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Lin Gao

Publications and source records attributed to Lin Gao.

At least 19 recordsLinked to original sources
PubMed2005

[Determination of oxidation-reduction level in patients with rheumatoid arthritis].

OBJECTIVE: To examine the oxidation or reduction products in patients with rheumatism arthritis (RA), and investigate the relationship between oxidation or reduction products and occurrence and development of RA. METHODS: The serum levels of total ascorbic acid (TAA), dehydroascorbic acid (DHAA)/TAA, vitamin E, advanced oxidation protein products (AOPP) and malondialdehyde (MDA) were detected by high-performance liquid chromatography with electrochemical detection in 83 RA patients and 30 healthy adults. Correlation analysis of AOPP, MDA and hs-CRP was performed. RESULTS: Compared with normal control group, significantly higher serum MDA, DHAA/TAA, and AOPP levels were detected in RA patients (P<0.05), but vitamin E showed no significant difference (P<0.05). Linear regression analysis showed that MDA (P<0.01) was positively but AOPP (P>0.05) negatively correlated to hs-CRP. CONCLUSIONS: Oxidation or reduction products in serum of RA patients increases significantly, which may be an important mechanism for the occurrence and development of RA. Serum AOPP and MDA levels can reflect the oxidation status in RA patients.

Adult↗
PubMed2004

Cross-linked polyacrylamide coating for capillary isoelectric focusing.

Polyacrylamide has been used for capillary wall coating for decades. The coating chemistry includes two main steps: (i) attachment of a bifunctional reagent containing a vinyl group to the silica surface and (ii) extension of the anchored vinyl groups through acrylamide polymerization. Since the introduction of this method, many modifications and improvements have been made. However, few of them are successful for routine capillary isoelectric focusing. One of the major problems is the presence of some microspots in which the silica surfaces are poorly coated. Cross-linking the polyacrylamide molecules anchored around these poorly coated spots seems to be a straightforward solution to this problem. Attempts have been made toward this direction, but cross-linked polyacrylamide coatings have not been demonstrated to be much superior over linear polyacrylamide ones. In this report, we have reexamined this approach and demonstrated that cross-linked polyacrylamide could be excellent for capillary isoelectric focusing. A simple device and a new coating protocol have been developed to produce this coating reliably and reproducibly. Compared to the commercial linear polyacrylamide and hydroxypropyl cellulose coatings for CIEF, the cross-linked polyacrylamide coating is much more stable and robust although their initial performances are comparable.

Acrylic Resins↗
PubMed2004

Isoforms of the polarity protein par6 have distinct functions.

PAR-6 is essential for asymmetric division of the Caenorhabditis elegans zygote. It is also critical for cell polarization in many other contexts throughout the Metazoa. The Par6 protein contains a PDZ domain and a partial CRIB (Cdc42/Rac interactive binding) domain, which mediate interactions with other polarity proteins such as Par3, Cdc42, Pals1, and Lgl. A family of mammalian Par6 isoforms (Par6A-D) has been described, but the significance of this diversification has been unclear. Here we demonstrate that Par6 family members localize differently when expressed in Madin-Darby canine kidney epithelial cells and have distinct effects on tight junction (TJ) assembly. Par6B localizes to the cytosol and inhibits TJ formation, but Par6A co-localizes predominantly with the TJ marker ZO-1 at cell-cell contacts and does not affect junctions. These functional differences correlate with differences in Pals1 binding; Par6B interacts strongly with Pals1, whereas Par6A binds weakly to Pals1 even in the presence of active Cdc42. Pals1 has a low affinity for the isolated CRIB-PDZ domain of Par6A, but analysis of chimeras showed that in addition Pals1 binding is blocked by an inhibitory property of the N terminus of Par6A. Unexpectedly, the localization of Par6A to cell-cell contacts is Cdc42-independent.

Amino Acid Sequence↗
PubMed2004

Induction of the glucose-6-phosphate dehydrogenase gene expression by chronic hypoxia in PC12 cells.

