纤维结合蛋白诱导下成纤维细胞增殖与成骨活性表达的研究

[背景]探讨纤维结合蛋白(fibronectin,FN)诱导下的成纤维细胞(fibroblast,FB)增殖与成骨活性,以及它们之间可能存在的剂量依赖关系.[方法]以组织块粘附法获得FB,经不同浓度FN(10,20,40,60,80μg/ml)诱导培养14 d.设立空白对照组(FN 0 μg/ml),以3H胸腺嘧啶脱氧核苷掺入试验、3H脯氨酸掺入试验分别检测细胞DNA、胶原合成含量,以四环素标记矿化结节,以123Ⅰ标记竞争免疫分析法检测细胞上清液中骨钙素合成量.[结果]FN诱导下,FB DNA与胶原合成含量显著增加,具有统计学意义(P＜0.05),其中以FN40μg/ml组最为明显(P＜0.05).与其他各组比较,FN 40 μg/ml组骨钙素含量显著增加,具有统计学意义(P＜0.05).经四环素标记,诱导培养的FB形成矿化结节,在荧光显微镜下呈现金黄色的小梁样结构.[结论]FN诱导下,FB具有显著的增殖与成骨活性,而且与FN剂量相关.FB可能具有原位多能干细胞潜能,有望成为组织工程学研究中的种子细胞.     

人工神经制备修复兔长段神经缺损的实验研究

目的：研究静脉（V）、基底膜（BM）、雪旺细胞（SC）和神经生长因子（NGF）制备人工神经桥接15mm兔坐骨神经缺损的作用。方法：用日本大耳白兔70只，分为7组（A、B、C、D、E、F、H），每组10条坐骨神经。各组均修复坐骨神经15mm缺损，以自体神经移植作为对照组（A）。术后观察3d,2,4,8和12周肢体运动，肌电图动作电位波幅，潜伏期和传导速度，记录电生理结果。术后12周取材，肉眼观察标本。比较近侧吻合口段（a)，移植体中段（b)，远侧吻合口段（c)神经再生情况，随机半数组织行HE染色，免疫组化髓磷脂碱性蛋白（MBP）染色，另半数组织做Cajar吡啶神经银染色，镜下观察神经再生情况，断端连接情况，并对各组再生神经束面积，有髓神经纤维密度，有髓神经纤维直径和轴突直径进行测量分析。结果：在实验组中以H组（V＋BM＋NGF＋SC组）修复情况最佳，上述各指标与自体神经移植组比较，差异无显著性意义（P＞0．05）。结论：V＋BM＋NGF＋SC组修复神经缺损，效果最好，符合神经组织学结构，能达到自体神经移植的效果，很有可能成为周围神经缺损修复的一种新的方法。     

偏振光显微观察不同胶原纤维在骨折愈合过程中的动态变化

背景：天狼星红是一种强酸性阴离子染料,染色后不褪色且具特异性,是目前胶原染色的最佳染料.胶原作为细胞外基质的主要成分之一,具有许多特殊的生理功能,机体通过胶原的合成和改建使骨折修复得以完善.目的：采用苦味酸天狼星红偏振光显微观察骨折愈合过程各型胶原的比例、分布的动态变化过程.设计：观察对比实验.单位：武汉大学人民医院骨外科,天津医院创伤骨科,北京大学医学部积水潭医院创伤骨科,解放军军事医学科学院基础医学研究所组织工程中心.材料：实验于2002-03/2003-09在解放军军事医学科学院基础医学研究所组织工程中心完成.选取健康成年中国绵羊3只,雄性,体质量25～35 kg.方法：全部动物麻醉消毒后在跖骨干中段截取1 cm长的骨缺损,骨折端用6孔加压钢板固定.术后1,3,6个月取骨折部位标本,乙二胺四乙酸脱钙制备切片,苦味酸天狼星红染色,偏振光显微镜观察胶原的类型和分布.主要观察指标：骨折愈合不同时期骨缺损区胶原的类型及其分布.结果：实验纳入3只绵羊,全部进入结果分析.①术后不同胶原纤维偏振光显微镜下的形态学表现：Ⅰ型胶原纤维紧密排列,显示很强的双折光性,呈黄色、橙色和红色的粗纤维;Ⅱ型胶原纤维显示弱的双折光,呈各种不同颜色的疏松网状;Ⅲ型胶原纤维疏网状,显示弱的双折光,呈绿色的细纤维.②术后不同胶原纤维偏振光显微镜下的数量观察：术后1个月,骨折处红色和黄色的Ⅰ型胶原极少见,绿色的Ⅲ型胶原纤维占大多数,胶原排列杂乱;术后3个月,骨折处红色和黄色的Ⅰ型胶原纤维明显增加,Ⅲ型胶原纤维的比例下降,胶原纤维的排列方向开始向有序的方向发展;术后6个月,粗大而鲜艳的红黄色Ⅰ型胶原成分占据了绝大部分,细小的绿色Ⅲ型胶原纤维的数量急剧减少,呈现明显的斜行螺旋性交叉的三维排列.结论：苦味酸天狼星红偏振光显微观察不仅能区分骨折局部Ⅰ、Ⅲ型胶原的类型,还可以清晰显示Ⅰ、Ⅲ型胶原的形态、分布和比例关系,具有操作简便、特异性强、灵敏度高的特点.     

