Li-Ping Xie

Tsinghua University, Beijing, Beijing Shi, China

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Publications (18)41.55 Total impact

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    ABSTRACT: Decitabine is a synthesized cytosine analog that is a potent inhibitor of DNA methylation. There have been a few reports on the in vitro anti-melanoma effect of decitabine or its functional mechanisms. We investigated the anti-proliferation effect of decitabine on the cultured murine melanoma cell line K1735M2. MTT assay showed that decitabine had strong inhibition on melanoma K1735M2 in a time- and dose-dependent manner in vitro. Morphological observation showed that decitabine could induce melanoma K1735M2 cells to produce dendrite-like structures with the increase of decitabine concentration and incubation time. Decitabine could effectively induce K1735M2 cells to differentiate in vitro. Additionally, decitabine could induce a dose-dependent G2/M cell cycle arrest in K1735M2 cells. We provided experimental evidences that the anti-proliferation effect of decitabine on murine K1735M2 melanoma cells was associated predominately with G2/M cell cycle arrest and the induction of differentiation rather than apopotosis.
    No preview · Article · Nov 2011
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    ABSTRACT: The nacreous layer of molluskan shells, which consists of highly oriented aragonitic crystals and an organic matrix (including chitin and proteins), is a product of biomineralization. This paper briefly introduces the recent research advances on nacre biomineralization of shells from bivalves and gastropods, which mainly focus on analysis of the micro- and nano-structure and components of shell nacreous layers, and investigations of the characteristics and functions of matrix proteins from nacre. Matrix proteins not only participate in construction of the organic nacre framework, but also control the nucleation and growth of aragonitic crystals, as well as determine the polymorph specificity of calcium carbonate in nacre. Moreover, the inorganic aragonite phase also plays an active role in organizing nacre microstructure. Based on these studies, several models to illustrate the formation mechanism related to lamellar nacre in bivalves, and columnar nacre in gastropods are introduced.
    No preview · Article · Jan 2011 · Progress in molecular and subcellular biology
  • Yu-Juan ZHOU · Li-Ping XIE · Rong-Qing ZHANG
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    ABSTRACT: The DNA fragment coding the third P region of α1B subunit of N-type voltage-gated calcium channel of Rattus norvegicus (Cav22P for short) was amplified by high fidelity PCR, inserted into vectors pET28b and pGEX-4T-1 respectively, and expressed in Escherichia coli Rosetta. The expressed product HIS-Cav22P mostly deposited in an inclusive body. The inclusive body of HIS-Cav22P dissolved in urea buffer. After dilution and dialysis, the refolded protein HIS-Cav22P was finally purified via Histrap chelating HP column. Ultraviolet spectroscopy results demonstrated that HIS-Cav22P protein can bind to calcium ions reversely. The expressed product GST-Cav22P was purified from the supernatant of cell lysate using Glutathione Sepharose 4B column. However, GST-Cav22P degraded severely, which caused it difficult to purify the protein Cav22P and the GST pull-down assay. All the results suggest that the active recombinant proteins of Cav22P may act as a molecular target for high through screen of non-narcotic analgesic drugs.
