孢子丝菌复合体分子分型研究进展

孢子丝菌复合体属于双相真菌,全球分布,可引起人类及动物的慢性深部感染。不同地域的菌株在致病力、传播途径及药物敏感性等方面均存在差异。孢子丝菌病作为一种人兽共患病,其发病率逐年上升,出现多次暴发流行。分子分型不仅是明确感染源和传播途径、预防和控制疾病流行的有力手段,同时有助于了解孢子丝菌基因型与表型的相关性,在研究其致病机制以及临床诊治过程中都具有十分重要的意义。本文对孢子丝菌的分子分型方法的研究进行综述。     

Attitudes in China about Crops and Foods Developed by Biotechnology

Transgenic Bt cotton has been planted in China since 1997 and, in 2009, biosafety certificates for the commercial production of Bt rice and phytase corn were issued by the Chinese government. The public attitude in China toward agricultural biotechnology and genetically modified (GM) crops and foods has received considerable attention worldwide. We investigated the attitudes of consumers, Bt cotton farmers and scientists in China regarding GM crops and foods and the factors influencing their attitudes. Data were collected using interview surveys of consumer households, farmer households and scientists. A discrete choice approach was used to elicit the purchase intentions of the respondents. Two separate probit models were developed to examine the effect of various factors on the choices of the respondents. Bt cotton farmers had a very positive attitude because Bt cotton provided them with significant economic benefits. Chinese consumers from developed regions had a higher acceptance and willingness to pay for GM foods than consumers in other regions. The positive attitude toward GM foods by the scientific community will help to promote biotechnology in China in the future. Our survey emphasized that educational efforts made by government officials, the media and scientists can facilitate the acceptance of GM technology in China. Further educational efforts will be critical for influencing consumer attitudes and decisions of government agencies in the future. More effective educational efforts by government agencies and public media concerning the scientific facts and safety of GM foods would enhance the acceptance of GM crops in China.     

KfoE encodes a fructosyltransferase involved in capsular polysaccharide biosynthesis in Escherichia coli K4

Escherichia coli K4 synthesizes a capsular polysaccharide (CPS) consisting of a fructose-branched chondroitin (GalNAc-GlcA(fructose)n), which is a biosynthetic precursor of chondroitin sulfate. Here, the role of kfoE in the modification of the chondroitin backbone was investigated using knock-out and recombinant complementation experiments. kfoE disruption and complementation had no significant effect on cell growth. CPS production was increased by 15 % in the knock-out strain, and decreased by 21 % in the knock-out strain complemented with recombinant kfoE. CPS extracted from the knock-out strain was chondroitin, whereas CPS extracted from the complemented strain was a fructose-branched chondroitin. The results demonstrated that the kfoE gene product altered the fructose group at the C3 position of the GlcA residue during production of K4CPS.     

羽叶金合欢的DNA提取和SSR引物筛选

利用SSR分子标记对羽叶金合欢( Acacia pennata )的野生种和栽培种进行了分析,利用改良的CTAB方法优化了其总DNA的提取方法,并优化了PCR扩增条件.从已有的金合欢属植物的82对SSR引物中筛选出多态性高,稳定性好的12对引物作为羽叶金合欢的SSR分析引物,为进一步对其进行遗传多样性研究提供了依据.     

Fluorescent labeling for clonal selection of Marc 145 cells secreting high levels of recombinant protein PBD-1

Despite the powerful impact gene expression markers like the green fluorescent protein (GFP) or enhanced GFP (EGFP) exert on linking the expression of recombinant protein for selection of high producers in recent years, there is still a strong incentive to develop more economical and efficient methods for isolating mammalian cell clones secreting high levels of recombinant proteins. Here we present a new method based on the co-expression of EGFP that allows clonal selection in standard 96-well cell culture plates. The genes encoding the EGFP protein and the related protein are linked by an internal ribosome entry site and thus are transcribed into the same mRNA in an independent translation process. Since both proteins arise from a common mRNA, the EGFP expression level correlates with the expression level of the therapeutic protein in each clone. By expressing recombinant porcine β-defensin 1 in Marc 145 cells, we demonstrate the robustness and performance of this technique. The method can be served as an alternative to identify high-producer clones with various cell sorting methods.     

Heparan sulfate proteoglycans regulate autophagy in Drosophila

Heparan sulfate-modified proteoglycans (HSPGs) are important regulators of signaling and molecular recognition at the cell surface and in the extracellular space. Disruption of HSPG core proteins, HS-synthesis, or HS-degradation can have profound effects on growth, patterning, and cell survival. The Drosophila neuromuscular junction provides a tractable model for understanding the activities of HSPGs at a synapse that displays developmental and activity-dependent plasticity. Muscle cell-specific knockdown of HS biosynthesis disrupted the organization of a specialized postsynaptic membrane, the subsynaptic reticulum (SSR), and affected the number and morphology of mitochondria. We provide evidence that these changes result from a dysregulation of macroautophagy (hereafter referred to as autophagy). Cellular and molecular markers of autophagy are all consistent with an increase in the levels of autophagy in the absence of normal HS-chain biosynthesis and modification. HS production is also required for normal levels of autophagy in the fat body, the central energy storage and nutritional sensing organ in Drosophila. Genetic mosaic analysis indicates that HS-dependent regulation of autophagy occurs non-cell autonomously, consistent with HSPGs influencing this cellular process via signaling in the extracellular space. These findings demonstrate that HS biosynthesis has important regulatory effects on autophagy and that autophagy is critical for normal assembly of postsynaptic membrane specializations.     

