一种新型免疫层析技术在临床乳腺癌Her-2蛋白定量检测中的应用

建立一种靶点蛋白质快速定量检测方法。在原有侧向流动免疫层析技术的基础上,通过优化层析材料和纳米微球的均一性、改进检测区的检测方法,经逐点扫描技术,建立标准浓度曲线,以达到对临床靶点蛋白质的定量检测。以乳腺癌组织中的Her2表达为例,通过对已知浓度样品的检测,验证本技术方法的准确度大于96%。另外,以蛋白质免疫印迹作为组织中特定蛋白质检测金标准,分析临床肿瘤组织中Her2蛋白的含量,其准确率也达到95.5%,而免疫组织化学方法检测准确率仅为69.58%。新型免疫层析法检测结果与靶向治疗患者的愈后密切相关(P<0.01)。改进后的新型免疫层析方法能够准确地对临床靶点蛋白质进行定量检测,而且结合侧向流动技术的简单、快速和易用性,这种新型检测方法可以广泛应用于临床组织标本、血液标本和体液标本中靶点蛋白质的临场定量检测,在一定程度上可以替代免疫组化技术。     

De novo organogenesis and plant regeneration in <Emphasis Type="Italic">Pongamia pinnata</Emphasis>, oil producing tree legume

Role of Thidiazuron (TDZ) in inducing adventitious organogenesis in Pongamia was studied. TDZ at different concentrations (0, 0.45, 2.27, 4.54, 6.71, 9.08, 11.35, 13.12 and 22.71 μM) were used for induction of caulogenic bud formation in deembryonated cotyledon explants. Each cotyledon was cut into three segments and identified as proximal, middle and distal. Duration of TDZ exposure, influence of the segment and orientation of the explant were studied. TDZ at 11.35 μM concentration was optimum for the induction of shoots and rapid elongation. Shoots induced at higher concentration elongated after several passages in growth regulator free medium, thereby extending the period of differentiation. Exposure of the explant for 20 days yielded more number of buds than 10 days. Proximal segment of the cotyledon was more responsive. Contact of abaxial surface in the medium was more effective and generated more buds than the adaxial side. Buds differentiated and elongated on transfer to MS basal medium for 8–12 passages of 15 days each. Rooting and elongation of shoots was achieved in charcoal supplemented half-strength MS medium. Rooted plantlets survived on transfer to sand soil mixture. The plants were hardened and transferred to green house. This is the first report on in vitro regeneration of Pongamia pinnata via adventitious organogenesis using TDZ. This protocol may find application in studies in genetic transformation, isolation of somaclonal variants and in induction of mutants. It also provides a system to study the inhibitory role of TDZ on shoot differentiation.     

利用大孔微载体体外制备工程化组织

组织工程从诞生至今已有二十年的历史.微载体在组织工程领域的应用研究近年来才日趋升温.利用大孔微载体Cytopore体外构建工程化微构组织.并基于其聚团特性,进行灌注再装配大组织的尝试.结果20 d内微构组织达最高细胞密度16.4×107 cells/cm3;32 d后.所形成的微构组织整体均质性好、活性高、基质生成丰富;培养后期微构组织发生明显聚团,经灌注再装配得到细胞和基质分布均匀的厘米级(12 mm×6 mm)大组织.说明利用大孔微载体构建工程化组织是一种行之有效、富有潜力的体外组织构建方案,有望构建尺寸>5 mm、细胞和基质分布均匀的大组织.为体外组织构建提供新的思路.     

Administration of signalling molecules dictates stem cell homing for in situ regeneration

Ex vivo‐expanded stem cells have long been a cornerstone of biotherapeutics and have attracted increasing attention for treating intractable diseases and improving tissue regeneration. However, using exogenous cellular materials to develop restorative treatments for large numbers of patients has become a major concern for both economic and safety reasons. Advances in cell biological research over the past two decades have expanded the potential for using endogenous stem cells during wound healing processes, and in particular, recent insight into stem cell movement and homing has prompted regenerative research and therapy based on recruiting endogenous cells. Inspired by the natural healing process, artificial administration of specific chemokines as signals systemically or at the injury site, typically using biomaterials as vehicles, is a state‐of‐the‐art strategy that potentiates stem cell homing and recreates an anti‐inflammatory and immunomodulatory microenvironment to enhance in situ tissue regeneration. However, pharmacologically coaxing endogenous stem cells to act as therapeutics in the field of biomedicine remains in the early stages; its efficacy is limited by the lack of innovative methodologies for chemokine presentation and release. This review describes how to direct the homing of endogenous stem cells via the administration of specific signals, with a particular emphasis on targeted signalling molecules that regulate this homing process, to enhance in situ tissue regeneration. We also provide an outlook on and critical considerations for future investigations to enhance stem cell recruitment and harness the reparative potential of these recruited cells as a clinically relevant cell therapy.     

Can generic knee joint models improve the measurement of osteoarthritic knee kinematics during squatting activity?

Knee joint kinematics derived from multi-body optimisation (MBO) still requires evaluation. The objective of this study was to corroborate model-derived kinematics of osteoarthritic knees obtained using four generic knee joint models used in musculoskeletal modelling – spherical, hinge, degree-of-freedom coupling curves and parallel mechanism – against reference knee kinematics measured by stereo-radiography. Root mean square errors ranged from 0.7° to 23.4° for knee rotations and from 0.6 to 9.0 mm for knee displacements. Model-derived knee kinematics computed from generic knee joint models was inaccurate. Future developments and experiments should improve the reliability of osteoarthritic knee models in MBO and musculoskeletal modelling.     