We studied the regulation of glucose-6-phosphate dehydrogenase (G6PD) gene expression by chronic hypoxia. G6PD mRNA level and activity were increased in PC12 cells by hypoxia in a dose- and time-dependent manner. Cobalt chloride and dimethyloxalylglycine, which can mimic hypoxia, also activated G6PD gene expression. Interestingly, hypoxia-induced G6PD expression followed a time course much slower than that of phosphoglycerate kinase 1 (PGK1), a hypoxia-inducible factor (HIF)-dependent glycolytic enzyme. Hypoxic-G6PD induction was almost negligible in non-excitable Buffalo rat liver cells, although in these cells PGK1 was strongly upregulated by low PO(2). Furthermore, G6PD but not PGK1 induction was blocked by the antioxidants glutathione and N-acetylcysteine. These results suggest the dependence of G6PD gene expression on HIF and intracellular redox status and the differential hypoxic regulation of glucose-metabolizing enzymes.

Animals↗
PubMed2004

Selective growth inhibition of tumor cells by a novel histone deacetylase inhibitor, NVP-LAQ824.

We have synthesized a histone deacetylase inhibitor, NVP-LAQ824, a cinnamic hydroxamic acid, that inhibited in vitro enzymatic activities and transcriptionally activated the p21 promoter in reporter gene assays. NVP-LAQ824 selectively inhibited growth of cancer cell lines at submicromolar levels after 48-72 h of exposure, whereas higher concentrations and longer exposure times were required to retard the growth of normal dermal human fibroblasts. Flow cytometry studies revealed that both tumor and normal cells arrested in the G(2)-M phase of the cell cycle after compound treatment. However, an increased sub-G(1) population at 48 h (reminiscent of apoptotic cells) was observed only in the cancer cell line. Annexin V staining data supported our hypothesis that NVP-LAQ824 induced apoptosis in tumor and transformed cells but not in normal cells. Western blotting experiments showed an increased histone H3 and H4 acetylation level in NVP-LAQ824-treated cancer cells, suggesting that the likely in vivo target of NVP-LAQ824 was histone deacetylase(s). Finally, NVP-LAQ824 exhibited antitumor effects in a xenograft animal model. Together, our data indicated that the activity of NVP-LAQ824 was consistent with its intended mechanism of action. This novel histone deacetylase inhibitor is currently in clinical trials as an anticancer agent.

Animals↗
PubMed2004

Leaky termination at premature stop codons antagonizes nonsense-mediated mRNA decay in S. cerevisiae.

The Nonsense-Mediated mRNA Decay (NMD) pathway mediates the rapid degradation of mRNAs that contain premature stop mutations in eukaryotic organisms. It was recently shown that mutations in three yeast genes that encode proteins involved in the NMD process, UPF1, UPF2, and UPF3, also reduce the efficiency of translation termination. In the current study, we compared the efficiency of translation termination in a upf1Delta strain and a [PSI(+)] strain using a collection of translation termination reporter constructs. The [PSI(+)] state is caused by a prion form of the polypeptide chain release factor eRF3 that limits its availability to participate in translation termination. In contrast, the mechanism by which Upf1p influences translation termination is poorly understood. The efficiency of translation termination is primarily determined by a tetranucleotide termination signal consisting of the stop codon and the first nucleotide immediately 3' of the stop codon. We found that the upf1Delta mutation, like the [PSI(+)] state, decreases the efficiency of translation termination over a broad range of tetranucleotide termination signals in a unique, context-dependent manner. These results suggest that Upf1p may associate with the termination complex prior to polypeptide chain release. We also found that the increase in readthrough observed in a [PSI(+)]/upf1Delta strain was larger than the readthrough observed in strains carrying either defect alone, indicating that the upf1Delta mutation and the [PSI(+)] state influence the termination process in distinct ways. Finally, our analysis revealed that the mRNA destabilization associated with NMD could be separated into two distinct forms that correlated with the extent the premature stop codon was suppressed. The minor component of NMD was a 25% decrease in mRNA levels observed when readthrough was >/=0.5%, while the major component was represented by a larger decrease in mRNA abundance that was observed only when readthrough was </=0.5%. This low threshold for the onset of the major component of NMD indicates that mRNA surveillance is an ongoing process that occurs throughout the lifetime of an mRNA.