Combination of bone tissue engineering and BMP-2 gene transfection promotes bone healing in osteoporotic rats

OBJECTIVE: The aim of this study was to develop a feasible approach to promote bone healing in osteoporotic rats using autogenous bone tissue-engineering and gene transfection of human bone morphogenetic protein 2 (hBMP-2). METHODS: Bone marrow stromal cells (BMSCs) from the left tibia of osteoporotic rats were transfected with the hBMP-2 gene in vitro which was confirmed by immunohistochemistry, in situ hybridization and Western blotting. Autogenous transfected or untransfected BMSCs were seeded on macroporous coral hydroxyapatite (CHA) scaffolds. Each cell-scaffold construct was implanted into a defect site which was created in the ramus of the mandible of osteoporotic rats. Four or eight weeks after implantation in situ hybridization was performed in BMSCs transfected with hBMP-2, X-ray examinations, histological and histomorphological analyses were used to evaluate the effect of tissue-engineered bone on osseous defect repair. RESULTS: Newly formed bone was observed at the margin of the defect 4 weeks after implantation with BMSCs transfected with BMP-2. Mature bone was observed 8 weeks after treatment. In the control group there was considerably less new bone and some adipose tissue was observed at the defect margins 8 weeks after implantation. CONCLUSIONS: Autogenous cells transfected with hBMP-2 promote bone formation in osteoporotic rats. BMSC-mediated BMP-2 gene therapy used in conjunction with bone tissue engineering may be used to successfully treat bone defects in osteoporotic rats. This method provides a powerful tool for bone regeneration and other tissue engineering.     

山西阳泉老窑水硫酸盐还原菌的分离鉴定及其还原功能驯化

【背景】含硫煤矿开采后,地表水/地下水回流至采空区形成酸性老窑水,含有高浓度重金属离子和硫酸盐,严重危害生态系统健康。利用微生物自身生长处理老窑水具有成本低、环境友好等特点,具有良好的应用前景。目前利用的硫酸盐还原菌大多只在适宜温度和中性pH条件下具有较高活性,在北方低温和酸性条件下难以发挥作用。【目的】本研究旨在从山西阳泉山底河流域的老窑水环境中分离硫酸盐还原菌,并调节温度和pH进行驯化,从而得到高效耐低温耐酸菌株,为北方老窑水微生物治理提供可用菌种资源。【方法】对山底河流域典型老窑水样品中的微生物进行富集培养,并筛选硫酸盐还原菌。通过革兰氏染色、扫描电镜对菌株形貌特性进行表征,利用16SrRNA基因序列比对进行菌种鉴定,探究其生长特性和硫酸盐还原性能。在此基础上降低温度和pH,对高效还原硫酸盐菌株进行驯化,探讨其在北方老窑水污染治理中的应用潜力。【结果】本研究筛选得到2株硫酸盐还原菌,命名为YQ-1和YQ-2,分别属于革兰氏阴性瘤胃解蛋白质菌属(Proteiniclasticum)和脱硫弧菌属(Desulfovibrio)。在30°C、pH 7.5条件下,YQ-1和YQ-2对1 1...     

Anti-angiogenic active immunotherapy: a new approach to cancer treatment

Tumor angiogenesis plays an important role in tumor growth, aggression and metastasis. Many molecules have been demonstrated as positive regulators of angiogenesis, including vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), epidermal growth factor (EGF), and others. In recent years, significant progress has been made in the research on anti-angiogenic strategies for tumor therapies. In this review, anti-angiogenic active immunotherapies for tumors based on vaccination with xenogeneic homologous molecules and non-xenogeneic homologous molecules are discussed.     

Molecular Cloning of Novel Cellulase Genes <Emphasis Type="Italic">cel</Emphasis>9A and <Emphasis Type="Italic">cel</Emphasis>12A from <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> GXN151 and Synergism of Their Encoded Polypeptides

A thermophilic bacterial strain GXN151 which could degrade Avicel efficiently was isolated and identified as Bacillus licheniformis. A genomic library of GXN151 was constructed and two novel endoglucanase genes designated cel9A and cel12A were isolated by screening the library on carboxylmethyl cellulase indicator plates. The analysis of amino acid sequences deduced from the genes indicated that Cel9A consisted of a catalytic domain belonging to glycosyl hydrolase family 9, a linker domain, and a carbohydrate binding module family 3 from N-terminal to C-terminal; Cel12A had only one catalytic domain belonging to glycosyl hydrolase family 12. The combinations of Cel9A and Cel12A produced by the recombinant E. coli exhibited synergistic action against substrates of carboxylmethyl cellulose as well as Avicel.