Large-scale de novo transcriptome analysis reveals specific gene expression and novel simple sequence repeats markers in salinized roots of the euhalophyte <Emphasis Type="Italic">Salicornia europaea</Emphasis>

Glycophytic plants suffer from severe stress and injury when roots are exposed to high salinity in the rhizosphere. In contrast, the euhalophyte Salicornia europaea grows well at 200 mM NaCl and can withstand up to 1000 mM NaCl in the root zone. Analysis of gene expression profiles and the underlying molecular mechanisms responsible for this tolerance have been largely overlooked. Using the Illumina sequencing platform and the short-reads assembly programme Trinity, we generated a total of 40 and 39 million clean reads and further 140,086 and 122,728 unigenes from the 200 mM NaCl and 0 mM NaCl treated tissues of S. europaea roots, respectively. All unigenes in this study were functionally annotated within context of the COG, GO and KEGG pathways. Unigene functional annotation analysis allowed us to identify hundreds of ion transporters related to homeostasis and osmotic adaptation as well as a variety of proteins related to cation, amino acid, lipid and sugar transport. We found significant enrichment in response to stress including the functional categories of “antioxidant activity”, “catalytic activity” and “response to stimuli”. These findings represent for a useful resource for the scientific community working on salt tolerance mechanisms. Conversely, a total of 8639 EST-SSRs from 131,594 unigenes were identified and 4539 non-redundant SSRs primers pairs were developed. These data provide a good foundation for future studies on molecular adaptation mechanisms of euhalophytes roots under saline environments and will likely facilitate the identification of critical salt tolerance traits to be transferred in economically important crops.     

云南地方猪种血液蛋白多态性研究

采用蛋白电泳技术研究了云南地方猪种血液蛋白多态性。共分析了3个云南地方猪种32-36个遗传位点,其中AKP、CAT、DIA、ES、G6PD、PA、6PGD、PHI、TF等9个 位点检测到多态性,多态位点百分比为0.1875-0.2121,平均杂合度为0.0712-0.1027。结果表明,云南地方猪种血液蛋白多态程度较高,反映在蛋白水平上的遗传多样性较为丰富。 Abstract:In this paper,protein electrophoresis was used to analyze the blood protein polymorphism in Yunnan local pig breeds and 32~36 genetic loci in the Yunnan local pig breeds were surveyed,Nine of them,such as AKP、CAT、DIA、ES、G6PD、PA、6PGD、PHI and TF,were found to be polymorphic,the mean heterozygosit(H)was 0.0712~0.1027.The results indicated that the blood protein polymorphism in the Yunnan local pig breeds is high,the Yunnan local pig breeds are wealthy in genetic diversity in point of their protein level.     

缺氧对右心室最大心肌血流量的影响

为了探讨缺氧对冠状血管贮备力的影响,我们观察了缺氧时大鼠血流动力学及右心室最大心肌血流量的变化。结果表明,急性缺氧引起ＰａＯ２、心输出量及氧运送量降低,但右心室心肌血流量增加,右心室最大与安静血流量比值降低。慢性缺氧时ＰａＯ２降低,血球比积和右心室重量指数增加,氧运送量和右心室血流量正常,但最大血流量降低,小动脉增厚、外膜胶元增加。以上结果提示,慢性缺氧对冠状血管贮备减少可能是小动脉壁增厚、外膜胶元增加和血液粘滞性增加及右心室肥大的结果。     

p33 Enhances UVB-Induced Apoptosis in Melanoma Cells

The biological functions of the tumor suppressor ING1 have been studied extensively in the past few years since it was cloned. It shares many biological functions with p53 and has been reported to mediate growth arrest, senescence, apoptosis, anchorage-dependent growth, chemosensitivity, and DNA repair. Some of these functions, such as cell cycle arrest and apoptosis, have been shown to be dependent on the activity of both ING1 and p53 proteins. Two recent reports by Scott and colleagues demonstrate that p33^ING1 (one of the ING1 isoforms) translocates to the nucleus and binds to PCNA upon UV irradiation. Here we report that p33^ING1 mediates UV-induced cell death in melanoma cells. We found that overexpression of p33^ING1 increased while the introduction of an antisense p33^ING1 plasmid reduced the apoptosis rate in melanoma cells after UVB irradiation. We also demonstrated that enhancement of UV-induced apoptosis by p33^ING1 required the presence of p53. Moreover, we found that p33^ING1 enhanced the expression of endogenous Bax and altered the mitochondrial membrane potential. Taken together, these observations strongly suggest that p33^ING1 cooperates with p53 in UVB-induced apoptosis via the mitochondrial cell death pathway in melanoma cells.     

Isolation of microsatellite markers from Pinus densiflora Sieb. et Zucc. using a dual PCR technique

We developed seven microsatellite loci from Pinus densiflora using a dual polymerase chain reaction (PCR) technique. Of 186 clones from a library based on suppression PCR, 127 contained microsatellite sequences. Of these, 43 candidates were determined sequences of both flanking regions, and 16 regions from this group were chosen as development markers. Seven of these primer pairs successfully amplified polymorphic single loci among 83 resistant trees against pine wood nematode. The observed heterozygosity of the seven microsatellite markers ranged from 0.247 to 0.843. Mendelian inheritance was confirmed using megagametophytes.     