Codon, Nonsense↗
PubMed2003

Structure of Cdc42 in a complex with the GTPase-binding domain of the cell polarity protein, Par6.

Cdc42 is a small GTPase that is required for cell polarity establishment in eukaryotes as diverse as budding yeast and mammals. Par6 is also implicated in metazoan cell polarity establishment and asymmetric cell divisions. Cdc42.GTP interacts with proteins that contain a conserved sequence called a CRIB motif. Uniquely, Par6 possesses a semi-CRIB motif that is not sufficient for binding to Cdc42. An adjacent PDZ domain is also necessary and is required for biological effects of Par6. Here we report the crystal structure of a complex between Cdc42 and the Par6 GTPase-binding domain. The semi-CRIB motif forms a beta-strand that inserts between the four strands of Cdc42 and the three strands of the PDZ domain to form a continuous eight-stranded sheet. Cdc42 induces a conformational change in Par6, detectable by fluorescence resonance energy transfer spectroscopy. Nuclear magnetic resonance studies indicate that the semi-CRIB motif of Par6 is at least partially structured by the PDZ domain. The structure highlights a novel role for a PDZ domain as a structural scaffold.

Amino Acid Sequence↗
PubMed2003

[The synthesis of MT in rat myocardial fibroblasts].

OBJECTIVE: To study the influence on the synthesis of metallothionein (MT) induced by lipopolysaccharide (LPS), basic fibroblast growth factor (bFGF), oxidized low-density lipoproteins (oxLDL) and ZnCl2 in rat myocardial fibroblasts. METHODS: The content of MT was measured by 109Cd-Hb saturation method and RT-PCR was used to determine the expression of MT mRNA. RESULTS: All of the stimulus in this experiment increased the synthesis of MT in myocardial fibroblasts. Compared with control, 1 and 5 mg.L-1 LPS increased the content of MT by 94% and 130% (P < 0.01), 0.2 and 1.0 mg.L-1 bFGF by 41% and 59% (P < 0.01), 2 and 10 mg.L-1 oxLDL by 23% (P < 0.05) and 41% (P < 0.01) respectively. 1,2 and 5 mg.L-1 ZnCl2 increased the content of MT by 66%, 99% and 149% (P < 0.01) in a concentration-dependent manner. 2-8 mg.L-1 ZnCl2 increased the content of MT1 mRNA by 69%-129%, MT2 mRNA by 31%-60% (P < 0.01). But 10 mg.L-1 ZnCl2 didn't stimulate the synthesis of MT more significantly, probably because ZnCl2 of high concentration damaged myocardial fibroblasts and decreased the survival rate of the cells. CONCLUSION: Many stimulus such as LPS, bFGF, oxLDL and ZnCl2 can induce the synthesis of MT, and ZnCl2 can increase the expression of MT mRNA in rat myocardial fibroblasts.

Animals↗
PubMed2003

Inhibition of taurine transport by high concentration of glucose in cultured rat cardiomyocytes.

