中国乌头属属下等级数量分类的初步研究

本文取中国乌头属50种(含变种)植物,选取形态学性状46个、化学性状8个、组织学性状5个、生态学性状1个,运用数量分类方法,对该属进行属下等级的分类研究。根据数学分析结果,确定了树系图中变种、种、系、亚属的分级界线,并在聚类分析的基础上,进一步探讨了各等级类群的划分。     

过硫酸铵-N,N,N′,N′-四甲基乙二胺体系产生的氧自由基及其清除的研究

应用ESR和自旋捕集相结合的技术直接测定了过硫酸铵—N,N,N′,N′-四甲基乙二胺(AP-TEMED)体系产生的氧自由基,经计算机波谱模拟和计算波谱参数证实该体系产生的氧自由基是O_2~-和·OH。并用维生素C、茶多酚、超氧化物歧化酶等氧自由基清除剂,从聚丙烯酰胺凝胶法、化学发光法和脂质过氧化法不同角度研究了AP-TEMED体系在自由基研究方面的应用意义。     

用2DNMR鉴定倍半萜多元酯的结构

利用2DNMR结合其他波谱方法从苦皮藤种子油中鉴定了2个新倍半萜多元酯,它们分别是1α-苯甲酰氧-8β-烟酰氧-9α、11β-2乙酰氧-β-二氢沉香呋喃,1β-苯甲酰氧-2α、8β、9α、11β-4乙酰氧-β-二氢沉香呋喃,命名为苦皮藤酯2和3。     

毛枝南蛇藤根皮中南蛇藤素的分离鉴定及含量测定

从卫矛科南蛇藤属植物毛枝南蛇藤(Celastrus hookeri Prain)根皮中首次分离出一个红色立方体形结晶,经紫外、红外、高分辨质谱和~(13)C核磁共振谱分析,并与有关文献数据对照,确认是南蛇藤素。作者用薄层扫描和高效液相两种方法初步测定了毛枝南蛇藤根皮和根的木栓化表皮的南蛇藤素含量,结果表明南蛇藤素主要集中在根的木栓化表皮,含量在12%以上,而在整个根皮中的含量大约为1%。     

咽喉部高频喷射通气调节各参数对狗肺通气效率的影响

用相关和回归处理方法,研究了8条正常狗咽喉部高频喷射通气时,调节驱动压、呼吸比和频率对喷气量、吸入气氧浓度、动脉血气及气道内压的作用。结果显示,驱动压和呼吸此对各观察指标几乎有同等重要的作用,频率的影响很小,喷气量与吸入气氧浓度、动脉血气、气道内压间存在显著的正相关关系。说明调节参数的意义主要在于改变了喷气量。     

Semisynthetic des-(B27-B30)-insulins with modified B26-tyrosine

Semisynthetic des-(B27-B30)-insulins containing modified B26-tyrosine residues were prepared to refine the understanding of the importance of position B26 with regard to biological and structural properties of the hormone. The following shortened insulin analogues were synthesized by trypsin-catalysed peptide-bond formation between the C-terminal amino acid ArgB22 of des-(B23-B30)-insulin and synthetic tetrapeptides as amino components: des-(B27-B30)-insulin, des-(B27-B30)-insulin-B26-methyl ester, -B26-carboxamide with varying C-terminal hydrophobicity of the B-chain, and [Tyr(NH2)B26]-, [Tyr(NO2)B26]-, [Tyr(I2)B26]-, [D-TyrB26]des-(B27-B30)-insulin-B26-carboxamide containing non-proteinogenic amino acids in position B26. Starting from insulin and an excess of synthetic Gly-Phe-Phe-Tyr-OMe as nucleophile, des-(B27-B30)-insulin-B26-methyl ester--the formal transpeptidation product at ArgB22--was formed in one step. Biological in vitro properties (binding to cultured human IM-9 lymphocytes, relative lipogenic potency in isolated rat adipocytes) of all semisynthetic analogues are reported, ranging from slightly decreased to two-fold receptor affinity and nearly three-fold biopotency relative to insulin. If the C-terminal tetrapeptide B27-B30 is removed, full relative insulin activity is still preserved, while the shortening results in the loss of ability to associate in solution. Only after carboxamidation or methyl esterification of TyrB26 the self-association typical of native insulin can be observed, and the CD-spectral effects in the near UV spectrum related to association and hexamerization of the native hormone are qualitatively reestablished. The results of this investigation underline the importance of position B26 to the modulation of hormonal properties and solution structure of the shortened insulins.     

