环化腺苷酸对细菌生长的影响

用大肠杆菌(Escherichia coli AS 1.797)、北京棒状杆菌(Corynebacterium pekinense AS1.299)和巨大芽孢杆菌(Bacills megatertum AS 1.217)研究了细胞内环化腺苷酸(cAMP)浓度和外源cAMP对细胞生长的影响。结果表明,大肠杆菌在不同碳源中生长时,细胞的生长量随细胞内cAMF’浓度升高而降低。在以葡萄糖作碳源时,细胞内cAMP浓度低,外源cAMP。对生长有抑制作用,而cAMP的类似物5'-AMP则无抑制作用。在以乳糖、麦芽糖和甘油分别作碳源时,细胞内cAMP浓度高,外源cAMP对生长无影响。北京捧状杆菌以葡萄糖作碳源时,细胞生长也受外源cAMP的抑制,但cAMP的抑制作用不是专一的,它的作用可用类似物5’-AMP来代替。自身不合cAMP的巨大芽孢杆菌在不同碳原(包括葡萄糖)中生长时,生长不受外源cAMP抑制,也不受5’-Amt’的影响。因此认为,cAMP不是细菌生长的必需物,而是生长调节物,但这种调节物对巨大芽孢杆菌无效。     

Cloning and nucleotide sequence analysis of genes coding for the major chlorophyll-binding protein of the moss Physcomitrella patens and the halotolerant alga Dunaliella salina

Two clones have been isolated from a genomic library of the moss Physcomitrella patens and a cDNA library of the halotolerant green alga Dunaliella salina. The isolates contain genes coding for the major light-harvesting chlorophyll-a/b-binding protein (CAB) in the photosystem II (PSII) light-harvesting complex (LHCII). The 2544-bp insert of the moss genomic clone contains the complete CAB-coding region and 5' and 3' flanking sequences. The coding region contains an intron of 359 bp which is spanned by a pair of 9-bp perfect direct repeats. There are two CCAAT boxes and five enhancer-like elements related to (G)TGGTTTAAA(G) (Weiher et al., 1983) residing in the intron. Comparisons of the moss cab gene with sequences of light-inducible genes of higher plants reveal homologous and repeated sequences similar to the enhancer element in the 5' region upstream from the TATA and CCAAT boxes thought to be responsive to light inducibility. The 1256-bp algal cDNA contains the complete CAB-coding sequence, a 170-bp 5'-nontranslated region, and a 264-bp 3'-nontranslated region. While the overall homology in the nontranslated regions is low between the cab gene of the moss and that of the alga, the 3'-nontranslated regions of the two contain some sequences that are conserved among the cab genes in higher plants. The deduced amino acid sequences of these two clones are highly conserved except for the N-terminal region. Their hydropathic plots are very similar and both possess three hydrophobic segments that are likely alpha-helical transmembrane segments. The proposed CAB transit peptide sequence of the alga is divergent from that of the moss or higher plants, suggesting that they may have evolved from different origins. Southern blot analysis shows that the cab genes in the moss and the alga, as in higher plants, are encoded by a number of homologous genes constituting a multigene family.     

Amplification of multicistronic plasmids in the human 293 cell line and secretion of correctly processed recombinant human protein C

We have constructed multicistronic vectors containing the cDNAs for murine dihydrofolate reductase (DHFR), hygromycin phosphotransferase (HyPR), and human protein C (HPC), an antithrombotic factor. Using a sequential selection protocol with hygromycin (Hy) and methotrexate (MTX), we demonstrate the selective amplification of the murine dhfr cDNA in the adenovirus-transformed human kidney cell line 293, and the coamplification of the cDNA for HPC. Such recombinant 293 cell lines secreted HPC at levels as high as 25 micrograms/10(6) cells/day. In addition, we found that the complex vitamin K-dependent posttranslational modification of gamma-carboxylation of glutamate was not limiting at these high secretion levels, although the proteolytic processing of the protein was slightly reduced. Further, the HPC secreted from the gene-amplified cell lines had full anticoagulant activity when compared to plasma-derived HPC.     

心肌细胞间的电耦联

心脏是由无数个独立的心肌细胞通过缝隙连接形成的一个功能性合胞体。心肌细胞间的电耦联是心脏“全或无”性动作电位传导和机械收缩的先决条件。缺氧、缺血、药物、细胞内外离子浓度的改变以及激素等因素可直接或间接地影响心肌细胞间的电耦联,从而导致心脏功能的改变。     

千金藤属植物小叶地不容中的生物碱(Ⅷ)

