胶电泳检测Rho—A mRNA的表达，进行大鼠后肢运动功能评分，并作相关性分析。结果（1）A、B、C组在各时间点均有Rho—A mRNA表达，但同时间点A、B组的表达明显高于C组，A组同时间点的表达高于B组，C组各时间点表达维持在较恒定水平；脊髓损伤后8hRho-A mRNA表达增加，伤后3d达高峰，持续高水平表达28d。（2）C组各时间点大鼠后肢运动功能评分明显高于A、B组，A组同时间点的评分低于B组。结论脊髓损伤后Rho—A mRNA的表达明显升高，且与脊髓损伤程度呈正相关。     

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。     

脊髓型减压病大鼠脊髓组织内NGF和Trk A的蛋白表达

目的 观察大鼠脊髓减压病后脊髓组织内神经生长因子（NGF）及其受体酪氨酸激酶A（Trk A）蛋白表达随疾病进程的变化过程，探讨减压病脊髓损伤后自身的神经保护机制。方法 采用1.0MPa暴露5.5min后极快速减压的方法制备SD大鼠脊髓减压病模型，在出舱0，6，24，48，72h应用免疫组织化学染色和半定量图象分析方法检测脊髓组织NGF及其受体Trk A的蛋白表达。结果 极快速减压后6h开始有少量NGF和Trk A蛋白表达，在24h表达明显增强并达到高峰，此后Trk A表达逐渐减弱，NGF表达直到72h仍比基础值明显增加。结论减压病脊髓损伤诱导了脊髓神经元和胶质细胞的NGF及其受体的合成，提示脊髓型减压病在损伤神经组织的同时也激活了自身的神经保护机制，通过Trk A受体途径NGF可能在保护受损神经细胞中起作用。     

依达拉奉对大鼠急性脊髓损伤后bcl-2/bax表达及细胞凋亡的实验研究

目的观察依达拉奉对大鼠急性脊髓损伤后bcl-2/bax表达及细胞凋亡率的影响,探讨其对急性脊髓损伤后脊髓组织的保护作用。方法成年SD雄性大鼠60只,随机分成对照组和依达拉奉组,均采用改良Allen’s撞击法,致伤力为50gcf（10g×5cm）损伤T9～10制作动物模型,术后实验组给予依达拉奉3mg/kg,对照组给予等量生理盐水。每组分别于6h、12h、24h、72h、7d5个时间点处死动物取材,每个时间点6只大鼠,对受损脊髓部位进行免疫组织化学检测神经细胞凋亡因子bcl-2和bax基因的表达及用原位末端标记法（TUNEL法）标记神经元凋亡细胞。结果与对照组相比,依达拉奉组bcl-2蛋白表达水平显著提高,bax蛋白表达水平明显下降。对照组检测到较高的细胞凋亡率。依达拉奉组细胞凋亡率较对照组明显下降。结论依达拉奉可能通过清除氧自由基、上调bcl-2和下调bax蛋白表达来抑制大鼠脊髓损伤后神经细胞凋亡,从而保护大鼠神经组织。     

脑创伤后nNOS和iNOS对大鼠海马神经细胞凋亡的影响

目的 探讨大鼠脑创伤后神经元型一氧化氮合酶(nNOS)、诱导型一氧化氮合酶(iNOs)的神经毒性作用机制及7-硝基吲唑(7-NI)和氨基胍(AG)的治疗作用.方法 雄性成年Wistar大鼠250只,随机分为5组(假手术组、创伤组、AG治疗组、7-NI治疗组、AG+7-NI联合治疗组),每组又分为伤后1、3、6、12h及1、3、7、14d共8个时相点.采用Marmatou法制作大鼠重型弥漫性颅脑创伤模型,免疫组织化学法检测海马CA1区nNOS和iNOS表达情况,TUNEL法观察大鼠海马CA1区神经细胞凋亡情况,双标染色观察细胞凋亡与nNOS、iNOS的关系.结果 创伤组海马CA1区nNOS表达在伤后6h达高峰,iNOS表达及细胞凋亡在伤后3d达高峰,各治疗组凋亡细胞均有不同程度的减少.伤后6h创伤组TUNEL与nNOS共阳性细胞较多,应用7-M后共阳性细胞明显减少.伤后3d创伤组TUNEL与iNOS共阳性细胞较多,应用AG后共阳性细胞明显减少.结论 大鼠脑创伤后nNOS和iNOS的过度表达对大脑神经组织具有毒性作用,是海马CA1区神经细胞凋亡的原因之一.7-NI和AG可抑制nNOS和iNOS的活性,减少神经细胞凋亡,从而起到神经保护作用.     

