Hepatoprotective and TNF-α inhibitory activity of Zosima absinthifolia extracts and coumarins

Zosima absinthifolia (ZA) extracts and the coumarins (+)-columbianadin and (-)-deltoin were evaluated for their potential hepatoprotective and antiinflamatory effects in a CCl(4)-induced hepatotoxicity assay in rats and by the inhibition of TNF-α production on LPS-stimulated THP-1 macrophages, respectively. Both the ZA extracts and the coumarins showed hepatoprotective activity confirmed by monitoring the ALT/AST levels and by histopathological examination. The antiinflamatory activity, proved by the inhibition of TNF-α production, was found to be higher for the n-hexane root extract than for coumarins, suggesting synergic potential of the extract. The concentration of (-)-deltoin and (+)-columbianadin in extracts was determined by HPLC analysis.     

Chemical composition and free radicals restraining activity of extracts from three Manglietia species leaves

The extracts from leaves of Manglietia insignis （Wall） Blume, Manglietia chingii Dandy and Manglietia yuyuanensis Law were prepared by organic solvent extraction and their components were analyzed by GC/MS and quantified. Meanwhile, the free radicals restraining activities were detected. The 21 compounds in M. insignis, 36 compounds in M. chigii and 20 compounds in M. yuyuanensis were identified. There were 11 common components in the extracts from three Manglietia species, and 12 components in two Manglietia species. The results of relative contents of every component in three extracts showed that the main constituents of M. insignis were terpenoids and alkene, amounting to 38.93%, followed by alkane （28.18%）, the nitrogen containing compounds （15.73%） and aromatic compounds （7.23 %）. The main constituents of leaf extract from M. chingii were the terpenoids and alkene, carboxylic acid, alkane and aromatic compounds, amounting to 30.22%, 14.17%, 13.87% and 13.29%, respectively, The main constituents of M. yuyuanensis were alcohol compounds, the terpenoids and alkene, and aromatic compounds, amounting to 28.00%, 25.38% and 18.00% respectively. The results showed that the three extracts had strong function of restraining oxygen free radicals. The ultra oxygen anions activity was restrained at the highest level, when the three extracts were diluted by hundred-fold, whereas the restraining capacity of hydroxyl free radicals reached maximum, when the three extracts were diluted by twenty-fold. The above results provide scientific evidences for further approaching the ecological healthy function of three MangUetia species     

DNA Extraction from Eriocaulon Plants and Construction of RAPD System

There have been many arguments on the classification of Eriocaulon Linn. by morphology so far, and little is known about the use of molecular marker for genetic for genetic diversity of Eriocaulon plants. To apply the technique of molecular marker to the research of genetic diversity of Eriocaulon plants, the study of the extraction method of DNA from the Eriocaulon plants and the RAPD system are essential for researchers. In this paper, the extraction of genome DNA from the silica-gel-dried leaves of several species of Eriocaulon distributed in China was studied, and the best RAPD analysis technique condition of Eriocaulon plants was analyzed.     

Do tannins in leaves of trees and shrubs from African and Himalayan regions differ in level and activity?

Mature leaves of trees and shrubs from sub-humid tropical regions of Benin (Acioa barteri, Cassia sieberiana, Dialium guineense, Dichrostachys cinerea, Guiera senegalensis, Milletia thonningii, Piliostigma reticulatum) and arid and semiarid regions of Zimbabwe and Niger (Acacia holosericea, A. nilotica, Dichrostachys cinerea, Securidaca longepedunculata, Parinari cuvetelio, Ziziphus mucronata) in Africa, and from sub-tropical region in foot-hills of North-West Humid Himalayan range (Albizia stipulata, Bauhenia variegata, Cedrela toona, Celtis australis, Dendrocalamus hamiltonii, Grewia optiva, Leucocephala leucocephala, Morus alba, Papulus ciliata, Quercus incana, Q. semecarpifolia, Q. glauca, Q. serrata, Q. ilex, Robinia pseudoacacia, Salix tetrasperma) were analysed for crude protein, total phenols (TP), protein precipitation capacity (PPC) and operational activity of tannins (values are as mean ± SE). There was no significant difference in the crude protein values of forages obtained from the Himalayan and African region (15.2 ± 1.16 and 14.1 ± 1.19%, respectively), however the levels of TP and biological value of tannins as PPC were significantly higher for the African forages (TP 15.7 ± 4.27 vs 6.0 ± 1.0%; PPC 327.2 ± 113.6 vs 56 ± 15.9 mg bovine serum albumin precipitated/g). The operational activity of tannins expressed as mg protein precipitated per unit of phenols was also significantly higher in forages from the African regions (1.97 ± 0.47 vs 0.66 ± 0.17). For a small set of leaves from arid and semiarid zones of Middle East (Syria, A. cyanophylla; Israel, A. saligna) and India (Eugenia jambolana, Eucalyptus punctata, Prosopis cineraria and Shorea robusta) TP, PPC and tannin activity were closer to those for the African forages.This revised version was published online in November 2005 with corrections to the Cover Date.     

