基于流式细胞术的桫椤基因组大小测定

以买麻藤(Gnetum montanum)为内参,测定5种常用裂解液(Tris·MgCl_2、LB01、WPB、Otto’s、Galbraith’s)对桫椤(Alsophila spinulosa)细胞核的裂解效果,并通过比较桫椤和买麻藤流式细胞仪测定的荧光峰值,计算出桫椤的基因组大小。结果表明:5种裂解液中,WPB为桫椤最佳裂解液;桫椤基因组大小为6 003.25Mb,即2C DNA含量为6.138 pg。     

Abnormal Sysmex XT‐2000iV DIFF scattergram in a cat with a prominent mastocytemia

A 2‐year‐old neutered male domestic shorthair cat was presented to the emergency service of the National Veterinary School of Toulouse (France) for acute vomiting and diarrhea with lethargy, inappetence, and adypsia for the past 48 hours. Complete blood counts were performed with the ProCyte DX at the emergency department and with the Sysmex XT‐2000iV at the laboratory 2 weeks later. The scattergrams from the two analyzers revealed similar unusual and abnormal dot plots. The Sysmex XT‐2000iV DIFF scattergram also showed no clear separation between different leukocyte populations. The eosinophil cluster was in an abnormal location compared with that of the “typical” location in a normal cat. A blood smear evaluation revealed the presence of numerous mast cells. Thus, we hypothesized that the Sysmex XT‐2000iV had detected the mast cell population, and this led to errors in the differential counts. To explore this hypothesis, we manually gated on the DIFF scattergram and performed a manual differential on the blood smear. With this new gating strategy, the Sysmex XT‐2000iV and manual differentials were similar. Thus, in the case of systemic mastocytosis, mast cells can be located between the lymphocyte, monocyte, and eosinophil clusters on scattergrams.     

猪霍乱沙门菌菌蜕的制备及免疫保护力研究

为制备猪霍乱沙门菌菌蜕并研究其免疫保护力,本实验克隆噬菌体phiX174裂解基因E,与pBV220连接,构建温控溶菌表达载体,将该载体转入猪霍乱沙门菌TTB1中,制备菌蜕并对其裂解率、安全性以及对小鼠的免疫保护力进行了研究。结果显示：成果克隆了裂解基因E、片段大小274bp,构建了温控溶菌质粒载体pBVE,制备了猪霍乱沙门菌菌蜕,其裂解率最高为99.46%、经冻干灭活残余活菌,安全性试验检测证实其安全后,对小鼠进行了免疫保护试验,其保护率为70%、与弗氏佐剂灭活苗保护率相当、优于甲醛灭活苗。研究结果为猪霍乱沙门菌引发疾病的防治奠定了基础。     

Is the decline of soil microbial biomass in late winter coupled to changes in the physical state of cold soils?

During winter when the active layer of Arctic and alpine soils is below 0 °C, soil microbes are alive but metabolizing slowly, presumably in contact with unfrozen water. This unfrozen water is at the same negative chemical potential as the ice. While both the hydrostatic and the osmotic components of the chemical potential will contribute to this negative value, we argue that the osmotic component (osmotic potential) is the significant contributor. Hence, the soil microorganisms need to be at least halotolerant and psychrotolerant to survive in seasonally frozen soils. The low osmotic potential of unfrozen soil water will lead to the withdrawal of cell water, unless balanced by accumulation of compatible solutes. Many microbes appear to survive this dehydration, since microbial biomass in some situations is high, and rising, in winter. In late winter however, before the soil temperature rises above zero, there can be a considerable decline in soil microbial biomass due to the loss of compatible solutes from viable cells or to cell rupture. This decline may be caused by changes in the physical state of the system, specifically by sudden fluxes of melt water down channels in frozen soil, rapidly raising the chemical potential. The dehydrated cells may be unable to accommodate a rapid rise in osmotic potential so that cell membranes rupture and cells lyse. The exhaustion of soluble substrates released from senescing plant and microbial tissues in autumn and winter may also limit microbial growth, while in addition the rising temperatures may terminate a winter bloom of psychrophiles.Climate change is predicted to cause a decline in plant production in these northern soils, due to summer drought and to an increase in freeze-thaw cycles. Both of these may be expected to reduce soil microbial biomass in late winter. After lysis of microbial cells this biomass provides nutrients for plant growth in early spring. These feedbacks, in turn, could affect herbivory and production at higher trophic levels.     

