Neurofibromatosis： The role of guanosine triphosphatase activating proteins in sensory neuron function

Neurofibromatosis type 1 (NF1) is a common autosomal dominant disease characterized by formation of multiple benign and malignant tumors. People with this disorder also experience chronic pain, which can be disabling. Neurofibromin, the protein product of the Nfl gene, is a gnanosine triphosphatase activating protein (GAP) for p21Ras (Ras). Loss of Nfl results in an increase in activity of the Ras transduction cascade. Because of the growing evidence suggesting involvement of downstream components of the Ras transduction cascade in the sensitization of nociceptive sensory neurons, we examined the stimulus-evoked release of the neuropeptides, substance P (SP) and calcitonin gene-related peptide (CGRP), from primary sensory neurons of mice with a mutation of the Nfl gene (NfI 1-). Measuring the levels of SP and CGRP by radioimmunoassay, we demonstrated that capsaicin-stimulated release of neuropep-tides is 3-5 folds higher in spinal cord slices from Nfl 1-mice than that from wildtype mouse tissue. In addition, the potassium- and capsaicin-stimulated release of CGRP from the culture of sensory neurons isolated from Nfl 1- mice was more than double that from the culture of wildtype neurons. Using patch-clamp electrophysiological techniques, we also examined the excitability of capsaicin-sensitive sensory neurons. It was found that the number of action potentials generated by the neurons from Nfl 1- mice, responsing to a ramp of depolarizing current, was more than three times of that generated by wildtype neurons. Consistent with that observation, neurons from Nfl 1- mice had lower firing thresholds, lower rheobase currents and shorter firing latencies compared with wildtype neurons. These data clearly demonstrate that GAPs, such as neurofihromin, can alter the excitability of nociceptive sensory neurons. The augmented response of sensory neurons with altered Ras signaling may explain the abnormal pain sensations experienced by people with NFI and suggests an important role of GAPs in the mechanism of sensory neuron sensitization.     

Root Pressure of Tropical Lianas and Their Relationships with Phylogeny and Environments

Lianas usually possess large vessels, which are vulnerable to cavitation. Root pressure may play an important role in embolism repair of vessels. However, little is known about the generality of root pressure in tropical lianas. To characterize root pressure of lianas in tropical rainforests, we used pressure transducers to measure root pressure in the rainy and dry seasons for a total of 32 lianas from 14 families common in Xishuangbanna. We further analyzed the associations of maximum root pressure with phylogeny and of transient root pressure with environmental factors. We found that all lianas we selected had root pressure, with maximum root pressure ranging from 2-138kPa. In the dry season, about 72% (23 species) of the lianas had relatively low root pressure (<15kPa) and maintained positive throughout the day. This may be important for water balance for roots and basal stems of lianas. There were three types of diurnal changes in liana root pressure. In Type I, root pressure had obvious diurnal variation in the dry and rainy seasons. In Type II, root pressure did not show obvious diurnal variation in the dry and rainy seasons. In Type III, either in the dry or in the rainy season, root pressure showed obvious diurnal variation. Root pressure varied substantially among lianas, with lianas from two families, Fabaceae and Vitaceae, usually having relatively higher root pressure, suggesting that phylogeny may influence root pressure. Transient root pressure closely responded to photosynthetically active radiation. In most cases, however, it was not related to rainfall and vapour pressure deficit. These results suggest that the associations of liana root pressure with environments need further investigation.     

车八岭山地常绿阔叶林冰灾后林木受损的生态学评估

从植物生态学的角度对2008年初南方冻雨冰雪灾害对典型的亚热带山地常绿阔叶林造成的损害情况做了研究。对12个优势种和亚优势种的受灾情况做了对比分析, 对不同径级和不同地形因子下林木抵抗冻雨灾害的差异做了对比。统计检验结果显示林木受损程度存在显著的种间差异、径级差异和地形级差异。优势种米槠(Castanopsis carlesii)比多数亚优势种受害更严重; 各树种抗冻雨灾害能力以山茶科和樟科的种较强, 而壳斗科的多个种, 如米槠、栲(C. fargesii)受灾较为严重; 林木的受损比例随着胸径(diameter at breast height, DBH)增加而增大; 在未受灾的各级林木中, 超过70%的个体集中在最小一级径阶(1-5 cm)。χ²检验显示坡度、坡向、坡位等地形因子对林木受损状况有显著的影响, 随着坡位上升, 林木受损程度逐渐加重, 上坡位的林木受损比例最大, 下坡位最小, 这可能与随着坡位升高, 迎风面降温效果更迅速有关。对于坡向和坡度而言, 位于半阳坡的林木受灾比例显著高于半阴坡, 位于坡度级III (15°-25°)、IV (25°-35°)、V (35°-45°)上的林木受灾最严重。该研究结果对于亚热带常绿阔叶林的保育和应对未来可能再次发生的冰雪灾害有重要意义; 同时对于森林恢复和演替、乡土阔叶树种的选育及森林经营等方面的研究和应用也有重要的参考价值。     

