氯喹与氟康唑联合对新生隐球菌的体外药敏实验

目的研究氯喹与氟康唑体外联合应用对隐球菌生长作用的影响。方法参考M27-A方案,检测10μmol/L,100μmol/L与1 000μmol/L 3种浓度氯喹与氟康唑联合后对20株菌株的氟康唑对隐球菌最小抑菌浓度与最小杀菌浓度的变化。结果与10μmol/L氯喹组相比,100μmol/L以上浓度的氯喹组可以显著降低氟康唑对隐球菌的M IC与MFC。结论氯喹在体外可以提高氟康唑抗隐球菌的作用,与氟康唑具有协同抗隐球菌的作用。     

The role of PilZ domain proteins in the virulence of Xanthomonas campestris pv. campestris

Cyclic di‐GMP [(bis‐(3′–5′)‐cyclic di‐guanosine monophosphate)] is an almost ubiquitous second messenger in bacteria that is implicated in the regulation of a range of functions that include developmental transitions, aggregative behaviour, adhesion, biofilm formation and virulence. Comparatively little is known about the mechanism(s) by which cyclic di‐GMP exerts these various regulatory effects. PilZ has been identified as a cyclic di‐GMP binding protein domain; proteins with this domain are involved in regulation of specific cellular processes, including the virulence of animal pathogens. Here we have examined the role of PilZ domain proteins in virulence and the regulation of virulence factor synthesis in Xanthomonas campestris pv. campestris (Xcc), the causal agent of black rot of crucifers. The Xcc genome encodes four proteins (XC0965, XC2249, XC2317 and XC3221) that have a PilZ domain. Mutation of XC0965, XC2249 and XC3221 led to a significant reduction of virulence in Chinese radish. Mutation of XC2249 and XC3221 led to a reduction in motility whereas mutation of XC2249 and XC0965 affected extracellular enzyme production. All mutant strains were unaffected in biofilm formation in vitro. The reduction of virulence following mutation of XC3221 could not be wholly attributed to an effect on motility as mutation of pilA, which abolishes motility, has a lesser effect on virulence.     

Genetic polymorphism of mitochondrial DNA HVS-I and HVS-Ⅱ of Chinese Tu ethnic minority group

We analyzed the two hypervariable segments HVS-Ⅰ and HVS-Ⅱ of 108 Chinese Tu ethnic minority group samples for forensic and population genetics purposes.Comparing with Anderson sequence,79 polymorphic loci in HVS-Ⅰ and 40 in HVS-Ⅱ were found in Chi-nese Tu ethnic minority group mtDNA sequences,and 90 and 64 haplotypes were then defined.Haplotype diversity and the mean pair-wise differences were 0.9903:±0.0013 and 5.7785 in HVS-Ⅰ,and 0.9777±0.0013 and 3.5819 in HVS-Ⅱ,respectively.By analyzing the hypervariable domain from nucleotide 1,6180 to 1,6193 in HVS-Ⅰ,we defined some new types of sequence variations.We also compared the relationship between Tu population and other populations using mtDNA HVS-Ⅰ sequences.According to Rst genetic distances,the phylogenetic tree showed that the Tu population,the Xi'an Han population,the Chinese Korean,and the Mongol ethnic group were in a clade.This indicated a close genetic relationship between them.There were far relations between the Tu population and other Chinese southern Han populations,Siberian,European,African,and other foreign populations.The results suggest that Tu population has a multi-origin and has also merged with other local populations.     

体外重组的人促黄体素对小鼠早期胚胎发育的影响

为了研究人的重组促黄体素（r-hLH)对小鼠早期胚胎发育的作用,我们用一定剂量的r-hFSH(人促卵泡素,20IU）结合不同剂量的r-hLH（0－20IU）,或先与不同剂量r-hLH配比（5－20IU）,再与一定剂量,r-hLH（15IU）结合,对性成熟前的小鼠进行超数排卵;并对从其输卵管冲出的早期胚胎进行体外培养,通过观察这些早期胚胎体外发育情况,再与PMSG－hCG超排小鼠的早期胚胎体外培养结果进行比较。我们发现用重组激素处理的性成熟前小鼠,无论在平均冲胚数或桑囊率,囊胚率上,都以结合r-hLH或配比r-hLH的剂量为15IU时最好,但各重组激素处理组在桑囊率和囊胚率上都极显著低于PMSG－hCG对照组的相应指标。上述结果表明,r-hLH对小鼠早期胚胎的发育（尤其是囊胚的形成）有明显的抑制作用,且与r-hLH呈一定的剂量依赖性关系。     

Regeneration of plants from callus tissues of hairy roots induced by Agrobacterium rhizogenes on Alhagi pseudoalhagi

INTRoDUCTIONThe hairy root disease is a patholOgical syndromeof dicotyledonous plants fOllowing wounding and in-fection with Agrobacterum rhjZOgenesI1]. The rhi-zogenicity is conferred to p1ant cel1s by a fragmentof DNA (Ri T-DNA), which is transferred from thelarge root--inducting (Ri) plasmid, haJrboured by thebacterium, to the genome, where it is stably inte-grated and expressed. Illtegration of a DNA segment(T-DNA) of pRi into the host genome 1eads to ac-tive proliferation of…     

