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P Liang

Publications and source records attributed to P Liang.

At least 37 records · Page 2Linked to original sources
PubMed2001

[Effect of mm-LDL on NF-kB activation in endothelial cell].

OBJECTIVE: To investigate the signal transduction pathway of NF-kB activated by minimally modified low density lipoprotein (mm-LDL) in endothelial cells and the effect of NF-kB on platelet derived growth factor b (PDGFb) mRNA expression. METHODS: mm-LDL was prepared through iron oxidation by dialyzing the native LDL against FeSO4 in PBS. Endothelial cells were incubated in a medium containing mm-LDL, TNF, and IL-1 respectively and electrophoretic mobility shift assay (EMSA) was displayed to check on the activation of NF-kB. Luciferase reporter gene was analysed to investigate the effect of nuclear factor inducing kinase (NIK), inhibitor of NF-kB kinase alpha (IKK alpha) and inhibitor of NF-kB kinase beta (IKK beta) on NF-kB activation. In addition, endothelial cells were transfected using PDGFb promoter-luciferase for reporter gene analysis or transfected with mut-NIK for slot blot analysis to study the effect of NF-kB on PDGFb mRNA expression. RESULTS: mm-LDL was able to activate NF-kB in endothelial cells. mut-NIK and mut-IKK beta inhibited luciferase activity induced by mm-LDL. mm-LDL could also enhance luciferase activity controlled by upstream sequence of PDGFb promoter which contains element interacting with NF-kB. Result of slot blot showed inhibition of PDGFb mRNA expression by mut-NIK in the endothelial cells stimulated by mm-LDL. CONCLUSION: mm-LDL may activate NF-kB through NIK-IKK beta pathway and promote PDGFb mRNA expression in endothelial cells.

Cells, Cultured↗
PubMed2001

[Relationship between proteoglycans and apoE on the HepG2 cell surface].

OBJECTIVE: To characterize the associations and functions of apoE with the HepG2 cell surface through the study of the relationship between proteoglycans and apoE on HepG2 cell surface. METHODS: The specific binding of 7C9, a monoclonal antibody against the N-terminal domain of apoE was employed to demonstrate the effects of glycosaminoglycans, heparinase, chondroitinase, xyloside, and chlorate on the apoE of HepG2 cell surface. RESULTS: Growth of cells in beta-D-xyloside decreased cell surface apoE by 45% with a concomitant increase in apoE secretion (4.3-fold), underlining the importance of glycosaminoglycan association of apo E. Incubations with heparinase (3 U/ml) and heparin (1 mg/ml) decreased apoE by 25.0% and 30.5% respectively indicating association through cell surface haparin sulfate proteoglycans. Incubations with chondroitinase ABC (1.5 U/ml) reduced cell surface apoE by 40.0%. Chondroitin sulfate A and chondroitin sulfate B reduced cell surface apoE by 23.6% and 15.3% respectively while incubations with chondrointin sulfate C not effective. Decreasing the levels of cell surface apoE with haparin or chondrointin sulfates A and B increased the subsequent binding of LDL to the HepG2 cell surface. CONCLUSIONS: ApoE associates with the cell surface mainly through chondrointin sulfate proteoglycans and to a lesser extent through heparan sulfate proteoglycans, decreased levels of cell surface apoE increase the binding of LDL to the cell surface.

Apolipoproteins E↗
PubMed2001

[Characterization of macrophage cell (THP-1) surface apoE and it's role in metabolism of cell lipids].

