Sensitive HPLC-APCI-MS method for the determination of cyclovirobuxine D in human plasma

A sensitive high performance liquid chromatography-atmospheric pressure chemical ionisation-mass spectrometry (HPLC-APCI-MS) assay for determination of cyclovirobuxine D (CVB-D) in human plasma using mirtazapine as internal standard (I.S.) was established. After adjustment to a basic pH with sodium hydroxide, plasma was extracted by ethyl acetate and separated by high performance liquid chromatography (HPLC) on a reversed-phase C(18) column with a mobile phase of 30 mM ammonium acetate buffer solution containing 1% formic acid-methanol (48:52, v/v). CVB-D was determined with atmospheric pressure chemical ionisation-mass spectrometry (APCI-MS). HPLC-APCI-MS was performed in the selected-ion monitoring (SIM) mode using target ions at [M+H](+)m/z 403.4 for CVB-D and [M+H](+)m/z 266.2 for I.S. Calibration curves were linear over the range 10.11-4044 pg/ml. The lower limit of quantification was 10.11 pg/ml. The intra- and inter-run variability values were less than 9.5 and 12.4%, respectively. The mean plasma extraction recovery of CVB-D was in the range of 85.3-92.8%. The method was successfully applied to determine the plasma concentrations of CVB-D in Chinese volunteers.     

Proteome analysis of up-regulated proteins in the rat spinal cord induced by transection injury

The inability of the CNS to regenerate in adult mammals propels us to reveal associated proteins involved in the injured CNS. In this paper, either thoracic laminectomy (as sham control) or thoracic spinal cord transection was performed on male adult rats. Five days after surgery, the whole spinal cord tissue was dissected and fractionated into water-soluble (dissolved in Tris buffer) and water-insoluble (dissolved in a solution containing chaotropes and surfactants) portions for 2-DE. Protein identification was performed by MS and further confirmed by Western blot. As a result, over 30 protein spots in the injured spinal cord were shown to be up-regulated no less than 1.5-fold. These identified proteins possibly play various roles during the injury and repair process and may be functionally categorized as several different groups, such as stress-responsive and metabolic changes, lipid and protein degeneration, neural survival and regeneration. In particular, over-expression of 11-zinc finger protein and glypican may be responsible for the inhibition of axonal growth and regeneration. Moreover, three unknown proteins with novel sequences were found to be up-regulated by spinal cord injury. Further characterization of these molecules may help us come closer to understanding the mechanisms that underlie the inability of the adult CNS to regenerate.     

High cell density and high expression of recombinant human ApoA-IMilano in Escherichia coli by twice temperature-shifted induction

The effect of temperature on the formation of recombinant protein, apolipoprotein A-I_Milano was investigated in the present study. The temperature of the initial growth phase was set at 30°C, while temperature variation in induction phase was arranged in three modes. High cell-density culture of Escherichia coli and high expression of recombinant human by twice temperature-shifted induction were carried out. Experimental results showed that ApoA-I_Milano reached 4.8 g/L with the final cell density of OD₆₀₀, 150. It was found that twice temperature-shifted induction could successfully avoid the effect of acetic acid on cell density and the expression of the product. The present study provides a basic procedure for the production of recombinant ApoA-I_Milano.     

类风湿关节炎与白细胞介素-18 的研究进展

类风湿关节炎发病机制不明,许多细胞因子参与了RA的发病机制。近年发现,白细胞介素-18可作用于RA发病的多个参与因素,成为研究RA的新热点。     

Functional differentiation of human brain progenitor cells

Stem cells and progenitor cells derived from the developing human brain have been shown to differentiate into neurons and astrocytes. However, few studies have examined the functional, physiological properties of these differentiated neurons and astrocytes. In this study we have used immunocytochemistry in combination with electrophysiology to examine protein machinery and functional properties of neurons and astrocytes differentiated from human brain progenitor cells (hBPCs).Our results show that serum induces mainly astrocytic phenotype cells that express GFAP and have physiological properties that are typical of astrocytes. hBPCs differentiated with BDNF and PDGF develop mainly into neurons expressing mature neuronal proteins MAP-2, synaptobrevin II and vesicular glutamate transporter I in the process, plus a small population of GFAP-positive radial cells. Based on electrophysiology of BDNF/PDGF-treated cells two classes of cell were identified. Class I cells have functional neuronal properties, including functional voltage-gated Na(+) and K(+) currents, functional AMPA receptors and the ability to generate action potentials. A smaller subpopulation of cells (Class II cells) expresses GFAP and exhibit functional properties of astrocytes, including linear current-voltage relationship and dye-coupling.     