NGF 与Neuritin 在脑外伤伴四肢骨折病人血清中的表达及意义

目的:通过检测脑外伤患者、脑外伤合并骨折患者、骨折患者及正常人外周血中NGF,neuritin不同时间段的表达,根据其含量的变化,判断与骨折愈合速度的相关性,寻找骨折愈合的关键因子。方法:收集单纯脑外伤、单纯骨折患者各80例、脑外伤合并骨折患者60例、健康体检人群20例。选取外伤后3天、10天、2周抽取所有实验对象的静脉血,应用ELISA技术测定标本中NGF与Neuritin的含量。结果:损伤后每个时间段里,患者血清中NGF、neuritin含量有不同程度升高,均高于正常对照组,其中又以脑外伤合并骨折组最高。血清NGF在骨折合并脑外伤组伤后第3天含量明显升高,为(0.86±0.21),伤后10天为(1.47±0.29),14天为(2.0 7±0.21),脑外伤合并四肢骨折组neuritin血清含量在伤后第3天略有升高为(83.47±18.85),10天(108.50±31.65),2周(91.86±21.12).脑外伤合并骨折病人血清NGF、neuritin表达明显高于其他对照组,差别均有统计学意义(P<0.05)。结论:脑外伤合并骨折病人血清NGF、neuritin表达明显高于其他对照组,说明与骨折愈合有着密切的相关性,两种因子可能在骨折愈合修复过程中共同起作用,从而,推测可能是脑外伤后骨折愈合加速的重要因素。     

Partial purification and characterization of a cellular protein that binds to the SV40 core origin of DNA replication

A monkey cell factor that interacts specifically with double- and single-stranded DNA sequences in the early domain of the simian virus 40 (SV40) core origin of replication was identified using gel-retention assays. The protein was enriched over 1200-fold using ion-exchange and affinity chromatography on single-strand DNA cellulose. Binding of protein to mutant origin DNA restriction fragments was correlated with replication activity of the mutant DNAs. Exonuclease footprint experiments on single-stranded DNA revealed prominent pause sites in the early domain of the core origin. The results suggest that this cellular protein may be involved in SV40 DNA replication.     

Proteomic analysis of phytopathogenic fungus <Emphasis Type="Italic">Botrytis cinerea</Emphasis> as a potential tool for identifying pathogenicity factors,therapeutic targets and for basic research

Botrytis cinerea is a phytopathogenic fungus causing disease in a substantial number of economically important crops. In an attempt to identify putative fungal virulence factors, the two-dimensional gel electrophoresis (2-DE) protein profile from two B. cinerea strains differing in virulence and toxin production were compared. Protein extracts from fungal mycelium obtained by tissue homogenization were analyzed. The mycelial 2-DE protein profile revealed the existence of qualitative and quantitative differences between the analyzed strains. The lack of genomic data from B. cinerea required the use of peptide fragmentation data from MALDI-TOF/TOF and ESI ion trap for protein identification, resulting in the identification of 27 protein spots. A significant number of spots were identified as malate dehydrogenase (MDH) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The different expression patterns revealed by some of the identified proteins could be ascribed to differences in virulence between strains. Our results indicate that proteomic analysis are becoming an important tool to be used as a starting point for identifying new pathogenicity factors, therapeutic targets and for basic research on this plant pathogen in the postgenomic era.     

In situ hybridization of corticotropin-releasing factor-encoding messenger RNA in the hypothalamus of the white sucker,Catostomus commersoni

Summary In situ hybridization procedure with a ³²P-labelled synthetic oligonucleotide probe was used to detect corticotropin-releasing factor-encoding messenger RNA (CRF mRNA) in the hypothalamus of the white sucker, Catostomus commersoni. Adjacent sections were immunostained by a sucker CRF-specific antiserum. CRF mRNA-containing neurons were identified by autoradiography in the magnocellular and parvocellular subdivisions of the preoptic nucleus (PON). Many of these neurons were also immunostained by sucker antiserum, showing the same distribution patterns. These results confirm the presence of CRF mRNA and CRF peptide in the white sucker hypothalamus and support the view that the magnocellular and parvocellular neurons of the PON may be involved in the control of adrenocorticotropic hormone secretion from the pituitary in the white sucker.