多西紫杉醇修饰的人工晶体与眼组织相容性分析

目的：探讨经多西紫杉醇修饰的人工晶体对眼组织相容性的影响。方法：按照随机数字表法将32 只日本大耳兔分为两组： 实验组通过手术植入表面经多西紫杉醇修饰处理后的疏水性人工晶体，对照组植入疏水性人工晶体。比较两组人工晶体亲水角、 术后24 小时光耀斑块计数以及人工晶体周围组织炎症浸润数。结果：实验组的亲水角小于对照组，差异有统计学意义（P<0.05）。 实验组光耀斑块计数低于对照组，差异有统计学意义（P<0.05）。实验组家兔人工晶体周围组织炎症浸润计数低于对照组，差异有 统计学意义（P<0.05）。结论：人工晶体表面经多西紫杉醇修饰后，其亲水性、与眼组织的组织相容性增加，且可缓解光耀斑炎症感 染和降低并发症的发生，有重要的临床参考价值。     

A novel micro-to-macro approach for cardiac tissue mechanics

For studying cardiac mechanics, hyperelastic anisotropic computational models have been developed which require the tissue anisotropic and hyperelastic parameters. These parameters are obtained by tissue samples mechanically testing. The validity of such parameters are limited to the specific tissue sample only. They are not adaptable for pathological tissues commonly associated with tissue microstructure alterations. To investigate cardiac tissue mechanics, a novel approach is proposed to model hyperelasticity and anisotropy. This approach is adaptable to various tissue microstructural constituent’s distributions in normal and pathological tissues. In this approach, the tissue is idealized as composite material consisting of cardiomyocytes distributed in extracellular matrix (ECM). The major myocardial tissue constituents are mitochondria and myofibrils while the main ECM’s constituents are collagen fibers and fibroblasts. Accordingly, finite element simulations of uniaxial and equibiaxial tests of normal and infarcted tissue samples with known amounts of these constituents were conducted, leading to corresponding tissue stress–strain data that were fitted to anisotropic/hyperelastic models. The models were validated where they showed good agreement characterized by maximum average stress-strain errors of 16.17 and 10.01% for normal and infarcted cardiac tissue, respectively. This demonstrate the effectiveness of the proposed models in accurate characterization of healthy and pathological cardiac tissues.     

Nitrate induced iron deficiency chlorosis in Juncus acutiflorus

Chlorosis caused by iron deficiency is commonly associated with high bicarbonate levels in the soil. However, in rare cases such chlorosis has been observed in soils with high nitrate levels. In a dutch rich-fen, chlorosis has been noted in stands of Juncus acutiflorus at locations where groundwater containing high levels of nitrate reached the surface. Experiments revealed that the chlorosis could be attributed to iron deficiency although iron levels in the shoots were well above the known physiological threshold values for iron deficiency. It is postulated that increased nitrate assimilation leads to an increased apoplastic pH and to a concomitant immobilisation of iron and/or lower iron (III) reduction. Moreover free amino acid levels were markedly higher in the iron deficient plants in the field. It was found, however, that the percentage of nitrogen present as free amino acids was not influenced directly by low iron levels but mainly by the C/N ratios in the shoots. Nowadays, nitrate concentrations in ground water as high 1000 µM are no longer an exception in the Netherlands. We propose that strongly increased nitrate inputs may cause iron stress in natural vegetations, especially in wet habitats.     

Direct somatic embryogenesis from axes of mature peanut embryos

Summary Plant regeneration via somatic embryogenesis was obtained in peanut (Arachis hypogaea L.) from axes of mature zygotic embryos. The area of greatest embryogenic activity was a 2-mm region adjacent to and encircling the epicotyl. Somatic embryogenesis was evaluated on Murashige and Skoog media supplemented with a variety of auxin treatments. Maximum production occurred on medium supplemented with 3 mg · liter⁻¹ 4-amino-3,5,6-trichloropicolinic acid. Explant cultures were transferred to half-strength medium supplemented with 1 mg · liter⁻¹ gibberellic acid for somatic embryo germination and early plantlet growth. Plantlets, transferred to soil, were placed in a greenhouse and grown to maturity.     

Tree-ring dating of “brush structures” in Kluane National Park and Reserve,Yukon, Canada

“Brush structures” are temporary wooden structures built with unmodified local materials and used as shelters by First Nation Peoples in the forests of the Yukon prior to European contact. This paper reports a preliminary attempt to date these structures using dendrochronology. Investigations were carried out of four njel (“teepee like”) structures and eight män-ku (low 2–3 sided wall structures) at four main sites. The primary material cored was poles (dead spruce trunks), often only 10–20 cm diameter, with narrow, sometimes extremely suppressed ring sequences. These structures are dated between 1865 and 1887, based on the latest (outermost) ring in the sampled material. The limited sampling and use of old wood in these structures (whether fire-kill, standing dead or reused from previous features) makes it difficult to give precise dates for the initial evidence of First Nation activity at these sites: more extensive sampling could provide further insight into the settlement history and construction techniques used. The sites investigated date from the latter half of the nineteenth century shortly before the first European gold rush to this region.