    No preview · Article · Dec 2010 · Progress in Biochemistry and Biophysics
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    ABSTRACT: Molluscan engrailed proteins and bone morphogenetic proteins have attracted lots of interests in their potential important roles during embryonic shell compartment boundary formation. Engrailed is also proposed to be one of the important regulators of matrix proteins regional expressions in oyster mantle tissue, therefore determination of the regulation mechanisms on the particular expression patterns of engrailed in mollusks would be of great scientific value. However the study has been bogged down because the whole genomes of the oysters have not been sequenced completely and no oyster cell lines are available at present so that many genes of the regulators and mediators need to be cloned and classic signal pathways assays are hard to be applied. In previous study, the bone morphogenetic protein 2 (BMP2) and the oyster mothers against decapentaplegic 3 homolog (Smad3) have been identified from the pearl oyster Pinctada fucata. It is interesting to investigate the relationships among the oyster engrailed, BMP2 and Smad3 expressions in the oyster. Thereby a partial fragment of an engrailed homolog has been amplified from the genome template of the pearl oyster Pinctada fucata. Alignments with the deduced amino acids sequence show that the DNA-binding homeobox (EH4) domain of the oyster engrailed shares extremely high similarities with other engrailed proteins. Shell notching experiment and semi quantitative polymerase chain reaction results suggest that the change pattern of the oyster engrailed mRNA level is similar to that of the oyster mothers against decapentaplegic 3 homolog (Smad3) in shell repair process. Then the Smad3, the engrailed and the BMP2 mRNA expression levels in primary cultured oyster mantle tissue cells that are treated with dexamethasone (DXM) and hydrogen peroxide (H2O2) respectively have been examined. Real time quantitative polymerase chain reaction results show that the engrailed, the Smad3 and the BMP2 mRNA levels in the oyster mantle cells exhibit significant correlations with each other. All the results provide important clues and basis for further study on developmental and signal transduction mechanisms on oyster biomineralisation.
    No preview · Article · Jul 2010 · Progress in Biochemistry and Biophysics
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    ABSTRACT: We investigated possible mechanism(s) where honokiol induces apoptosis in human hepatocellular carcinoma SMMC-7721 cells. MTT assay showed that honokiol has strong inhibition on SMMC-7721 cells in a dose-dependent manner. SMMC-7721 cells after honokiol treatment display morphological characteristics such as cell shrinkage, detachment from the culture plate, formation of apoptotic bodies, change to a round shape, and marked nuclear condensation and fragmentation after 32258 staining. Cell apoptosis was measured by Annexin-V/PI staining and alternatively, by the subG0/G1 percentage of the cell cycle analysis followed by FACS. An obvious loss of ΔΨ(m) and a quick burst of ROS was detected when honokiol reached 4μg/ml, which was coincident with the high apoptosis percentage in our previous research. Up-regulation of Bax and down-regulation of Bcl-2 were observed, suggesting that honokiol-induced apoptosis was associated with reactive oxygen species (ROS) production and an increase of Bax/Bcl-2 ratios.
    No preview · Article · Jul 2009 · Environmental Toxicology and Pharmacology
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    ABSTRACT: Alkaline phosphatases are ubiquitous enzymes found in most species including the pearl oyster, Pinctada fucata, where it is presumably involved in nacreous biomineralization processes. In the present study, we have purified alkaline phosphatases from the pearl oyster and modified the tryptophan residues using N-bromosuccinimide (NBS). We show that the resulting inactivation of purified alkaline phosphatase by NBS is dependent on modification of only one of five tryptophan residues in the enzyme. Substrate protection experiments showed that the tryptophan residue was not located at the substrate-binding site but was involved in the catalytic activity.
    No preview · Article · Feb 2008 · Biochemistry (Moscow)
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    ABSTRACT: SO3 belongs to the O-superfamily of conotoxins and is known to have analgesic effects in experimental animals. In order to explore the mechanism of its potential pharmacological actions, the effect of SO3 on synchronized spontaneous calcium spikes was examined in cultured hippocampal networks by calcium imaging. Spontaneous oscillations of intracellular concentrations of calcium (Ca(2+)) in the form of waves and spikes are found in cultured hippocampal networks. Exposure to increasing concentrations of SO3 resulted in a progressive decrease in synchronized spontaneous calcium spikes. The higher concentrations (0.1 micromol/L and 1 micromol/L) of SO3 showed the strongest inhibition. The rank order of inhibition was 1 micromol/L > 0.1 micromol/L > 10 micromol/L > 0.01 micromol/L. This action of SO3 in reducing synchronized calcium spikes suggests a possible application for therapeutic treatment of epilepsy.