骨质疏松与骨性关节炎关系的研究进展

骨质疏松(osteoporosis,OP)与骨关节炎(osteoarthritis,OA)是临床常见病多发病,骨质疏松症是以骨量减少、骨组织微细结构破坏导致骨骼脆性和骨折危险性增加为特征的一种系统性全身性骨骼疾病。而骨性关节炎是以软骨改变为主要特征,进而造成软骨下骨坏死,囊性变,骨密度增加和伴发骨赘形成的一种非特异性炎症。骨质疏松在骨关节炎发病过程中到底扮演什么角色,是否为骨关节炎的病因之一,国内外有大量文献对此进行了报道,其观点也不甚一致,目前存在3种:①OA与OP正相关,OP是OA的病因之一;②OA与OP负相关,OP不是OA的病因;③OA与OP不相关。通过查阅文献,笔者认为OP与OA不相关的可能性不大。本文就两者之间的正负关系作一综述,以期能对以后的临床工作有一定的指导作用。     

Open reduction and internal fixation of ankle fracture using wide-awake local anaesthesia no tourniquet technique

IntroductionAnkle fractures frequently occur and must be treated with open reduction for long-term stability. The existing anaesthesia methods include general anaesthesia, spinal and epidural anaesthesia, peripheral nerve block and local anaesthesia with IV sedation. However, each method has its inherent risks and potential costs, and the use of a tourniquet is inevitable. Therefore, the wide-awake local anaesthesia no tourniquet (WALANT) technique provides an alternative method for equivalent haemostasis and pain control without the use of a tourniquet.Patients and methodsWe prospectively enrolled 13 consecutive patients (9 males and 4 females) who presented ankle fractures and required ORIF from January 2017 to December 2017. The fracture types of the 13 patients included lateral malleolar fracture (three patients), bimalleolar fracture (two patients), bimalleolar equivalent fracture (three patients), medial malleolar fracture (two patients) and trimalleolar fracture (three patients; articular surface involvement <25%). We used a solution of 1% lidocaine mixed with 1:40,000 epinephrine for WALANT.ResultsAll patients underwent surgery if they exhibited an initial numerical pain rating scale (NPRS) score of 0 without using a tourniquet. Only two patients required an additional 5 ml of local anaesthesia due to NPRS score elevation during the surgery; no dose exceeded the safe limit of 7 mg/kg. No local complications occurred, and no shifts to other anaesthesia methods were required due to the failure of WALANT.ConclusionsWALANT simplified surgical preparations and provided a safe and reliable method for ankle fracture management. Because the use of a tourniquet was not required, reduced postsurgical pain was observed. Moreover, the use of local anaesthesia resulted in more satisfied patients and facilitated easier recovery.     

腰椎损伤疝出型椎间盘组织JNK信号通路激活情况的研究

目的:探究c-Jun氨基末端激酶（JNK）信号通路在腰椎损伤疝出型椎间盘组织中激活情况。方法:随机选择24例行手术治疗的腰椎间盘突出症（LDH）患者,其中12例损伤疝出型和12例退变突出型,分别为损伤疝出组和退变突出组。收集术中摘除的椎间盘组织,HE染色观察髓核组织病理变化;TUNEL染色分析髓核细胞凋亡情况;免疫蛋白印迹技术检测MKK4、JNK、pJNK、JunD蛋白表达水平。结果:损伤疝出组出现炎性细胞浸润的比例、髓核细胞凋亡计数以及MKK4、JNK、pJNK、JunD蛋白表达水平高于退变突出组（P<0.05）。结论:损伤疝出型髓核组织炎症反应更明显;JNK信号通路异常激活与髓核细胞凋亡增加可能是损伤疝出型LDH病理机制之一;JNK信号通路激活可能是LDH病理分型的标志之一。     

A posterior anchoring method decreases pullout suture translation of the medial meniscus posterior root repair during knee flexion

BackgroundThe medial meniscus (MM) translates posteriorly and extrudes severely from the medial tibial plateau (MTP) during knee flexion in the MM posterior root tear (PRT) knee. Transtibial pullout repair of the MMPRT has been performed to regulate MM extrusion. This study aimed to evaluate pullout suture translation during knee flexion before and after posterior anchoring during pullout repair. We hypothesized that suture translation after posterior anchoring would be significantly decreased relative to that before posterior anchoring.MethodsThirty-five patients who underwent MM posterior root repair were prospectively investigated. Pullout repair was performed using two cinch sutures (outer and inner sutures) and posterior anchoring through the MM posterior horn and an additional bone tunnel on the MTP. The translation of the outer suture from 0° to 90° of knee flexion was measured and compared before and after posterior anchoring intraoperatively. The MM morphologic features were measured using preoperative magnetic resonance imaging, and the correlation between these values and outer suture translation was evaluated.ResultsThe average outer suture translation after posterior anchoring (1.6 ± 1.5 mm) was significantly decreased relative to that before posterior anchoring (2.5 ± 1.7 mm, P < 0.01). No significant correlations were observed between the MM morphological features and outer suture translation.ConclusionsThe posterior anchoring method with an MM posterior root repair is useful in decreasing posterior translation of the pullout suture during knee flexion, which might have an advantage in preventing suture pullout from the repaired MM, leading to good clinical outcomes.     

