Apofix内固定系统治疗创伤性寰枢椎不稳

目的 探讨Apofix内固定系统治疗创伤性寰枢椎不稳的临床效果。方法 回顾分析2002年6月至2006年4月共24例创伤性寰枢椎不稳患者行后路Apofix系统内固定。结果 经6～18个月随访，所有患者均达到骨性愈合，术后局部症状及脊髓功能均有不同程度的改善。     

几种经后路手术治疗创伤性寰枢椎不稳的方法比较

目的：对经后路手术治疗创伤性寰枢椎不稳的几处方法进行回顾总结,评价APOFIX椎板钩内固定并后路．融合术的效果。方法：对25例创伤性寰枢椎不稳患者行后路手术,包括齿突骨折20例,横韧带断裂5例,其中Gallie钢丝并后路融合术12例,Brooks钢丝并后路融合术9例,APOFIX椎板钩并后路融合术4例,结果：术后随访6－40个月（平均21个月）,全部行颈椎侧位X线片检查,部分行MRI,有性融合分别为Gallie法10例（83％）,Brooks法8例（89％）,APOFIX法4例（100％）,结论：APOFIX椎板钩并后路融合术操作简便,稳定,效果好,并发症少且不影响MRI成像而有利随访,是治疗创伤性寰枢椎不委较为理想的一种方法。     

经后路寰枢椎椎弓根螺钉内固定治疗陈旧性齿突骨折

目的探讨经后路寰枢椎椎弓根螺钉内固定治疗陈旧性齿突骨折的临床疗效。方法 7例患者中,AndersonⅡ型陈旧性齿突骨折6例,不稳定Ⅲ型1例,术前日本骨科协会（JOA）评分为6~13分,平均9.6分,影像学检查均伴有寰枢椎不稳或脱位表现。采用经寰枢椎椎弓根螺钉进行固定,并在寰枢椎后弓间植入自体颗粒状骨。结果 7例患者全部得到随访,平均随访15个月,所有患者螺钉植入位置和复位均满意。寰枢椎植骨于术后6~9个月达骨性融合。术后3个月JOA评分12~17分,平均14.6分。所有患者颈部运动功能恢复良好,但轴向旋转活动部分丧失。结论应用寰枢椎椎弓根螺钉内固定治疗陈旧性齿突骨折可使寰枢椎获得即刻的坚强固定,有利于寰枢间植骨融合,疗效满意。     

寰枢椎钩内固定治疗齿状突骨折并寰枢椎脱位

目的 评估寰枢椎钩内固定治疗齿状突骨折并寰枢椎脱位的手术疗效。方法 回顾分析一年半 来6例手术病人的手术及疗效。结果 手术方法符合生理要求,内固定可靠。结论 寰枢椎钩操作 简便、安全,内固定坚强牢靠,并发症少。     

寰椎椎弓和枢椎椎板的应用解剖

目的：为颈后路寰枢椎钛缆固定术和相关器械设计提供解剖学依据。方法：在100例中国成人干燥标本上,观察寰椎后弓和枢椎椎板的形态学特点并进行解剖学测量。结果：(1)寰椎后弓较纤弱,上面与椎管不平行,从后上方向内下方倾斜;(2)枢椎椎板较粗大,上窄下宽,内面基本平行椎管,内面下缘多形成一骨嵴。结论：(1)寰椎后弓和枢椎椎板穿绕钛缆时宜自上向下操作,为方便操作,可以咬除部分枢椎椎板上缘;(2)设计枢椎椎板穿绕钛缆的引导器械时需考虑到其内面的骨嵴。     

多普勒超声观察创伤性寰枢椎不稳手术前后椎动脉血流的变化

目的探讨创伤性寰枢椎不稳患者内固定手术前后椎动脉血流的变化。方法32例创伤性寰枢椎不稳患者,其中男性22例,女性10例;年龄22～57岁,平均年龄38岁。手术前及内固定术后,均应用彩色多普勒超声测量颈椎1、2间和颈椎5、6间椎动脉血流速度、阻力指数。并对检查结果进行对比分析。结果32例创伤性寰枢椎不稳患者术前椎动脉血流39侧异常,25侧正常。内固定术后39侧异常者中35侧恢复正常,4侧无变化,3侧术前正常变为术后异常。结论创伤性寰枢椎不稳可导致椎动脉血流变化,手术复位内固定后增加了寰枢椎稳定性,可改善椎动脉血液供应。     