Molecular cloning of the mouse 46-kDa mannose 6-phosphate receptor (MPR 46).

A cDNA clone for the mouse 46-kDa mannose 6-phosphate receptor (MPR 46) was isolated from an embryonic mouse cDNA library. Its single open reading frame codes for a protein of 278 residues. It shows an over-all amino-acid identity of 93% with the human receptor. Nine non-conservative amino-acid exchanges are found in the luminal domain, one non-conservative exchange of hydrophobic amino acids is in the transmembrane domain, while the cytoplasmic receptor tails are identical. All five potential N-glycosylation sites are conserved as well as amino acids that are important for ligand binding (Arg 137 and His 131) and disulfide pairing (Cys 32 and 78, Cys 132 and Cys 167, Cys 145 and Cys 179). The absolute identity in the cytoplasmic MPR 46 tail suggests the importance of this amino-acid sequence for the intracellular routing of the MPR 46.     

N-terminal amino-acid sequence of pig kidney dipeptidyl peptidase IV solubilized by autolysis.

The N-terminal amino-acid sequence of the intrinsic membrane protein dipeptidyl peptidase IV (DP IV) was determined. The protein was isolated from pig kidney and solubilized by autolysis at pH 3.8. The first 34 amino acids were sequenced and indicated approximately 78% identity to the N-terminal sequence of rat liver DP IV.     

Influence of tissue preparation techniques on p53 expression in bronchial and bladder carcinomas, assessed by immunofluorescence staining and flow cytometry.

In a series of bronchial and bladder carcinomas, p53 protein expression was examined. Samples from formalin-fixed, paraffin-embedded tissue (routine-treated) were compared with parallel samples of fresh tissue and tissue fixed in paraformaldehyde and ethanol. The expression of p53 was measured by immunofluorescence staining and dual parameter flow cytometry, with simultaneous monitoring of DNA content. For each tumor, p53 fluorescence with different fixatives was expressed relative to fresh tissue. The p53 fluorescence signals were on average brighter from routine-treated tissue than from fresh tissue. The tissue fixed in paraformaldehyde showed no difference from fresh tissue. In the ethanol-fixed tissue, however, fluorescence signals were reduced by nearly 70%, and the fraction of detectable p53 positive cells in tumor tissue was reduced by more than 50%. This loss of fluorescence was probably due to a leakage of the antigen from nucleus to cytoplasm. Pepsin treatment did not influence p53 fluorescence. Within the same tumor, the S-phase fraction in p53 positive cells was significantly higher than in p53 negative cells (13.1 +/- 2.0% vs. 6.5 +/- 0.8%). This pattern was not influenced by formalin fixation or pepsin treatment. Our study demonstrates that in measuring a nuclear antigen, tissue handling may influence the results, and care should be taken to optimize the preparation procedure. Using the antibody PAb 1801, p53 expression measured in archival material is not reduced as compared to fresh tissue.     

Comparison of two cytochromes P-450 from Candida maltosa: primary structures, substrate specificities and effects of their expression in Saccharomyces cerevisiae on the proliferation of the endoplasmic reticulum

cDNAs were cloned, sequenced and expressed which encode two different cytochrome P-450 forms of the alkane-assimilating yeast Candida maltosa, designated as P-450Cm1 and P-450Cm2. The amino acid sequences deduced were about 55% identical. Expression in Saccharomyces cerevisiae resulted in the formation of intact microsomal P-450 systems catalyzing the hydroxylation of n-hexadecane and lauric acid with significantly different substrate preferences. A massive proliferation of the endoplasmic reticulum was observed in the S. cerevisiae cells which produced P-450. Depending on the P-450 form expressed, distinctly organized stacks of paired membranes appeared and occupied considerable areas of the cytoplasm. As shown by immunoelectron microscopy for P-450Cm1, the protein expressed was highly concentrated within these newly formed membrane structures.