2019年《OARSI非手术治疗膝、髋及多关节骨关节炎临床实践指南》方法学解读和推荐意见阐释

2019年国际骨关节炎学会(osteoarthritis research society international,OARSI)发布的《非手术治疗膝、髋及多关节骨关节炎临床实践指南》涉及膝关节、髋关节及多关节骨关节炎(osteoarthritis,OA)非手术治疗相关的多项干预措施。与国内的关节炎诊疗共识相比,OARSI制订的这部指南在方法学的选择和推荐意见的提出方面存在一定的优势且具有很好的代表性。因此,我们对该指南进行解读,以期达到最终目的:加强国内医师对OA非手术治疗的规范和理解;加强OA临床实践者对本指南制订方法学的理解,从而更好地结合推荐意见和证据开展临床实践和加快指南的实施;加快指南制订方法学在我国的传播,帮助临床工作者理解指南的内涵和价值;为制订基于我国现状的、涉及其他临床学科的临床实践指南提供方法学指引和参考。     

Bilateral Low-Dose Computed Tomography Assessment for Post-Operative Rotational Malalignment After Intramedullary Nailing for Tibial Shaft Fractures: Reliability of a Practical Imaging Technique

IntroductionThe purpose of this study is to evaluate the intra- and inter-observer reliability of low-dose protocolled bilateral postoperative Computed Tomography (CT)-assessment of rotational malalignment after intramedullary nailing (IMN) of tibial shaft fractures.Materials and methods156 patients were prospectively included with tibial shaft fractures that were treated with IMN in a Level-I Trauma Centre. All patients underwent post-operative bilateral low-dose CT-assessment (effective dose of 0.03784 – 0.05768 mGy) as per hospital protocol. Four observers performed the validated reproducible measurements of tibial torsion in degrees, based on standardized techniques. The Intra-Class Coefficient (ICC) was calculated to evaluate intra- and inter-observer reliability. The intra- and inter-observer reliability was categorized according to Landis and Koch.ResultsIntra-observer reliability for quantification of rotational malalignment on post-operative CT after IMN of tibial shaft fractures was excellent with 0.95 (95% CI = 0.92-0.97).The overall inter-observer reliability was 0.90 (95% CI = 0.87-0.92), also excellent according Landis and Koch.ConclusionFirstly, bilateral post-operative low-dose –similar radiation exposure as plain chest radiographs– CT assessment of tibial rotational alignment is a reliable diagnostic imaging modality to assess rotational malalignment in patients following IMN of tibial shaft fractures and it allows for early revision surgery. Secondly, it may contribute to our understanding of the incidence-, predictors- and clinical relevance of post-operative tibial rotational malalignment in patients treated with IMN for a tibial shaft fracture, and facilitates future studies on this topic.     

Using Naive Bayes Classifier to predict osteonecrosis of the femoral head with cannulated screw fixation

Predictive models permitting personalized prognostication for patients with cannulated screw fixation for the femoral neck fracture before operation are lacking. The objective of this study was to train, test, and cross-validate a Naive Bayes Classifier to predict the occurrence of postoperative osteonecrosis of cannulated screw fixation before the patient underwent the operation. The data for the classifier model were obtained from a ambispective cohort of 120 patients who had undergone closed reduction and cannulated screw fixation from January 2011 to June 2013. Three spatial displaced parameters of femoral neck: displacement of centre of femoral head, displacement of deepest of femoral head foveae and rotational displacement were measured from preoperative CT scans using a 3-dimensional software. The Naive Bayes Classifier was modelled with age, gender, side of fractures, mechanism of injury, preoperative traction, Pauwels angle and the three spatial parameters. After modelling, the ten-fold cross-validation method was used in this study to validate its performance. The ten-fold cross-validation method uses the whole dataset to be trained and tested by the given algorithm. Two of the three spatial parameters of femoral neck (displacement of center of femoral head and rotational displacement) were included successfully in the final Naive Bayes Classifier. The Classifier achieved good performance of the accuracy (74.4%), sensitivity (74.2%), specificity (75%), positive predictive value (92%), negative predictive value (42.9%) and AUC (0.746). We showed that the Naive Bayes Classifier have the potential utility to be used to predict the osteonecrosis of femoral head within 5 years after surgery. Although this study population was restricted to patients treated with cannulated screws fixation, Bayesian-derived models may be developed for application to patients with other surgical procedures at risk of osteonecrosis.