寰枢椎不稳经后路椎弓根钉固定融合手术治疗

目的 探讨寰枢椎不稳经后路椎弓根螺钉固定融合手术治疗的方法及临床疗效.方法 对15寰枢椎不稳的患者应用直视下椎弓根钉固定并行自体骼骨植骨融合.结果 15例60枚螺钉均成功植入,复位固定满意,全组病例未发生椎动脉、脊髓损伤,术后临床症状得到不同程度的改善,所有患者均获得6～24个月随访,平均7.5个月;患者均在3～6个月寰枢椎骨性融合,未发现螺钉松动、断裂现象.结论 直视下寰枢椎椎弓根螺钉植入技术具有置钉相对安全、准确、融合率高等特点,是治疗寰枢椎不稳的一种有效手术技术.     

寰枢椎的解剖学测量及其临床意义

目的 :为寰枢椎区病损机制和手术治疗提供解剖学依据。方法 :在 15 0例中国成年人干燥寰枢椎标本上 ,对具有临床意义的数据进行解剖学测量。结果 :寰椎上关节面内倾角、下关节面外倾角及侧块内倾角两侧不对称分别占 19.3 %、10 .7%和 7.3 % ;椎动脉沟处形成沟环者占 16 .0 % ;所有横突孔的前后径和横径均大于 4mm ;后弓内侧半距仅为外侧半距的 1/2。枢椎椎弓根内倾角变异较大 (-3 .5°～2 1.5°) ;齿突腰部宽度小于 9mm者占 71.2 % ,齿突后倾角的变异较大 (0°～ 2 2°) ;2 7.5 %的椎动脉在经过枢椎侧块下方时会形成一动脉压迹沟 ,致使侧块外端和椎弓根变薄。结论 :①寰枢椎的解剖学形态与其生物力学性质及损伤机制密切相关 ;②手术时寰椎前弓向外显露不宜超过 2 0mm ,后弓切除向外不宜超过 10mm ,经寰枢外侧关节融合或内固定术应选择在关节的内侧 2 /3 ;③大部分中国人不适宜于两枚齿突螺钉内固定术 ;④ 2 7.5 %的中国人不适宜于经椎弓根螺钉固定术 ,枢椎椎弓根变异较大 ,手术前应作CT检测。     

个体化设计解剖定位椎弓根镙钉置入内固定治疗寰枢椎不稳

背景：经寰枢椎椎弓根螺钉置入内固定常用于治疗寰枢椎不稳。但是,目前寰枢椎椎弓根螺钉的进钉点和进钉角度多以统计处理的解剖数据确定,解剖差异的存在使目前置钉方法难以保证椎弓根螺钉准确置入到每一例患者。 目的：评估个体化设计解剖定位寰枢椎椎弓根螺钉置入内固定技术治疗寰枢椎不稳的安全性和可靠性。 方法：对16例寰枢椎不稳患者分别以术中寰椎椎弓根下壁、内侧壁以及枢椎上壁、内侧壁为定位依据,结合置入前影像学测量确定寰、枢椎椎弓根镙钉的进钉点、进钉角度以及镙钉长度,置入寰、枢椎椎弓根螺钉内固定系统。 结果与结论：16例患者均成功置入螺钉,置入后X射线片及CT显示寰枢椎椎弓根螺钉位置良好。置入后随访9~24个月,所有患者均获得骨性融合、症状改善,置入后脊髓功能改善优良率为84.6%,其中优6例(46.2%),良5例(38.5%),可2例(15.4%),无神经血管损伤等并发症发生。提示个体化设计解剖定位寰枢椎椎弓根螺钉置入内固定治疗寰枢椎不稳的进钉点和进钉角度定位准确,螺钉置入安全、可靠。 关键词：寰枢椎不稳;椎弓根螺钉;个体化设计;解剖定位;植入体 doi:10.3969/j.issn.1673-8225.2012.09.026      

