固定化虫荧光素酶光纤传感器

固定化虫荧光素酶光纤传感器蔡谨,王顺光,杨歧生,吉鑫松（浙江大学化工系生化教研室,杭州３１００２７;中国科学院上海生物化学研究所,２０００３１）关键词虫荧光素酶,ＡＴＰ,固定化酶,光纤生物传感器ＡＴＰ是生物体内极为重要的能量物质。如何准确快速地定量Ａ...     

反相胶束体系中辣根过氧化物酶的活力和动力学性质

本文系统研究辣根过氧化物酶在ＣＴＡＢ／Ｈ２Ｏ／ＣＨＣ．３－ｉｓｏｏｃｔａｎｅ（１∶１,Ｖ／Ｖ）反相胶束体系中的催化行为。在一定条件下酶反符合Ｍｉｃｈａｅｌｉｓ－Ｍｅｎｔｅｎ动力学。研究水含量、底物浓度、ＰＨ、温度、表面活性剂的浓度等对酶反应的影响,结果表明表面活性剂对酶表现非竞争性抑制作用,高浓度的过氧化氢抑制酶活,最适ＰＨ为７．０。在低水含量（Ｗ０＜５）的胶束体系中保温后,酶的活力发生不可逆的改     

Rescue of the fission yeast snRNA synthesis mutant snm1 by overexpression of the double-strand-specific Pac1 ribonuclease

The Schizosaccharomyces pombe temperature-sensitive mutant snm1 maintains reduced steady-state quantities of the spliceosomal small nuclear RNAs (snRNAs) and the RNA subunit of the tRNA processing enzyme RNase P. We report here the isolation of the pac1 ⁺ gene as a multi-copy suppressor of snm1. The pac1 ⁺ gene was previously identified as a suppressor of the ran1 mutant and by its ability to cause sterility when overexpressed. The pac1 ⁺ gene encodes a double-strand-specific ribonuclease that is similar to RNase III, an RNA processing and turnover enzyme in Escherichia coli. To investigate the essential structural features of the Pac1 RNase, we altered the pac1 ⁺ gene by deletion and point mutation and tested the mutant constructs for their ability to complement the snm1 and ran1 mutants and to cause sterility. These experiments identified four essential amino acids in the Pac1 sequence: glycine 178, glutamic acid 251, and valines 346 and 347. These amino acids are conserved in all RNase III-like proteins. The glycine and glutamic acid residues were previously identified as essential for E. coli RNase III activity. The valines are conserved in an element found in a family of double-stranded RNA binding proteins. Our results support the hypothesis that the Pac1 RNase is an RNase III homolog and suggest a role for the Pac1 RNase in snRNA metabolism.     

SUBSTITUTION PROCESSES IN MOLECULAR EVOLUTION. II. EXCHANGEABLE MODELS FROM POPULATION GENETICS

Substitution processes are of two sorts: origination processes record the times at which nucleotide mutations that ultimately fix in the population first appear, and fixation processes record the times at which they actually fix. Substitution processes may be generated by combining models of population genetics—here the symmetrical-neutral, overdominance, underdominance, TIM, and SAS-CFF models—with the infinite-sites, no-recombination model of the gene. This paper is mainly concerned with a computer simulation study of these substitution processes. The rate of substitution is shown to be remarkably insensitive to the strength of selection for models with strong balancing selection caused by the genealogical drift of mutations through alleles held in the population by selection. The origination process is shown to be more regular than Poisson for the overdominance, TIM, and SAS-CFF models but more clustered for the underdominance model. A class of point processes called Sawyer processes is introduced to help explain these observations as well as the observation that the times between successive originations are nearly uncorrelated. Fixation processes are shown to be more complex than origination processes, with regularly spaced bursts of multiple fixations. An approximation to the fixation process is described. One important conclusion is that protein evolution is not easily reconciled with any of these models without adding perturbations that recur on a time scale that is commensurate with that of molecular evolution.     

中国摇蚊亚科记述Ⅳ．拟隐摇蚊属（双翅目：摇蚊科）

本文为中国摇蚊亚科系列报道之四,记述了采自海南省的拟隐摇蚊属一新种：宽尖拟隐摇蚊Ｄｅｍｉｃｒｙｐｔｏｃｈｉｒｏｎｏｍｕｓ（Ｉｒｍａｋｉａ）ｓｐａｔｕｌａｔｕｓｓｐ．ｎｏｖ．及１中国新记录种：缺损拟隐摇蚊Ｄｅｍｉｃｒｙｐｔｏｃｈｉｒｏｎｏｍｕｓ（Ｉｒｍａｋｉａ）ｖａｌｅｒａｔｕｓ（Ｚｅｔｔｅｒｓｔｅｄｔ）．模式标本存南开大学生物系．     

Epidermal growth factor stimulates vascular endothelial growth factor production by human malignant glioma cells: a model of glioblastoma multiforme pathophysiology.