    No preview · Article · Feb 2008 · Cell Biology and Toxicology
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    ABSTRACT: Alkaline phosphatases are ubiquitous enzymes involved in many important biological processes. Mammalian tissue-nonspecific alkaline phosphatase (TNAP) has long been thought to play an important role in bone mineralization. In this study, we identified a full-length cDNA encoding a potential alkaline phosphatse from pearl oyster Pinctada fucata by RT-PCR and RACE and designated the encoded protein as PFAP. The sequence of PFAP shares an overall similarity of 67% with that of human TNAP. Prediction and analysis of its secondary and tertiary structure revealed that the PFAP contains two mammalian-specific regions, the crown domain, involved in collagen binding, and the calcium binding domain, which hint its potential ability to participate in biomineralization. RT-PCR and in situ hybridization showed that the PFAP mRNA distributes specifically in the hepatic duct of the digestive diverticula. These findings implied its possible role in calcium absorption and transportation. In vivo, PFAP could be specifically released by phosphatidylinositol-specific phospholipase C (PIPLC), suggesting it is glycophosphatidylinositol-anchored to the plasma membrane. Therefore, a human growth hormone-PFAP fusion was constructed to locate the cleavage/attachment site. Immunofluorescent labeling and immunoblotting showed that Asn-477 is the cleavage/attachment site and the 25-residue peptide COOH-terminal to Asn-477 is removed during glycophosphatidylinositol anchoring. This research will hopefully pave the way to illustrate the role PFAP plays in calcium transportation related to pearl biomineralization.
    No preview · Article · Oct 2007 · Marine Biotechnology
  • Yue-bin Ge · Da-wei Chen · Li-ping Xie · Rong-qing Zhang
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    ABSTRACT: A spherical symmetric design-response surface methodology was applied to optimize the preparation of daidzein-loaded chitosan microspheres by the emulsification/chemical cross-linking technique. The influence of polymer concentration, ratio of drug to polymer, and the stirring speed on the encapsulation efficiency, particle size, particle size distribution, and accumulative drug release percent in microspheres were evaluated. Scan electron microscopy of the optimized microspheres showed spherical particles, loading with drug microcrystal uniformly on the surface of and inside the microspheres. In vivo pharmacokinetic characteristics were evaluated after intramuscular injection of the microspheres in rats. The time-resolved fluoroimmunoassay method was used to determine plasma concentrations of daidzein. The data showed that the release of daidzein in the microspheres in vitro and in vivo almost lasted for 35 days. The bioavailability of daidzein in the microspheres by intramuscular injection increased up to 39% in rats, suggesting that the cross-linked chitosan microspheres are a valuable system for the long-term delivery of isoflavones.
    No preview · Article · Jul 2007 · International Journal of Pharmaceutics
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    ABSTRACT: To elucidate the mechanism of nacre biomineralization, the mantle of Pinctada fucata (P. fucata) from the South China Sea was used. Using the mantle cDNA library and the ESTs we have cloned through suppression subtractive hybridization (SSH), ten novel genes including PFMG1 were obtained through nested PCR. Bioinformative results showed that PFMG1 had a high homology (40%) with Onchocerca volvulus calcium-binding protein CBP-1 and had two EF-hand calcium-binding domains from the 81st to the 93rd amino acid and from the 98th to the 133rd amino acid in the deduced amino acid sequence. The results of multitissue RT-PCR and in situ hybridization demonstrated the high expression of PFMG1 in the mantle of P. fucata and confirmed the SSH method. The results of GST-PFMG1 on CaCO3 crystallization showed significant effects on nucleation and precipitation of CaCO3. PFMG1 was cloned into the pcDNA.3.1/myc-HisA vector and was subsequently transfected into MC3T3-E1 cells. RT-PCR revealed upregulation of the marker genes related to cell growth, differentiation, and mineralization, and BMP-2, osterix, and osteopontin were upregulated as a result. This research work suggests that PFMG1 plays an important role in the nacre biomineralization, and the SSH method can pave the way for the bulk cloning and characterization of new genes involved in biomineralization in P. fucata and may accelerate research on the mechanism of pearl formation.