三维CT重建参照下徒手寰枢椎椎弓根螺钉置入内固定治疗外伤性寰枢椎不稳

背景：寰枢椎椎弓根螺钉内固定在三维CT重建参照下比传统方法能提高置钉的准确率,减少置入并发症。 目的：探讨以寰枢椎三维CT重建为参照,进行寰枢椎椎弓根螺钉内固定治疗外伤性寰枢椎不稳的方法,明确其手术指导意义以及临床治疗效果。　 方法：对30例因外伤导致寰枢椎不稳需行寰枢椎椎弓根螺钉内固定治疗的患者内固定置入前行三维CT重建。 结果与结论：与螺钉置入前设计钉道内倾度、设计钉道测得进钉点与中线的距离比较,经C1、C2椎弓根螺钉实际钉道内倾角及进钉点与中线的距离差异均无显著性意义。30例患者观察到的C1后弓及C2椎弓表面解剖特征与置入前CT容积再现的影像一致。说明根据三维CT重建图像为参照进行寰枢椎椎弓根螺钉内固定,徒手置入,节省时间,并减少术中接收X线辐射,个性化置钉,精确、安全性高、疗效优良。     

Contribution of cysteine residues to the structure and function of herpes simplex virus gH/gL

In HSV types 1 and 2, gH forms a noncovalent heterodimer with gL. Previous studies demonstrated that the first 323 amino acids of gH1 and the first 161 amino acids of gL1 are sufficient for gH/gL binding. For gL1, substitution of any of its four cysteine (C) residues (all located within the gH/gL binding region) destroyed gH binding and function. Although gH1 contains 8 cysteines in its ectodomain, gH 2 contains 7 (C3 of gH1 is replaced by arginine in gH2). We found that mutation of any of the four C-terminal cysteines led to a reduction or loss of gH/gL function. Mutation of C5 or C6 in gH1 or gH2 rendered the proteins non-functional. However, substitution of C7 and/or C8 in gH1 has a definite negative impact on cell-cell fusion, although these mutations had less effect on complementation. Remarkably, all four gH1 N-terminal cysteines could be mutated simultaneously with little effect on fusion or complementation. As gH2 already lacks C3, we constructed a triple mutant (gH2-C1/2/4) which exhibited a similar phenotype. Since gH1 is known to bind gL2 and vice versa, we wondered whether binding of gH2 to the heterologous gL1 would enhance the fusion defect seen with the gH2-C2 mutant. The combination of mutant gH2-C2 with wild-type gL1 was nonfunctional in a cell-cell fusion assay. Interestingly, the reciprocal was not true, as gH1-C2 could utilize both gL1 and gL2. These findings suggest that there is a structural difference in the gH2 N-terminus as compared to gH1. We also present genetic evidence for at least one disulfide bond within gH2, between cysteines 2 and 4.     

Cervical spine in the Apert syndrome.

Radiographs of the cervical spine--in many cases longitudinal--were available for study in 68 cases of Apert syndrome. Autopsy material was available in one of these cases, and a 3-dimensional reconstruction from a CT scan was also studied in one case. Variable degrees of fusion were observed, involving the articular facets, the neural arch or transverse processes, or block fusion of the vertebral bodies. Ossification may not always be evident in some early radiographs. However, early radiographic signs of impending fusion may be irregularity in vertical orientation of the vertebral bodies and narrowing of the involved intervertebral spaces. Cervical fusions occurred in 68%, single fusions being found in 37%, and multiple fusions in 31%. C5-C6 fusion was most common, alone or in combination with other fusions. In contrast, cervical fusions are known to occur in 25% of Crouzon patients, most commonly involving C2-C3 only. It appears that when fusions are present, C5-C6 involvement in the Apert syndrome and C2-C3 involvement in the Crouzon syndrome separate the 2 conditions in most cases. Because cervical anomalies may complicate an already compromised airway in any form of acrocephalosyndactyly, it is imperative to initiate radiographic study of the cervical spine before undertaking anesthesia for surgery.     

