A modified chorioallantoic membrane (CAM) assay for qualitative and quantitative study of growth factors

Summary The application of Thermanox tissue culture coverslips to the day 9 CAM of the chick causes constant effects beneath the carrier after 3 days, and these are associated with a change in the blood vessel pattern. Histological sections show enormous thickening of the CAM in the reactive areas. The stroma of the CAM shows fibrocyte proliferation, leucocyte infiltration, and clusters of dispersed ectodermal epithelial cells exhibiting signs of necrosis. The latter obviously cause a strong vascular response. The same effects are seen when the Thermanox discs are applied at day 11. Following application on day 12 a positive or negative response to the carrier is observed, whereas on day 13 no such carrier effects are seen. The only remaining effect is compression of the intra-ectodermal capillary plexus of the CAM. This can macroscopically be seen after peroxidase staining of the blood vessels. The effect of 5 l PBS dried on the Thermanox disc and applied to the day 13 CAM is to cause, after 3 days, hyperosmotic damage to the ectodermal epithelium, which becomes overgrown by fibrocytes. We found dose-dependent effects of salt-free human bFGF applied to the day 13 CAM. The first and main effect is fibrocyte proliferation (0.5 g). New capillaries appear with higher doses, but are not as frequent as would be expected for an angiogenic substance (1.25–2.5 g). Also with higher doses additional hyperplasia of the endodermal (3.75 g) and ectodermal (5 g) epithelium can be seen. The latter might be a non-specific hyperosmotic effect. Leucocytes are regularly present within the reactive areas. When salt-free angiogenin is applied to the day 13 CAM, some effects appear with doses of 4.6 g and more. The ectodermal epithelium of the reactive areas is discontinuous, exhibiting signs of necrosis. It is overgrown by parallel fibrocytes. Whether this is a non-specific hyperosmotic effect, or indicates enhancement of invasive growth, calls for further investigation.     

Agrobacterium-mediated DNA transfer

Summary Agrobacterium tumefaciens naturally transfers DNA into plant cells and is clearly one of the most effective methods of directed DNA transfer presently available. Two kinds of vectors are commonly used. Cointegrative vectors have the foreign genes incorporated directly into the Ti plasmid. Binary vectors carry two plasmids; the main Ti plasmid where most of the T-DNA has been removed, and a second plasmid containing the foreign genes between the usual border sequences. The vir genes on the main plasmid function to mobilize the foreign genes into a plant cell. Most plant transformation methods follow the procedure of cocultivating wounded tissue with vir-gene-induced bacteria. The cocultivation step is followed by transfer to a selective medium containing antibiotics to kill the bacterium and to allow only growth of transformed tissue. Several selectable markers are available that include resistance to antibiotics, herbicides, or drugs. In addition, several scorable markers such as the bacterial glucuronidase, chloramphenicol acetyl transferase, and the Agrobacterium opine genes are used to verify transformation. Southern blotting and inheritance of transferred genes are ultimately used to demonstrate stable transformation.     

肩锁关节及关节盘的相关解剖学特征

背景:目前临床上多数认为,肩部疾病如肩锁关节脱位、肩峰下撞击综合征等与肩锁关节的解剖形态有很大的关系,但国内外文献很少有关于肩锁关节的形态学数据支持,特别对于肩锁关节关节盘的解剖学研究。同时肩锁关节属于微动关节,参与肩关节的联合运动,现临床上有关肩锁关节脱位的手术方式繁多,较流行的手术方式多为刚性固定,并未保留其微动特点以及锁骨和肩胛骨的运动特点,其中关节盘的取舍目前国内外并无大样本多中心对照研究。目的:对肩锁关节及关节盘行相关解剖学研究,用于更好地指导肩部疾病的诊治。方法:对58具肩锁关节尸体标本(同济大学解剖教研室提供)进行形态学及组织结构研究,共获得58个肩峰、58个锁骨和36个关节盘(包括完全型和不完全型)。采用BIGLIANI肩峰形态分型、DEPALMA锁骨形态分型、EMURA肩锁关节盘分型,获得各肩峰与锁骨组合下关节盘出现的频率并行苏木精-伊红染色(上海市普陀区人民医院提供)。结果与结论:①在58个肩锁关节尸体中,肩锁关节盘type1型共15例,所占比例最高,为25.9%;type3a型共4例,所占比例最少,为6.9%。②Ⅰ型肩峰32个,所占比例最高,为55.2%;Ⅲ型肩峰3个,所占比例最低,为5.2%。③Ⅰ型锁骨27个,所占比例最高,为46.6%;Ⅲ型锁骨9个,所占比例最低,为15.5%。④以Ⅰ型及Ⅱ型肩峰与Ⅰ型锁骨组合所占比例较大,分别为24.1%和22.4%,以Ⅲ型肩峰对Ⅰ型锁骨所占比例最小,为0%。⑤而在Ⅰ型肩峰与Ⅰ型锁骨组合下type1型和type2b型关节盘所占比例最大,分别为35.7%和28.6%;Ⅱ型肩峰与Ⅰ型锁骨组合下type2a型和type1型关节盘所占比例最大,分别为38.5%和23.1%。⑥苏木精-伊红染色在组织切片中可以发现,关节盘中的细胞外基质清晰可辨,细胞形态饱满,含有丰富的细胞质,提示为软骨细胞,即形成纤维软骨的主要成分。此外,从关节囊顶端和底端两侧可见部分骨膜纤维层移行至关节盘内侧,考虑共同组成关节盘结构,即关节盘由近骨面的软骨细胞及远离骨面的纤维层共同构成。     

经皮穿刺椎间盘髓核摘除术入路的研究

为提供椎间盘髓核摘除术形态学依据，在7具成人横断面标本上观察测量了L1～L5椎间盘平面经皮穿刺椎间盘髓核摘除术的进针点、角度和深度。发现在L1～L3、L4～L5椎间盘髓核的穿刺点以距后正中线8～10cm，9～11cm，穿刺角度以46．7°～59．5°，穿刺深度为110．2～128．9mm为最宜。     

Methods for culture ofBombyx mori wing discs

Summary A culture method forBombyx wing discs is described. Grace's insect medium,Bombyx hemolymph, and moderate doses of ecdysterone (0.02 to 0.1 µg/ml) were necessary for complete imaginal differentiation ofBombyx wing discs. This procedure is useful for understanding ecdysteroid action on insect imaginal differentiation and metamorphosis.     

