Biomedical subjects

G Xiao

Publications and source records attributed to G Xiao.

At least 37 records · Page 2Linked to original sources

PubMed2001

[Prognostic influence of parapharyngeal space involvement in nasopharyngeal carcinoma].

OBJECTIVE: To explore the prognostic influence of parapharyngeal space involvement in nasopharyngeal carcinoma (NPC). METHODS: From Jan. 1987 to Dec. 1994, 197 previously untreated NPC patients were examined by CT scan. The prognostic factors ware evaluated by univariate and multivariate analyses on SPSS 8.0 software. RESULTS: The actual survival rate, disease-free survival rate and local failure-free rate were negatively affected by the presence of parapharyngeal space involvement (P = 0.0115, P = 0.0035, P = 0.0367). The 5-year actual survival rate, disease-free survival rate and local failure-free rate in patients without tumor invasion to parapharyngeal space was 87.9%, 88.0% and 96.9%, respectively. The survival rate in patients with tumor invasion to prestyloid space was 75.0%, 71.2% and 93.7%, respectively. That in patients with tumor invasion to poststyloid space was 60.5%, 57.7% and 86.8%, respectively. The 5-year distant metastasis-free survival rate was 88.1% in patients without poststyloid involvement as compared to 73.8% in those with poststyloid involvement. CONCLUSION: The survival and local tumor control are affected by parapharyngeal space involvement. Distant metastasis is more likely to occur when the tumor growth involves poststyloid space.

Adolescent↗

PubMed2001

[Realization of a new segmental bioelectrical impedance method for assessing body composition: correlation analysis of human body parameters].

The method of segmental bioelectrical impedance analysis for assessing body composition requires measurement of segmental lengths and cross-sectional areas; thus its complexity is increased. This paper reports the correlation analysis of human body parameters and the linear regressions for predicting segmental parameters by use of gender, height and weight. Accordingly, segmental impedance analysis is simplified.

Adolescent↗

PubMed2001

[An experimental study on the release of endotoxin from gram negative bacteria induced by antibiotics].

OBJECTIVE: To explore the characteristics and possible mechanism of LPS released from Gram negative bacteria induced by antibiotics, so as to improve clinical management of endotoxemia and sepsis. METHODS: Cultures containing PA103 subtype of Pseudomonas aeruginosa (PA) and E coli 25922 subtype of E coli were treated with four kinds of antibiotics as Imipenam (IMP), ceftazidime (CTZ), amikacin (AMN) and pefloxacine (PFX) in four concentrations of 0.5, 1, 5 and 10 MIC for 8 hours. The changes in the bacterial quantity and morphology and the supernatant levels of free LPS of the culture media were observed at different time points. RESULTS: All the four kinds of antibiotics could kill the tested bacteria in similar degree, but lead to the different types of morphological changes of the bacteria. In detail, IMP could convert the bacteria into spherical shape, while CTZ and PFX made the bacteria to filamentous shape. But AMN could induce lysis of bacterial thallus. Under same condition, the ability of different kinds and concentrations of antibiotics to induce LPS release ranked as CTZ > PFX > IMP > AMN, 0.5MIC > 1MIC > 5MIC > 10MIC. Along with the prolongation of the action time, the LPS release increased. Furthermore, PA103 released less endotoxin than E. coli after the action of antibiotics. CONCLUSION: All of the four antibiotics, i,e, IMP, CTZ, AMN and PFX could induce PA103 and E coli 25922 to release different levels of LPS, which was related to bacterial morphological changes. The LPS release from the bacteria was correlated to the antibiotics applied, concentrations, action time and the bacterial features. Antibiotics with less ability of inducing LPS release were recommended for clinical management of the sepsis and/or septic shock caused by Gram negative bacteria.

Amikacin↗

PubMed2001

[42 years' experience on the prevention and treatment of systemic infection after severe burn].