正常青少年及青少年特发性脊柱侧凸患者枕颈部矢状面形态相关研究

目的比较正常青少年与青少年特发性脊柱侧凸（AIS）患者枕颈矢状面形态,探讨其枕颈矢状面形态与颈椎矢状面形态相关性。方法收集2012年3月—2014年3月在南京鼓楼医院脊柱外科入院治疗并符合入选标准的AIS患者80例（男14例,女66例）作为AIS组;正常青少年志愿者100名（男17名,女83名）作为对照组。在枕颈部侧位X线片上测量并记录枕骨入射角（OI）、枕骨斜率（OS）、枕骨倾斜角（OX）、上颈椎前凸角（C0-C2）、下颈椎前凸角（C2-C7）和颈椎前凸角（C0-C7）,比较对照组与AIS组枕颈参数差异及其与年龄、性别的关系,并分析枕颈参数与颈椎矢状面形态相关性。结果对照组OI、OS和OT分别为36．12°±2．55°（30°～44°）、26．34°±8．41°（15°-46°）和-10．06°±7．51°（-22°～11°）,AIS组OI、OS和OT分别为35．62°±3．01°（31°～42°）、24．27°±8．49°（7°-42°）和-11．52°±9．23°（-28°-10°）,两组枕颈部形态参数差异均无统计学意义（t分别为0．878、1．014、1．306,P值均〉0．05）,且不受年龄（≤14岁,〉14～18岁）、性别影响（P值均〉0．05）。对照组OI与C0-C2角和C0-C7角显著相关（r=0．307和r=0．298,P值均〈0．05）,OS和OT分别与C2-C7角和C0-C7角显著相关（r=0．402和r=0．560、r=0．428和r=0．550,P值均〈0．05）;而AIS组仅OI和OS与C0-C2角存在显著相关性（r=0．532和r=0．620,P值均〈0．05）。结论正常青少年和AIS患者的枕颈参数无显著差异,且不受年龄与性别影响。正常青少年OI、OS和OT与颈椎矢状面形态显著相关,而AIS患者枕颈部矢状面形态仅与其上颈椎矢状面形态密切相关。     

Biomechanical effect of anterior cervical spine fusion on adjacent segments.

The biomechanical effects of superior (C4-C5) and inferior (C5-C6) level fusions with different graft materials on the adjacent unaltered components were quantified using an anatomically accurate and experimentally validated C4-C5-C6 finite element model. Smith-Robinson and Bailey-Badgley fusion procedures were analyzed with five different types of inter-body fusion materials with varying stiffnesses. Intact and surgically altered finite element models were subjected to physiologic compression, flexion, extension and lateral bending. The external axial and angular stiffness, and the internal unaltered intervertebral disc (C5-C6 for the superior and C4-C5 for inferior fusion) and C5 vertebral body stresses were determined. The superior level fusion resulted in the highest increase in external response in lateral bending for all implant materials in both surgical procedures. In contrast, the inferior level fusion produced a higher increase in the C4-C5 disc and C5 vertebral body stresses in compression than the superior level fusion in both surgical procedures. The increased internal stress responses reflecting the changes in the load-sharing following inferior level fusion may explain clinical observations such as enhanced degeneration subsequent to surgery. Because of the inclusion of three levels in the present multi-segment finite element model, it was possible to determine these responses in the unaltered adjacent components of the cervical spine.     

单链抗体-碱性磷酸酶融合蛋白的制备

目的：制备单链抗体-碱性磷酸酶融合蛋白。方法：把碱性磷酸酶编码区插入pSTE2-8E5质粒DNA，用scFvC66的重链和轻链可变区DNA取代scFv-8E5的重链和轻链可变区DNA，构成表达载体pSTE2-C66-Ap在大肠杆菌中表达，用聚丙烯酰胺凝胶电泳和免疫印迹法分析产物活性。结果：获得一个分子量为75kD的scFvC66-Ap融合蛋白，可结合来自KGla细胞裂解物中的一个分子量约60kD的蛋白质带，其结合功能可通过其碱性磷酸酶活性直接测定。结论：建立了一个用大肠杆菌制备单链抗体-碱性磷酸酶融合蛋白的方法，它可能取代常规的碱性磷酸酶标记方法而用于免疫检测。     

Characterization of the neutralizing activity of three anti-human TNF monoclonal antibodies and prediction of their TNF epitopes by molecular modeling and mutant protein approach

