Simultaneous determination of geniposide, baicalin, cholic acid and hyodeoxycholic acid in rat serum for the pharmacokinetic investigations by high performance liquid chromatography-tandem mass spectrometry

A simple, rapid, and specific analytical method for simultaneous determination of geniposide, baicalin, cholic acid and hyodeoxycholic acid in 50 microL samples of rat serum was developed by high performance liquid chromatography-tandem mass spectrometry. The quantification of the target compounds was determined by multiple reaction monitoring (MRM) mode using electrospray ionization (ESI). The correlation coefficients of the calibration curves were better than 0.997. The intra- and inter-day accuracy, precision, and linear range had been investigated in detail. This method was subsequently applied to pharmacokinetic studies of geniposide, baicalin, cholic acid and hyodeoxycholic acid in rats successfully.     

Molecular imprinted polymer with cloned bacterial protein template enriches authentic target in cell extract

Here we describe a new method for preparing a protein-imprinted polymer with a cloned bacterial protein template, which recognizes/adsorbs authentic target protein present at a relatively low level in cell extract. In this work, cloned pig cyclophilin 18 (pCyP18) was used as a template. The template protein was selectively assembled with memory molecules from their library, which consists of numerous limited length polymer chains with randomly distributed recognition sites and immobilizing sites. These assemblies of protein and memory molecules were adsorbed by porous polymeric beads and immobilized by cross-linking polymerization. After removing the template, binding sites that were complementary to the target protein in size, shape and the position of recognition groups were exposed, and their confirmation was preserved by the cross-linked structure. The synthesized imprinted polymer was used to adsorb authentic pCyP18 from cell extract, and its proportional content was enriched 300 times.     

Analysis of heat-induced disassembly process of three different monomeric forms of the major light-harvesting chlorophyll a/b complex of photosystem II

The temperature-dependent disassembly process of three monomeric isoforms, namely Lhcb1, Lhcb2, and Lhcb3, of the major light-harvesting chlorophyll (Chl) a/b complexes of photosystem II (LHCIIb) were characterized by observing the changes of absorption spectra, circular dichroism (CD), and dissociation processes of the bound pigments to the in vitro reconstituted complexes subjected to high temperatures. Our results suggest that the three isoforms of LHCIIb undergo conformational rearrangements, structural changes, and dissociations of the bound pigments when the ambient temperature increases from 20 to 90°C. The conformation of the complexes changed sensitively to the changing temperatures because the absorption peaks in the Soret region (436 and 471?nm) and the Qy region (650-660 and 680?nm) decreased immediately upon elevating the ambient temperatures. Analyzing temperature-dependent denaturing and pigment dissociation process, we can divide the disassembly process into three stages: The first stage, appeared from 20°C to around 50-60°C, was characterized by the diminishment of the absorption around 650-660 and 680?nm, accompanied by the blue-shift of the peak at 471?nm and disappearance of the absorbance at 436?nm, which is related to changes in the transition energy of the Chl b cluster, and the red-most Chl a cluster in the LHCIIb. The second stage, beginning at about 50-60°C, was signified by the diminishment of the CD signal between (+)483?nm and (-)490?nm, which implied the disturbance of dipole-dipole interaction of pigments, and the onset of the pigment dissociation. The last stage, beginning at about 70-80°C, indicates the complete dissociation of the pigments from the complex. The physiological aspects of the three stages in the denaturing process are also discussed.     

Purification and antioxidant activity of a polysaccharide from bulbs of Fritillaria ussuriensis Maxim

A water-soluble polysaccharide (FUP-1) was obtained from Fritillaria ussuriensis Maxim through warm water extraction, ethanol precipitation, anion-exchange and gel-permeation chromatography. Structural analysis of this compound revealed that the average molecular weight of FUP-1 was 4.1×10(4)Da, and the major monosaccharide component was xylose, followed by glucose and galactose, at low levels. In vitro antioxidant assays showed that FUP-1 exhibited strong hydroxyl, superoxide anion and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activities. During in vivo antioxidant testing, the administration of FUP-1 could largely attenuate the increase in d-gal-induced malondialdehyde (MDA) and renew the activities of superoxide dismutase (SOD) in serums as well as glutathione peroxidase (GSH-Px) and the total antioxidant capacity (T-AOC) in the livers of d-gal-treated mice. Overall, the results of the present study indicate that FUP-1 exerts antioxidant activity not only through its own radical-scavenging activity but also by boosting the enzymatic and nonenzymatic antioxidant defense system of the host.     

