脏器微血管对荧光素钠通透性的实验方法

大鼠颈动脉注射１％ＦｌＮａ,荧光显微镜下活体观察肠系膜微血管血流状态及ＦｌＮａ的渗出情况,并在不同时间点经股动脉采血,测定血浆内ＦｌＮａ浓度随时间的变化,利用组织匀浆测定不同脏器中ＦｌＮａ的分布,再辅以冰冻切片进行观察。活体观察发现,ＦｌＮａ注入体内后,经微血管迅速向周围组织渗出,最后汇集于淋巴管,血浆及组织匀浆ＦｌＮａ浓度的测定表明,ＦｌＮａ浓度随时间的变化呈指数衰减,各脏器ＦｌＮａ的分布极不相同。冰冻切片也显示了同样的分布差别。这些结果表明,我们所建立的方法可直观、定量地反映ＦｌＮａ在微血管的通透情况。     

地高辛精标记酪氨酸羟化酶RNA探针的制备研究

为制备用地高辛精（Ｄｉｇｏｘｉｇｅｎｉｎ,Ｄｉｇ）标记的酶氨酸羟化酶（ＴｙｒｏｓｉｎｅＨｙｄｒｏｘｙｌａｓｅ,ＴＨ）ＲＮＡ探针,本研究用分子生物学技术重组质粒ＰＧＥＭＴＨＩ,即分别将携带Ｔ７和Ｓｐ６启动子的ＰＧＥＭ－３ｚｆ质粒和携带ＴＨ基因的ＰＫＳＴＨ质粒用限制性内切酶消化并行分离纯化,得到带Ｔ７和ｓｐ６启动子的ＤＮＡ片段和ＴＨ基因片段;经Ｔ４ＤＮＡ连接酶连接后转入大肠杆菌,得到ｐＧＥＭＴＨ１重组克隆。经小量提取质粒井用酶切分析检测,证实其确有ＴＨ基因且方向正确。将此质粒再经限制性内切酶消化则得到线状ＤＮＡ片段,用Ｔ７ＲＮＡ聚合酶转录合成带有Ｄｉｇ标记的高比活度的单链ＲＮＡ探针;经斑点杂交试验证实该探针具有较高的可靠性。这将为基因治疗帕金森氏病动物模型的疗效检测提供有效手段。     

不同蔬菜防癌抗促活性的比较研究

本研究采用EBV-EA诱导抑制实验的方法,对40种蔬菜,60个品种,共150个样品的防癌抗促作用效果进行了筛选与比较,其中具有中等以上抑制活性的样品117个,占样品总数的78％,尤其以非洲野苋菜、辣椒、羽衣甘蓝、山药芋头、苦瓜及紫苏、罗勒等一些芳香莱的效果较好。不同品种、不同植株部位、不同提取方法以及不同产地,对蔬菜的抗促活性也有影响,值得进一步研究。     

蜂花粉抗脑衰老的实验动物研究

蜂花粉抗脑衰老的实验动物研究蒋滢,杨炳华,黄美英苏州医学院生化教研室苏州２１５００７２探索衰老机制,寻求延缓衰老的有效途径是生命科学中的重大问题,也是亟待解决的实际问题。脑是指挥全身一切活动的中枢,脑组织特别容易遭受自由基及活性氧的损伤,因此防治脑衰...     

植物种质资源的超低温保存研究进展（综述）

植物种质资源的超低温保存研究进展（综述）殷晓辉,舒理慧（武汉大学生命科学学院,武汉４３００７２）ＡＤＶＡＮＣＥＳＩＮＣＲＹＯＰＲＥＳＥＲＶＡＴＩＯＮＲＥＳＥＡＲＣＨＯＮＰＬＡＮＴＧＥＲＭＰＬＡＳＭ￥ＹｉｎｇＸｉａｏｈｕｉ;ＳｈｕＬｉｈｕｉ（Ｓｃｈｏｏ...     

甘露醇对Lowry法测定冻干注射用胸腺肽中多肽含量的影响

用Ｌｏｗｒｙ法测定冻干注射用胸腺肽中多肽含量时,由于冻干前所加赋形剂甘露醇的干扰,其测定结果明显偏高（Ｐ＜０．０００５）。本文经试验证明,用等量于检品中的甘露醇作空白对照,可以消除这种干扰。     

Intraspecific molecular variation inHieracium sect.Alpina (Asteraceae), an apomictic group

Intraspecific variation of four agamospecies ofHieracium sect.Alpina was studied using RAPD and isozyme techniques. No variation in either multiprimer RAPD or multi-enzyme phenotypes was observed withinH. holosericeum, suggesting that this widespread species consists of only a single genotype. A low level of within-population isozyme variation was seen inH. tenuifrons andH. calenduliflorum, the origin of which appears to be consistent with somatic mutation. Most isozyme and all RAPD variation in these two species was partitioned between populations. A strong correlation with geography suggests that its cause may be due to polytopic (-polyphyletic?) origin or perhaps to mutation and dispersal. The most variable species wasH. alpinum, in which isozyme variation occurred mostly within populations rather than between them, suggesting occasional sexual events or that the parents ofH. alpinum were heterozygous. RAPD variation in this species, in contrast, was partitioned between Scottish and Swiss populations, suggesting the existence of geographical races.     