Cultured rat cardiomyocytes were treated with 10, 20, and 30 mmol/L glucose and 30 mmol/L glucose plus protein kinase C (PKC) inhibitor, Chelerythrine. In the 20 and 30 mmol/L glucose-treated cells, taurine contents reduced by 15% and 27% (P<.05), respectively, taurine transporter (TAUT) mRNA levels reduced by 47% and 64% (P<.05), respectively, and cysteine sulfinate decarboxylase (CSD) mRNA reduced slightly, but not significantly. Time-dependent taurine uptakes reduced in the 10, 20, and 30 mmol/L glucose-treated cells, and time-dependent taurine release reduced in the 30 mmol/L glucose-treated cells. The Vmax of taurine transport decreased by 18%, 30%, and 35% (P<.05) in the 10, 20, and 30 mmol/L glucose-treated cells, respectively, while Km of taurine transport remained unchanged. When PKC inhibitor, Chelerythrine, combined with 30 mmol/L glucose was treated with the cells, the lowered taurine content, taurine uptake, taurine release, and Vmax of taurine transport caused by 30 mmol/L glucose were eliminated. These results demonstrate that high glucose considerably and specifically decreases intracellular taurine content, taurine transport activity, and TAUT mRNA, possibly through PKC-mediated transcriptional and posttranslational pathways.

Alkaloids↗
PubMed2003

Direct interaction of two polarity complexes implicated in epithelial tight junction assembly.

Tight junctions help establish polarity in mammalian epithelia by forming a physical barrier that separates apical and basolateral membranes. Two evolutionarily conserved multi-protein complexes, Crumbs (Crb)-PALS1 (Stardust)-PATJ (DiscsLost) and Cdc42-Par6-Par3-atypical protein kinase C (aPKC), have been implicated in the assembly of tight junctions and in polarization of Drosophila melanogaster epithelia. Here we identify a biochemical and functional link between these two complexes that is mediated by Par6 and PALS1 (proteins associated with Lin7). The interaction between Par6 and PALS1 is direct, requires the amino terminus of PALS1 and the PDZ domain of Par6, and is regulated by Cdc42-GTP. The transmembrane protein Crb can recruit wild-type Par6, but not Par6 with a mutated PDZ domain, to the cell surface. Expression of dominant-negative PALS1-associated tight junction protein (PATJ) in MDCK cells results in mis-localization of PALS1, members of the Par3-Par6-aPKC complex and the tight junction marker, ZO-1. Similarly, overexpression of Par6 in MDCK cells inhibits localization of PALS1 to the tight junction. Our data highlight a previously unrecognized link between protein complexes that are essential for epithelial polarity and formation of tight junctions.

Animals↗
PubMed2002

Dysfunction of myocardial and vascular taurine transport in spontaneously hypertensive rats.

The alterations of taurine transport and the expression of taurine transporter (TAUT) mRNA in myocardium and aortic wall were investigated in spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. It was demonstrated that plasma taurine concentration and taurine release from myocardium and aortic wall in SHR were higher than those in WKY rats, whereas taurine content, taurine uptake and TAUT mRNA in myocardium and aortic wall of SHR were lower than those of WKY rats. In SHR, the maximal velocity (V(max)) of taurine transportation in myocardium and aortic wall was lower by 24% (P<0.05) and 35% (P<0.05) than that in WKY, their michaelis constants (Km) values were higher by 16% (P<0.05) and 39% (P<0.05), respectively. The results suggest that there is dysfunction of taurine transport in myocardium and aortic wall in SHR, which may be partly resulted from the decrease of TAUT activity and affinity, and down-regulation of TAUT gene expression.

Animals↗
PubMed2002

[Changes in adrenomedullin and receptor activity-modifying protein 2 mRNA in myocardium and vessels during L-NNA-induced hypertension in rats].