人工髓核材料-聚乙烯醇水凝胶/聚乙烯纤维复合物的生物相容性研究*

目的评估人工髓核材料聚乙烯醇水凝胶/聚乙烯纤维复合物的生物相容性。方法根据ISO10993-1标准,采用细胞毒性试验(琼脂扩散法)、皮内刺激试验、Ame's致突变试验、微核试验和体内植入(360天)试验对聚乙烯醇水凝胶/聚乙烯纤维复合物的生物相容性进行评估。结果聚乙烯醇水凝胶/聚乙烯纤维复合物的细胞毒性评分小于Ⅰ级,细胞生长无明显抑制现象,对皮内无刺激作用,Ames致突变试验为阴性,微核出现率为3.48‰,无致突变反应。体内植入符合植入材料生物学评价要求。结论聚乙烯醇水凝胶/聚乙烯纤维复合物具有良好的生物安全性,是一种无毒、对皮肤及肌肉、椎间隙无刺激作用的生物医用材料,在动物体内不引起排异反应,可应用于临床。     

后路腰椎间盘镜治疗多节段腰椎间盘突出症

目的探讨多节段腰椎间盘突出症的后路腰椎间盘镜(MED)手术治疗方法。方法回顾性分析2000年至2004年经MED治疗的45例多节段椎间盘突出症的临床资料。结果对45例患者共93个椎间盘进行了手术，平均住院时间14．8d，平均手术时间92min，平均手术出血量145ml，术后随访3个月至40个月，手术优良率为91．6％。结论多节段腰椎间盘突出症是MED手术治疗的相对适应证，相对于单节段的MED手术，术前定位和术中操作更困难，严格的病例选择和全面的术前分析，辅以熟练的手术操作，仍可获得较为满意的疗效。     

不同位置植入SMH人工腰椎间盘对小关节应力分布的影响

目的:通过三维有限元方法研究正常椎间盘、SMH人工腰椎间盘前置、中置和后置四组模型中小关节的应力分布情况,探讨不同位置植入SMH人工腰椎间盘对小关节应力分布的影响。方法:建立正常腰椎间盘、人工SMH腰椎间盘前置、中置和后置的三维模型,然后模拟腰椎节段的运动,进行小关节应力分布的比较研究。结果:与正常L4/5椎间盘模型相比,除SMH人工腰椎间盘前置模型前屈时比正常模型小关节应力小外,其余运动时SMH人工腰椎间盘前置、中置和后置模型中小关节的应力均高于正常腰椎间盘组,但后置SMH人工腰椎间盘组则相对较小。结论:SMH人工腰椎间盘后置可以更好地预防小关节的退变;人工椎间盘植入位置的不同对人工腰椎间盘术远期疗效的预测具一定的作用。     

终板蛋白多糖的变化对颈椎间盘力学性能的影响

目的:通过测定退变颈椎间盘压缩性能的改变,同时观测软骨终板基质蛋白多糖(proteoglycan PG)的变化,为退变的椎间盘显现异常力学特性作一分子机制上的探讨。方法:将24只家兔随机分为对照组和模型组,模型组家兔保持45°低屈曲住5小时/次·天,取C5-6椎间盘进行生物力学测定;同时生化测定椎间盘终板糖胺多糖(glycosaminoglycan,GAG)总量、硫酸软骨素(chongdroitin sulphate,CS)和硫酸角质素(keratan sulphate,KS)比值和透明质酸(hyaluronic acid,HA)含量;总结分析颈椎间盘退变时基质各成分的系统变化及其对椎间盘力学性能的影响。结果:模型组终板抗压强度、断裂时的最大形变、终板基质蛋白多糖总含量、硫酸软骨素/硫酸角质素比值、透明质酸含量都低于对照组(P<0.05),并随造模进程的深入而减小。结论:长时间的应力不均状态,造成椎间盘终板材料力学特性改变;颈椎间盘基质蛋白多糖含量和成分改变是颈椎间盘生物力学性能减退的主要原因之一。     

磷酸钙人工骨(CPC)在颈前路椎间融合的临床应用

目的探讨磷酸钙人工骨(CPC)在颈椎前路椎间融合手术中的应用效果。方法2001年4月至2003年10月颈前路手术中应用磷酸钙人工骨栓椎间融合结合钛钢板固定治疗颈椎病17例,颈椎间盘突出症5例,颈椎外伤脱位2例,共24例35个节段。采用JOA评分评价神经功能,X线片判定融合效果。结果随访18±6.5个月,术后无感染,无过敏或毒性反应。JOA评分由术前9.28±2.15分增加到14.65±2.18分(P<0.001)。术后X线片未见CPC骨栓塌陷或移位,钛板和螺钉无松动及折断。术后16.5±6.8个月均获得椎间融合。结论颈椎前路椎间融合手术应用磷酸钙人工骨替代自体骨,经济、安全、简便、效果可靠。