GABAB Receptor-Mediated Effects in Synaptosomes of Lethargic (lh/lh) Mice

Abstract: Previously, we have shown a significant increase in number of GABA_B receptor binding sites in neocortex and thalamus of lethargic ( lh/lh ) mice, a mutant strain exhibiting absence seizures. This study was performed to test our hypothesis that presynaptic GABA_B receptors would inhibit [³H]GABA release to a greater degree in lh/lh mice compared with their nonepileptic littermates (designated +/+). Synaptosomes isolated from neocortex and thalamus of age-matched male lh/lh and +/+ mice were similar in uptake of [³H]GABA. In the neocortical preparation, baclofen dose-dependently inhibited [³H]GABA release evoked by 12 m M KCl, an effect mediated by GABA_B receptors. The maximal inhibition ( I _max) value was significantly greater (80%) in lh/lh than +/+ mice, whereas the IC₅₀ (3 µ M ) was unchanged. In the thalamic preparation, the effect of baclofen (50 µ M ) was 58% less robust in lh/lh mice. Other effects mediated by GABA_B receptors (inhibitions in Ca²⁺ uptake and cyclic AMP formation) were also significantly reduced in thalamic synaptosomes from lh/lh mice. These data suggest a greater presynaptic GABA_B receptor-mediated effect in neocortex and a reduced effect in thalamic nuclei of lh/lh mice. It is possible that selective effects of presynaptic GABA_B receptors on GABA release in neocortex and thalamic nuclei of lh/lh mice may contribute to mechanisms underlying absence seizures.     

线粒体DNA缺失与神经肌肉性疾病的研究

应用PCR技术对13例神经肌肉疾病患者的线粒体DNA缺失进行了研究。结果表明,其中二例肢带型肌营养不良症患者骨骼肌组织和一例帕金森氏病患者的血细胞线粒体DNA中存在至少526bp的缺失。提示线粒体突变在一些神经肌肉性疾病的发生中起一定的作用。 Abstract: Using PCR technique,we analysed the skeletal muscle and blood of 13 patients with neuromuscular disaeases.The results show that a mutant mitochondrial DNA with at least 526bp deletion exits in the skeletal muscles of 2 patients with Erb muscular dystrophy and in the blood of a patients with Parkinson’s disease.From our results,mitochondrial DNA mutations could be an important contributory factor to neuromuscular diseases.     

Expression of Bcl-2 protein in the epiphyseal plate cartilage and trabecular bone of growing rats

 The protooncogene protein, Bcl-2, protects cells from apoptosis and ensures their survival in vitro by inhibiting the action of the apoptosis-inducer, Bax. Its expression in proliferative and long-lived cells in vivo also indicates that it protects against cell death. The chondrocytes of the epiphyseal plate cartilage undergo a series of maturation steps and deposit mineral in the cartilage matrix before dying. The possibility that Bcl-2 helps protect chondrocytes until mineral deposition is completed was investigated by determining the distribution of Bcl-2 immunoreactivity in the epiphyseal plate cartilage of growing rats and its subcellular localization, using a specific antibody. The involvement of Bax in the triggering of chondrocyte death was checked by immunocytochemistry. Bcl-2 expression in the osteoblasts and the final result of their evolution, the osteocytes, was also examined in trabecular bone. Bcl-2 immunoreactivity was non-uniformly distributed throughout the epiphyseal cartilage. It was maximal in proliferative chondrocytes, decreased in mature chondrocytes, and low in hypertrophic chondrocytes, whereas there was Bax immunoreactivity in all chondrocytes examined. Immunolabeling was intense in osteoblasts but considerably lower in fully differentiated osteocytes. Bcl-2 immunoreactivity was mainly in the cytoplasm of chondrocytes, osteoblasts, and early osteocytes; the nuclei appeared clear. The subcellular distribution of Bcl-2 immunolabeling in chondrocytes, revealed by gold particles in the electron microscope, showed that gold particles were frequently concentrated in the mitochondria in all the cartilage zones and lay mainly within the organelles, not at their periphery. The endoplasmic reticulum contained moderate immunoreactivity and there were few gold particles in the cytoplasm and nuclei. The number of gold particles decreased in all the subcellular compartments from proliferative to hypertrophic chondrocytes. In contrast, Bax immunoreactivity changed little during chondrocyte terminal evolution, and its subcellular distribution mirrored that of Bcl-2. These immunocytochemical data indicate that Bcl-2 helps maintain chondrocytes and osteoblasts until their terminal maturation. Accepted: 19 February 1997