To explore the changes in adrenomedullin (ADM) and receptor activity-modifying protein 2 (RAMP2) mRNA in myocardium and vessels in hypertension, a hypertensive rat model was prepared by administering L-NNA. Contents of ADM in plasma, myocardium and vessels were measured by radioimmunoassay (RIA). The levels of pro-ADM mRNA of myocardium and vessels were determined by competitive quantitative RT-PCR. The results showed that L-NNA induced hypertension and cardiomegaly. The ratio of heart to body weight increased by 35.5% (P<0.01). In hypertensive rats the ir-ADM in plasma, myocardium and vessels was increased by 80%, 72% and 57% (P<0.01), respectively compared with the control. The amounts of ADM mRNA in myocardium and vessels were increased by 50% and 109.2% (P<0.05), respectively, and the amounts of RAMP2 mRNA was increased by 132% and 87% (P<0.01), respectively, compared with control. The levels of ADM in myocardium and vessels were positively correlated with RAMP2 mRNA, the correlation coefficients were 0.741 and 0.885 (P<0.01), respectively. The results obtained indicate that in hypertensive rats, ADM is elevated in plasma, myocardium and ves-myocardium and vessel, and ADM and RAMP2 mRNA are up-regulated in myocardium and vessel. The ADM/RAMP2 system may play an important role in the pathogenesis of hypertension.

Adrenomedullin↗
PubMed2002

Multiple splice variants of Par3 and of a novel related gene, Par3L, produce proteins with different binding properties.

The partitioning-defective 3 (par3) gene encodes a protein with three postsynaptic density90/DiscslargeA/ZO-1 (PDZ) domains that is required for cell polarity establishment in metazoans. Par3 is a component of a protein complex that can include Cdc42-GTP, Par6 and atypical protein kinase Cs (aPKCs). We now describe the identification of a related human gene, Par3L. Both Par3L and Par3 are expressed as numerous alternatively spliced variants. Although Par3 expression appears to be ubiquitous, that of Par3L is more restricted. Multiple variants are often expressed simultaneously within a specific cell type or tissue. Although all of the Par3L/Par3 isoforms can associate with tight junctions in epithelial cells, they show different binding properties. No Par3L isoforms and only a subset of Par3 isoforms detectably bind aPKCs. These data suggest that aPKC binding or phosphorylation is not required for targeting of Par3/Par3L to cell-cell contacts. Par3L isoforms also show differential binding to Par6. Despite these differences, the N-terminal region of Par3L, like that of Par3, can disrupt the formation of tight junctions when ectopically expressed in Madin-Darby canine kidney (MDCK) cells.

Adaptor Proteins, Signal Transducing↗
PubMed2002

Cloning and functional characterization of HDAC11, a novel member of the human histone deacetylase family.

We have cloned and characterized a human cDNA that belongs to the histone deacetylase family, which we designate as HDAC11. The predicted HDAC11 amino acid sequence reveals an open reading frame of 347 residues with a corresponding molecular mass of 39 kDa. Sequence analyses of the putative HDAC11 protein indicate that it contains conserved residues in the catalytic core regions shared by both class I and II mammalian HDAC enzymes. Putative orthologues of HDAC11 exist in primate, mouse, Drosophila, and plant. Epitope-tagged HDAC11 protein expressed in mammalian cells displays histone deacetylase activity in vitro. Furthermore, HDAC11's enzymatic activity is inhibited by trapoxin, a known histone deacetylase inhibitor. Multiple tissue Northern blot and real-time PCR experiments show that the high expression level of HDAC11 transcripts is limited to kidney, heart, brain, skeletal muscle, and testis. Epitope-tagged HDAC11 protein localizes predominantly to the cell nucleus. Co-immunoprecipitation experiments indicate that HDAC11 may be present in protein complexes that also contain HDAC6. These results indicate that HDAC11 is a novel and unique member of the histone deacetylase family and it may have distinct physiological roles from those of the known HDACs.

Amino Acid Sequence↗
PubMed2002

Assembly of epithelial tight junctions is negatively regulated by Par6.

