扁蓄总黄酮提取方法的比较

The factors which effected the extraction of total flavones from Polygonum aviculare L.var.aviculore were discuseed with respects to solvent,concentration and the extracting temperature.The rutin was used as a standard compound,the total flavones extracted by the different extracting procedure were quantitated by measurement of absorbance at 510 nm.The results show that 80℃ and 35% alcohol are optimal and the extracting rate of the total flavones in as high as 4.349%.     

Inflorescence development in the Vitis–Ampelocissus clade of Vitaceae: the unusual lamellate inflorescence of Pterisanthes

Pterisanthes (Vitaceae) is a genus of c. 20 species of scandent climbers endemic to Southeast Asia with unusual lamellate inflorescences. Molecular phylogenetic analysis supports its relationship in the well‐supported Vitis–Ampelocissus–Nothocissus–Pterisanthes clade (i.e. the Ampelocissus–Vitis clade). Shoot tips and floral buds were collected from wild and greenhouse‐grown P. eriopoda at different developmental stages and were examined using epi‐illumination, light and scanning electron microscopy. Inflorescence and floral ontogeny was studied to discover how the lamellate inflorescence evolved and to make morphological comparisons to infer relationships with closely related members of Vitaceae. The second‐order branches in P. eriopoda are racemose and develop helically around the inflorescence axis in a similar fashion to Vitis and Ampelocissus. Inflorescence branching is restricted to the second order in P. eriopoda, whereas in Vitis and most Ampelocissus species subsequent branching orders culminate in the typical vitaceous determinate dichasium. In P. eriopoda subsequent lateral growth of the second‐order branches combined with the inhibition of peduncle or pedicel formation and loss of dichasial branching results in the unique lamellae in Pterisanthes, on which the floral primordia arise directly in a helical pattern. Floral development in P. eriopoda is the same as in other genera of Vitaceae examined to date with initiation of floral whorls centripetally, the calyx ring developing first and calyx lobes fused to cover the petals and stamen primordia. Given the recent phylogenetic results that placed Pterisanthes firmly within Ampelocissus, the most likely scenario is that the Pterisanthes inflorescence is derived from the thyrse of an Ampelocissus‐like ancestor and that the thyrse is a morphological synapomorphy of the Ampelocissus–Vitis clade. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 725–741.     

JINKENITES——A STRANGE FOSSIL FROM EARLIEST CAMBRIAN IN W. HUBEI

A strange fossil described and illustrated in the present paper was collected by the writer in 1978 from the Huangshandong Member of the Lower Cambrian Tongying Formation in western Hubei. The material was preserved in greyish brown phosphoritic dolomites in association with elements of the Yangtze Micromolluscan Fauna, such as Tchangsichiton notabilus Yu and Sinuconus clypeus, Yu of Class Polyplacophora; Yangtzemerisma rarum Yu and Y.? cancellatum Yu of Class Merismoconchia; Yangtzeconus priscus Yu, Huangshandongoconus pileus Yu, Obtusoconus paucicostatus Yu, O. rostriptutea (Qian) and Spatuloconus rudis Yu of Class Monoplacophora; Bemella simplex Yu, Latouchlla of. memmorabilis Missarzhevsky, L. sanxiaesis Yu and L. lauta Yu of Class uncertain; Archaeospira ornata Yu, A. imbricata Yu and Cambrospira sinensis Yu of Class Gastropoda; Heraultipegma yunnanense He et Yang of Class Rostroconcbia: hyolithids and some other uncertain skeletal fossils. Although the systematic position of this peculiar fossil is unknown at present, it is inferred that this genus may be one of the typical primitive animal groups based on the characters of the shell and the spe cial spines. The shell is bilaterally symmetrical, elliptical in apical view; the dorsal side is roundly convex with three different forms of dorsal spines and several pairs of marginal spines on the anterior margin. Judged from the general morphological characters of the bilaterally symmetrical shell, these lower animals, generally speaking, have adapted themselves readily to different circumstances, and reduced resistance to any directional movement necessary for varied physiological activities. As to the function of the shell’s characteristics, the narrowly rounded side may serve as the anterior. Such a model of the body helps these animals go on with their benthonic creeping or swimming life. On the other hand, the dorsal spines are different from each other in shape, size, mode of arrangement and in number; especially, the end of the hook-shaped spines is generally curved outward, while the geniculate spines are oblique outward, and curved posteriorly all of a sudden. These structures of mechanism also provide us with the evidence that the narrowly rounded side serves as the anterior, while the opposite end as the posterior.It is of particular interest that this genus is somewhat similar to Phylum Mollusca, especially to the tryblidiids of Class Monoplacophora, in some morphological characters, such as: 1) the bilateral symmetry of the shell, 2) the outline elliptical in apical view and cap shaped in lateral view and 3) the dorsal side generally roundly convex. After careful study, it has been found that since this genus is strikingly different from monoplacopharans, may be it belongs to another interesting group of skeletal fossils. The reasons for this are: 1) in the former, there are many pairs of marginal spines on the anterior margin, while in monoplacophorans, no marginal spines can be seen; and 2) in this genus, the sculptures are different from each other in different areas, not only in shape and number, but also in the mode of arrangement, whereas in monoplacophorans, they are identical with each other in the same type and shape. Judged from the different forms of dorsal sculptures, this genus is comparatively similar to such uncertain shelly fossils as Lepidites Zhong, 1977 emend. Yu, 1987 and Xiadongoconus Yu, 1979 from the same horizon in the Yangtze Region of China, but differs from the latter two genera in the presence of marginal spines on the anterior margin. In addition, this new genus also differs from brachiopods and other skeletal fossils in the peculiar type of the shell. According to the above comparison, this genus is entirely different from all the classes of Phylum Mollusca, possibly representing another interesting group of skeletal fossils. It is impossible to determine the exact taxonomic position of this genus until more materials are available.     