OBJECTIVE: To study the characterization of THP-1 cell surface apoE in order to learn the further role of macrophage in development of atherosclerosis. METHODS: Using monoclonal antibodies (7C9,3H1) against apoE to analyze the effects of AcLDL, heparin, chondroitinase, and Liposyn II on the apoE of THP-1 cell surface. RESULTS: Cholesterol (AcLDL) loaded cell demonstrated reduced binding of 125I-7C9 to the cell surface by 19% at 4 degrees C, but if the THP-1 cell incubated at 37 degrees C for 2 hours after the removal of AcLDL there was a increase (25%) in 7C9 bound to loaded cells when compared to the 4 degrees C controls; prior loading with AcLDL also increased the secretion of apoE to the extracellular medium in a time-dependent manner; heparinase or chondroitinase ABC did not reduce the surface apoE associated with unloaded cells; the apoE on the surface of loaded macrophage was only marginally affected (9.3%) by incubation with heparinase but was efficiently reduced (50%) by incubation with chondroitinase ABC; Liposyn II dramatically increased the binding of apoA1 to the cell surface (1.5-1.8 fold), the Liposyn II mediated increased binding of apoA1 was antagonized by pre-incubation of THP-1 cells with 3H1 indicating that it was an apoE-dependent process. CONCLUSIONS: Cell surface apoE of THP-1 macrophage relates to the process of cholesterol loading of macrophage cells and lipid mediated binding of apoA1 to the cell surface. It may affect the process of atherosclerosis development.

Apolipoprotein A-I↗
PubMed2000

E2F1 and p53 are dispensable, whereas p21(Waf1/Cip1) cooperates with Rb to restrict endoreduplication and apoptosis during skeletal myogenesis.

We describe temporal and genetic analyses of partially rescued Rb mutant fetuses, mgRb:Rb-/-, that survive to birth and reveal specific defects in skeletal muscle differentiation. We show that in the absence of Rb, these fetuses exhibit increased apoptosis, bona fide endoreduplication, and incomplete differentiation throughout terminal myogenesis. These defects were further augmented in composite mutant fetuses, mgRb:Rb-/-:p21-/-, lacking both Rb and the cyclin-dependent kinase inhibitor p21(Waf1/Cip1). Although E2F1 and p53 mediate ectopic DNA synthesis and cell death in several tissues in Rb mutant embryos, both endoreduplication and apoptosis persisted in mgRb:Rb-/-:E2F1-/- and mgRb:Rb-/-:p53-/- compound mutant muscles. Thus, combined inactivation of Rb and p21(Waf1/Cip1) augments endoreduplication and apoptosis, whereas E2F1 and p53 are dispensable during aberrant myogenesis in Rb-deficient fetuses.

Animals↗
PubMed2000

Genome sequence of Halobacterium species NRC-1.

We report the complete sequence of an extreme halophile, Halobacterium sp. NRC-1, harboring a dynamic 2,571,010-bp genome containing 91 insertion sequences representing 12 families and organized into a large chromosome and 2 related minichromosomes. The Halobacterium NRC-1 genome codes for 2,630 predicted proteins, 36% of which are unrelated to any previously reported. Analysis of the genome sequence shows the presence of pathways for uptake and utilization of amino acids, active sodium-proton antiporter and potassium uptake systems, sophisticated photosensory and signal transduction pathways, and DNA replication, transcription, and translation systems resembling more complex eukaryotic organisms. Whole proteome comparisons show the definite archaeal nature of this halophile with additional similarities to the Gram-positive Bacillus subtilis and other bacteria. The ease of culturing Halobacterium and the availability of methods for its genetic manipulation in the laboratory, including construction of gene knockouts and replacements, indicate this halophile can serve as an excellent model system among the archaea.

Biological Evolution↗
PubMed2000

MGSA/GRO-mediated melanocyte transformation involves induction of Ras expression.

The MGSA/GRO protein is endogenously expressed in almost 70% of the melanoma cell lines and tumors, but not in normal melanocytes. We have previously demonstrated that over-expression of human MGSA/GROalpha, beta or gamma in immortalized murine melanocytes (melan-a cells) enables these cells to form tumors in SCID and nude mice. To examine the possibility that the MGSA/GRO effect on melanocyte transformation requires expression of other genes, differential display was performed. One of the mRNA's identified in the screen as overexpressed in MGSA/GRO transformed melan-a clones was the newly described M-Ras or R-Ras3 gene, a member of the Ras gene superfamily. Over-expression of MGSA/GRO upregulates M-Ras expression at both the mRNA and protein levels, and this induction requires an intact glutamine-leucine-arginine (ELR)-motif in the MGSA/GRO protein. Western blot examination of Ras expression revealed that K- and N-Ras proteins are also elevated in MGSA/GRO-expressing melan-a clones, leading to an overall increase in the amount of activated Ras. MGSA/GRO-expressing melan-a clones exhibited enhanced AP-1 activity. The effects of MGSA/GRO on AP-1 activation could be mimicked by over-expression of wild-type M-Ras or a constitutively activated M-Ras mutant in control melan-a cells as monitored by an AP-1-luciferase reporter, while expression of a dominant negative M-Ras blocked AP-1-luciferase activity in MGSA/GRO-transformed melan-a clones. In the in vitro transformation assay, over-expression of M-Ras mimicked the effects of MGSA/GRO by inducing cellular transformation in control melan-a cells, while over-expression of dominant negative M-Ras in MGSA/GROalpha-expressing melan-a-6 cells blocked transformation. These data suggest that MGSA/GRO-mediated transformation requires Ras activation in melanocytes.

