蛇毒类凝血酶calobin在毕赤酵母中的表达

蛇毒类凝血酶是临床上防治血栓栓塞性疾病的有效药物。参照朝鲜蝮蛇(Agkistrodon caliginosus,Korean Viper)类凝血酶calobin基因序列(GenBank AccessionNo.U32937.1),将人工合成的calobin基因克隆到酵母表达载体pPICZαA,于毕赤酵母中表达,得到了分子量约为32kD的重组calobin蛋白,经甲醇诱导培养,表达产物可获得3.5g/L的高表达量。重组蛋白经过阴离子交换柱Q-Sepharose Fast Flow和分子筛Sephacryl-S-100凝胶过滤层析等纯化步骤进行了初步纯化。纯化后的重组calobin可以在纤维蛋白原平板上形成水解圈,经SDS-PAGE实验显示,重组蛋白能水解纤维蛋白原的Aα链,产生一条约40kD左右的降解带。在实验中未能发现重组calobin对纤维蛋白原的凝固作用。     

Switching of the Bacterial Flagellar Motor Near Zero Load

Flagellated bacteria, such as Escherichia coli, are able to swim up gradients of chemical attractants by modulating the direction of rotation of their flagellar motors, which spin alternately clockwise (CW) and counterclockwise (CCW). Chemotactic behavior has been studied under a variety of conditions, mostly at high loads (at large motor torques). Here, we examine motor switching at low loads. Nano-gold spheres of various sizes were attached to hooks (the flexible coupling at the base of the flagellar filament) of cells lacking flagellar filaments in media containing different concentrations of the viscous agent Ficoll. The speeds and directions of rotation of the spheres were measured. Contrary to the case at high loads, motor switching rates increased appreciably with load. Both the CW → CCW and CCW → CW switching rates increased linearly with motor torque. Evidently, the switch senses stator-rotor interactions as well as the CheY-P concentration.     

Mechanisms of pre‐apoptotic calreticulin exposure in immunogenic cell death

Dying tumour cells can elicit a potent anticancer immune response by exposing the calreticulin (CRT)/ERp57 complex on the cell surface before the cells manifest any signs of apoptosis. Here, we enumerate elements of the pathway that mediates pre‐apoptotic CRT/ERp57 exposure in response to several immunogenic anticancer agents. Early activation of the endoplasmic reticulum (ER)‐sessile kinase PERK leads to phosphorylation of the translation initiation factor eIF2α, followed by partial activation of caspase‐8 (but not caspase‐3), caspase‐8‐mediated cleavage of the ER protein BAP31 and conformational activation of Bax and Bak. Finally, a pool of CRT that has transited the Golgi apparatus is secreted by SNARE‐dependent exocytosis. Knock‐in mutation of eIF2α (to make it non‐phosphorylatable) or BAP31 (to render it uncleavable), depletion of PERK, caspase‐8, BAP31, Bax, Bak or SNAREs abolished CRT/ERp57 exposure induced by anthracyclines, oxaliplatin and ultraviolet C light. Depletion of PERK, caspase‐8 or SNAREs had no effect on cell death induced by anthracyclines, yet abolished the immunogenicity of cell death, which could be restored by absorbing recombinant CRT to the cell surface.     

加拿大一枝黄花不同居群叶表皮和茎结构特征及其与环境因子的CCA分析

对采自华东地区45个居群并移栽于同一生境条件下的加拿大一枝黄花(Solidago canadensis L.)叶表皮和茎横切面结构特征进行了比较观察,并对叶表皮和茎结构特征与环境因子、居群分布与环境因子的相关性进行了CCA分析.结果表明,加拿大一枝黄花叶片上、下表皮毛均为单列多细胞型非腺毛,下表皮毛密度高于上表皮;上表皮细胞为多边形且垂周壁平直或波状,下表皮细胞为不规则形或多边形且垂周壁较平直或呈波状、浅波状;叶片上表皮几无气孔分布,下表皮均有气孔分布,气孔器为无规则型,气孔多近圆形;不同居群间表皮毛密度、气孔密度、气孔长度和宽度有显著差异,在较干燥环境下的居群表皮毛密度较大、气孔较小,在较湿润生境下的居群表皮毛较少、气孔较大.茎横切面均由表皮、皮层和中柱组成;各居群表皮和皮层结构没有明显差异,维管束数目差异明显(8.05～13.39 mm-1);髓部面积百分比也有明显差异(25.7%～49.5%).叶表皮和茎结构特征的可塑性指数存在居群间差异,其中气孔密度的平均可塑性指数最大(0.38),髓部面积百分比的平均可塑性指数最小(0.13).CCA分析结果表明,茎和叶表皮结构特征与土壤湿润度和纬度的关系较大,气孔密度、长度和宽度与土壤湿润度呈正相关,维管束数目、表皮毛密度及髓部面积百分比与土壤湿润度和纬度呈负相关;各居群的分布与分布地的土壤湿润度和纬度的相关性较大.结果显示, 加拿大一枝黄花居群间的形态特征适应性分化利于其入侵不同的生境,而采用无性繁殖方式可保持其个体形态特征上的变异,从而增加其入侵的成功率.     