高温对昆虫影响的生理生化作用机理研究进展

温度是影响昆虫生命活动的重要因素，将其与某一时间的种群数量结合，可用于对昆虫未来种群数量进行预测预报。过高的环境温度常使昆虫的生长发育、生殖及存活等受到严重影响，对这种影响缺乏了解降低了害虫测报的准确性。探明高温对昆虫生理生化的作用机理是了解高温对昆虫生命活动影响的根本途径。总结了高温对昆虫生理生化的重要影响。高温使昆虫表皮的蜡质层瓦解，油脂融化，表皮渗透性增加，虫体大量失水。引起昆虫体内重要离子的浓度发生变化，改变许多重要大分子的电荷状态，使生物大分子的动力学能量增大，离子键、氢键和范德华力降低，分子间疏水作用增强，大分子保持形状的能力降低，空间构象发生改变，从而影响生物大分子行使其功能。高温使昆虫细胞骨架瓦解，细胞遭到破坏；细胞膜内磷脂组分比例改变，细胞膜流动性下降。虫体内重要遗传物质DNA和RNA复杂的二级结构和三级结构在高温下发生改变，对昆虫性状的稳定遗传造成严重影响。细胞内蛋白质的数量和种类组成均发生改变，原有常温下的蛋白质合成系统关闭，空间构象及功能发生变化，而产生耐热性物质（如热激蛋白）的蛋白质合成系统则开启。高温影响酶及酶促反应速率，对昆虫体内神经传导关键酯酶——乙酰胆碱酯酶的影响使昆虫无法进行正常的神经传递，丧失躲避不良环境的能力。高温影响脂质、低聚糖等物质的代谢。最后梳理了高温作用下昆虫各种生理生化指标发生变化之间可能存在的关联性，并提出高温对昆虫造成伤害的顺序过程假设。讨论了不同程度的高温对昆虫造成死亡的机理可能不同。指出了未来该领域研究的重点内容，如高温对昆虫造成损害的最初作用位点；高温伤害的完整生理生化路径；耐热性产生的生化基础；高温对昆虫不同发育阶段或生命过程的具体作用机制等。     

重组胰高血糖素样肽-1与人血清白蛋白融合蛋白的纯化及活性研究

利用构建的重组菌株Pichia pastoris GS115/GLP-1/HSA,在10L发酵罐中表达了胰高血糖素样肽-1与人血清白蛋白融合蛋白(GLP-1/HSA),表达量为63.6mg/L。发酵液经中空纤维柱浓缩、疏水层析、阴离子交换层析和凝胶过滤分离纯化,获得了较高纯度的GLP-1/HSA,经HPLC分析纯度达95.8%。进一步的体内活性分析结果表明,GLP-1/HSA不仅具有天然GLP-1的生物活性,而且在给药后4h仍能发挥显著性降血糖作用。以上结果表明,利用Pichia pastoris分泌型表达系统和建立的分离纯化方法,能获得大量较高纯度的GLP-1/HSA,为进一步研究和开发能够用于糖尿病临床治疗的长效GLP-1类似物奠定了基础。     

Molecular cloning of a novel rat gene Tsarg1, a member of the DnaJ/HSP40 protein family.

Beginning with a mouse gene mTSARG3, which was related to apoptosis of spermatogenic cells, bioinformatics was applied and a predicted novel rat gene full-length cDNA sequence was attained. Gene-specific primers were designed for PCR in rat testis cDNA library. A new gene Tsarg1 (GenBank Accession No. AY380804) was cloned, which is related to apoptosis in rat spermatogenic cells. The gene whose full cDNA length is 1176 bp containing 8 exons and 7 introns is located in rat chromosome 1q32-1q33, which encoded a protein containing 316 amino acid residues and being a new member of HSP40 protein family since the sequence contains the highly conserved J domain, which is present in all DnaJ-like proteins and is supported to have a critical role in DnaJ-DnaK protein-protein interactions. The results of RT-PCR and Northern blot analysis showed that Tsarg1 was specifically expressed in rat testis, which probably inhibits rat testis spermatogenic cell apoptosis.     

Detection of blob objects in microscopic zebrafish images based on gradient vector diffusion.

The zebrafish has become an important vertebrate animal model for the study of developmental biology, functional genomics, and disease mechanisms. It is also being used for drug discovery. Computerized detection of blob objects has been one of the important tasks in quantitative phenotyping of zebrafish. We present a new automated method that is able to detect blob objects, such as nuclei or cells in microscopic zebrafish images. This method is composed of three key steps. The first step is to produce a diffused gradient vector field by a physical elastic deformable model. In the second step, the flux image is computed on the diffused gradient vector field. The third step performs thresholding and nonmaximum suppression based on the flux image. We report the validation and experimental results of this method using zebrafish image datasets from three independent research labs. Both sensitivity and specificity of this method are over 90%. This method is able to differentiate closely juxtaposed or connected blob objects, with high sensitivity and specificity in different situations. It is characterized by a good, consistent performance in blob object detection.