OBJECTIVE: To summarize the experience of the management of 329 patients with major burn accompanied by severe systemic infection during the past 42 years, so as to help improve the treatment standard. METHODS: One thousand and one hundred twenty seven cases with major burns (TBSA > 50%) were admitted to our hospital during the past 42 years. The incidence of postburn systemic infection and change in the mortality were analyzed in terms of 3 periods, i.e. 1958 to 1980, 1981 to 1990 and 1991 to 2000. RESULTS: Postburn systemic infection developed in 329 out of 1127 cases with the morbidity of 29.4%. The incidences of the infection were 48.5%, 29.3% and 13.1% in the 3 periods, consecutively, with significant differences between them (P < 0.01 similar 0.05). During the 3 periods of past 42 years, the overall cure rate and cure rates in patients with different TBSA increased gradually, especially in those with TBSA Larger than 70%. CONCLUSION: The management of pastburn systemic infection was satisfactorily achieved by early comprehensive clinical measures such as immediate fluid resuscitation, early feeding, appropriate application of antibiotics and early escharectomy en masse, etc. which were established on the basis of experimental results.

Anti-Bacterial Agents↗

PubMed2001

[The effects of gut -- origin lipopolysaccharide translocation on the apoptosis of lymphocytes in scalded rats].

OBJECTIVE: To investigate the relationship between lymphocyte apoptosis in spleen and mesenteric lymph nodes (MLNs) and gut -- origin lipopolysaccharide (LPS) translocation after severe scalding in rats. METHODS: Wistar rats inflicted 30% TBSA III degree scalding were employed as the model. Serum LPS levels in portal vein and vena cava were determined by limulus amebocyte lysate (LAL) test. Apoptotic lymphocytes in paraffin -- embedded spleen and MLNs sections were detected with TdT -- mediated and dUTP nick end labeling (TUNEL) and peroxidase (POD) staining. The images were taken by cooldccd camera system, and the count and optical density (OD) value of apoptotic lymphocytes were analyzed with software Spot and Imagine proplus 4.10a (IPP4.10a). RESULTS: Serum LPS levels in portal vein and vena cava increased sharply in rats postburn (P < 0.01) and reached a peak value at 6 postburn hour (PBH). LPS level in portal vein was higher than that in vena cava (P < 0.01) in the period of 3 to 48 PBHs. The results of TUNEL -- POD staining and the count of apoptotic lymphocytes showed that the apoptotic cells were few in normal MLNs, but increased obviously postburn (P < 0.01), especially at 3 PBH. On the contrary, apoptotic lymphocytes were concentrated in spleen cortex before injury and decreased obviously postburn (P < 0.01), though a slight recovery was found after 24 PBH. Optical density of apoptotic lymphocytes was significantly reduced in spleen at 24 PBH, But the OD value of apoptotic lymphocytes in MLNs increased significantly within 48 PBHs when compared with that before injury (P < 0.01). CONCLUSION: Gut -- origin endotoxemia in rat could occur in early postburn period. The LPS translocation might induce lymphocyte apoptosis in MLNs. The decreased apoptotic lymphocytes in spleen implied abnormal immune function in spleen.

Animals↗

PubMed2000

Domain-specific interaction with the I kappa B kinase (IKK)regulatory subunit IKK gamma is an essential step in tax-mediated activation of IKK.

The human T-cell leukemia virus type 1 Tax oncoprotein deregulates the NF-kappa B signaling pathway by persistently stimulating a key signal transducer, the I kappa B kinase (IKK). Tax physically associates with the IKK regulatory subunit, IKK gamma, although the underlying biochemical mechanism and functional significance remain unclear. We show that the Tax-IKK gamma interaction requires two homologous leucine zipper domains located within IKK gamma. These leucine zipper domains are unique for the presence of a conserved upstream region that is essential for Tax binding. Site-directed mutagenesis analysis revealed that a leucine-repeat region of Tax is important for IKK gamma binding. Interestingly, all the Tax mutants defective in IKK gamma binding failed to engage the IKK complex or stimulate IKK activity, and these functional defects can be rescued by fusing the Tax mutants to IKK gamma. These results provide mechanistic insights into how Tax specifically targets and functionally activates the cellular kinase IKK.

Amino Acid Motifs↗

PubMed2000

Activation of I-kappaB kinase by the HTLV type 1 Tax protein: mechanistic insights into the adaptor function of IKKgamma.

