小鼠肾组织体外培养的研究

目的通过观察胚龄12、14、16天胎鼠和生后1天的仔鼠的后肾组织体外培养的存活发育情况,与同期在体比较,建立能模拟体内生存环境稳定的肾组织培养模型。方法采用体外培养倒置显微镜观察,光镜连续切片技术结合体视学定量分析方法,观察、检测培养后肾组织的存活情况以及培养前后肾脏皮质的发育状况。结果对同一胚（日）龄小鼠后肾组织随培养时间逐渐发育,直至成熟;且随胚（日）龄的增加,肾组织块存活比率逐渐下降,肾脏皮质的发育呈下降趋势。结论小鼠后肾组织体外培养胚（日）龄越小其存活发育情况越好,E12天取材培养的肾组织与在体发育一致,利用微孔膜肾组织培养是一种简便有效的神经组织体外培养方法。     

组织工程小血管摩擦力测量方法的研究与应用

目的设计一套用于组织工程小血管内壁摩擦力测量的装置并对组织工程血管的材料进行初步的测量,以验证装置的可靠性和相关结论。方法根据血液动力学的原理,对血管的力学模型进行分析,确定组织工程血管内皮细胞摩擦力的测量方法,从而设计完成整套装置并对弹性元件进行测量,并进行组织工程降解材料聚羟基乙酸PGA构成的管状支架进行摩擦力的测量。结果设计完成了测量组织工程血管内壁摩擦力的实验装置,弹性元件的牵拉力与位移的关系呈良好的线性关系。聚羟基乙酸PGA管状支架的摩擦力随流速的增加而增加。结论测量组织工程血管内壁摩擦力的实验装置是可行的,聚羟基乙酸PGA管状支架的摩擦力与流速基本呈现线性关系。     

连续浅缝法修补膜周型室间隔缺损

目的探讨连续浅缝法修补单纯膜周型室间隔缺损(pVSD)法的效果。方法选取2002年1月至2004年10月我院收治的体重10kg以内(≤10kg)50例单纯膜周型室间隔缺损患者,均采用自体心包连续浅缝法修补。结果主动脉阻断时间32±21min(14-52min),体外循环时间56±35min(29-69min)。无°房室传导阻滞发生,9例发生右束支传导阻滞,2例结性心律。2例膜周偏流出道型VSD患者因前上缘残余分流分别为0.4cm和0.3cm,再次手术修补;1例后下缘残留细束样分流,直径0.15cm,随访6个月后自愈;10例膜周偏流出道型VSD患者剪开三尖瓣,8例垂直瓣环,2例平行瓣环;1例三尖瓣中度反流,随访无加重;5例轻度反流,4例轻微反流,随访均无加重;1例心包积液;1例再次进入手术室止血。结论连续浅缝法是修补单纯膜周型室间隔缺损的有效外科纠治方法。     

不同持续时间的周期性压力对构建组织工程软骨的影响

Objective To determine the best duration for exerting the cyclic pressure under which the tissue-engineered cartilage is constructed. Methods Free chondrocytes isolated from rabbit articular cartilage were seeded into polylactic acid-co-glycolic acid(PLGA) scaffolds after expansion in vitro, and ran-domized into 4 groups. In Groups 1 to 3, chondrocytes were cultured under daily cyclic pressure (0 ～ 200 kPa, 0.1Hz) for 4 hours, 8 hours, 12 hours respectively; Group 4 was a control in which no pressure was exerted. In each group, after 2 weeks of culture, the tissue engineered cartilages were observed in vitro and assessed by his-tological staining of liE. Next, the content of DNA and the secretion of type Ⅱ collagen and GAG in cartilages were detected quantitatively. Results Under the daily cyclic pressure (0 ～200 kPa, 0.1 Hz), the scaf-fold-chondrocytes complex in the group of 8 hours got the largest volume, smooth, lucidus, and elastic surface, the most queuing chondrocytes, and the highest content of type Ⅱ collagen and GAG (P < 0.01). Conclusions Since chondrecytes are baro-senstive, the metabolism of chondrocytes can be affected by the time of cyclic pressure. Under the effect of 0 ～200 kPa, 0.1Hz, the daily cyclic pressure of 8 hours may be optimal for chondrocytes to multiply and synthesize extracellular matrixes such as type Ⅱ collagen and GAG.     

关节镜下应用“微骨折”方法修复关节软骨缺损

目的:探讨关节镜下应用“微骨折”技术对膝关节全层关节软骨缺损修复的效果。方法:对68例全层关节软骨缺损患者进行随机分组:实验组(35例),男17例,女18例,平均年龄35.1岁,采用关节清理后应用“微骨折”技术进行处理,即利用骨刀设计的特性和适度的锤击力量造成软骨下的骨组织微小骨折,刺激软骨生长。对照组(33例),男17例,女16例,平均年龄31.6岁,仅作关节清理术。结果:术后随访6~18个月,平均8.6个月,按Lysholm评分标准,实验组明显优于对照组(P<0.01)。结论:关节镜下应用“微骨折”技术能够显著减轻关节疼痛,增加关节活动度,改善关节功能,是一种简单有效的修复全层关节软骨缺损方法。     

Safety of, and biological and functional response to, a novel metallic implant for the management of focal full-thickness cartilage defects: Preliminary assessment in an animal model out to 1 year.