Caffeoylquinic acid derivatives isolated from the aerial parts of Gynura divaricata and their yeast α-glucosidase and PTP1B inhibitory activity

The phytochemical investigation of natural products of Gynura divaricata led to the isolation of eleven caffeoylquinic acid derivatives. They were characterized by spectrometric methods as 5-O-caffeoylquinic acid (1), 5-O-p-coumaroylquinic acid (2), 5-O-feruloylquinic acid (3), methyl 5-O-caffeoylquinate (4), 3,4-dicaffeoylquinic acid (5), 3,5-dicaffeoylquinic acid (6), 4,5-dicaffeoylquinic acid (7), methyl 3,4-dicaffeoylquinate (8), methyl 3,5-dicaffeoylquinate (9), methyl 4,5-dicaffeoylquinate (10) and ethyl 4,5-dicaffeoylquinate (11). The individual compounds were screened for the inhibition of yeast α-glucosidase and Protein Tyrosine Phosphatase 1B (PTP1B) using in vitro assays. Among the isolated compounds, 3,4-dicaffeoylquinic acid (5), 4,5-dicaffeoylquinic acid (7), methyl 3,4-dicaffeoylquinate (8) and methyl 4,5-dicaffeoylquinate (10) exhibited significant inhibitory activities against α-glucosidase. In addition, 5-O-p-coumaroylquinic acid (2), 3,5-dicaffeoylquinic acid (6) and 4,5-dicaffeoylquinic acid (7) had considerable inhibitory effect against PTP1B. Based on these findings, the caffeoylquinic acid derivatives were deduced to be potentially responsible for the anti-diabetic activity of G. divaricata. The preliminary structure–activity relationship study suggests that the number and positioning of caffeoyl groups in the quinic acid derivatives are important for both α-glucosidase and PTP1B inhibitory potency. Moreover, the corresponding methyl esters of some dicaffeoylquinic acids have enhanced inhibitory activity against yeast α-glucosidase.     

Comparisons of extraction and purification methods of soil microorganism DNA from rhizosphere soil

Microorganism DNA of rhizosphere soil from Pinus koraiensis and Pinus sylvestriformis were extracted by proteinase K based on SDS method, CTAB method, PVP （polyvinylpolypyrrolidone） method, and freezing and thawing method and the crude DNA from rhizosphere soil were purified by dialysis method, silver beads absorption method, and squeezing DNA gel method. The results of different extracting and purifying methods were compared and evaluated. Results indicated that the best method of extraction for microorganism DNA in rhizosphere soil was proteinse K based on SDS method with high salt concentration of 1.0% （w/v） NaCl, which could effectively eliminate humic acids and other impurities. The dialysis method was suitable to purify DNA from rhizosphere soil because of effectively removing brown matters and humic acids and the purified products were suited to PCR amplification. Squeezing DNA gel method was also a good purification method with the advantage of inexpensive in cost and efficient in use.     

Bioassay-guided isolation of urease and α-chymotrypsin inhibitory constituents from the stems of Lawsonia alba Lam. (Henna)

Seven constituents were isolated from the stems of Lawsonia alba Lam., following an activity-guided isolation, which include two new constituents, namely lawsorosemarinol (1) and lawsofructose (2), one known compound 2-(β-d-glucopyranosyloxy)-1, 4-naphthoquinone (3) and four compounds, 4-hydroxy coumarine (4), 3-(4-hyroxyphenyl)-triacontyl-(Z)-propenoate (5), 3-(4-hydroxy-3-methoxyphenyl)-triacontyl-(Z)-propenoate (6) and 7-hydroxy-4-methyl coumarin (7) first time isolated from Lawsonia alba. Their structure elucidation was based on spectroscopic data analyses. Compounds 3 and 7 showed a moderate inhibition of urease activity, while rest of them showed less than 50% inhibition. These compounds did not show any significant inhibition against α-chymotrypsin.     

Effect of epoxides and α-methylene-γ-lactone skeleton of sesquiterpenes from yacon (Smallanthus sonchifolius) leaves on caspase-dependent apoptosis and NF-κB inhibition in human cercival cancer cells

The present study investigated the cytotoxicity of enhydrin (1), uvedalin (2) and sonchifolin (3) in cervical cancer cells. We have found that SLs 1–3 in doses in range of 0.22–10 μM inhibited cell proliferation and induced apoptosis in both a dose- and time-dependent fashion. A significant cell death induction was supported by morphological studies. The apoptotic effect is associated with caspase-3/7 activation and NF-кB inhibition. Interestingly, enhydrin possessing two epoxide units was found to be the most cytotoxic compound. Therefore it can be assumed that number of epoxides and existence of α-methylene-γ-lactone moiety are essential for the acceleration of apoptosis.     

