Acid Ethanol Fixation and Polyester Wax Embedding Combines Preservation of Antigenic Determinants with Good Morphology and Enables Simultaneous Bromodeoxyuridine (BRDU) Labeling

Lymphoid tissue, and/or isolated peripheral mononuclear blood cells were fixed in acid ethanol and embedded in polyester wax (melting point 37 C). The excellent cytomorphol-ogy obtained allowed distinguishing different types of individual lymphoid and nonlymphoid cells. Furthermore, this procedure was satisfactory in the immunophenotyping of histiocytes, endothelial, mesenchymal, epithelial cells, different (sub-) types of lymphocytes and also of lym-phokine activated killer (LAK) cells. The staining patterns obtained with the different poly- and monoclonal antibodies on polyester wax sections were not only analogous to those obtained on frozen sections, but cells which had incorporated bromodeoxyuridine could be double labeled with specific antiserum.     

Enhanced protoplast division by encapsulation in droplets: An advance towards somatic hybridisation in recalcitrant white lupin

Lupins are highly nutritious fodder and pulse crops but the greatest challenge in their genetic enhancement is the difficulty in obtaining hybrids through conventional sexual approaches. To bypass this, a procedure for the culture of hitherto recalcitrant lupin protoplasts is now being developed so that the somatic hybrids can be regenerated. This study provides a basis for a regime to culture lupin protoplasts. Cotyledonary protoplasts of white lupin (Lupinus albus) were plated in two diverse media for the evaluation of various plating regimes. The protoplasts divided in agarose as well as in Gelrite? but embedding in agarose at 6 g L^?1 concentration resulted in a higher rate of mitosis. Sodium alginate embedding inhibited protoplast division. Protoplast plating in the form of liquid suspension was significantly inferior to embedding. A filter paper substratum was clearly noxious to protoplast division. Vis‐à‐vis other designs of plating, a 400% improvement in protoplast elongation and division was achieved by plating in the form of 25 μL droplets at the base of 60 mm × 15 mm Nunclon? dish and overlaying with liquid medium. Better results in terms of protoplast elongation and division were obtained with K8p medium as compared to the AS medium. This report on lupin protoplast culture represents a significant breakthrough in the genus in which morphogenesis has not been described to date.     

水稻原生质体培养及植株再生的研究

由粳稻77-170品系及籼稻品种IR-50的细胞悬浮培养物游离的原生质体,用琼脂糖包埋于RY-2培养基中,发生了持续分裂。前者植板率达2.5％以上,二者最后都再生出植株。对游离和培养方法做了如下改进:1)采用两步法,即先用果胶酶,再用果胶酶和纤维素酶的混合酶进行游离,可避免原生质体发生融合并获得高质量的原生质体;2)悬浮细胞培养基中加入ABA有利于原生质体的存活和分裂;3)琼脂糖包埋培养可大大提高植板率;4)用较高渗透压的培养基培养原生质体再生的细胞团及愈伤组织,可提高植株再生频率。由于这两个品种(系)的培养物都已继代一年半之久,再生植株均为白化苗。这是迄今第一个由籼稻原生质体再生植株的报道。     

Immunohistochemistry on Resin Sections: A Comparison of Resin Embedding Techniques for Small Mucosal Biopsies

We have modified resin embedding methods to provide optimal information from en-doscopic biopsies. Mucosal biopsies were fixed either in buffered formalin and processed for embedding in Araldite or in acetone containing protease inhibitors and embedded in glycol meth-acrylate (GMA). GMA embedding generated an im-munophenotypic profile similar to that obtained in frozen sections while yielding far superior morphology and greater numbers of sections from small biopsies. The phenotypic markers included those for T cells, macrophages, mast cells, eosin-ophils and neutrophils. We have also demonstrated collagens, cell adhesion molecules and integrin molecules. Sections of similar quality were obtained with Araldite but the repertoire of antibodies was restricted to those which can be applied to formalin fixed, paraffin embedded tissues. We suggest that for optimal results, small biopsies to be subjected to immunochemistry are fixed in acetone at -20 C with the inclusion of protease inhibitors and embedded in GUIA with careful temperature control.     