现存被子植物原始类群及其植物地理学研究

在吸取各学派对被子植物原始类群界定的基础上,根据八纲系统,提出被子植物原始类群有60科的新见解,并以分子系统学提出的狭义的基部类群32科为对比,进行了植物地理学研究。以科为性状,以Takhtajan 划分的世界植物区系的“区”为OTU,UPGMA分析显示：(1)东亚区确是一个十分特殊的区,它既与北美东、西部(北美大西洋区、马德雷区)有密切关系,但更接近印度支那区;(2)环太平洋的4个地区集中了较多的原始被子植物的科,它们是东亚地区,北美东部和西部地区,部分热带亚洲、澳大利亚东部和西南太平洋岛屿地区,中、南美热带地区。这种分布格局显然和被子植物起源地与扩散以及太平洋的形成历史有关。     

Dynamic changes of interleukin-8 network along the colorectal adenoma–carcinoma sequence

The interleukin-8 (IL-8) network is involved in the colorectal cancer (CRC) progression. However, its role during the adenoma–carcinoma transition to date has not been fully investigated. To evaluate the dynamic changes of IL-8 network along the colorectal adenoma–carcinoma sequence, we examined the tissue IL-8 mRNA level in colorectal biopsies from 53 colorectal adenomas, 44 CRCs and 18 controls by quantitative real-time PCR (Q-PCR), and the expressions of IL-8 and its receptors (IL-8RA and IL-8RB) in the tumor microenvironment by immunohistochemistry (IHC) and double IHCs. The results showed that the tissue IL-8 mRNA level began to increase in the precancerous lesions (adenomas) as compared with the controls and became even higher in the CRCs. Significantly, the increase of IL-8 mRNA levels was associated with the increase of dysplastic grades in the adenomas, and also paralleled to the increase of Duke’s stages in the CRCs. IHC results revealed that IL-8 and its receptors, IL-8RA and IL-8RB, were observed both in the stroma and in the adenomatous/cancerous cells. By double IHCs, the IL-8 expression was characterized in macrophages, lymphocytes and myofibroblasts in the tumor stroma. Further double IHC identified the co-expression of IL-8 receptors (IL-8RA and IL-8RB) with CD34 positive tumor-associated microvessels in both the adenomas and CRCs. We, therefore, conclude that activated IL-8 network in the tumor microenvironment may function as a significant regulatory factor for the adenoma progression and the adenoma–carcinoma transition.     

Temperature Changes between Neighboring Days and Mortality in Summer: A Distributed Lag Non-Linear Time Series Analysis

Background

Many studies have shown that high temperatures or heat waves were associated with mortality and morbidity. However, few studies have examined whether temperature changes between neighboring days have any significant impact on human health.

Method

A distributed lag non-linear model was employed to investigate the effect of temperature changes on mortality in summer during 2006–2010 in two subtropical Chinese cities. The temperature change was defined as the difference of the current day’s and the previous day’s mean temperature.

Results

We found non-linear effects of temperature changes between neighboring days in summer on mortality in both cities. Temperature increase was associated with increased mortality from non-accidental diseases and cardiovascular diseases, while temperature decrease had a protective effect on non-accidental mortality and cardiovascular mortality in both cities. Significant association between temperature changes and respiratory mortality was only found in Guangzhou.

Conclusion

This study suggests that temperature changes between neighboring days might be an alternative temperature indicator for studying temperature-mortality relationship.     

游离及固定化果糖基转移酶部分酶学性质的比较研究

 从诱变、筛选的米曲霉GX0 0 10菌株所产生的果糖基转移酶 ,经过纯化和固定化操作分别制备游离酶和固定化酶 ,对两者的酶学性质进行了比较研究 .结果表明 ,两者在蔗糖转化为蔗果低聚糖的酶促反应中 ,最适pH为 5 5,在pH5 0～ 7 5之间酶活性相对稳定 .游离酶和固定化酶的适宜温度范围分别是 4 5～ 52℃和 4 0～ 55℃ .在 55℃保温 60min ,酶活性保存率分别是 61 6%和 87 5% .固定化酶的热稳定性提高 .0 1mmol LHg2 +和 1mmol LAg+能完全抑制游离酶的活性 ,但只能部分抑制固定化酶的活性 ,1mmol L的Ti2 +能完全抑制两者的活性 .以蔗糖为底物时 ,游离酶的米氏常数Km=2 15mmol L ,而固定化酶Km =386mmol L .游离酶只能使用一次 ,固定化酶反复使用 54次后 ,剩余活力为 55 2 % .用 55% (W V)蔗糖溶液与固定化酶在pH5 0 ,4 6℃下作用 12h ,可获得61 5% (总低聚糖 总糖 )产物 ,其中蔗果五糖含量达到 7 2 % .