Individualized medicine 2010

Molecular and cell biology have revolutionized not only diagnosis, therapy and prevention of human diseases but also greatly contributed to the understanding of their pathogenesis. Based on modern molecular and biochemical methods it is possible to identify on the one hand point mutations and single nucleotide polymorphisms. On the other hand, using high throughput array technologies, it is possible to analyse thousands of genes or gene products simultaneously, resulting in an individual gene or gene expression profile (signature). These data increasingly allow to define the individual risk for a given disease and to predict the individual prognosis of a disease as well as the efficacy of therapeutic strategies (individualized medicine). In the following sections some of the recent advances of predictive medicine and their clinical relevance will be addressed.     

虫草素生理功效的研究进展

综述了虫草素生理功效的研究进展。虫草素的分子结构为3′-脱氧腺苷,它具有抗菌、抗病毒、抗肿瘤、调节免疫等作用,在医药、食品等领域具有广阔的应用前景。     

Neurotrophins improve synaptic transmission in the adult rodent diaphragm

Neurotrophins are usually viewed as secreted proteins that control long-term survival and differentiation of neurons. However, recent studies have established that among the most important functions of neurotrophins is their capacity to regulate synaptic functions and plasticity. When altering synaptic function, neurotrophins are able to produce two types of outcomes, an immediate effect on synaptic transmission and long-term control of synaptic structure and function. The first effect occurs within seconds or minutes after the neurotrophic factor has been applied and usually involves acute modification of synaptic transmission. The second effect takes hours and days, as protein synthesis is required to complete the structural changes. Neurotrophins and their receptors are expressed within the neuromuscular system, making these agents ideal candidates for the short-and long-term regulation of skeletal muscle function. For instance, neurotrophins can alter neuromuscular function acutely, by modulating the amount of neurotransmitter released with each nerve impulse, or chronically, by changing postsynaptic properties or the content and size of synaptic vesicles. It is obvious that the effects of neurotrophins depend on the specific neurotrophin involved (four neurotrophins have been found in mammals; these are nerve growth factor, brain-derived neurotrophic factor, and neurotrophins-3 and-4) and on the specific synapse being studied. Growing evidence highlights the role of neurotrophins in the development and function of neuromuscular synapses. This review will examine the role of neurotrophins in the regulation of neuromuscular transmission. Neirofiziologiya/Neurophysiology, Vol. 39, Nos. 4/5, pp. 327–337, July–October, 2007.     

Novel recombination system using Cre recombinase alpha complementation

A major limitation for the use of Cre recombinase is its toxicity and a lack of temporal control over its activity. We have developed a new recombination system using Cre recombinase α-complementation. Cre recombinase was divided and one fragment (β) was introduced into cells between two loxP sites with a CMV promoter in the upstream. The gene of interest (EGFP) was positioned just downstream of this construct. Cre recombinase activity was recovered by adding the other part of the molecule (α) to cells as a protein fragment, as evidenced by the expression of EGFP under the control of the CMV promoter. The activity of fragmented cre reached 68% of that of the wild type enzyme at 1 μM α-protein.     

Effect of high oxygen on placental function in short-term explant cultures

Ex situ culture of human gestational tissues has been routinely used as a model to investigate tissue function. The objective of this study was to determine the effect of varying oxygen concentrations on human term placental explants over a 24-h time period. Specifically, the effect of incubating placental explants in oxygen concentrations of 8%, 21% or 95% on tissue viability, metabolism and cell death was measured by assessing glucose consumption, lactate production, release of lactate dehydrogenase, parathyroid hormone-related protein (PTHrP), tumour necrosis factor-alpha (TNF-α) and 8-isoprostane, immunoreactivity for cleaved-caspase-9 and immunohistochemistry for the caspase-3-cleaved cytokeratin-18 neoepitope, M30. Exposure to higher oxygen concentrations significantly increased the rates of glucose consumption and lactate production. Apoptosis was significantly increased under conditions of higher oxygen as evidenced by increased M30 in placental explant sections. Similarly, hyperoxia significantly increased the releases of PTHrP, TNF-α and 8-isoprostane. Thus, incubation of placental explants with oxygen concentrations of 95% and, to a lesser extent, 21% oxygen was associated with the modulation of multiple cellular response pathways including those associated with tissue viability and cell death. These data are consistent with the hypothesis that hyperoxia activates pathways and mechanisms involved in cellular metabolism, necrosis and apoptosis, thereby shifting the balance from a steady state towards cell death.     