The inhibitory effect of a polysaccharide from Codonopsis pilosula on tumor growth and metastasis in vitro

In this study, we prepared an acidic polysaccharide (CPPA) from the roots of Codonopsis pilosula. The effects of CPPA on tumor cell growth, invasion, and migration were examined in vitro. The CPPA not only induced a potent inhibitory effect on the invasion and migration potential of human epithelial ovarian cancer HO-8910 cells in vitro, evaluated by wound healing, transwell and cell adhesion assays, but also had an efficient anti-proliferation effect on tumor cells. Moreover, the CD44 expression on the HO-8910 cells was also attenuated by CPPA treatment. Therefore, our results indicate that CPPA may be a potential candidate compound for the prevention of tumor metastasis, presumably by inhibiting invasion, migration and adhesion of tumor cells, as well as the CD44 expression on the tumor cells.     

Purification and antitumor activity of two acidic polysaccharides from the roots of Polygala tenuifolia

Two acidic polysaccharide fractions (PTPa and PTPb) extracted from the roots of Polygala tenuifolia, were obtained by DEAE-Sephacel anion-exchange, and Sephadex G-100 gel-permeation chromatography. High-performance liquid chromatography (HPLC) identified that PTPa and PTPb was composed of Ara, Glc, Gal, Man and GlcUA in the proportion of 2.4:1.2:0.6:0.4:1.1 and 2.1:1.7:0.5:0.6:1.7, respectively. Their molecular weight was evaluated to be 5.9×10(4) (PTPa) and 2.5×10(4)Da (PTPb) as determined by high performance size exclusion chromatography (HPSEC). Pharmacological studies revealed PTPa and PTPb significantly inhibited the growth of A549 cells in vitro and exhibited significantly higher antitumor activity against solid tumor A549 in vivo than did a blank control. Moreover, treatment with two acidic polysaccharides caused an enhancement of superoxide dismutase (SOD) and catalase (CAT) activities in tumor-bearing mice and a reduction in thiobarbituric acid reactive substances (TBARS) level. Taken together, these results indicated that two acidic polysaccharides from the roots of P. tenuifolia may be useful as potent antitumor agents for the prevention of lung tumorigenesis.     

Pyrazole and pyrimidine phenylacylsulfonamides as dual Bcl-2/Bcl-xL antagonists

5-Butyl-1,4-diphenyl pyrazole and 2-amino-5-chloro pyrimidine acylsulfonamides were developed as potent dual antagonists of Bcl-2 and Bcl-xL. Compounds were optimized for binding to the I88, L92, I95, and F99 pockets normally occupied by pro-apoptotic protein Bim. An X-ray crystal structure confirmed the proposed binding mode. Observation of cytochrome c release from isolated mitochondria in MV-411 cells provides further evidence of target inhibition. Compounds demonstrated submicromolar antiproliferative activity in Bcl-2/Bcl-xL dependent cell lines.     

Genetic analysis of Down syndrome facilitated by mouse chromosome engineering

Human trisomy 21 is the most frequent live-born human aneuploidy and causes a constellation of disease phenotypes classified as Down syndrome, which include heart defects, myeloproliferative disorder, cognitive disabilities and Alzheimer-type neurodegeneration. Because these phenotypes are associated with an extra copy of a human chromosome, the genetic analysis of Down syndrome has been a major challenge. To complement human genetic approaches, mouse models have been generated and analyzed based on evolutionary conservation between the human and mouse genomes. These efforts have been greatly facilitated by Cre/loxP-mediated mouse chromosome engineering, which may result in the establishment of minimal critical genomic regions and eventually new dosage-sensitive genes associated with Down syndrome phenotypes. The success in genetic analysis of Down syndrome will further enhance our understanding of this disorder and lead to better strategies in developing effective therapeutic interventions.     

光系统Ⅱ的结构与功能以及光合膜对环境因素的响应机制

光合膜是地球上捕获、转换和利用太阳能的关键场所,光合膜的活动所提供的能源、粮食及氧气,是人类世界赖以生存的基础。经过35亿年的进化,光合膜已经进化成了一个高度精密的结构,色素分子高密度结合并合理排列,具有高精度的能级耦联网络和高效率的能量传递系统,这使得光合膜成为自然界中能够最高效地吸收和传递太阳能、并能在常温常压下高效地将太阳能转换成化学能和还原势的色素蛋白复合体体系。由于这一特性,光合膜被认为是最有潜力的固定太阳能的新材料,并为研究新型光电转换器件提供了新思路和新理论。因此,长期以来,光合膜的结构-功能关系研究及其功能模拟,特别是执行固定和转化太阳能第一步的光系统Ⅱ,在新能源的利用中吸引了大量的研究力量,取得了突飞猛进的进展。本文总结了近年来关于光系统Ⅱ的结构与功能,以及光合膜对环境的感应和功能调节机制等方面的研究进展。