Mutational analysis of the SARS virus Nsp15 endoribonuclease: identification of residues affecting hexamer formation

The severe acute respiratory syndrome (SARS) coronavirus virus non-structural protein 15 is a Mn2+-dependent endoribonuclease with specificity for cleavage at uridylate residues. To better understand structural and functional characteristics of Nsp15, 22 mutant versions of Nsp15 were produced in Escherichia coli as His-tagged proteins and purified by metal-affinity and ion-exchange chromatography. Nineteen of the mutants were soluble and were analyzed for enzymatic activity. Six mutants, including four at the putative active site, were significantly reduced in endoribonuclease activity. Two of the inactive mutants had unusual secondary structures compared to the wild-type protein, as measured by circular dichroism spectroscopy. Gel-filtration analysis, velocity sedimentation ultracentrifugation, and native gradient pore electrophoresis all showed that the wild-type protein exists in an equilibrium between hexamers and monomers in solution, with hexamers dominating at micromolar protein concentration, while native gradient pore electrophoresis also revealed the presence of trimers. A mutant in the N terminus of Nsp15 was impaired in hexamer formation and had low endoribonuclease activity, suggesting that oligomerization is required for endoribonuclease activity. This idea was supported by titration experiments showing that enzyme activity was strongly concentration-dependent, indicating that oligomeric Nsp15 is the active form. Three-dimensional reconstruction of negatively stained single particles of Nsp15 viewed by transmission electron microscopic analysis suggested that the six subunits were arranged as a dimer of trimers with a number of cavities or channels that may constitute RNA binding sites.     

Identification of a Giardia krr1 homolog gene and the secondarily anucleolate condition of Giaridia lamblia

Giaridia lamblia was long considered to be one of the most primitive eukaryotes and to lie close to the transition between prokaryotes and eukaryotes, but several supporting features, such as lack of mitochondrion and Golgi, have been challenged recently. It was also reported previously that G. lamblia lacked nucleolus, which is the site of pre-rRNA processing and ribosomal assembling in the other eukaryotic cells. Here, we report the identification of the yeast homolog gene, krr1, in the anucleolate eukaryote, G. lamblia. The krr1 gene, encoding one of the pre-rRNA processing proteins in yeast, is actively transcribed in G. lamblia. The deduced protein sequence of G. lamblia krr1 is highly similar to yeast KRR1p that contains a single-KH domain. Our database searches indicated that krr1 genes actually present in diverse eukaryotes and also seem to present in Archaea. However, only the eukaryotic homologs, including that of G. lamblia, have the single-KH domain, which contains the conserved motif KR(K)R. Fibrillarin, another important pre-rRNA processing protein has also been identified previously in G. lamblia. Moreover, our database search shows that nearly half of the other nucleolus-localized protein genes of eukaryotic cells also have their homologs in Giardia. Therefore, we suggest that a common mechanism of pre-RNA processing may operate in the anucleolate eukaryote G. lamblia and in the other eukaryotes and that like the case of "lack of mitochondrion," "lack of nucleolus" may not be a primitive feature, but a secondarily evolutionary condition of the parasite.     