The neutralizing activity of three anti-human TNF monoclonal antibodies, designated D2, E6, and F6 were investigated by three experimental systems. The results from the systems showed that all the three mAbs could neutralize TNF-mediated cytotoxicity in L929 cells, TNF-induced NF-kappaB activation in ECV304 cells, and TNF-upregulated ICAM-1 surface expression on ECV304 cells in dose-dependent manners. D2 had the highest neutralizing activity of the three mAbs, and F6 had higher level of neutralizing activity than E6. We also cloned the VH and VL cDNAs and obtained their cDNA sequences. The sequences were used in molecular modeling to establish the complex structures of TNF with variable regions of the three mAbs, respectively. In the structures, the TNF epitopes of D2, E6, and F6 were predicted at amino acids of (A109, A111-A112, C19, C21-C29, C44-C46, C66-C75, C77, C79, C90, C101, C103, C105, C114, C134-C148), (C18-C19, C21-C30, C32, C37, C43-C47, C67-C75, C83, C105-C106, C131, C135-C141), and (C21-C32, C45-C47, C65, C67-C72, C74, C81, C83, C90-C95, C105-C113, C133-C147), respectively, and the affinities of D2, E6, and F6 to TNF were predicted as -252.69, -232.83, and -299.92 kcal, respectively. Moreover, we proved the binding ability of F6 to the epitopes of amino acids of 141-146 in TNF molecule was better than that of E6, and that of D2 was the best of the three mAbs by Western blot and ELISA, in which the mutant TNF deleted the amino acids of 141-146 in TNF molecule was employed. These results make a basic foundation for selecting candidate mAbs for various purposes, such as construction of chimeric or humanized mAbs for therapeutic purpose, establishment of ELISA kits for determination of TNF, and production of affinity columns to purify TNF.     

Spinal cord stimulation modulates intraspinal colorectal visceroreceptive transmission in rats

Previous studies have shown that spinal cord stimulation (SCS) of upper lumbar segments decreases visceromotor responses to mechanical stimuli in a sensitized rat colon and reduces symptoms of irritable bowel syndrome in patients. SCS applied to the upper cervical spinal dorsal column reduces pain of chronic refractory angina. Further, chemical stimulation of C1-C2 propriospinal neurons in rats modulates the responses of lumbosacral spinal neurons to colorectal distension. The present study was designed to compare the effects of upper cervical and lumbar SCS on activity of lumbosacral neurons receiving noxious colorectal input. Extracellular potentials of L6-S2 spinal neurons were recorded in pentobarbital anesthetized, paralyzed and ventilated male rats. SCS (50 Hz, 0.2 ms) at low intensity (90% of motor threshold) was applied to the dorsal column of upper cervical (C1-C2) or upper lumbar (L2-L3) ipsilateral spinal segments. Colorectal distension (CRD, 20 mmHg, 40 mmHg, 60 mmHg, 20s) was produced by air inflation of a latex balloon. Results showed that SCS applied to L2-L3 and C1-C2 segments significantly reduced the excitatory responses to noxious CRD from 417.6+/-68.0 to 296.3+/-53.6 imp (P<0.05, n=24) and from 336.2+/-64.5 to 225.0+/-73.3 imp (P<0.05, n=18), respectively. Effects of L2-L3 and C1-C2 SCS lasted 10.2+/-1.9 and 8.0+/-0.9 min after offset of CRD. Effects of SCS were observed on spinal neurons with either high or low-threshold excitatory responses to CRD. However, L2-L3 or C1-C2 SCS did not significantly affect inhibitory neuronal responses to CRD. C1-C2 SCS-induced effects were abolished by cutting the C7-C8 dorsal column but not by spinal transection at cervicomedullary junction. These data demonstrated that upper cervical or lumbar SCS modulated responses of lumbosacral spinal neurons to noxious mechanical stimulation of the colon, thereby, proved two loci for a potential therapeutic effect of SCS in patients with irritable bowel syndrome and other colonic disorders.     