地中海弧菌Vibrio mediterranei 117-T6的溶藻活性

很多藻类致病菌具有广谱溶藻活性。本研究发现紫菜黄斑病致病菌地中海弧菌Vibrio mediterranei 117-T6（Vm 117-T6）对赤潮异湾藻、颗石藻、叉编金藻和东海原甲藻等微藻也具有较强的溶藻活性，可使其叶绿素含量显著降低，是一株广谱性溶藻细菌。Vm 117-T6能在40 min内使赤潮异弯藻的活动能力下降，光合作用受抑，感染120 min即可导致藻体裂解并产生白色絮状沉淀。Vm 117-T6能降解卡拉胶，不能降解果胶、纤维素和几丁质。菌体、胞内及胞外提取物均具有较强的溶藻活性，其胞外溶藻物质易溶于水、不溶于石油醚和乙酸乙酯，具较强极性；耐高温、不易被活性炭吸附、乙醇处理会降低溶藻活性。Vm 117-T6在常规与易感条件下的关键差异蛋白包括大量的ABC转运蛋白、外膜蛋白、鞭毛蛋白及少量Toxin。这些研究结果提示Vm 117-T6毒力效应物中含有极性非蛋白质类物质，且与毒力因子转运、分泌以及细胞黏附相关的蛋白可能在其毒力作用中发挥重要作用。上述研究结果表明：多种溶藻机制参与了Vm 117-T6的溶藻过程。     

连续热裂解法大规模提取质粒DNA

选择质粒保有率高及最适合发酵的TOP10菌株,在传统的煮沸裂解法提取质粒的基础上进行了提取方法的改进,使用恒流泵和热交换螺旋管,使菌体在流动过程中完成热裂解,释放质粒DNA。改进后的方法裂解温度可降到70",提取工艺流程简单,缩短了提取时间,提高了质粒DNA的提取效率。利用本方法提取的质粒大小正确、纯度合格,每升发酵液得率平均为100mg质粒DNA。     

应用碱性抽提法和羟基磷灰石柱层析法提纯质粒DNA

用碱性抽提法和羟基磷灰石柱层析法,加以改进,总结出一套简便易行、低花费的质粒提纯方法。此法适用于一般条件试验室,可进行质粒的批量制备,所提纯质粒经限制性内切酶消解和转化试验,其产量(900μg/500 ml)、纯度和生物学活性均可满足重组DNA的需要。     

鲫血清转铁蛋白推导序列分析及空间结构预测

用DNA Club分析软件推导出鲫血清转铁蛋白序列,并对该序列进行分析。推导的蛋白质长度为807个氨基酸,MW约为90 kD。其中半胱氨酸43个,可以组成21对二硫键。疏水性和二级结构几率分析表明：有2个非常相似的结构域分别存在于1～390和390-807区域。旋管结构和螺旋结构的分布区域,均分别集中在上述2个区段中,同样位于2个结构域中,结构也非常类似。证实了Tf由2个同源性较高的结构域(N结构域和C结构域)组成。Tf的三级结构呈“V”字形,每个结构域又各自分为3个小的结构小区(即N1、N2、N3和C1、C2、C3)。2个糖基位于N1和C1区,呈“Y”字型结构;铁链位于N2和C2区;受体结合位点R1和R2位于N3和C3区。     

青饲切碎机动刀片受力对比分析

对PCCⅣ15．0S型青饲切碎机的两种动刀片进行受力分析,深入探讨了动刀片在切碎物料过程中各种参数的变化规律,从而得出圆弧刃口动刀片在综合切碎性能上较直刃口动刀片优越。     

Transient hypoparathyroidism following successful treatment of hypercalcaemia of malignancy in a dog

A 4-year-old, entire female, English Cocker Spaniel was presented for treatment of lymphosarcoma and secondary hypercalcaemia. After induction chemotherapy the dog became severely hypocalcaemic and showed signs of weakness, muscle fasciculation and facial pruritus. Hormone analysis confirmed inadequate production of parathyroid hormone. Although hypocalcaemia has been previously reported as a component of tumour lysis syndrome, it has not been associated with transient parathyroid hormone deficiency.