实现PCR热启动的一种简便实验室技术

将Taq酶加入热的低熔点琼脂糖凝胶中,趁热将液态的凝胶加入扩增管底部,待凝胶冷却凝固后,Toq酶便得以包埋。扩增过程中只有当温度升高至70℃以上时,凝胶才能融化释放Taq酶,从而实现PCR过程中的热启动。这一方法简单方便、节约经费,是一种值得在实验室大力推广的实用技术。     

膝关节镜辅助微创手术治疗复杂性胫骨平台骨折的疗效分析

目的:探讨膝关节镜辅助微创手术治疗复杂性胫骨平台骨折的疗效。方法:搜集2013年2月-2015年1月期间我院收治的确诊为复杂性胫骨平台骨折患者104例,按照随机数字表法分为微创组和对照组,每组各52例。对照组采用传统切开复位钢板内固定术治疗,微创组采用膝关节镜辅助微创手术治疗;观察两组患者临床各项指标、膝关节功能HSS评分以及术后并发症发生率。结果:术后微创组下床活动时间、完全负重下地时间和骨折愈合时间显著低于对照组(P0.05);三个月后的关节活动度、一年后的膝关节功能优良率显著高于对照组(P0.05);术后微创组并发症发生率为9.62%(5/52),显著低于对照组的23.08%(12/52),差异具有统计学意义(P0.05)。结论:膝关节镜辅助微创手术治疗复杂性胫骨平台骨折,临床疗效显著,术后膝关节功能恢复好,并发症发生率低,值得临床推广应用。     

Distal trisomy 10q and limb defects

Analysis of the literature showed that hypoplasia (or aplasia) of tibiae was found at least in six persons with trisomy 10q25.2-qter. Therefore, these defects should be considered as a characteristic manifestation of the distal trisomy 10q. In most of these patients, tibial abnormalities were associated with other defects of the lower extremities (hypoplastic femora, ectrodactyly, preaxial polydactyly). Upper limbs were affected in one patient (as well as in her sib without tibial defects). Most likely, segment 10q25.2-qter contains a gene which (when triplicated) leads to maldevelopment of the limbs, and tibial malformations are only one manifestation of this field defect.     

锁定加压钢板接骨术治疗胫骨Pilon 骨折

目的:评估锁定加压钢板接骨术治疗胫骨Pilon骨折的应用价值。方法:2006年4月-2010年12月,对48例胫骨Pilon骨折患者,采用锁定加压钢板(LCP)治疗的结果进行总结。手术前后运用AOFAS(美国足与踝关节协会)踝与后足功能评分(Ankl-e-Hindfoot Scale)系统进行治疗效果的评估。结果:本组按AOFAS踝与后足功能评分系统:术前平均为(67.6±7.4)分,可18例,差30例;术后评分改善到平均为(90.9±7.2)分,优19例,良21例,可5例,差3例,踝与后足功能评分术前与术后比较,差异有统计学意义(P<0.05)。结论:用锁定加压钢板接骨术治疗胫骨Pilon骨折,有利于踝关节功能恢复,疗效确切,适应症广泛,较其他内固定器而言,锁定加压钢板在临床上更适用。     

Load along the tibial shaft during activities of daily living

External load at the tibia during activities of daily living provides baseline measures for the improvement of the design of the bone–implant interface for relevant internal and external prostheses. A motion analysis system was used together with an established protocol with skin markers to estimate three-dimensional forces and moments acting on ten equidistant points along the tibial shaft. Twenty young and able-bodied volunteers were analysed while performing three repetitions of the following tasks: level walking at three different speeds, in a straight-line and with sudden changes of direction to the right and to the left, stair ascending and descending, squatting, rising from a chair and sitting down. Moment and force patterns were normalised to the percentage of body weight per height and body weight, respectively, and then averaged over all subjects for each point, about the three tibial anatomical axes, and for each task. Load patterns were found to be consistent over subjects, but different among the anatomical axes, tasks and points. Generally, moments were higher in the medio/lateral axis and influenced by walking speed. In all five walking tasks and in ascending stairs with alternating feet, the more proximal the point was the smaller the mean moment was. For the remaining tasks the opposite trend was observed. The overall largest value was observed in the medio/lateral direction at the ankle centre in level walking at high speed (9.1% body weight * height on average), nearly three times larger than that of the anterior/posterior axis (2.9) during level walking with a sidestep turn. The present results should be of value also for in-vitro mechanical tests and finite element models.