Animals↗
PubMed2000

Identification of a novel ligand-receptor pair constitutively activated by ras oncogenes.

The Ras signaling pathway is thought to control the expression of a subset of yet to be defined genes that are crucial for cell growth and differentiation. Here we have identified by differential display a novel oncogenic Ras target, mob-5, encoding a 23-kDa cytokine-like secreted protein. Mob-5 expression could be induced by oncogenic Ha-ras and Ki-ras, but not by normal ras activation. Inhibitors of both Ha-Ras and mitogen-activated protein kinase kinase completely abolished the mob-5 expression in ras transformed cells, with concomitant loss of the transformation phenotype. Using an alkaline phosphatase-tagged Mob-5 as ligand, a putative Mob-5 receptor was identified on the cell surface of oncogenic ras transformed cells. Thus, the Mob-5/Mob-5 receptor may represent a novel putative autocrine loop coordinately activated by ras oncogenes.

Amino Acid Sequence↗
PubMed2000

NF-kappa B is required for H-ras oncogene induced abnormal cell proliferation and tumorigenesis.

Oncogenic mutations in ras lead to constitutive activation of downstream signaling pathways that modulate the activities of transcription factors. In turn, these factors control the expression of a subset of genes responsible for neoplastic cell transformation. Recent studies suggest that transcription factor NF-kappa B contributes to cell transformation by inhibiting the cell death signal activated by oncogenic Ras. In this study, inhibition of NF-kappa B activity by forced expression of a super-repressor form of I kappa B alpha, the major inhibitor of NF-kappa B, markedly decreased the growth rate, saturation density and tumorigenicity of oncogenic H-Ras transformed rat embryo fibroblasts. Such clonally isolated cells overexpressing I kappa B alpha super-repressor not only were viable but also exhibited no sign of spontaneous apoptosis. Inhibition of NF-kappa B in these cells was functionally demonstrated by both the loss of cytokine induced DNA binding activity and a profoundly increased sensitivity to cell death in response to TNF-alpha treatment. In contrast, inhibition of NF-kappa B activity in non-transformed fibroblasts had minimal effect on growth, but rendered the cells resistant to a subsequent transformation by H-ras oncogene. Similar results were also obtained with rat intestinal epithelial cells harboring an inducible ras oncogene. Taken together, these findings suggest that NF-kappa B activity is essential for abnormal cell proliferation and tumorigenicity activated by the ras oncogene and highlight an alternative functional role for NF-kappa B in oncogenic Ras-mediated cell transformation that is distinct from its anti-apoptotic activity. Oncogene (2000) 19, 841 - 849.

Animals↗
PubMed2000

Study of the adsorption behavior of heavy metal ions on nanometer-size titanium dioxide with ICP-AES.

A new method using nanoparticle TiO2 as solid-phase extractant coupled with ICP-AES was proposed for simultaneous determination of trace elements. The adsorption behavior of nanometer TiO2 towards Cu, Cr, Mn and Ni was investigated by ICP-AES, and the adsorption pH curves, adsorption isotherms and adsorption capacities were obtained. It was found that the adsorption rates of the metal ions studied were more than 90% in pH 8.0-9.0, and 2.0 mol L-1 HCl was sufficient for complete elution. Nanometer TiO2 possesses a significant capacity for the sorption of the metal ions studied which is higher than the capacity of silica, the commonly used extractant. The method has been applied to the analysis of some environmental samples with satisfactory results.