Focal full-thickness cartilage lesions of the human medial femoral condyle (MFC) can cause pain and functional impairment. Affected middle-aged patients respond unpredictably to existing treatments and knee arthroplasty may be required, prompting risk of revision. This study assesses the safety of, and biological and functional response to, a metallic resurfacing implant which may delay or obviate the need for traditional arthroplasty. The anatomic contour of the surgically exposed MFC of six adult goats was digitally mapped and an 11 mm diameter full-thickness osteochondral defect was created. An anchor-based Co-Cr resurfacing implant, matching the mapped articular contour, was implanted. Each goat's contralateral unoperated femorotibial joint was used as a control. Postoperative outcome was assessed by lameness examination, radiography, arthroscopy, synoviocentesis, necropsy, and histology up to 26 (n = 3) or 52 (n = 3) weeks. By postoperative week (POW) 4, goats demonstrated normal range of motion, no joint effusion, and only mild lameness in the operated limb. By POW 26 the animals were sound with only occasional very mild lameness. Arthroscopy at POW 14 revealed moderate synovial inflammation and a chondral membrane extending centrally across the implant surface. Radiographs at POWs 14 to 52 implied implant stability in the operated joints, as well as subchondral bone remodeling and mild exostosis formation in the operated and contralateral unoperated joints of some goats. By POW 26, histology revealed new trabecular bone abutting the implant. At POWs 26 and 52 MFC cartilage was metachromatic and intact in the operated and unoperated femorotibial joints. Proximal tibiae of some operated and unoperated limbs demonstrated limited subchondral bone remodeling and foci of articular cartilage fibrillation and thinning. The chondral membrane crossing the prosthesis possessed a metachromatic matrix containing singular and clustered chondrocytes. Our data imply the safety, biocompatibility, and functionality of the implant. Focal articular damage was documented in the operated joints at POWs 26 and 52, but lesions were much reduced over those previously reported in untreated defects. Expanded animal or preclinical human studies are justified.     

管状膨体聚四氟乙烯用于组织工程支架材料的动物实验研究

目的：研究膨体聚四氟乙烯(ePTFE)植入动物体内后与机体的关系。方法：管状膨体聚四氟乙烯两端闭合，内充DMEM培养液，埋入家兔皮下，观察宿主生命情况，于术后1、2、4、6周取出标本，行大体及光镜下观察。结果：家兔在植入膨体聚四氟乙烯材料及DMEM培养液后存活情况良好，DMEM培养液在1周时即已完全无色透明；4周时膨体聚四氟乙烯与周围组织有粘连，6周时粘连紧密；光镜下观察，各时间点材料间隙均可见着色，1周时可见少量淋巴细胞浸润，无血管及组织细胞。材料内壁未见细胞附着。结论：植入膨体聚四氟乙烯材料及少量DMEM培养液对宿主动物存活无影响；管内外液体可相互交通，管状膨体聚四氟乙烯可作为组织工程化尿道的支架材料。     

快速眼动睡眠剥夺后大鼠脑内神经元骨架蛋白及超微结构的变化

目的探讨经历不同时间快速眼动（REM）睡眠剥夺对大鼠皮质及海马各区神经元形态结构的影响。方法选择微管相关蛋白（MAP2）和神经丝（NF）作为正常神经元结构的标识物，利用免疫组织化学法和Western blot技术观察REM睡眠剥夺1、3、5、7 d4个时间点大鼠皮质及海马MAP2和NF表达的时空变化规律。同时运用电镜技术观察睡眠剥夺后神经元超微结构的变化。我们的实验是用改良的多平台睡眠剥夺模型进行REM睡眠剥夺，结合免疫组织化学染色技术和蛋白质电泳以及电镜超微结构分析。结果REM睡眠剥夺后5d大鼠皮质、海马CA1及齿状回神经元结构蛋白MAP2和NF表达较对照组明显减少（P〈0．05）；电镜神经元核仁偏位，胞质中出现少量肿胀的线粒体和内质网；部分神经轴突的髓鞘溶解与浓集。环境对照组、REM睡眠剥夺5d和7d组，皮质中超微结构改变的神经元所占比例分别为1．2％、3．6％和5．8％。结论REM睡眠剥夺能够导致大鼠脑内神经元的超微结构发生异常变化。     

Supragaleal placement of tissue expander for post-burn alopecia.

Supragaleal placement of tissue expander for management of alopecia of scalp excludes tough galeal layer so that easier and faster expansion of scalp may be achieved.