表达禽流感病毒特异性siRNA分子减毒鼠伤寒沙门氏菌的构建

为了寻找一种新的短干扰RNA(short interfering RNA,si RNA)片段的表达和递送系统,并进行评价,根据禽流感病毒核苷酸序列,设计了3条分别针对A型禽流感病毒核衣壳蛋白、聚合酶A和H5亚型禽流感病毒血凝素的特异性si RNA分子,人工合成相应DNA片段,退火后形成双链,与阴性对照片段分别连接到构建的pSIREN-ASD ZsGreen载体鼠源U6启动子下游,测序鉴定证实获得了阳性质粒,分别命名为PAZ-NP、PAZ-PA、PAZ-HA和PAZ-NC,1ng的质粒可以完全抑制50个PFU的禽流感病毒在MDCK细胞上的复制,细胞不产生任何病变,细胞上清的血凝效价为20,而对照组细胞在病毒感染36~48h后细胞变圆、死亡、脱落,其细胞上清的血凝效价可达24。将重组质粒转化减毒鼠伤寒沙门氏菌X4550得到重组沙门氏菌X4550(PAZ-NP)、X4550(PAZ-PA)、X4550(PAZ-HA)和X4550(PAZ-NC)。对4日龄雏鸡口服1×109菌落形成单位(CFU)的重组菌后第1d、7d、15d用105ELD50的H5亚型AIV进行攻毒,最后一次攻毒后观察15d,实验结束时采泄殖腔棉拭子对存活鸡进行病毒分离。结果表明,表达不同的si RNA分子对雏鸡有一定的保护力,保护率为27%~50%不等,存活鸡泄殖腔棉拭子未分离到病毒,说明重组减毒沙门氏菌携带质粒在体内产生的si RNA能够对机体产生一定保护力。     

艾迪注射液对急性白血病患者化疗减毒增效作用及肠道菌群影响的研究

艾迪注射液可扶植肠道正常菌群生长,减轻化疗药副作用,提高白血病患者化疗疗效。方法将选择病例随机分为艾迪注射液治疗组(试验组)及常规化疗组(对照组)。观察其化疗前后肠道常驻菌(肠杆菌、肠球菌、双歧杆菌、乳酸杆菌)指标,骨髓象检查及化疗后毒副作用观察。结果各项指标同单纯化疗组相比,差异均有显著性(P<0.05或P<0.01)。结论艾迪注射液具有扶植肠道正常菌群生长的作用,与化疗合用,减轻了化疗药的毒副作用,保证了化疗的顺利进行,提高了急性白血病患者的化疗疗效。     

红蚁净防治红火蚁的药效试验

以含1%氟虫腈、植物淀粉、引诱物质及粘附剂等自制的红蚁净(Pyragne)粉剂,在室内测定红火蚁Solenopsis invicta Buren各虫态的毒力。结果显示:红蚁净粉剂对红火蚁工蚁LT50=20.106h、LT90=23.78h,对有翅繁殖蚁LT50=20.951h、LT90=25.64h;工蚁能将红蚁净传递给蛹和卵引起中毒死亡。     

Seed bank persistence of clonal weeds in contrasting habitats: implications for control

The ability of weeds to form a seed bank is important for their population dynamics and management because it provides a refuge enabling reinvasion after established target plants have died. However, knowledge of the differential seed behaviour of individual species over multiple years and varying environmental conditions is surprisingly rare but necessary for effective control of diverse weed populations. We established a seed burial experiment in alpine habitats differing in management regime (i.e., forest, hay meadow and pasture) to determine whether seeds of the unpalatable perennial weeds, Veratrum album (white hellebore) and Gentiana lutea (yellow gentian) were able to delay germination and remain viable over 3 years. Our study shows that both species formed a short-term persistent seed bank; in the third-year, the soil seed banks of both species were nearly depleted, having declined to <5% of their original size. Both species had strikingly different germination strategies: G. lutea seeds mainly germinated in their first-year, whilst the majority of V. album seeds germinated in their second-year. The fraction of dormant G. lutea seeds increased with seed age, indicating that seeds remained viable after forgoing germination in the previous year. Habitat-specific differences in seed germination increased with seed age, with germination fractions being lowest in moist hay meadows. This suggests that the negative effects of anoxic conditions became more pronounced as seeds aged in hay meadows. Conversely, seed dormancy was equal among habitats. The absence of a long-term persistent seed bank has important implications for the management of both nuisance and endangered-plant populations. In the case of V. album and G. lutea, re-colonization of habitats from the seed bank is unlikely after established plants have been removed.