Biomimetic zinc oxide replica with structural color using butterfly (Ideopsis similis) wings as templates

Nano-structured colorful zinc oxide (ZnO) replicas were produced using the wings of the Ideopsis similis butterfly as templates. The ZnO replicas we obtained exhibit iridescence, which was clearly observed under an optical microscope (OM). Field emission scanning electron microscope analysis shows that all the microstructure details are maintained faithfully in the ZnO replica. A computer model was established to simulate the diffraction spectral results, which agreed well with the OM images.     

L-lactic acid production by Lactobacillus casei fermentation with corn steep liquor-supplemented acid-hydrolysate of soybean meal

Batch and fed-batch fermentation studies were performed to evaluate the potential of corn steep liquor (CSL)-supplemented acid-hydrolysate of soybean meal (AHSM) as an alternative to yeast extract (YE) for the production of L-lactic acid by Lactobacillus casei LA-04-1. The CSL-supplemented AHSM gave an outstanding result in supporting L-lactic acid production from glucose. In the exponential fed-batch fermentation, the concentration, yield and productivity of L-lactic acid were 162.5 g/L, 89.7% and 1.69 g/L per h, respectively, which were lower than those with 20 g/L YE (180 g/L, 90.3%, 2.14 g/L per h) after 96 h of fermentation. However, the raw material cost of the nitrogen resource was estimated as only 25% of that using the YE.     

Effect of Hillslope Gradient on Vegetation Recovery on Abandoned Land of Shifting Cultivation in Hainan Island, South China

In the present study, we investigated the effect of hillslope gradient on vegetation recovery on abandoned land of shifting cultivation In Hainan Island, south China, by measuring community composition and structure of 25-year-old secondary forest fallows along a hillslope gradient （up-, middle-, and down-slope position）. A total of 49 733 free-standing woody plant stems higher than 10 cm and belonging to 170 species, 112 genera, and 57 families was found in the three l-hm2 investigation plots. Stem density was highest in the down-slope stand and lowest in the up-slope stand. Species richness and the Shannon-Wiener Index were both highest in the middle-slope stand, and lower In the down-slope and up-slope stands. The recovery forest fallows on different hiUslope positions were all dominated by a few species. The five most abundant species accounted for 70.1%, 58.8%, and 72.9% of total stem densities in stands in the up-, middle-, and down-slope positions, respectively. The five species with the greatest basal areas accounted for 74.5%, 84.3%, and 74.7% of total stem basal area for the up-, middle-, and down-slope positions, respectively. The number of low-density species （stem abundance less than five） Increased from the up-slope position downward. Of the nine local common species among three different functional groups, the short-lived pioneer species dominated the up-slope position, but long-lived pioneer species dominated the middle- and down-slope positions. The climax species of primary tropical lowland rain forest was found in the downslope position. Both the mean diameter at breast height （DBH） and mean height of the trees Increased with decreasing hillslope gradient. The stem density and basal area in different size classes were significantly different in stands In different slope positions. Our results indicated that the rate of secondary succession varies, even over small spatial scales caused by the hlllslope gradient, in early vegetation recovery.     

Bird community patterns in response to the island features of urban woodlots in eastern China

Many studies have demonstrated the changes in the spatial patterns of plant and animal communities with respect to habitat fragmentation. Insular communities tend to exhibit some special patterns in connection with the characteristics of island habitats. In this paper, the relationships between richness, assemblage, and abundance of bird communities with respect to island features were analyzed in 20 urban woodlots in Hangzhou, China. Field investigations of bird communities, using the line transect method, were conducted from January to December, 1997. Each woodlot was surveyed 16 times during the year. Results indicated that bird richness was higher, per unit area, in the smaller woodlots than the larger ones, and overall bird density decreased with the increase in the size of woodlot. However, the evenness of species abundance increased with the area, and small woodlots were usually dominated by higher density species and large woodlots by medium density species. Most species occurring in the small woodlots also occurred in larger woodlots. Also, bird communities among urban woodlots showed a nestedness pattern in assemblage. These patterns implied that the main impacts of woodland habitat fragmentation are: (1) species are constricted and thus species number will increase at a given sample size; (2) as surface area decreases, the proportion of forest edge species as to interior species will increase; (3) community abundance will therefore increase per unit area but most individuals will be from a few dominant species; and (4) overall species diversity will decrease at a habitat level as well as at a region level. These patterns of community in response to the island features were therefore summarized as “island effects in community”. The underlying processes of such observations were also examined in this paper. Woodlot area, edge ratio, isolation, and habitat nestedness were considered as the important factors forming the island effects in community. High heterogeneity between habitats usually contributed most to the maintenance of regional biodiversity, especially in urban woodlots. __________ Translated from Acta Ecologica Sinica, 2005, 25(4): 657–663 [译自: 生态学报, 2005, 25(4): 657–663]