基因治疗在周围神经损伤修复中的应用

随着分子生物学研究的飞速发展，基因治疗在基础和临床得到了广泛的应用，在周围神经损伤修复领域表现出尤为广阔的前景。目前，基因治疗修复周围神经损伤的基本策略是利用载体工具将编码神经营养因子的基因转入靶细胞，使之在体内持续表达，进而达到治疗效果。本文着重就神经营养因子、靶细胞、载体工具3个方面介绍当前国际上基因治疗修复周围神经损伤的研究情况。     

基因修饰的组织工程骨联合血管束植入修复节段性骨缺损

目的 评价骨形态发生蛋白2（BMP-2）基因修饰的组织工程骨联合血管束植入修复节段性骨缺损效果。方法 分离培养兔骨髓基质干细胞，经BMP-2基因转染后复合异种骨支架体外构建基因修饰的组织工程骨。建立兔双侧桡骨缺损（2．0cm）模型，采用4种方法修复：A组：基因修饰的组织工程骨＋血管束植入；B组：基因修饰的组织工程骨；C组：无基因修饰的组织工程骨＋血管束植入；D组：单纯支架。术后4、8、12周行X线检查、计量组织学检查、生物力学测定和微血管墨汁灌注等观察血管形成及成骨情况。结果 A组缺损区在成骨活跃程度、再生骨量等方面均显著优于其他组，血管束发出分支向移植骨内长入，血管再生旺盛，其骨缺损得到了较彻底的修复。B组在BMP-2基因诱导下成骨速度和质量优于C组；而D组缺损内主要形成纤维组织。结论 BMP-2基因治疗联合血管束植入具有良好的成骨及血管化效果，对骨不连、长段骨缺损的治疗具有重要意义。     

人骨形态发生蛋白2重组腺病毒在动物模型中活性观测

目的利用基因工程技术构建人骨形态发生蛋白2重组腺病毒,并进行体内表达,测定活性,为深入开展的基因治疗研究创造条件。方法同源重组构建重组腺病毒,将30只SD大鼠随机分为治疗组和对照组,将纯化后的重组腺病毒50μl注入SD大鼠股部肌肉陷窝病损处,通过组织学染色、X线片对动物模型的组织变化进行观测。结果重组腺病毒治疗组有明显的诱导成骨活性,治疗组碱性磷酸酶活性明显高于对照组。结论应用于动物实验中目的基因表达,并具有生物学活性。本研究是在骨缺损基因治疗方面的一次尝试,且人骨形态发生蛋白2重组腺病毒的构建成功,也为我国骨科疾病的基因治疗进一步研究奠定了基础。     

成肌细胞在肌肉骨骼系统疾病基因治疗中的应用研究进展

基因治疗 ,就是利用分子生物学的方法将目的基因转入靶细胞内 ,通过靶细胞表达目的基因产物治疗疾病的方法。基因治疗主要有两种途径 :即invivo及exvivo方式。invivo途径即体内基因治疗 ,是将外源基因装配于特定的真核细胞表达载体 ,直接导入体内。exvivo途径也称“回体基因治疗” ,是将含有外源基因的载体在体外导入自体或异体 (异种 )细胞 ,经体外细胞扩增后 ,输回自体。此方法易于操作 ,并且使用自体细胞有利于解决安全性问题。自从 80年代初以来 ,靶细胞的选择问题一直伴随着基因治疗研究。目前研究较多的有骨髓干细胞、成纤维细胞、肌…     

高压氧辅助带蒂筋膜瓣为膜诱导骨修复兔尺骨缺损的实验研究

目的 研究高压氧辅助带蒂筋膜瓣为膜诱导的非细胞型组织工程化骨修复骨缺损的效果,为临床应用提供依据.方法 研究对象为24只成年新西兰大白兔左侧尺骨造骨缺损模型.取白体红骨髓与含BMP的骨诱导活性材料混合成非细胞型组织工程骨,利用显微外科技术在骨缺损邻近制备一个带有无名血管蒂所属毛细血管网的筋膜瓣,使其包裹组织工程骨并充填骨缺损,术后将造模兔按数字随机表随机分为对照组(A组)与实验组(B组)各12只,A组术后呼吸常压空气,B组术后高压氧治疗,术后4、8、12周进行X线检查、大体形态观察、组织学检查、修复区内骨形态计量分析和交界区血管图像分析.结果 X线片、大体形态观察和组织学切片多种方法,检测2组各时间段同一时间点植入物内部血管的长入、骨小梁及软骨组织形成的数量和速度,成熟骨结构的形成、骨干结构的重塑、骨髓腔的再通、植入物的吸收降解,并对修复区内骨形态计量分析,发现2组差异有统计学意义(P＜0.05),B组体内成骨的能力和修复骨缺损的效果明显优于A组.交界区血管图像分析仪动态测量2组各时间段同一时间点单位面积内血管的数量及质量,经统计学比较差异均有统计学意义(P＜0.05),B组较A组有明显的优势,提示高压氧对组织工程骨构建血管化进程有明显的促进作用.结论 高压氧辅助带蒂筋膜瓣为膜诱导的非细胞型组织工程化骨修复骨缺损,具有促进膜诱导骨再生和血管化双重作用,对缩短骨缺损修复时间,提高成骨的质和量有显著效果.     