The Tax protein encoded by human T cell leukemia virus type 1 (HTLV-1) induces constitutive nuclear expression of the transcription factor NF-kappaB, causing aberrant expression of a large array of cellular genes. Tax activates NF-kappaB by stimulating the activity of the I-kappaB kinase (IKK), which in turn leads to phosphorylation and degradation of the NF-kappaB inhibitor I-kappaBalpha. In normal T cells, IKK activation occurs transiently on cellular stimulation through the T cell receptor (TCR) and the CD28 costimulatory molecule. However, this inducible kinase is constitutively activated in Tax-expressing and HTLV-1-infected T cells, which contributes to the deregulated nuclear expression of NF-kappaB. As a genetic approach to dissect the pathways mediating IKK activation by Tax and T cell activation signals, somatic cell mutagenesis was performed to isolate signaling-defective mutant Jurkat T cell lines. One of the mutant cell lines was shown to have a defect in NF-kappaB activation by both T cell mitogens and Tax. Interestingly, this mutant cell line lacks expression of the IKK regulatory protein, IKKgamma. Expression of exogenous IKKgamma in the mutant cells restored NF-kappaB activation, thus confirming the essential role of this regulatory factor in IKK activation by the cellular and viral stimuli. Mechanistic studies have shown that Tax physically interacts with IKKgamma via specific domains, including two homologous leucine zipper motifs present in IKKgamma. The Tax/IKKgamma interaction serves to recruit Tax to the IKK catalytic subunits, IKKalpha and IKKbeta, and this recruitment appears to be an essential mechanism by which Tax stimulates the activity of IKK.

Amino Acid Sequence↗

PubMed2000

Activation of IKKalpha and IKKbeta through their fusion with HTLV-I tax protein.

Human T-cell leukemia virus type I (HTLV-I) Tax protein persistently stimulates the activity of IkappaB kinase (IKK), resulting in constitutive activation of the transcription factor NF-kappaB. Tax activation of IKK requires physical interaction of this viral protein with the IKK regulatory subunit, IKKgamma. The Tax/IKKgamma interaction allows Tax to engage the IKK catalytic subunits, IKKalpha and IKKbeta, although it remains unclear whether this linker function of IKKgamma is sufficient for supporting the Tax-specific IKK activation. To address this question, we have examined the sequences of IKKgamma required for modulating the Tax/IKK signaling. We demonstrate that when fused to Tax, a small N-terminal fragment of IKKgamma, containing its minimal IKKalpha/beta-binding domain, is sufficient for bringing Tax to and activating the IKK catalytic subunits. Disruption of the IKKalpha/beta-binding activity of this domain abolishes its function in modulating the Tax/IKK signaling. We further demonstrate that direct fusion of Tax to IKKalpha and IKKbeta leads to activation of these kinases. These findings suggest that the IKKgamma-directed Tax/IKK association serves as a molecular trigger for IKK activation.

Binding Sites↗

PubMed2000

Stressing-out DNA? The contribution of serine-phosphodiester interactions in catalysis by uracil DNA glycosylase.

The DNA repair enzyme uracil DNA glycosylase (UDG) pinches the phosphodiester backbone of damaged DNA using the hydroxyl side chains of a conserved trio of serine residues, resulting in flipping of the deoxyuridine from the DNA helix into the enzyme active site. We have investigated the energetic role of these serine-phosphodiester interactions using the complementary approaches of crystallography, directed mutagenesis, and stereospecific phosphorothioate substitutions. A new crystal structure of UDG bound to 5'-HO-dUAAp-3' (which lacks the 5' phosphodiester group that interacts with the Ser88 pinching finger) shows that the glycosidic bond of dU has been cleaved, and that the enzyme has undergone the same specific clamping motion that brings key active site groups into position as previously observed in the structures of human UDG bound to large duplex DNA substrates. From this structure, it may be concluded that glycosidic bond cleavage and the induced fit conformational change in UDG can occur without the 5' pinching interaction. The S88A, S189A, and S192G "pinching" mutations exhibit 360-, 80-, and 21-fold damaging effects on k(cat)/K(m), respectively, while the S88A/S189A double mutant exhibits an 8200-fold damaging effect. A free energy analysis of the combined effects of nonbridging phosphorothioate substitution and mutation at these positions reveals the presence of a modest amount of strain energy between the compressed 5' and 3' phosphodiester groups flanking the bound uridine. Overall, these results indicate a role for these serine-phosphodiester interactions in uracil flipping and preorganization of the sugar ring into a reactive conformation. However, in contrast to a recent proposal [Parikh, S. S., et al. (2000) Proc Natl. Acad. Sci. 94, 5083], there is no evidence that conformational strain of the glycosidic bond induced by serine pinching plays a major role in the 10(12)-fold rate enhancement brought about by UDG.