Journal Article↗
PubMed2000

Progesterone synthesized by Schwann cells during myelin formation regulates neuronal gene expression.

Previously, progesterone was found to regulate the initiation and biosynthetic rate of myelin synthesis in Schwann cell/neuronal cocultures. The mRNA for cytochrome P450scc (converts cholesterol to pregnenolone), 3beta-hydroxysteroid dehydrogenase (3beta-HSD, converts pregnenolone to progesterone), and the progesterone receptor were found to be markedly induced during active myelin synthesis. However, the cells in the cocultures responsible for these changes were not identified. In this study, in situ hybridization was used to determine the localization of the enzymes responsible for steroid biosynthesis. The mRNA for cytochrome P450scc and 3beta-HSD were detected only in actively myelinating cocultures and were localized exclusively in the Schwann cells. Using immunocytochemistry, with minimal staining of the Schwann cells, we found the progesterone receptor in the dorsal root ganglia (DRG) neurons. The progesterone receptor in the neurons translocated into the nuclei of these cells when progesterone was added to neuronal cultures or during myelin synthesis in the cocultures. Additionally, a marked induction of the progesterone receptor was found in neuronal cultures after the addition of progesterone. The induction of various genes in the neurons was also investigated using mRNA differential display PCR in an attempt to elucidate the mechanism of steroid action on myelin synthesis. Two novel genes were induced in neuronal cultures by progesterone. These genes, along with the progesterone receptor, were also induced in cocultures during myelin synthesis, and their induction was blocked by RU-486 (a progesterone receptor antagonist). These genes were not induced in Schwann cells cultured alone after the addition of progesterone. These results suggest that progesterone is synthesized in Schwann cells and that it can indirectly regulate myelin formation by activating transcription via the classical steroid receptor in the DRG neurons.

3-Hydroxysteroid Dehydrogenases↗
PubMed2000

[Effect of percutaneous microwave coagulation therapy on local immune response in patients with hepatocellular carcinoma].

OBJECTIVE: To assess the local immune response to percutaneous microwave coagulation therapy (PMCT) for hepatocellular carcinoma (HCC). METHODS: Seventy-eight patients with HCC underwent PMCT. Samples of cancer and adjacent liver tissues were taken at pre-therapy, and 3, 17 and 30 days after therapy by ultrasound-guided biopsy and were stained by immunohistochemical technique. The CD3, CD45RO, CD56, CD68 and CD20 positive cells were detected in the tissue section and counted under a microscope. RESULTS: There were some CD3, CD45RO, CD56, CD68 and CD20 positive cells scattered among the cancer stroma and sinusoids and surrounding liver tissue pre-therapy. Those cells increased significantly at the third day post-therapy compared with those of pre-therapy both in cancer and adjacent liver tissues, except CD20 positive cells. The number of infiltrating immunocytes was related to the basic immune state of patients (r = 0.56, P = 0.013). Those immunocytes scattered among the parenchymal area of the tumor, the lumen of small blood vessels and the stroma closing to small vessels. The number of CD45RO and CD56 cells reached their highest point at the day 17 and CD68 positive cells at day 3 and sustained to the day 30 after PMCT. Those immunocytes were seen among the cancer cells and the lumen of small blood vessels after. CONCLUSION: The number of immunocytes in the lesion area is increased and the local immune function is enhanced by PMCT.

Adult↗
PubMed1999

Deletion of the C-terminal domain of apolipoprotein A-I impairs cell surface binding and lipid efflux in macrophage.