Molecular analysis of the O-antigen gene cluster of Escherichia coli O86:B7 and characterization of the chain length determinant gene (wzz)

Escherichia coli O86:B7 has long been used as a model bacterial strain to study the generation of natural blood group antibody in humans, and it has been shown to possess high human blood B activity. The O-antigen structure of O86:B7 was solved recently in our laboratory. Comparison with the published structure of O86:H2 showed that both O86 subtypes shared the same O unit, yet each of the O antigens is polymerized from a different terminal sugar in a different glycosidic linkage. To determine the genetic basis for the O-antigen differences between the two O86 strains, we report the complete sequence of O86:B7 O-antigen gene cluster between galF and hisI, each gene was identified based on homology to other genes in the GenBank databases. Comparison of the two O86 O-antigen gene clusters revealed that the encoding regions between galF and gnd are identical, including wzy genes. However, deletion of the two wzy genes revealed that wzy in O86:B7 is responsible for the polymerization of the O antigen, while the deletion of wzy in O86:H2 has no effect on O-antigen biosynthesis. Therefore, we proposed that there must be another functional wzy gene outside the O86:H2 O-antigen gene cluster. Wzz proteins determine the degree of polymerization of the O antigen. When separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the lipopolysaccharide (LPS) of O86:B7 exhibited a modal distribution of LPS bands with relatively short O units attached to lipid A-core, which differs from the LPS pattern of O86:H2. We proved that the wzz genes are responsible for the different LPS patterns found in the two O86 subtypes, and we also showed that the very short type of LPS is responsible for the serum sensitivity of the O86:B7 strain.     

The structure-anticoagulant activity relationships of sulfated lacquer polysaccharide: effect of carboxyl group and position of sulfation

Regiospecific oxidation of the primary hydroxyl groups in lacquer polysaccharide (LPL, Mw 6.85 x 10(4)) and its NaIO4 oxidation derivatives (LPLde) to C-6 carboxy groups was achieved with NaOCl in the presence of Tempo and NaBr. Sulfate groups were incorporated into the oxidated polysaccharides using Py.SO3 complex as a reagent. Reactivity of polysaccharide hydroxyl group was C-6 > C-2 > C-4. Sulfate groups were mainly linked to the second hydroxy at C-2 in the products. The results of APTT assay showed after incorporation of carboxyl groups into lacquer polysaccharides, the intrinsic coagulation pathway was promoted, and all sulfated polysaccharides had very weak anticoagulant activity within the scope of studied DS (0.39-1.11). These indicated that carboxyl groups and sulfate groups had the synergistic action. At the same time, the anticoagulant activity increased very slowly with the DS in the second hydroxy. This indicated that 6-O-SO3- in the side chains took an important role in the anticoagulant activity.     

Profound difference of metabolic pharmacokinetics between pure glycyrrhizin and glycyrrhizin in licorice decoction

To investigate the difference of metabolic pharmacokinetics between pure glycyrrhizin (GZ) and GZ in licorice decoction, six New Zealand White rabbits were orally given pure GZ and licorice decoction containing equivalent content of GZ in a randomized crossover design. HPLC methods were used for the quantitation of GZ and glycyrrhetic acid (GA) in serum. The results indicated that the areas under curves (AUCs) of GZ and GA after administration of licorice decoction were significantly higher than those after pure GZ. This result was contradictory with that obtained in rats. To explore the mechanism of the pharmacokinetic difference, feces of rabbits, rats, pigs and humans were used to investigate the presystemic metabolism of pure GZ and GZ in licorice decoction. The results indicated that pure GZ was hydrolyzed to GA more rapidly and to a greater extent than that in licorice decoction by various feces. In addition, when pure GZ was fermented, the metabolic profiles of GA and 3-dehydroGA in rabbit feces were quite different from other feces. In conclusion, the bioavailabilities of GZ and GA are significantly better from licorice than from pure GZ in rabbits but the presystemic metabolism of pure GZ in rabbit is rather different from that in rat, pig and human.