    No preview · Article · Feb 2007 · Biochemistry
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    ABSTRACT: To prepare an inclusion complex of daidzein and hydropropyl-beta-cyclodextrin to enhance the solubility of daidzein. The inclusion complex of daidzein was prepared by the solution stirring method. The binary system of daidzein and HP-beta-CD was confirmed by differential thermal, thermogravimetry analysis, infrared spectroscopy and X-ray diffractometry. The drug content in the inclusion complex was 6. 76% and the solubility was 13.68 mg x mL(-1). The identification results showed that the inclusion complex was formed. The preparation method of the inclusion complex of daidzein and hydropropyl-beta-cyclodextrin is simple and available, with a increased solubility of daidzein.
    No preview · Article · Jan 2007 · Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica
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    ABSTRACT: In this study, we report that the steroid extract 5alpha, 8alpha-epidioxycholest-6-ene-3beta-ol (MME) from Meretrix meretrix has the ability to inhibit growth of hepatoma cells and to induce G1-phase cell cycle arrest in two human hepatoma cell lines, HepG2 and Hep3B. HepG2 cells were more sensitive than Hep3B to MME. The extract markedly up-regulated the expression of p53 and p21WAF1/CIP1 in HepG2, suggesting that MME-induced G1 phase cell cycle arrest in HepG2 might be p53-dependent. Therefore, the up-regulation of p27KIP1and p16INK4A in both cell lines indicates that a p53-independent pathway might be involved in the mechanism of MME inducing cell cycle arrest. In conclusion, MME induces G1 phase cell cycle arrest via both p53-dependent and p53-independent pathways.
    No preview · Article · Mar 2006 · Cancer Letters
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    ABSTRACT: Oridonin, a diterpenoid isolated from Rabdosia rubescens (Hemsl.) Hara, is currently one of the most important traditional Chinese herbal medicines. We investigated the anti-proliferation effect of oridonin on the cultured murine melanoma cell line K1735M2. The growth inhibition activity of oridonin for K1735M2 cells occurred in a time- and dose-dependent manner (IC50 was 7.4+/-0.6 microM). Further studies showed that these inhibition effects were associated with dose-dependent G2/M phase arrest and differentiation induction. Detection of morphological observation showed that oridonin could induce K1735M2 cells to produce dendrite-like structures. The results of the migration indicated that oridonin affected motility of K1735M2 cells in a dose-dependent manner. These results suggested that oridonin is a potential candidate for melanoma cancer therapy.
    Preview · Article · Feb 2006 · Journal of Ethnopharmacology
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    ABSTRACT: Alkaline phosphatases (ALP, EC are ubiquitous enzymes found in most species. ALP from a pearl oyster, Pinctada fucata (PALP), is presumably involved in nacreous biomineralization processes. Here, chemical modification was used to investigate the involvement of basic residues in the catalytic activity of PALP. The Tsou's plot analysis indicated that the inactivation of PALP by 2,4,6-trinitrobenzenesulfonic acid (TNBS) and phenylglyoxal (PG) is dependent upon modification of one essential lysine and one essential arginine residue, respectively. Substrate reaction course analysis showed that the TNBS and PG inactivation of PALP followed pseudo-first-order kinetics and the second-order inactivation constants for the enzyme with or without substrate binding were determined. It was found that binding substrate slowed the PG inactivation whereas had little effect on TNBS inactivation. Protection experiments showed that substrates and competitive inhibitors provided significant protection against PG inactivation, and the modified enzyme lost its ability to bind the specific affinity column. However, the TNBS-induced inactivation could not be prevented in presence of substrates or competitive inhibitors, and the modified enzyme retained the ability to bind the affinity column. In a conclusion, an arginine residue involved in substrate binding and a lysine residue involved in catalysis were present at the active site of PALP. This study will facilitate to illustrate the role ALP plays in pearl formation and the mechanism involved.