Catalysis↗

PubMed2000

Low-frequency magnetic noise in micron-scale magnetic tunnel junctions.

We have observed low-frequency noise due to quasiequilibrium thermal magnetization fluctuations in micron-scale magnetic tunnel junctions (MTJs). This strongly field-dependent magnetic noise occurs within the magnetic hysteresis loops, either as 1/f or Lorentzian (random telegraph) noise. We attribute it to the thermally excited hopping of magnetic domain walls between pinning sites. Our results show that magnetic stability is a crucial factor in reducing the low-frequency noise in small MTJs.

Journal Article↗

PubMed2000

The NF-kappa B signaling pathway is not required for Fas ligand gene induction but mediates protection from activation-induced cell death.

Stimulation of T cells by antigens or mitogens triggers multiple signaling pathways leading to activation of genes encoding interleukin-2 and other growth-regulatory cytokines. The same stimuli also activate the gene encoding an apoptosis-inducing molecule, Fas ligand (FasL), which contributes to activation-induced cell death. It has been proposed that the signaling pathways involved in cytokine gene induction also contribute to activation-induced FasL expression; however, genetic evidence for this proposal is lacking. In the present study, the role of the NF-kappaB signaling pathway in FasL gene expression was examined using a mutant T cell line deficient in an essential NF-kappaB signaling component, IkappaB kinase gamma. These mutant cells have a blockade in signal-induced activation of NF-kappaB but remained normal in the activation of NF-AT and AP-1 transcription factors. Interestingly, the NF-kappaB signaling defect has no effect on mitogen-stimulated FasL gene expression, although it completely blocks the interleukin-2 gene induction. We further demonstrate that NF-kappaB activation is required for protecting T cells from apoptosis induction by mitogens and an agonistic anti-Fas antibody. These genetic results suggest that the NF-kappaB signaling pathway is not required for activation-induced FasL expression but rather mediates cell growth and protection from activation-induced cell death.

Annexin A5↗

PubMed2000

Negative regulation of the nuclear factor kappa B-inducing kinase by a cis-acting domain.

Nuclear factor kappaB (NF-kappaB)-inducing kinase (NIK) participates in the activation of NF-kappaB, a family of eukaryotic transcription factors that mediate cell growth and transformation. NIK activates the IkappaB kinase both in vivo and in vitro, although how the activity of NIK is regulated has remained unclear. Here we show that the N-terminal region of NIK contains a negative-regulatory domain (NRD), which is composed of a basic motif and a proline-rich repeat motif. Deletion of these motifs leads to a marked enhancement of NIK function. We further demonstrate that the N-terminal NRD interacts with the C-terminal region of NIK, thereby inhibiting the binding of NIK to its substrate IkappaB kinase. Consistently, when expressed alone, the NRD potently inhibits NIK-mediated NF-kappaB signaling. These results provide a new insight into the mechanism of NIK regulation.

Amino Acid Sequence↗

PubMed2000

Inhibition of InhA, the enoyl reductase from Mycobacterium tuberculosis, by triclosan and isoniazid.

