自发性树鼩乳腺肿瘤的特性(英文)

乳腺癌是严重危害女性健康的常见恶性肿瘤,建立合适的乳腺癌动物模型对于研究人类乳腺癌的生物学机制及发展新的防治方法至关重要。相对于常用的啮齿类动物,树鼩(Tupaia belangeri chinensis,tree shrew)因在进化层次上更接近于人类而可用于建立更适合的乳腺癌模型。该文详细了介绍一例树鼩自发性乳头状良性乳腺肿瘤。免疫组化结果显示该例肿瘤孕激素受体阳性且Ki-67阳性细胞比例显著增加;而活化的Caspase3阳性细胞比例较低;且肿瘤的形态和病理与人导管内乳头状肿瘤非常接近。提示利用树鼩建立乳腺肿瘤模型的可行性。     

扁蓄总黄酮含量的测定

以35％乙醇为提取溶剂,聚酰胺吸附提纯,芦丁为标准品,用分光光度法于510nm处测定了扁蓄各部位的总黄酮含量。结果表明：扁蓄全草总黄酮含量为4．349％,根为7．115％,茎为5．454％,叶为3．544％。     

OC-IΔD86 (Oryzacystatin-IΔD86)基因的原核表达及活性分析

根据OC-IΔD86基因序列, 设计合成了7条寡核苷酸片段, 通过重叠延伸PCR技术合成了OC-IΔD86基因, 利用设计好的BamH I/ Xho I酶切位点将OC-IΔD86基因克隆到原核表达载体pet21b中, 在1 mmol/L的IPTG 诱导后5 h, OC-IDD86融合基因在大肠杆菌中得到表达, 表达产物处于可溶状态, 其表达量占总蛋白的11.4%, 可溶性蛋白的16.4%; 利用Ni-NTA系统纯化该蛋白并经PEG20000浓缩后, 活性分析表明该蛋白酶抑制剂在体外表现出对木瓜蛋白酶明显的抑制作用。这为转OC-IΔD86基因的抗根结线虫植物基因工程抗体制备, 以及进一步体内的抗根结线虫研究奠定了基础。     

重组人血清白蛋白在Pichia pastoris中分泌表达影响因素的研究

为获得高产稳产重组人血清白蛋白的Ｐｉｃｈｉａ ｐａｓｔｏｒｉｓ细胞株用于高密度发酵,构建了多种分泌表达载体,在Ｐｉｃｈｉａ ｐａｓｔｏｒｉｓ中作表达,研究表达载体构建等因素对表达量的影响。发现采用酿酒酵母α性成熟因子前导肽比人血清白蛋白天然前导肽作为分泌信号肽的表达量要高１０％左右。而载体整合方式、整合拷贝数、５’非翻译区的改造和甲醇利用表型等对表达量无规律性影响。筛选到的高表达株经高密度发酵,细     

多级逆流工艺促进城市污泥厌氧发酵生产挥发性脂肪酸

采用一种新型的厌氧发酵工艺——多级逆流发酵工艺对城市污泥进行厌氧发酵, 实现高效产挥发性脂肪酸的目的。结果表明, 实验条件下应用多级逆流发酵工艺, 挥发性脂肪酸浓度与产率分别达到(10.5±0.5) g/L和0.20 gVFAs/ gVS, 与普通厌氧发酵工艺相比, 分别提高了31%和54%。此外, 在多级逆流工艺中, 底物有机质去除率可达50%, 较普通厌氧发酵提高了37%。进一步分析多级逆流工艺产酸的机制, 发现产酸效率的提高在于降低了发酵产物对厌氧产酸细菌的抑制效应, 并且工艺的VFAs产率以及有机质去除率分别取决于第一级和第三级厌氧发酵过程。因此, 城市污泥采用多级逆流工艺厌氧发酵不仅能够有效促进挥发性脂肪酸的生成, 而且能够较大程度上提高污泥中有机质的去除率。     

弱氧化醋杆菌阿拉伯糖醇脱氢酶基因克隆及其在大肠杆菌中的表达

The partial genomic library of Acetobacter suboxydans was constructed using Yeast\| E.coli shuttle plasmid YEp352 as vector.Two positive transformants,designated as DH5α(pAD91) and DH5α(pAD98),were obtained by screening the growth of transformants on the agar plate in which D\|arabitol was used as the sole carbon source.The results of Southern blot and restriction endonuclease analysis showed that the two recombinants are identical.The insert is about 2.3kb.Arabitol dehydrogenase activity assay indic…     

Dynamics of fertilizer-derived organic nitrogen fractions in an arable soil during a growing season

Background

Inorganic fertilizer is one of the most important anthropogenic inputs which influences soil nutrient turnover in agricultural ecosystems. However, as the key process involved in the maintenance, transformation and stability of soil nitrogen (N), the incorporation and allocation of fertilizer N between different soil organic N (SON) fractions in a growing season remains largely unknown.