Epithelial cells display apical-basal polarity, and the apical surface is segregated from the basolateral membranes by a barrier called the tight junction (TJ). TJs are constructed from transmembrane proteins that form cell-cell contacts-claudins, occludin, and junctional adhesion molecule (JAM)-plus peripheral proteins such as ZO-1. The Par proteins (partitioning-defective) Par3 and Par6, plus atypical protein kinase C (aPKC) function in the formation or maintenance of TJs and more generally in metazoan cell polarity establishment. Par6 contains a PDZ domain and a partial CRIB (Cdc42/Rac interactive binding) domain and binds the small GTPase Cdc42. Here, we show that Par6 inhibits TJ assembly in MDCK II epithelial cells after their disruption by Ca(2+) depletion but does not inhibit adherens junction (AJ) formation. Transepithelial resistance and paracellular diffusion assays confirmed that assembly of functional TJs is delayed by Par6 overexpression. Strikingly, the isolated, N-terminal fragment of PKCzeta, which binds Par6, also inhibits TJ assembly. Activated Cdc42 can disrupt TJs, but neither a dominant-negative Cdc42 mutant nor the CRIB domain of gammaPAK (p21-activated kinase), which inhibits Cdc42 function, observably inhibit TJ formation. These results suggest that Cdc42 and Par6 negatively regulate TJ assembly in mammalian epithelial cells.

Adherens Junctions↗
PubMed2002

CTLA-4 gene A/G polymorphism associated with diabetes mellitus in Han Chinese.

OBJECTIVE: To investigate the association of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) gene A/G polymorphism with susceptibility to diabetes mellitus in Han Chinese. METHODS: An A/G transition at position 49 of exon 1 was analyzed in 31 patients with type 1 diabetes, 31 patients with type 2 diabetes, and 36 controls were analyzed by polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: A highly significant increase in the frequency of the G allele was seen in patients with type 1 diabetes compared with controls (66.1 % vs. 34.7%, respectively; P < 0.0005; OR = 3.670) . This reflected an increase in the GG genotype in patients (48.4% vs. 22.2%, respectively; P =0.025; OR =3.281) and a significant decrease in the AA genotype (16.1 % vs. 52.8%, respectively; P = 0.002). The allele frequencies of A and G in patients with type 2 diabetes were not significantly different from controls(A/G, 50.0/50.0% vs. 65.3/34.7%; P = not significant) . The distribution of genotype, however, differed significantly. This difference reflected an increase in the AG genotype in patients (54.8% vs.25.0%, respectively; P=0.012; OR=3.643) and a decrease in the AA genotype (22.6% vs. 52.8%, respectively; P=0.011). CONCLUSIONS: CTLA-4 49 AA is protective from diabetes mellitus, whereas, CTLA-4 49 G allele (both as homozygotes and as heterozygotes ) confers an increased risk of diabetes mellitus.

Abatacept↗
PubMed2002

Dysfunction of myocardial taurine transport and effect of taurine supplement in rats with isoproterenol-induced myocardial injury.

AIM: To study the alterations of myocardial taurine transport function, taurine transporter (TAUT), and cysteine sulfinate decarboxylase (CSD) mRNA as well as effect of exogenous taurine in rats with isoproterenol (ISO)-induced cardiomegaly. METHODS: [3H]-Taurine uptake and release on myocardium were determined. Binding sites of [3H]-taurine for myocardial sarcolemma were measured. TAUT and CSD mRNA levels were assayed using competitive quantitative reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: ISO group as compared with control group, myocardial taurine uptake markedly reduced, taurine release obviously increased; Bmax value of [3H]-taurine binding on cardiac sarcolemma reduced by 42% (P<0.05); TAUT and CSD mRNA levels decreased by 40% and 38% (P<0.05), respectively. ISO+taurine group as compared with ISO-treated group, the amounts of taurine uptake and TAUT mRNA returned to normal; taurine release reduced; Bmax increased by 92% (P<0.01), and CSD mRNA content augmented by 23% (P<0.05). There were no statistical differences of Kd values among the four groups (P>0.05). CONCLUSION: The data indicate that the failure to generate sufficient TAUT on myocardial sarcolemma may be the fundamental abnormality in ISO-induced cardiac injury. The mechanism by which administration of taurine considerably improves ISO-induced cardiac damage is probably to increase the expression of TAUT gene and recover taurine transport function.

Animals↗