The contribution of the amphipathic alpha-helices of apoA-I toward lipid efflux from human skin fibroblasts and macrophage was examined. Four apoA-I mutants were designed, each by deletion of a pair of predicted adjacent helices. Three mutants lacked two consecutive central alpha-helices [Delta(100-143), Delta(122-165), and Delta(144-186)], whereas the final mutant lacked the C-terminal domain [Delta(187-243)]. When compared to recombinant wild-type apoA-I and mutants with central domain deletions, Delta(187-243) exhibited a marked reduction in its ability to promote either cholesterol or phospholipid efflux from THP-1 macrophages. This mutant also demonstrated a decreased ability to bind lipids and to form lipoprotein complexes. In contrast, the four mutants and apoA-I equally supported cholesterol efflux from fibroblasts, albeit with a reduced capacity when compared to macrophages. Delta(187-243) bound poorly to the macrophage cell surface when compared to apoA-I, and competitive binding studies with the central domain and C-terminal deletions mutants showed that only Delta(187-243) did not compete effectively with [(125)I]apoA-I. Omission of PMA during cholesterol loading enhanced cholesterol efflux to both apoA-I (1.5-fold) and the C-terminal deletion mutant (2.5-fold). Inclusion of the Sandoz ACAT inhibitor (58-035) during loading and, in the absence of PMA, increased and equalized cholesterol efflux to apoA-I and Delta(187-243). Surprisingly, omission of PMA during cholesterol loading had minimal effects on the binding of apoA-I or Delta(187-243) to the THP-1 cell surface. Overall, these results show that cholesterol efflux from cells such as fibroblasts does not require any specific sequence between residues 100 and 243 of apoA-I. In contrast, optimal cholesterol efflux in macrophages requires binding of the C-terminal domain of apoA-I to a cell surface-binding site and the subsequent translocation of intracellular cholesterol to an efflux-competent pool.

Amides↗
PubMed1999

The synthesis of a small heat shock/alpha-crystallin protein in Artemia and its relationship to stress tolerance during development.

Fertilized oocytes of the brine shrimp Artemia franciscana undergo either ovoviviparous or oviparous development, yielding free-swimming larvae (nauplii) or encysted gastrulae (cysts), respectively. Encystment is followed by diapause, wherein metabolism is greatly reduced; the resulting cysts are very resistant to extreme stress, including desiccation and long-term anoxia. The synthesis of p26, a small heat shock/alpha-crystallin protein produced only in oviparously developing Artemia, is shown in this paper to be transcriptionally regulated. A p26 mRNA of about 0.7 kb was detected on Northern blots in the second day after oocyte fertilization. It peaked as embryos encysted and declined rapidly when activated cysts resumed development. The appearance of p26 protein, as indicated by immunoprobing of Western blots, followed mRNA by 1 day; it also increased as encystment occurred but remained constant during postgastrula development of cysts. However, p26 underwent a marked reduction during emergence of nauplii and could not be detected in cell-free extracts of second-instar larvae. p26 entered nuclei of encysting embryos soon after synthesis and was localized therein as late as instar II, when it was restricted to a small set of salt gland nuclei. First-instar larvae derived from cysts were more thermotolerant than larvae that had developed ovoviviparously, but synthesis of p26 was not induced by heat under the experimental conditions employed. Additionally, transformed bacteria synthesizing p26 were more thermotolerant than bacteria that lacked the protein. The results support the proposal that p26, a developmentally regulated protein synthesized during embryo encystment, has chaperone activity in vivo and protects the proteins of encysted Artemia from stress-induced denaturation.

Animals↗
PubMed1999

ApoE of the HepG2 cell surface includes a major pool associated with chondroitin sulfate proteoglycans.

We have investigated the association of apolipoprotein E (apoE) with the HepG2 cell surface (i.e. plasma membrane and extracellular matrix) using domain specific monoclonal antibodies against apoE. Growth in beta-D-xyloside decreased the incorporation of 35S into glycosaminoglycans by 31% and cell surface apoE by 45% with a concomitant increase in apoE secretion (4.3-fold), underlining the importance of glycosaminoglycan association of apoE. Heparinase (3-10 U/mL) or heparin (1 mg/mL) decreased apoE by 25 and 30.5%, respectively, which suggests that some apoE is associated with cell surface heparan sulfate proteoglycans. Chondroitinase ABC (1.5 U/mL) reduced cell surface apoE by 40%, indicating that a major pool of apoE is associated with chondroitin sulfate proteoglycans. Further enzymatic and displacement analysis suggested that cell surface apoE associates specifically with GAGs containing chondroitin-4-sulfates. 3H1, a monoclonal antibody that recognizes an epitope within the lipid-binding C-terminal domain of apoE, decreased binding of apoE to chondroitin sulfate proteoglycans in solid-phase assays by 77% and to heparan sulfate proteoglycans by 46%, suggesting that this region is of increased importance for binding to chondroitin sulfate proteoglycans. Previous studies with 3H1 demonstrated that apoE of the extracellular matrix is lipid-poor (Burgess, J. W., Gould, D. R., and Marcel, Y. L. (1998) J. Biol. Chem. 273, 5645-5654), but we show here that apoE on the remaining cell surface is lipid-associated. In summary, lipidated apoE associates with the HepG2 plasma membrane through interactions with chondroitin-4-sulfate containing GAGs and, to a lesser extent, HSPG.