人IgE重链3-4区蛋白的表达、纯化及功能分析

目的通过基因重组的方法表达IgECε3-Cε4区,鉴定重组蛋白与天然细胞表面受体FcεRⅠ之间的相互作用．并用重组蛋白免疫小鼠制备血清。方法用RT—PCR方法从过敏性疾病病人外周血淋巴细胞调取IgECε3-Cε4cDNA片段,克隆至pET28a（＋）构建原核表达载体IgECε3-Cε4-pET28a（＋）,将重组质粒转化到大肠杆菌BL21（DE3）,诱导表达IgECε3-Cε4区蛋白,通过Ni-NTA亲和层析纯化融合蛋白后,用免疫荧光方法鉴定重组蛋白与天然细胞表面受体FcεRⅠ之间的结合能力,并用UniCAP100全自动分析检测仪对重组抗原进行定量检测与鉴定：用表达蛋白免疫BALB／c小鼠,制备抗鼠血清,用Western—blot法对多克隆抗体进行鉴定。结果成功调取的人IgECε3-Cε4区基因与已报道的序列相一致;获得的IgECε3-Cε4区蛋白相对分子质量（Mr）同预期的结果相一致;IgECε3-Cε4能特异性结合人嗜碱性粒细胞表面的FcεRⅠ受体;通过UniCAP 100全自动分析检测仪能够检测到重组抗原并精确到国际单位;免疫鼠血清多抗能特异性结合天然人血清IgE。结论成功构建了IgECε3-Cε4-pET28a（＋）表达载体,获得了能被人嗜碱性粒细胞表面的FcεRⅠα亚基特异性识别的IgECε3-Cε4区蛋白．并用天然人血清IgE鉴定了多克隆抗体,为下一步单克隆抗体的制备打下了基础。     

Responses and afferent pathways of C1-C2 spinal neurons to cervical and thoracic esophageal stimulation in rats

Because vagal and sympathetic inputs activate upper cervical spinal neurons, we hypothesized that stimulation of the esophagus would activate C(1)-C(2) neurons. This study examined responses of C(1)-C(2) spinal neurons to cervical and thoracic esophageal distension (CED, TED) and afferent pathways for CED and TED inputs to C(1)-C(2) spinal neurons. Extracellular potentials of single C(1)-C(2) spinal neurons were recorded in pentobarbital-anesthetized male rats. Graded CED or TED was produced by water inflation (0.1-0.5 ml) of a latex balloon. CED changed activity of 48/219 (22%) neurons; 34 were excited (E), 12 were inhibited (I), and 2 were E-I. CED elicited responses for 18/18 neurons tested after ipsilateral cervical vagotomy, for 12/14 neurons tested after bilateral vagotomy and for 9/11 neurons tested after bilateral vagotomy and C(6)-C(7) spinal cord transection. TED changed activity of 31/190 (16%) neurons (28E, 3 I). Ipsilateral cervical vagotomy abolished TED-evoked responses of 5/12 neurons. Bilateral vagotomy eliminated responses of 2/4 neurons tested, and C(6)-C(7) spinal transection plus bilateral vagotomy eliminated responses of 2/2 neurons. Thus inputs from CED to C(1)-C(2) neurons most likely entered upper cervical dorsal roots, whereas inputs from TED were dependent on vagal pathways and/or sympathetic afferent pathways that entered the thoracic dorsal roots. These results supported a concept that C(1)-C(2) spinal neurons play a role in integrating visceral information from cervical and thoracic esophagus.     

用于治疗难复位性寰枢椎脱位的新型经口前路寰枢椎侧块融合器的生物力学研究

目的　比较新型寰枢椎侧块融合器与传统内固定方式（TARP＋髂骨块内固定技术,后路椎弓根钉棒固定技术）的生物力学差异。 方法　收集40例TARP术后病人的CT扫描数据进行测量并设计新型融合器。挑选6具新鲜上颈椎标本。分别进行完整状态,失稳状态及进行3种内固定方式[（TARP+融合器（A1）;TARP+髂骨块（A2）;后路椎弓根钉棒技术（B）]处理。再测量不同状态下标本的屈伸,侧屈,旋转6组动作的活动范围（ROM）并进行统计学分析。 结果　新型融合器有三种型号：13/12/7,12/11/7,11/10/7（长／宽／高）;矢状面角设计为：16°/18°/20°。生物力学数据分析显示：Cage组（A1）与髂骨组（A2）在6个方向的活动度差异无统计学差异（P>0.05）;在屈伸及旋转活动中,TARP固定组（A1、A2）与后路固定组（B）不存在显著差异（P>0.05）;在侧屈活动中,TARP固定组（A1、A2）与后路固定组（B）存在显著差异（P<0.05）。 结论　配合TARP技术使用的新型融合器与TARP＋髂骨块的生物力学稳定性相仿,在侧屈方向优于寰枢椎后路椎弓根螺钉技术。且融合器相对于髂骨块而言,理论上具有以下优势：①简化手术步骤;②避免取髂骨相关并发症。