    No preview · Article · Aug 2005 · The International Journal of Biochemistry & Cell Biology
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    ABSTRACT: Mushroom tyrosinase (EC is a copper containing oxidase that catalyzes both the hydroxylation of tyrosine into o-diphenols and the oxidation of o-diphenols into o-quinones, and then forms brown or black pigments. In the present study, the effects of some flavonoids on the oxidation of L-3,4-dihydroxyphenylalanine (L-DOPA) have been studied. The results show that flavonoids can lead to reversible inhibition of the enzyme. A kinetic analysis showed that the flavonols are competitive inhibitors, whereas luteolin is an uncompetitive inhibitor. The rank order of inhibition was: quercetin > galangin > morin; fisetin > 3,7,4;-trihydroxyflavone; luteolin > apigenin > chrysin.
    No preview · Article · Apr 2003 · Biochemistry (Moscow)
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    ABSTRACT: Daidzein, a natural isoflavonoid found in Leguminosae, has received increasing attention because of its possible role in the prevention of osteoporosis. In the present investigation, primary osteoblastic cells isolated from newborn Wistar rats were used to investigate the effect of this isoflavonoid on osteoblasts. Daidzein (2-50 microM) increased the viability (P<0.05) of osteoblasts by about 1.4-fold. In addition, daidzein (2-100 microM) increased the alkaline phosphatase activity and osteocalcin synthesis (P<0.05) of osteoblasts by about 1.4- and 2.0-fold, respectively. Alkaline phosphatase and osteocalcin are phenotypic markers for early-stage differentiated osteoblasts and terminally differentiated osteoblasts, respectively. Our results indicated that daidzein stimulated osteoblast differentiation at various stages (from osteoprogenitors to terminally differentiated osteoblasts). We also investigated the effect of daidzein on bone morphogenetic protein (BMP) production in osteoblasts that display the mature osteoblast phenotype. The results indicated that BMP2 synthesis was elevated significantly in response to daidzein (the mRNA increased 5.0-fold, and the protein increased 7.0-fold), suggesting that some of the effects of daidzein on the cell may be mediated by the increased production of BMPs by the osteoblasts. In conclusion, daidzein has a direct stimulatory effect on bone formation in cultured osteoblastic cells in vitro, which may be mediated by increased production of BMPs in osteoblasts.
    No preview · Article · Mar 2003 · Biochemical Pharmacology
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    ABSTRACT: Genistein and daidzein are two major isoflavonoids in dietary soybean that have inhibition effect on the cell growth of different tumor cell lines. We previously reported the anti-tumor activities of genistein and daidzein in human co1on tumor (HCT) cells and their different ability to enhance the activation of murine lymphocytes. In the present study, the effect of genistein and daidzein on the cell growth, cell cycle progression, and differentiation of murine K1735M2 and human WM451 cel1s was investigated. It was found that genistein could inhibit the cell growth of two metastatic melanoma cell lines, murine Kl735M2 and human WM45l in a dose-dependent manner. Flow cytometry showed that genistein could cause arrest of both Kl735M2 and WM45l at G2/M phase, while daidzein increased the cell numbers at S phase, decreased the cell numbers at G1 phase. Detection of melanin and morphological observation showed that genistein can induce Kl735M2 and WM45l to produce dendrite-like structure and produce more melanin by 80%. In contrast, daidzein only retarded the growth of K1735M2 and did not induce differentiation in either K1735M2 or WM451. These results suggest that genistein and daidzein in soybean can inhibit certain malignant phenotype of melanoma via different mechanisms and be potential medical candidates for melanoma cancer therapy.
    No preview · Article · Jul 2002 · The Journal of Nutritional Biochemistry
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    ABSTRACT: An alkaline phosphatase was purified from Pinctada fucata, a kind of pearl oyster, by chromatography on DEAE-32 cellulose, Sephadex G-150 and DEAE A-25. The specific activity of the enzyme was 2040Umg−1. The kinetics characteristics of the enzyme have been studied. The product HPO42− and the product-analog WO43− competitively inhibited the enzyme activity. Positive monovalent cations had no effect on the enzyme activity, while positive bivalent cations had different effects on the enzyme: Mg2+, Ca2+, Co2+ and Mn2+ activated the enzyme while Zn2+, Cu2+ and Cd2+ inhibited the enzyme.
    No preview · Article · Apr 2002 · Journal of Molecular Catalysis B Enzymatic