Apolipoproteins E↗
PubMed1999

Aging and high concentrations of glucose potentiate injury to mitochondrial DNA.

Deletions of mitochondrial DNA (mtDNA) are associated with aging and several chronic diseases. We have reported heterogeneous mutations between base pair 8468 and 13446 in mtDNA, the region known as the "common" deletion, in muscle of older humans with impaired glucose tolerance or diabetes mellitus. To further characterize potential effects of age and glycemia on mtDNA integrity, we studied corpulent JCR:LA-cp rats that are characterized by insulin resistance, hyperinsulinemia, and hyperlipidemia, factors strongly associated with both aging and cardiovascular disease. In addition to skeletal muscle, we isolated vascular smooth muscle cells (VSMC) from aortas of 6-, 12-, and 17-month-old rats and exposed them to 5-, 25-, 62-, and 100-mM glucose or a combination of hypoxanthine (100 microM) and xanthine oxidase (0.025 U/ml) to generate reactive oxygen species in separate cultures. Long- and short-fragment and nested polymerase chain reaction was used to detect mutations in the common deletion region. The data demonstrate that aging and the cp genotype confer susceptibility to mtDNA deletions in vivo and that high glucose concentrations can induce mtDNA mutations in vitro. Accordingly, aging and glucose-related oxidative stress and possibly hyperinsulinemia may contribute to alterations in mitochondrial gene integrity and the cp genotype appears to increase the susceptibility of muscle to the age-related accumulation of mtDNA mutations.

Aging↗
PubMed1999

[A study on the role of contractile and relaxant reaction of umbilical vein in vitro and local changes of nitrix oxide synthase in the pathogenesis of pregnancy induced hypertension].

OBJECTIVE: To study the contractile and relaxant reaction of human umbilical veins in vitro from pregnancy induced hypertension (PIH) patients, pathophysiological significance and the role of local changes of nitric oxide synthase (NOS) in the pathogenesis of PIH. METHODS: The contractile and relaxant curves of umbilical vein in response to phenylephrine, isoprenaline and acetylcholine were recorded in the PIH group and normal pregnancy group (control group). The maximum effect (Emax) and affinity index (pD2) volume were calculated. Meanwhile, the NOS activity in the blood vessel tissue and the plasma concentration of nitric oxide metabolite (NO2-/NO3-) were examined. RESULTS: (1) Relaxtant of the Emax of umbilical vein in the control group in response to acetylcholine was (85.0 +/- 15.0)%, pD2 volume was 4.4 +/- 0.5; However, the Emax (70.0 +/- 13.0)% and the pD2 (3.7 +/- 0.6) were significantly lower in the PIH group (P < 0.01). No significant differences of Emax and pD2 were obtained in phenylephrine-induced vessel contraction and isoprenaline-induced relaxation between the groups (P > 0.05). (2) NOS activity of umbilicahich was significantly lower than that from the control group [(0.5 +/- 0.2) nmol.g-1.min-1] (P < 0.01). The plasma NO2-/NO3- concentration was (38.0 +/- 10.0) mumol/L in the PIH group, which was also lower than that in the control group [(56.0 +/- 14.0) mumol/L] (P < 0.01). CONCLUSIONS: The abnormal function of endothelium system and abnormal metabolism of nitric oxide in blood vessel, which resulted in the decreased endothelium dependent relaxation, might be possible mechanism involved in the pathogenesis of PIH.

Adult↗