黄土丘陵区坡面整地和植被耦合下的土壤水分特征

水分是干旱半干旱地区植被恢复的主要环境制约因子。在黄土高原小流域,合理整地能够有效截留降雨补给土壤水,进而促进植被恢复。选择地处甘肃定西的半干旱黄土小流域为研究区,基于野外实测数据,分析不同植被和整地方式(柠条水平阶、山杏水平沟、侧柏反坡台,油松反坡台)综合影响下的土壤水分特征。采用最优分割法将不同整地方式土壤水分垂直层次划分为活跃层,次活跃层和相对稳定层。结果表明:生长季不同整地方式土壤水分的变化与降水量的变化密切相关,不同月份以及不同深度各整地方式土壤水分之间差异显著(P0.05)。根据土壤水分垂直变化特征,山杏水平沟水分活跃层与次活跃层为0—80cm,其深度范围均大于其他3种整地方式,而柠条水平阶土壤水分均在30 cm以下较为稳定,其深度范围均小于其他3种整地方式。不同整地方式土壤水分含量具体表现为:山杏水平沟侧柏反坡台柠条水平阶油松反坡台。     

不同全血过滤方法用于去白细胞血液制备的临床对照研究

目的:观察不同全血过滤方法用于去白细胞血液制备的效果。方法:采用两种全血过滤方法进行对比研究,对照组采用常规法,将采集后全血混匀后直接与白细胞滤器连接直接过滤;实验组采用湿润滤盘法,血液采集完成混匀后静置,先用上层血清10～20 m L湿润滤盘,再混匀与白细胞滤器连接后进行过滤。比较两组制备方法所用的过滤时间、血液回收率、过滤前后血液指标情况及24小时内溶血的发生情况。结果:两组全血过滤方法过滤前后白细胞、红细胞、血红蛋白、血小板及血浆游离血红蛋白水平比较差异均无明显统计学意义(P0.05)。而实验组过滤时间短于对照组,血液回收率高于对照组,且24小时内溶血比例明显低于对照组(P0.05)。结论:常规法与湿润滤盘法均能达到去白细胞血液标准,但湿润滤盘法较常规法能有效的降低过滤时间、增加血液回收率,减少去白细胞悬浮红细胞因溶血造成的血液不合格率,值得临床推广应用。     

A Fast and Economical Sample Preparation Protocol for Interaction Proteomics Analysis

A simple and fast immunoprecipitation (IP) protocol is designed with the sample preparation incorporated, applicable to both low and high throughput. This new protocol combines two procedures based on magnetic beads in 96‐well plate format. Protein complexes are captured by antibodies and magnetic beads conjugated with protein A. Proteins are washed and on‐bead digested by using Single‐Pot solid‐phase sample preparation (SP3). The whole IP‐SP3 approach can be completed in one day, which is considerably faster compared to the classical approach. No major quantitative differences are found between SP3 and FASP (filter‐aided sample preparation) or a longer incubation protocol. Taken together, the IP‐SP3 protocol is a fast and economical approach easily applicable for large‐scale protein interactome analysis.     

Synthesis of the 'primer-adaptor' trisaccharide moiety of Escherichia coli O8, O9, and O9a lipopolysaccharide

Described is the synthesis of the trisaccharide alpha-D-Manp-(1-->3)-alpha-D-Manp-(1-->3)-beta-D-GlcpNAcO(CH2)8N3, the glycan portion of which corresponds to the 'adaptor-primer' moiety linking the O-chain and core oligosaccharide in the lipopolysaccharide of several Escherichia coli and Klebsiella pneumoniae serotypes. This report represents the first synthesis of this trisaccharide motif, and in the route involved, a key step is a [2+1] coupling of a protected Manp-(1-->3)-alpha-D-Manp glycosyl donor with a GlcpNAc acceptor. The azido group was included in the target to facilitate future preparation of neoglycoconjugates.     

Hydrolysis rates of 1-glucosyl-2-benzoylhydrazines in aqueous solution

A series of substituted 1-glucosyl-2-benzoylhydrazines were synthesized and their pseudo-first-order rate constants for hydrolysis were determined at pH 4, 5, 6 at 50 degrees C. All the compounds hydrolyzed quickly (t(1/2)<3h) at pH 4.0, but were increasingly stable as the pH approached neutrality.     

肠三叶因子在大肠杆菌中的表达、纯化及其抗体制备

目的:获得肠三叶因子(ITF)的原核表达产物及抗rITF抗体,为深入研究ITF的作用机制及其受体研究奠定基础。方法:常规提取人小肠组织总RNA,用RT-PCR获得ITF编码基因片段,克隆至质粒pET32a获得原核表达栽体,双酶切和测序后转化至Origami B(DE3)用IPTG诱导表达,优化条件获得最大表达产量;用SDS-PAGE、Western blot鉴定表达产物,亲和层析纯化获得的重组蛋白rITF皮下多点注射家兔,制备多克隆抗体,并用此抗体进行大鼠肠组织免疫组化研究。结果:测序证实PCR扩增获得ITF全长基因序列与基因文库中的完全一致,将该基因片段正确插入表达载体pET32a中、优化表达条件后,重组蛋白的表达量达到50mg/L;Western blot证明重组蛋白具有良好的抗原性和特异性;通过Ni-NTA亲和层析、超滤离心后,得到90%纯度的蛋白;收集兔血清,纯化后获得特异性良好的ITF抗体,免疫组化染色肠组织显示ITF表达的部位定位于杯状细胞。结论:成功构建了表达载体pET32a-ITF,在大肠杆菌中表达并纯化获得纯度较高的rITF,并获得了生物活性较高的ITF抗体,ITF主要在肠道杯状细胞分泌表达。     