从小叶地不容(Steghania succitera Lo.et Tsoong)的块根中分得11种生物碱,根据理化性质及波谱测定,鉴定为克班宁(crebanine)(Ⅰ),四氢巴马汀(tetrahydropalmatine)(Ⅱ),去氢克班宁(dehydrocrebanine)(Ⅲ),巴马汀(palmatine)(Ⅳ),去氢紫堇单酚碱(dehydro-corydalmiae)(Ⅴ),克班宁硝酸盐(Ⅵ),氧代克班宁(oxocrebanine)(Ⅶ),紫堇单酚碱(corydalmine)(Ⅷ),phanostenine(Ⅸ),离木明碱 discretamine(Ⅹ),及 corypalmine(Ⅺ),化合物(Ⅳ)、(Ⅴ)、(Ⅵ)、(Ⅶ)、(Ⅸ)、(Ⅹ)和Ⅺ为首次报道从该植物中分得。     

向日葵离体孤雌生殖的超微结构研究

本文是研究未受精胚珠培养诱导的孤雌生殖过程超微结构变化的首次报道。向日葵(Heliaanthus annuus L.)的卵细胞在离体条件下被激活,发生细胞核移位、极性丧失、细胞器增多并转变成活动状态、液泡化程度增大、合点端形成细胞壁等一系列变化,预示即将启动孤雌生殖。孤雌生殖的原胚具有若干显著特征,如极性颠倒、有自体吞噬活动、壁的自由生长、游离核分裂等。对这些现象作了初步的讨论。     

青海藏族青少年骨龄与生长发育关系研究

本文报告了青海省境内,世居在海拔3000-4000米地区的728名7-18岁健康藏族青少年学生的手、腕部骨骼发育情况,对骨化中心出现和骨骺愈合求出了50%出现年龄,并对骨龄与青春期身高突增的关系及与月经初衬潮的关系进行了分析。     

Prostaglandin E2 induced changes in renal blood flow, renal interstitial hydrostatic pressure and sodium excretion in the rat

Prostaglandin E2, when infused into the renal artery of the dog, is a vasodilator and increases both renal interstitial hydrostatic pressure and sodium excretion. Similar studies in the rat, however, have been inconclusive. The present study examined the effect of prostaglandin E2 infusion into the renal interstitium, by means of a chronically implanted matrix, on renal blood flow, renal interstitial hydrostatic pressure and sodium excretion in the rat. Prostaglandin E2 was continuously infused directly into the kidney interstitium to mimic endogenous prostaglandin E2 production by renal cells. The maximum change in each of these parameters occurred when 10(-5) M PGE2 was infused. Renal blood flow increased from 4.70 +/- 0.91 to 5.45 +/- 0.35 ml/min (p less than 0.05) while renal interstitial hydrostatic pressure decreased from 3.9 +/- 0.4 to 2.6 +/- 0.5 mmHg (p less than 0.05) and fractional excretion of sodium decreased from 1.02 +/- 0.20 to 0.61 +/- 0.12% (p less than 0.05). Thus, the present study demonstrates that renal interstitial infusion of prostaglandin E2 increases total renal blood flow but decreases both renal interstitial hydrostatic pressure and urinary sodium excretion in the rat.     

What is the relationship between the endothelium derived relaxant factor and nitric oxide?

Nitric oxide gas in solution (NO) relaxes blood vessels with similar actions and pharmacodynamics as the endothelium derived relaxant factor (EDRF) and has been proposed to be a component of the materials released from stimulated endothelial cells. Certain data however suggest that EDRF and NO may not be identical. In some non-vascular smooth muscles, NO and EDRF exhibit markedly different pharmacologic profiles. Furthermore the interaction of EDRF and NO with anion exchange resins differ. The hypothesis that EDRF is identical to nitric oxide gas in solution or a nitrogen oxide containing compound is discussed.     

Design and implementation of a particle concentration fluorescence method for the detection of HIV-1 protease inhibitors.

A critical step in the replicative cycle of the human immunodeficiency virus HIV-1 involves the proteolytic processing of the polyprotein products Prgag and Prgag-pol that are encoded by the gag and pol genes in the viral genome. Inhibitors of this processing step have the potential to be important therapeutic agents in the management of acquired immunodeficiency syndrome. Current assays for inhibitors of HIV-1 protease are slow, cumbersome, or susceptible to interference by test compounds. An approach to the generation of a rapid, sensitive assay for HIV-1 protease inhibitors that is devoid of interference problems is to use a capture system which allows for isolation of the products from the reaction mixture prior to signal quantitation. In this paper, we describe a novel method for the detection of HIV-1 protease inhibitors utilizing the concept of particle concentration fluorescence. Our approach involves the use of the HIV-1 protease peptide substrate Ser-Gln-Asn-Tyr-Pro-Ile-Val which has been modified to contain a biotin moiety on one side and a fluorescein reporter molecule on the other side of the scissile Tyr-Pro bond. This substrate is efficiently cleaved by the HIV-1 protease and the reaction can be readily quantitated. Known inhibitors of the protease were readily detected using this new assay. In addition, this approach is compatible with existing instrumentation in use for broad screening and is highly sensitive, accurate, and reproducible.