Hypervascularity, focal necrosis, persistent cerebral edema, and rapid cellular proliferation are key histopathologic features of glioblastoma multiforme (GBM), the most common and malignant of human brain tumors. By immunoperoxidase and immunofluorescence, we definitively have demonstrated the presence of vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFr) in five out of five human glioma cell lines (U-251MG, U-105MG, D-65MG, D-54MG, and CH-235MG) and in eight human GBM tumor surgical specimens. In vitro experiments with glioma cell lines revealed a consistent and reliable relation between EGFr activation and VEGF production; namely, EGF (1-20 ng/ml) stimulation of glioma cells resulted in a 25-125% increase in secretion of bioactive VEGF. Conditioned media (CM) prepared from EGF-stimulated glioma cell lines produced significant increases in cytosolic free intracellular concentrations of Ca2+ ([Ca2+]i) in human umbilical vein endothelial cells (HUVECs). Neither EGF alone or CM from glioma cultures prepared in the absence of EGF induced [Ca2+]i increases in HUVECs. Preincubation of glioma CM with A4.6.1, a monoclonal antibody to VEGF, completely abolished VEGF-mediated [Ca2+]i transients in HUVECs. Likewise, induction by glioma-derived CM of von Willebrand factor release from HUVECs was completely blocked by A4.6.1 pretreatment. These observations provide a key link in understanding the basic cellular pathophysiology of GBM tumor angiogenesis, increased vascular permeability, and cellular proliferation. Specifically, EGF activation of EGFr expressed on glioma cells leads to enhanced secretion of VEGF by glioma cells. VEGF released by glioma cells in situ most likely accounts for pathognomonic histopathologic and clinical features of GBM tumors in patients, including striking tumor angiogenesis, increased cerebral edema and hypercoagulability manifesting as focal tumor necrosis, deep vein thrombosis, or pulmonary embolism.     

大鼠杏仁核内注射八肽胆囊收缩素（CCK—8）拮抗吗啡镇痛的研究

大鼠双侧杏仁核内注射ＣＣＫ－８１ｎｇ（１μｌ）,能明显降低皮下注射４ｍｇ／ｋｇ吗啡产生的镇痛作用,并在０．１－１ｎｇ范围内呈量效关系。分别向双侧仁核注射ＣＣＫ－Ａ受体拮抗剂Ｄｅｖａｚｅｐｉｄｅ５０ｎｇ能部分翻转,２００ｎｇ则完全翻转ＣＣＫ－８的抗吗啡镇痛作用,１０ｎｇ无效;而ＣＣＫ－Ｂ受体拮抗剂Ｌ－３６５,２６０在５－８ｎｇ时即可完全番转ＣＣＫ－８的抗吗啡镇痛作用。杏仁核注射２００ｎｇ的Ｄｅｖａｚ     

红细胞抗高血压因子降压作用的进一步研究

我们曾报道原发性高血压患者(EHS)红细胞抗高血压因子(AHF)具有缓慢而持久的降压作用。本工作表明,AHF对卒中易感型自发性高血压大鼠(SHRsp)还具有快速短暂的降压作用,注射AHF后10—30s,SHRsp收缩压从原水平的26.8±1.7kPa降至20.1±1.5kPa(P<0.001)。正常人和大鼠红细胞AHF的降压作用明显强于EHS和高血压大鼠AHF。此外我们还发现EHS血浆中存在升压物质。以上结果提示,AHF缺乏和升压物质含量相对较高可能是原发性高血压发病的一个重要原因。     

一株C3形态多重耐药大肠杆菌噬菌体生物学特性和基因组分析

【目的】多重耐药菌的出现对公共卫生安全构成严重威胁,本研究分离多重耐药大肠杆菌噬菌体,研究其生物学特性和基因组特征,为耐药菌的噬菌体疗法提供理论依据。【方法】使用双层平板法从污水样本中分离纯化大肠杆菌噬菌体;磷钨酸染色后通过透射电镜观察形态;测定其宿主范围,测定温度和pH稳定性、一步生长曲线和体外抑菌效果等生物学特性;体内抑菌试验评估噬菌体对多重耐药大肠杆菌N1203-1Af感染的大蜡螟幼虫的保护作用;基于全基因组测序对其基因组特点进行分析。【结果】本研究分离共得到5株大肠杆菌噬菌体,分别命名为pEC-S163-2.1、pEC-S163-2.2、pEC-M1167-5Ar.1、pEC-m1291-2Dr.1和pEC-N1203-2Af.1;电镜结果显示噬菌体pEC-N1203-2Af.1属于短尾噬菌体中罕见的C3形态型,头部较长,长是宽的2–3倍;pEC-N1203-2Af.1可裂解受试15株大肠杆菌中的3株;感染10 min后进入指数增长期,–20-50℃、pH值为4.0–10.0的环境下均能够保持稳定活性;大蜡螟幼虫感染大肠杆菌N1203-2Af后噬菌体pEC-N1203-2Af....