正常人及肝细胞癌患者肝脏及血浆中丙酮酸激酶同工酶免疫测定

 分别从人肝及鼠肝中高度纯化L型和M_1型丙酮酸激酶(Pyk),制备相应的免抗血清。从抗血清中纯化IgG并水解成F(ab')_2,进而还原成Fab',后者与辣根过氧物酶(HRP)交联成Fab'-HRP,再利用这两种复合物建立了各自的高灵敏夹心ELISA来免疫定量L及M_2型(与鼠M_1-Pyk有交叉免疫)Pyk,结果发现:正常人肝中95％的Pyk为L型,M_2-Pyk仅占5％,而肝细胞癌中L-Pyk降至正常肝的3.5％,M_2-Pyk却增至正常的14.6倍,以致M_2-Pyk占总量的95.5％。免疫组化研究进一步证实定量的结果,正常人肝L-Pyk染色呈强阳性。M_2-Pyk染色较弱,而肝细胞癌恰好相反,L-Pyk染色明显减退,而M_2-Pyk不仅癌组织染色很深,癌周组织也明显深于正常,而且愈近癌巢,染色愈深。测定血浆Pyk,发现正常人L-Pyk占89.9％,M-Pyk(以M_2计)仅占10.1％。肝细胞癌患者血浆L-Pyk略见降低,为正常值的79.6％,p值在0.02～0.05之间,但血浆M型Pyk明显升高至正常的4.59倍,占Pyk总量的39.6％,并证明主要来源于肝癌组织中的M_2,故M_2-Pyk有可能成为肝细胞癌诊断的新指标。     

Deterministic versus Individualistic community structure: a test from invasion by Nothofagus menziesii in southern New Zealand

Abstract. The Clements' Deterministic model of plant communities implies that a change in one species, especially a change in the physiognomic dominant, would have profound effects on the remainder of the community. Gleason's Individualistic model suggests there would be little effect. These alternative models are tested by examining the forest composition on both sides of a boundary along which the tree Nothofagus menziesii is slowly invading. Classification of forest composition, excluding N. menziesii, gave little evidence of an effect of N. menziesii on the lower strata: understorey vegetation types were distributed across the boundary. Some differences were found at individual sites, but these were often inconsistent between sites. Ordination, also excluding N. menziesii, similarly showed that none of the first three understorey axes reflected any effect of N. menziesii. The fourth axis was correlated with the presence of N. menziesii, but only when canopy trees were included. It is concluded that specific composition of the canopy in these montane New Zealand evergreen forests has little effect on the understorey, supporting Gleason's Individualistic concept of the community.     

The biallelica mating type locus ofUstilago maydis: remnants of an additional pheromone gene indicate evolution from a multiallelic ancestor

Thea mating type locus ofUstilago maydis contains the structural genes for a pheromone-based cell recognition system that governs fusion of haploid cells. The locus exists in two alleles, termeda1 anda2. We have completed the analysis of the nucleotide sequences unique toa1 anda2. Within these dissimilar regions we find two short patches of DNA sequence similarity. Interestingly, one of these segments corresponds to the transcribed region of thea1 pheromone precursor. As a result of multiple nucleotide exchanges this sequence does not code for a functional product. The existence of a second pheromone gene in thea2 allele suggests that the present locus had a multiallelic ancestor. In addition, we describe the presence of two additional genes in thea2 allele. We have investigated the role of these genes during mating and pathogenic development and speculate that they might affect mitochondrial inheritance.     

The vegetation of the Puna Belt at laguna de Pozuelos Biosphere Reserve in northwest Argentina

The presence of species and their cover values together with soil and topographic characteristics were recorded at 40 sites along an altitudinal gradient of the Puna Belt at Pozuelos, in the High Andes in northwest Argentina. Classification and ordination of plant assemblages showed that soil and topography were the best predictors of the variation in species distribution. The different plant assemblages which occur along the topographic gradient follow the geographic variation in rainfall in this Andean region. Perennial bushes and grasses colonize the plains and slopes with fine textured soils which retain their water content. In contrast, deciduous bushes and cactuses grow on steep, rocky, dry slopes. Open woods also occur on east-facing steep, rocky slopes which are sheltered from winds.     

Does the energy equivalence rule apply to intertidal macrobenthic communities?

Energy equivalence assumes equal contribution of large and small species to production and energy flow in communities. As in a double logarithmic plot, physiological rates decline with body weight by –0.25, log biomass should increase by 0.25 and log abundance decline by –0.75 with log species weight, when this concept is valid. This was tested with annual data sets of the macrobenthos of 4 intertidal sites in the German Wadden Sea (Königshafen) and 3 sites in a south Portuguese lagoon (Ria Formosa). Only abundance data from two of these sites displayed significantly negative slopes with mean body size of the species. Biomass and secondary production data were significantly positively correlated with mean body size for all Ria Formosa sites and also for the biomass of a mussel bed in Königshafen. However, high variation in body size of the individuals of a species limits interpretation of these plots.It is preferable to test this concept by body weight classes regardless of its species composition. At Königshafen, biomass and production displayed two distinct peaks. One peak at small body size was caused by browsing species. The other peak at larger body size was caused by animals which potentially extract their food from the water column. This bimodality was only vaguely reflected at one station in the Ria Formosa, possibly because of a dominance of detritus feeding species. In a normalized form (log biomass or production / width of size classvs. log size class), these spectra imply a dominance of small individuals in biomass and production at all sites (except for a mussel bank at Königshafen). This is interpreted as a consequence of permanent disturbances.     