Tat-LK15介导siRNA干扰大鼠脊髓背角nNOS表达对神经病理性疼痛的治疗作用

目的 探讨细胞穿透肽Tat-LK15介导siRNA干扰大鼠脊髓背角神经元型一氧化氮合酶(nNOS)表达对神经病理性疼痛的治疗效应.方法 采用Tat-LK15介导FAM-siRNA转染原代大鼠脊髓背角神经元细胞(RN-dsc),于倒置荧光显微镜观察其转染效果.健康雄性SD大鼠50只,随机分为5组(n=10):正常组、假手术组、神经病理性疼痛组(SNL组)、Tat-LK15-nNOS siRNA组(TS组)和Tat-LK15-NC siRNA组(TN组).采用脊神经结扎法(SNL)制作神经病理性疼痛模型,正常组不予任何处理,假手术组仅暴露脊神经;SNL组、TS组、TN组构建SNL模型同时鞘内置管,于第7天开始每天分别鞘内注射10μl生理盐水、10μl含5μg siRNA的Tat-LK15-nNOS siRNA和Tat-LK15-NCsiRNA,持续7d.建模前1d及建模后3、7、10、14d测定各组大鼠机械缩足反应阈值(PWMT)及热缩足反应潜伏期(PWTL).第14天测定结束后取L4-6节段脊髓,采用q-PCR及Western blotting检测脊髓背角nNOS表达水平.结果 倒置荧光显微镜观察显示,Tat-LK15可成功介导siRNA转染原代大鼠脊髓背角神经元.与假手术组比较,SNL组建模后第3天起PWMT降低、PWTL缩短,脊髓背角nNOS mRNA及蛋白表达上调(P<0.01),正常组上述指标与假手术组比较差异无统计学意义;与SNL组比较,TS组第10、14天时PWMT增高、PWTL延长,脊髓背角nNOS mRNA及蛋白表达降低(P<0.01),TN组上述指标与SNL组比较差异无统计学意义.结论 Tat-LK15可成功介导nNOS siRNA转染脊髓背角神经元,沉默nNOS基因的过度表达,对SNL大鼠神经病理性疼痛具有一定的治疗作用.     

不同损伤条件下CIDE-B的变化与神经元凋亡的关系

目的通过对周围神经火器伤和切割伤CIDE-B基因及蛋白的表达检测，并与神经细胞的凋亡进行对比，探讨CIDE-B的变化与神经元凋亡的关系。方法113只大耳白兔随机分为火器伤组、切割伤组、正常对照组。在伤后不同时相点取材，以RT-PCR法检测CIDE-B mRNA表达水平，原位杂交检测CIDE-B mRNA的形态学分布，Western blot检测CIDE-B的蛋白水平。结果神经损伤后脊髓内CIDE-B mRNA表达增强。火器伤组增加的幅度较大，第1天开始增加，7d达到峰值，持续到2周，4周回落；切割伤组表达增加延迟，幅度也小。蛋白水平的变化与此相一致。正常对照组脊髓组织有少量的CIDE-B mRNA表达，分布于脊髓神经元细胞胞浆内。结论周围神经损伤后脊髓神经元CIDE-B在mRNA、蛋白水平的表达均增强，组织学分布在脊髓灰质内，火器性损伤改变更加明显。     

CX43在应激性溃疡发生发展过程中的表达与细胞凋亡、增殖的关系

探讨细胞间隙连接蛋白43（CX43）在胃粘膜遭受应激损伤的变化及与细胞凋亡，增殖发生的关系。制作大鼠水浸－束缚应激（WRS）模型，采用原位末端标记（TUNEL）法检测WRS结束前后不同的时间点细胞凋亡的发生；免疫组化ABC法原位检测增殖细胞核抗原（PCNA）蛋白的表达，原位杂交检测CS43 mRNA表达变化。结果发现，正常大鼠胃粘膜上皮散在TUNEL－细胞，PCNA蛋白主要在胃腺颈部近胃小凹处呈中等阳性表达，CX43 mRNA在胃腺区呈中等阳性表达，WRS 2h-WRS后5h TUNEL-细胞较对照组明显增多，并逐步达高峰，PCNA蛋白和CX43 mRNA的表达明显低于对照组；WRS后8－12h TUNEL-细胞逐渐减少，而PCNA表达高峰，CX43 mRNA表达恢复并明显增加，WRS后24hTUNEL－细胞仍高于正常水平，PCNA及CX43表达基本恢复至正常水平，相关分析，CX43 mRNA表达与PCNA蛋白表达呈显著正相关，结果提示，CX43 mRNA的表达可能与胃粘膜腺细胞区细胞稳定，增殖和分化有关，并影响上皮细胞凋亡发生。     

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。     

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.     

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).     

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.     

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.     

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）     

Die rechtsmedizinische Identifizierung — Bedeutung und Beweiswert —

Zusammenfassung Bei der rechtsmedizinischen Identifizierung kann die Identität im strengen Sinn allenfalls bei lebenden Personen festgestellt werden; sonst läßt sich nur von Teilen auf das Ganze (vom Untersuchungsobjekt auf die Person) schließen, wobei die verschiedenen Merkmale des Untersuchungsobjektes entsprechend der Hdufigkeit ihres Vorkommens eine unterschiedliche Beweiskraft haben. Bei der Schädelidentifizierung mit Hilfe moderner photographischer oder elektronischer Superprojektionsverfahren ergeben sich unter Berücksichtigung der Weichteildicken so viele (fiktive) Vergleichspunkte, daß bei geeignetem Vergleichsmaterial (Photographien) Identität wegen der Vielzahl übereinstimmender Bezugspunkte in den meisten Fällen evident ist.     

Imaging of abdominal nodal spread in malignant disease

This is a review of the role of imaging procedures for the assessment of abdominal and pelvic lymph nodes. The diagnosis of malignant lymphatic spread is rarely the sole purpose of imaging, because it is usually part of a general abdominal examination, most frequently with CT or US, or increasingly with MRI. These studies are often requested in order to obtain information about the situation to be encountered during surgery, or to alert the surgeon to irresectability or to unexpected metastases outside the initially planned area of exploration. In most surgically treated tumours the role of imaging for preoperative staging is limited, due either to its insufficient sensitivity or because the initial treatment is independent of the lymph node stage. Imaging is commonly used to verify treatment response to chemo- or radiotherapy and for follow-up.Correspondence to: S. Delorme