The occurrence and ecological importance of dissolved ATP in fresh water

SUMMARY. Dissolved ATP, defined as ATP which passes through 0.2 μm filters, was found in fresh water. During the spring diatom bloom in two eutrophic Danish lakes, concentrations of dissolved ATP varied between 0.1 and 3.8 μgl⁻¹, constituting 14–76% of the total ATP (particulate plus dissolved ATP). The kinetics of the light emission obtained from mixing firefly enzyme with dissolved ATP demonstrated that the major proportion of the dissolved ATP was in fact ATP. Despite some variations, the seasonal changes in dissolved ATP paralleled the changes in the increasing phytoplankton population during the rise of the diatom blooms. The dissolved ATP increased after the diatom peak, indicating that release of ATP from the phytoplankton due to mortality may be a major source of dissolved ATP.
Consumption of dissolved ATP was evaluated in uptake experiments using ³H-ATP. Rates of uptake of ³H-ATP by micro-organisms (diameter 0.2–0.6 μm) proved to be close to the rates for ³H-D-glucose uptake. The variations in ³H-ATP uptake during the diatom blooms showed non-systematic changes and ranged between 1.0 and 15.8% h⁻¹ (mean = 4.9% h⁻¹) of the quantity added. Turnover rates for dissolved ATP varied between 12 and 730 ng l⁻¹ h⁻¹ (mean = 175 ng l⁻¹). These rather high rates of turnover suggest that dissolved ATP is an important compound in the metabolism of freshwater bacteria.     

东亚钳蝎氯毒素通过下调PKM2介导的有氧糖酵解抑制神经胶质瘤细胞的增殖活性

东亚钳蝎氯离子通道毒素(Buthus martensii Karshch chlorion channel toxin,BmK CT)是一类短链神经毒素,在体内和体外均能有效抑制神经胶质瘤细胞侵袭和增殖。然而,BmK CT抑制胶质瘤细胞增殖活性的机制尚不清楚。本研究采用代谢组学的方法探索BmK CT抑制人脑神经胶质瘤细胞增殖的分子机制。首先,通过蛋白质表达纯化获得带有谷胱甘肽S转移酶(glutathione S-transferase, GST)标签的BmK CT蛋白和GST蛋白。利用圆二色谱检测两种蛋白质均具有α-螺旋二级结构,并且融合蛋白质GST-BmK CT热稳定性较好。通过噻唑蓝(methylthiazolyldiphenyl-tetrazolium bromide, MTT)法检测GST-BmK CT对神经胶质瘤细胞(U251和A172)增殖的影响。结果显示,与GST处理组相比,1μmol/L GST-BmK CT处理组对U251细胞的抑制率更明显(U251细胞19±1.1 vs. 2.25±0.95,P<0.001, 48 h; A172细胞12±1.4 vs. 2.25±1.25,P<0.001, 48 h)。利用气相色谱-质谱联用仪(gas chromatography-mass spectrometry, GC-MS)分析鉴定出细胞内的63种代谢物,通过通路分析和聚类分析证实,GST-BmK CT处理影响神经胶质瘤细胞的有氧糖酵解。根据峰面积统计数据显示,GST-BmK CT处理降低胞内丙酮酸的含量。GST-BmK CT处理组相比GST组,明显下调胞外培养基的乳酸分泌(16.33±3.78 vs. 35.6±2.31,P<0.001, 48 h),葡萄糖消耗(2.04±0.09 vs. 2.64±0.04,P<0.05, 48 h)和胞内腺苷三磷酸(ATP)含量(633.79±0.001 vs. 5392.71±266.67,P<0.05, 48 h)。结果表明,GST-BmK CT通过下调胶质瘤细胞有氧糖酵解水平降低ATP的生成。实时荧光定量PCR和蛋白质免疫印迹结果显示,GST-BmK CT降低U251细胞丙酮酸激酶(pyruvate kinase M2, PKM2)在转录水平和翻译水平的表达,进一步发现GST-BmK CT通过下调低氧诱导因子(hypoxia inducible factor-1α, HIF-1α)降低PKM2在转录水平的表达。以上结果表明,BmK CT通过下调HIF-1α的表达,降低PKM2介导的有氧糖酵解从而抑制神经胶质瘤细胞的增殖活性。     

小麦T型细胞质雄性不育系与相应保持系线粒体DNA的RAPD分析

利用RAPD技术对小麦T型细胞质雄性不育系75-3369A和相应保持系75-336 9B 的线粒体DNA(mtDNA)进行了81个单引物和14组双引物的多态性研究。结果有73个单引物和全部的双引物有扩增结果,表现出多态性的有10个单引物和一组双引物。 Abstract:Using 81 single-primers and 14 double-primers,T-type CMS(cytoplasmic male sterile)line 75-3369 A and is maintainer 75-3369B were studied by means of RAPD.73 single-primers and all double-primers gave amplified products in the experiment,among them 10 single-primers and one double-primer presented polymorphism.     

中脑导水管周围灰质内注射抗阿片肽血清对神经降压素增强电针镇痛的影响

本工作以钾离子透入引起大鼠甩尾反应作为痛反应指标,观察纳洛酮（ＮＸ）和抗阿片肽血清对大鼠中脑导水管周围灰质（ＰＡＧ）内微量注射神经降压素（ＮＴ）增强电针镇痛作用的影响。结果显示：（１）ＰＡＧ内微量注射ＮＴ后,大鼠电针镇痛效应明显提高;（２）ＰＡＧ内注射大剂量ＮＸ,可以显著减弱ＮＴ增强电针镇痛效应;（３）ＰＡＧ内微量注射抗甲脑啡肽血清和抗β内啡肽血清后,ＮＴ增强电针镇痛的作用明显降低;而ＰＡＧ内微量注射抗强啡肽Ａ１１３血清,对于ＮＴ增强电针镇痛的作用并无明显影响。提示：ＰＡＧ内ＮＴ在针刺镇痛过程中发挥重要作用,ＮＴ增强电针镇痛的作用与甲脑啡肽和β内啡肽有较密切的关系。