rhBMP-2和rhVEGF转染ADSCs后体外表达及诱导骨缺损成骨修复的实验研究

目的观察重组人骨形成蛋白2（rhBMP-2）和血管内皮生长因子（rhVEGF）转染脂肪源性干细胞（ADSCs）后的体外表达情况,并以含该转染体系的组织工程骨进行大动物负重骨缺损的修复研究。方法将外源性基因rhBMP-2和rhVEGF通过脂质体Lipofectamine^TM 2000介导的方式转染第四代ADSCs,以G418筛选出阳性克隆并扩大培养后,用RT-PCR和SABC免疫组化法在转录和蛋白质水平观察其转染后第2天和第4周表达情况。建立小香猪自体前肢尺骨中段1.5cm骨缺损模型,分3组进行对比试验：（A组）转染了生物活性因子的ADSCs与脱细胞骨基质材料（ACBM）复合而成新型组织工程骨植入修复组;（B组）未转染生物活性因子的ADSCs复合ACBM植入修复组;（C组）未处理组。以x线和组织切片评价其修复效果。结果转基因ADSCs在转染后瞬时和4周时都可表达rhBMP-2和rhVEGF。大动物骨缺损修复实验中,前2周内3组动物均未出现明显的急性炎症反应;连续x线观察和骨切片显示：A组在术后第3个月已完全修复,且修复效果和进程明显优于B组,C组缺损则主要由纤维结缔充填。结论rhBMP2及rh-VEGF转染ADSCs后可获得4周内稳定的表达。携带该缓释体系的新型组织工程骨是一种具有显著成骨能力的优良骨缺损修复材料。     

降解材料隔膜引导性骨再生的实验研究

目的 观察长管骨降解材料隔膜引导性骨再生现象,探讨其作用机制。方法 采用胶原隔膜、制做长管骨引导性骨再生动物模型。术后进行Ｘ线观察;不同时期处死取材,进行组织学观察及碱性成纤维细胞生长因子,转移生长因子－β（ＴＧＦ－β）,骨形态发生蛋白（ＢＭＰ）免疫组化染色,并对３种骨诱导因子的含量进行测量。结果 （１）胶原隔膜能够成功引导骨再生、１２周时修复骨缺损。（２）术后１周时,胶原隔膜即在骨缺损处形成密闭     

生物人工材料人工骨修复下肢骨缺损的临床观察

目的 观察生物人工材料(biological artificial material,BAM)人工骨修复下肢负重骨骨缺损的临床效果. 方法 选择2008年1月- 2010年12月收治的采用BAM人工骨进行手术植入修复骨缺损的32例下肢负重骨骨缺损患者,年龄21 ～ 77岁,平均32.5岁.骨缺损体积为1.0 cm×2.0 cm×2.5 cm～3.0 cm×3.5 cm ×5.0 cm,平均15.4 cm3.骨缺损病因主要为粉碎性骨折、骨囊肿、骨纤维结构不良、慢性骨髓炎等.术后即刻及术后1,2,3,5,7,9,12,18个月进行随访,观察术后全身及切口局部反应、血钙／磷改变情况、骨缺损修复及患肢负重功能恢复情况. 结果 所有患者均得到有效随访9 ～18个月,平均10.5个月.术后至末次随访所有患者均无局部或全身移植物排斥反应.随访X线片示术后3个月BAM人工骨植入区与骨缺损周围的骨组织之间界限模糊,有新生骨形成;术后6个月BAM人工骨植入区明显有新骨长入,人工骨材料与骨组织融为一体,骨缺损已基本修复.术后患肢完全负重时间为术后2.5 ～4个月,平均3.2个月. 结论 BAM人工骨具有良好的生物相容性和骨诱导作用,其内结构可以保持一定的刚度和强度,可用于下肢负重骨骨缺损的修复.     

腺病毒介导的人骨形态发生蛋白2基因修复兔桡骨缺损

目的 评价腺病毒介导的人骨形态发生蛋白2（Adv-hBMP-2)基因对兔骨髓间充质干细胞（BMSCs)的诱导成骨活性及修复长骨干骨缺损的效果。方法 （1）抽取兔骨髓行BMSCs培养，经Adv-hBMP-2转染后分别行免疫沉淀加Western印迹法和碱性磷酸酶（ALP）检测及von Kossa染色，并行裸鼠肌内诱导成骨试验。（2）修复兔桡骨缺损：15只兔30侧、1.5cm的骨缺损分为Adv-hBMP-2转染BMSCS加珊瑚及胶原载体组、β半乳糖苷酶（Adv-βgal)转染BMSCs加载体组、未转染BMSCs加载体组、载体组和未治疗组，术后行X线、组织学检查。结果 （1）Adv-hBMP-2组BMSCs表达hBMP-2，其ALP活性升高，并有钙结节形成,裸鼠肌内注射后有异位成骨。（2）在兔桡骨缺损处，Adv-hBMP-2转染组有明显骨痂形成，5个组的愈合率分别为4／5、2／5、2／5、0／5、0／5。结论 Adv-hBMP-2基因转染的BMSCs是修复骨缺损的好方法。     

骨组织工程在骨缺损修复中的应用研究进展

以往临床使用较多的植骨材料主要有自体骨、同种异体骨，以及经特殊处理的异种骨和人工骨材料等。但自体骨存在来源有限、术后取骨处疼痛等缺点，且术后并发症可达8％；同种异体骨亦存在免疫原性和致病性等缺点。随着组织工程学技术的发展，改变了治疗骨缺损的传统治疗模式。近20年来，应用生物学和组织工程学技术，将体外培养的高浓度种子细胞种植于人工或天然合成的细胞外基质载体上，并复合相关诱导因子，然后移植于体内，从而达到修复骨缺损的目的。骨组织工程是用最少量的组织细胞，经体外培养扩增后修复大块骨组织缺损，同时可依据缺损情况任意塑型，达到理想的形态修复，为最终实现无损伤修复创伤和真正意义上的功能重建开辟了新的途径。作者就骨组织工程中的3大要素（种子细胞、细胞生物支架、生长因子）的特点和作用作一综述。     

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.     

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）     

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.