一个恢复力受单基因控制的水稻CMS育性回复突变体

利用￣（６０）Ｃｏ－γ射线对具有印尼水田谷细胞质的籼稻细胞质雄性不育系Ⅱ－３２Ａ干种子进行诱变处理,获得了一育性回复突变体Ｔ２４。育性基因未纯合的突变体分离出可育株和完全不育株,比例为３∶１;其与Ⅱ－３２Ａ和珍汕９７Ａ测交,Ｆ１代分离出１∶１的可育株和不育株。育性稳定株系与Ⅱ－３２Ａ和珍汕９７Ａ杂交,Ｆ２分离成３∶１的可育株和不育株。表明其育性回复是由一对基因显性突变所致。这一突变体对不育系的育性恢复机制不同于明恢６３、２０９６４等恢复系,后者表现为两对显性恢复基因作用。未观察到Ｔ２４与亲本Ⅱ－３２Ａ除育性以外的其他性状的差异,因而两者构成育性恢复基因的近等基因系。本文还对不育系育性回复类型和Ｔ２４的理论意义与育种价值进行了讨论。     

An improved method for the isolation of type II and clara cells from mice

Identifying the causal events and temporal aspects of lung cancer development requires the ability to isolate target and nontarget cells for comparative analyses. Current methodology can either isolate only one pure specific cell population from a lung or multiple cell types at lower purity. Previous studies in our laboratory have identified the alveolar type II cell as the progenitor cell for tumor development in the A/J mouse. The purpose of this study was to develop new protocols for the isolation and culture of type II and Clara cells from the mouse lung. Both type II and Clara cells were obtained in high purity using a sequential centrifugal elutriation protocol. In the first elutriation, cell fractions were collected using a Standard chamber. The type II and Clara cell fractions were then elutriated separately (two different separations) using a Sanderson chamber. The final purity of the type II and Clara cell preparations was 73% and 76%, respectively. Colonies of 4 to 20 Clara cells exhibiting epithelial morphology were evident 1 wk after plating in low serum medium. The growth of type II cells required the addition of bronchioalveolar lavage fluid and acidic fibroblast growth factor to the medium. The isolation of viable mouse type II and Clara cells in high purity should facilitate the identification of cell-specific changes in gene expressions or in enzymatic pathways following in vivo or in vitro exposure to environmental carcinogens.     

The impact of type I diabetes on rat liver γ-glutamyltranspeptidase

The impact of type 1 diabetes mellitus on liver -glutamyltranspeptidase, a premalignant marker, was studied. Diabetes was induced in male Sprague Dawley and Fischer 344 rats by administration of Streptozotocin, which produced a stable and moderately severe diabetic state. In liver homogenates, -glutamyltranspeptidase was increased over control levels: 1.2, 8.1 and 13,2 fold in Strague-Dawley rats; 4.8, 58.4 and 84.7 fold in Fischer 344 rats; at 1, 3 and 6 weeks following Streptozotocin treatment. In plasma membranes isolated from the livers of Fischer 344 rats, -glutamyltranspeptidase was increased over control levels: 5.6, 75 and 127 fold at weeks 1, 3 and 6 following Streptozotocin treatment. The relative specific activity of 5-nuleohdase was found to be similar: 9–14, indicating comparable degrees of plasma membrane purity. Plasma glutamate-pyruvate transaminase levels were minimally and similarly affected at all time points indicating lack of association of increasing -glutamyltranspeptidase activity with overt liver damage. Thyroid hormone replacement, with both T₃ (0.6 g/Kg) once a day and T₄ (6.0 g/kg) twice a day for three days elicited a further 30% increment in enzyme activity. Insulin replacement (20–40 units/200 g body weight) twice a day for five days reduced enzyme activity 51% at week 6. This was associated with an increase in -glutamyltranspeptidase in the plasma from 14 fold over control levels in the diabetic state at week 6 to 53 fold ever control levels after insulin replacement at week 6. It is proposed that the diabetes-induced increase in -glutamyltranspeptidase is reduced by an insulin-directed shedding of the enzyme into the plasma.     

Classification of cultivated rices into indica and japonica types by the isozyme,RFLP and two milled-rice methods

Four methods for classifying cultivated rices (Oryza sativa L.) (including IR varieties) into indica and japonica types — waxy gene product in endosperm starch, glutelin ₃ molecular weight in milled rice, RFLP polymorphism at the Wx locus and Glaszmann's isozyme method — were compared. On the basis of the two endosperm traits and the RFLP method Glaszmann's group 1 (indica) was classified as mainly indica and intermediate groups 2, 3 and 4 as exclusively indica. However, the endosperm traits classified Glaszmann's group 5 as mainly indica, while the RFLP method classified it as japonica. The RFLP waxy gene probe was closest to the isozyme method in classifying group 6 as japonicas; the waxy gene product gave mainly indica reaction even in group 6, and the glutelin ₃ method was intermediate. All IR rices were classified as being indica on the basis of Wx gene product and by Glaszmann's method, but a few were classified as japonica by the glutelin ₃ method and by the RFLP waxy gene probe.