Methods

In this study, a field experiment was conducted in triplicate of micro-plots and a total of 200 kg N ha^?1 (¹⁵?N-labeled (NH₄)₂SO₄, 98 atom %) was applied as a basal dressing and two top dressings, at jointing and filling stages, respectively, to a maize crop during one growing season. The distribution and seasonal dynamics of fertilizer N in different SON fractions (i.e., amino acids, amino sugars, hydrolyzable ammonium N and acid insoluble-N) were measured by liquid/gas chromatography–mass spectrometry (LC/GC-MS) and element analysis-combustion-isotope ratio mass spectrometry (EA-C-IRMS) techniques. Path analysis was used to evaluate the transformation processes between organic N fractions derived from fertilizer and N supply strategy in soil-plant system.

Results

The accumulation of fertilizer-derived N in different organic fractions was season-specific. At jointing stage, preferential enrichment of ¹⁵?N was found in soil amino acids plus amino sugars, indicating the active biological immobilization of basal dressing fertilizer N. Nevertheless, there is still a small proportion of fertilizer N stabilized in the acid insoluble fraction. The accumulation of the residual fertilizer N in hydrolyzable ammonium N reached a maximum at filling stage and then declined significantly, implying the rapid release of the fertilizer N remained in mineral forms. The contents of amino acids changed slightly, but they played a very important role in mediating SON transformation.

Conclusion

The hydrolyzable ammonium N was a temporary pool for rapid fertilizer N retention and simultaneously was apt to release N for crop uptake in the current season. In contrast, the amino acids could serve as a transitional pool of available N in the soil-crop system, while the acid insoluble fraction was as a stable pool of fertilizer N. Importantly, there is an interim shift among different pools to maintain soil N turnover; hence N in the amino acid fraction mediates N supply and the depolymerization of SON constituents controls the proceeding of fertilizer N cycling in the soil-plant system.     

酸雨对温州三土羊湿地水体氮营养盐数量的影响

为了解酸雨对温州三羊湿地水体氮营养盐数量的影响,本文测定了三土羊湿地水体中不同形态氮的含量,其中氨氮2.90～10.75 mg·L^-1,平均538 mg·L^-1、硝态氮0.16～0.44 mg·L^-1,平均0.31 mg·L^-1;总氮34.04～63.20 mg·L^-1,平均55.75 mg·L^-1和水体的pH值,6.1～6.5,平均6.4;并测算了近两年通过降水输入到三羊湿地和三羊湿地水体中不同形态氮的数量及其占湿地现有水体中各形态氮的比例,其中输入到三羊湿地氨氮、硝态氮和总氮的量分别为2.48～2.86×10⁴ kg、2.87×10⁴～4.96×10⁴ kg和5.35×10⁴～7.82×10⁴ kg,分别占湿地水体现有量的56%～64%、11.21～19.38倍和12%～17%;直接输送到三羊湿地水体各形态氮的数量0.72×10⁴～0.84×10⁴ kg、0.83×10⁴～1.44×10⁴ kg和1.55×10⁴～2.27×10⁴,分别占现有量的16%～19%,3.24～5.63倍和3%～5%.结果表明,酸雨是三,羊湿地水体氮污染的主要来源之一,对三羊湿地水体富营养化产生重要影响.     

来源于铜绿假单胞杆菌(Pseudomonas aeruginosa)所产蛋白酶PT121克隆表达及其在小肽合成上的应用

【目的】基于前期筛选到的P.aeruginosa PT121所产有机溶剂耐受性蛋白酶,本研究对该蛋白酶进行克隆表达,研究了重组蛋白的酶学性质及小肽合成上的应用。【方法】参考文献中报道的相似蛋白酶pseudolysin设计引物从菌株PT121基因组中克隆到耐有机溶剂蛋白酶PT121基因las B。构建诱导表达重组质粒p ET22b-las B,于大肠杆菌中进行表达。考察蛋白酶PT121酶学性质及小肽合成上的应用。【结果】序列分析表明las B基因编码信号肽、前肽及成熟肽3个部分,成熟肽部分含有301个氨基酸,分子量约33 k Da,属于金属蛋白酶M4家族。通过破碎条件优化,一步法制备得到较为纯净的重组蛋白酶PT121,其比活力达7700 U/mg,该酶呈现了较高的热稳定性,p H稳定性及溶剂耐受性,与野生菌P.aeruginosa PT121所产蛋白酶性质一致。在50%DMSO体系中高效催化合成了多种小肽,其中阿斯巴甜前体(Cbz-Asp-Phe-NH2)产率高达91%。【结论】蛋白酶PT121基因在大肠杆菌中克隆表达为进一步研究相关催化机理及分子改造奠定了基础。