A hybrid swarm population of Pinus densiflora × P. sylvestris inferred from sequence analysis of chloroplast DNA and morphological characters

To confirm a hybrid swarm population of Pinus densiflora × P. sylvestris in Jilin, China, we used needles and seeds from P. densiflora, P. sylvestris, and P. densiflora × P. sylvestris collected from natural stands or experimental stations to study whether shoot apex morphology of 4- year old seedlings can be correlated with the sequence of a chloroplast DNA simple sequence repeat marker (cpDNA SSRs). Total genomic DNA was extracted and subjected to sequence analysis of the pine cpDNA SSR marker Pt15169. Results show that morphological characters from 4-year old seedlings did not correlate with sequence variants of this marker. Marker haplotypes from all P. sylvestris trees had a CTAT element that was absent from all sampled P. densiflora trees. However, both haplotype classes involving this insertion/deletion element were found in a P. densiflora × P. sylvestris population and its seedling progeny. It was concluded that the P. densiflora × P. sylvestris accessions sampled from Jilin, China resulted from bi-directional crosses, as evidenced by both species’ cpDNA haplotypes within the hybrid swarm population.     

Arabinogalactan protein from Jatropha curcas L. seeds as TGFβ1-mediated inductor of keratinocyte in vitro differentiation and stimulation of GM-CSF,HGF, KGF and in organotypic skin equivalents

Arabinogalactan protein JC from Jatropha curcas seed endosperm (mean molecular weight 140 kDa) was isolated by cold water extraction and characterized concerning sugar and amino acid composition. At 10 and 100 µg/mL JC stimulated mitochondrial activity (MTT test) of human skin cells (HaCaT keratinocytes, fibroblasts) and the ATP status of primary keratinocytes. JC did not influence the cellular proliferation, while primary keratinocytes were triggered into differentiation status. Investigations on a potential mode of action of JC were performed on complex organotypic skin equivalents. JC induced the production of HGF, KGF and TGFβ, with TGFβ being the main inductor for the differentiation-inducing effect of JC. Also the expression of GM-CSF was stimulated strongly by JC.     

Molecular Cloning of a Cellulose Synthase Gene PtoCesA1 from Populus tomentosa and Its Genetic Transformation in Tobacco

A 3 125 bp cellulose synthase gene, PtoCesA1, which has a 98% identity to PtrCesA1 from Populus tremuloides, was cloned from cDNA prepared from secondary xylem of P tomentosa. Four anti-expression vectors with different fragments of PtoCesAl, named as pBIPF, pBICC1, pBIPR and pBIBR, were constructed. Some traits of transformed tobacco of pBICC1, pBIPR and pBIBR differed from wild types, such as small leaves, ＂dwarf＂ phenotype and thinner xylem and fiber cell walls than wild plants consistent with a loss of cellulose. It indicated that the growth of transgenic tobacco was restrained by the expression of anti-PtoCesA1. Transgenic tobacco was obtained and the contents of cellulose and lignin were analyzed as well as the width and length of fiber cells, and xylem thickness for both transgenic and control plants. Transformed tobacco showed a different phenotype from control plants and it implied that PtoCesA1 was essential for the cellulose biosynthesis in poplar stems.     

In vitro anther culture and Agrobacterium-mediated transformation of the AP1 gene from Salix integra Linn. in haploid poplar(Populus simonii × P. nigra)

A reliable,efficient anther culture system,the dominant technique for generating haploid plants in breeding programs,that can be used for generating transgenic poplar plants has been needed.In the present study,therefore,an anther culture system was developed using isolated mid-and late-uninucleate anthers of poplar(Populus simonii x P.nigra).From a combination of SSR and ploidy analyses,six double haploid and two haploid lines were characterized from 86 plants grown from 16 regenerated anther cultured lines.After 48 months of development,two plant lines from the regenerated plants maintained their haploid level in vitro for over 2 years.A number of haploid plants from the different lines weretransferred to soil.The leaves of these transplants were then used as explants for transformation with the APETALA1(AP1) gene using Agrobacterium tumefaciens.Overexpression of AP1 in haploid poplar induced early flowering with obvious petals when ectopically expressed.To our knowledge,this is the first report on changes in flowering time in AP1-trangenic poplar,which is important for elucidating the regulatory mechanism of tree flower development.