组织工程支架制备中超临界CO2技术的应用*

作为组织工程研究中三大要素之一,组织工程支架可为细胞的附着、迁移和增殖提供理想的环境。传统的组织工程支架制备方法,如粒子沥滤法、相分离法及静电纺丝法等在理论和技术上已较为成熟,但由于大多需要有机溶剂的参与,在制备过程中仍存在有机溶剂难以去除,以及支架孔洞难以控制、连通性较差等问题。超临界二氧化碳(supercritical carbon dioxide,SC-CO₂)密度近似液体,黏度和扩散系数近似气体,具有流动性强、溶解能力大、传热效率高等特殊的理化性质,与传统工艺相结合,可在绿色温和的反应体系中有效规避上述问题,在组织工程支架制备及药物负载方面具有广阔前景。     

Effective field trips in nature: the interplay between novelty and learning

Educational field trips are common practice in environmental education and education for sustainable development, well recognised by researchers for their potential to achieve cognitive and affective educational outcomes. One of the factors that influences learning during field trips is their novelty. The current study focuses on the interplay between novelty, preparation and environmental learning outcomes of 5th and 6th grade students during a typical field trip in Flanders. Our dependent variables are Inclusion of Nature in the Self, the two major ecological values Preservation and Utilisation and ecosystem knowledge. The sample includes 484 students (10–12 years old) and their 24 teachers. Key questions addressed are: (1) What is learned during the field trip? (2) What is the level of novelty for students during a field trip? (3) How does the novelty effect relate to learning? Results show that participation in the field trip leads to a substantial increase in ecosystem knowledge, but fails in reaching the affective goals set out by the field trip organisers. Our results furthermore provide support for the hypothesised non-linear relationship between novelty and knowledge gain, showing that while a little novelty is positive, too much novelty can stand in the way of learning.     

Streptomycin-induced ribosome engineering complemented with fermentation optimization for enhanced production of 10-membered enediynes tiancimycin-A and tiancimycin-D

Tiancimycins (TNMs) are a group of 10-membered anthraquinone-fused enediynes, newly discovered from Streptomyces sp. CB03234. Among them, TNM-A and TNM-D have exhibited excellent antitumor performances and could be exploited as very promising warheads for the development of anticancer antibody-drug conjugates (ADCs). However, their low titers, especially TNM-D, have severely limited following progress. Therefore, the streptomycin-induced ribosome engineering was adopted in this work for strain improvement of CB03234, and a TNMs high producer S. sp. CB03234-S with the K43N mutation at 30S ribosomal protein S12 was successfully screened out. Subsequent media optimization revealed the essential effects of iodide and copper ion on the production of TNMs, while the substitution of nitrogen source could evidently promote the accumulation of TNM-D, and the ratio of produced TNM-A and TNM-D was responsive to the change of carbon and nitrogen ratio in the medium. Further amelioration of the pH control in scaled up 25 L fermentation increased the average titers of TNM-A and TNM-D up to 13.7 ± 0.3 and 19.2 ± 0.4 mg/L, respectively. The achieved over 45-fold titer improvement of TNM-A, and 109-fold total titer improvement of TNM-A and TNM-D enabled the efficient purification of over 200 mg of each target molecule from 25 L fermentation. Our efforts have demonstrated a practical strategy for titer improvement of anthraquinone-fused enediynes and set up a solid base for the pilot scale production and preclinical studies of TNMs to expedite the future development of anticancer ADC drugs.     

Cloning,overexpression and characterization of a thermostable β-xylosidase from Thermotoga petrophila and cooperated transformation of ginsenoside extract to ginsenoside 20(S)-Rg3 with a β-glucosidase

A thermostable β-xylosidase gene Tpexyl3 from Thermotoga petrophila DSM 13,995 was cloned and overexpressed by Escherichia coli. Recombinant Tpexyl3 was purified, and its molecular weight was approximately 86.7 kDa. Its optimal activity was exhibited at pH 6.0 and 90 °C. It had broad specificity to xylopyranosyl, arabinopyranosyl, arabinofuranosyl and glucopyranosyl moieties. The β-xylosidase activity of the recombinant Tpexyl3 was 6.81 U/mL in the LB medium and 151.4 U/mL in a 7.5 L bio-reactor. It was applied to transform ginsenoside extract into the pharmacologically active minor ginsenoside 20(S)-Rg3, which was combined with thermostable β-glucosidase Tpebgl3. After transforming under optimal condition, the 20 g/L of ginsenoside extract was transformed into 6.28 g/L of Rg3 within 90 min, with a corresponding molar conversion of 95.0% and Rg3 productivity of 1793.49 mg/L/h, respectively. This study is the highest report of a GH3 family glycosidase with arabinopyranosidase activity and also the first report on the high substrate concentration bioconversion of ginsenoside extract to ginsenoside 20(S)-Rg3 by using two thermostable glycosidases.