Structural and genetic studies indicate that the antibacterial compound triclosan, an additive in many personal care products, is an inhibitor of EnvM, the enoyl reductase from Escherichia coli. Here we show that triclosan specifically inhibits InhA, the enoyl reductase from Mycobacterium tuberculosis and a target for the antitubercular drug isoniazid. Binding of triclosan to wild-type InhA is uncompetitive with respect to both NADH and trans-2-dodecenoyl-CoA, with K(i)' values of 0.22+/-0.02 and 0.21+/-0.01 microM, respectively. Replacement of Y158, the catalytic tyrosine residue, with Phe, reduces the affinity of triclosan for the enzyme and results in noncompetitive inhibition, with K(i) and K(i)' values of 36+/-5 and 47+/-5 microM, respectively. Consequently, the Y158 hydroxyl group is important for triclosan binding, suggesting that triclosan binds in similar ways to both InhA and EnvM. In addition, the M161V and A124V InhA mutants, which result in resistance of Mycobacterium smegmatis to triclosan, show significantly reduced affinity for triclosan. Inhibition of M161V is noncompetitive with K(i)' = 4.3+/-0.5 microM and K(i) = 4.4+/-0.9 microM, while inhibition of A124V is uncompetitive with K(i)' = 0. 81 +/- 0.11 microM. These data support the hypothesis that the mycobacterial enoyl reductases are targets for triclosan. The M161V and A124V enzymes are also much less sensitive to isoniazid compared to the wild-type enzyme, indicating that triclosan can stimulate the emergence of isoniazid-resistant enoyl reductases. In contrast, I47T and I21V, two InhA mutations that occur in isoniazid-resistant clinical isolates of M. tuberculosis, show unimpaired inhibition by triclosan, with uncompetitive inhibition constants (K(i)') of 0.18+/-0.01 and 0.12+/- 0.01 microM, respectively. The latter result indicates that InhA inhibitors targeted at the enoyl substrate binding site may be effective against existing isoniazid-resistant strains of M. tuberculosis.

Antitubercular Agents↗

PubMed2000

Identification of a homeodomain binding element in the bone sialoprotein gene promoter that is required for its osteoblast-selective expression.

Bone sialoprotein is a 70-kDa extracellular matrix component that is intimately associated with biomineralization, yet the cis-acting elements of the Bsp gene that restrict its expression to mineralizing cells remain uncharacterized. To identify such elements, we analyzed a 2472-base pair fragment of the murine promoter that directs osteoblast-selective expression of a luciferase reporter gene and found that the region between -338 and -178 relative to the transcriptional start is crucial for its osteoblast-selective activity. We identified an element within this region that binds a protein complex in the nuclear extracts of osteoblastic cells and is required for its transcriptional activity. Introduction of a mutation that disrupts a homeodomain binding site within this sequence eliminates both its in vitro binding and nearly all of the osteoblastic-selective activity of the 2472-base pair promoter. We further found that the Dlx5 homeoprotein, which is able to regulate the osteoblast-specific osteocalcin promoter, can bind this element and stimulate its enhancer activity when overexpressed in COS7 cells. These data represent the first description of an osteoblast-specific element within the bone sialoprotein promoter and demonstrate its regulation by a member of a family of factors known to be involved in skeletogenesis.

Animals↗

PubMed2000

A DNA damage signal is required for p53 to activate gadd45.

We provide direct evidence that overexpression of p53 is not sufficient for robust p53-dependent activation of the endogenous gadd45 gene. When p53 was induced in TR9-7 cells in the absence of DNA damage, waf1/p21 and mdm2 mRNA levels were increased, but a change in gadd45 mRNA was barely detectable. Activation of the gadd45 gene was observed when camptothecin was added to cells containing p53 in the absence of a further increase in the p53 level. Phosphorylation of p53 at serine 15 and acetylation at lysine 382 were detected after drug treatment. It has been suggested that p53 posttranslational modification is critical during activation. However, inhibition of these modifications by wortmannin was not sufficient to block the transactivation of gadd45. Interestingly, after camptothecin treatment, increased DNase I sensitivity was detected at the gadd45 promoter, suggesting that an undetermined DNA damage signal is involved in inducing chromatin remodeling at the gadd45 promoter while cooperating with p53 to activate gadd45 transcription.

Acetylation↗

PubMed2000

Somatic mutagenesis studies of NF-kappa B signaling in human T cells: evidence for an essential role of IKK gamma in NF-kappa B activation by T-cell costimulatory signals and HTLV-I Tax protein.

NF-kappa B plays a pivotal role in normal T-cell activation and may also mediate human T-cell leukemia virus (HTLV)-induced T-cell transformation. Activation of NF-kappa B by both T-cell costimulatory signals and the HTLV Tax protein involves stimulation of I kappa B kinase (IKK). As a genetic approach to dissect the intermediate steps involved in NF-kappa B activation in human T cells, we performed somatic cell mutagenesis to isolate signaling-defective mutant Jurkat T-cell lines. One of the mutant cell lines was shown to have a specific blockade in the IKK signaling pathway but remained competent in the c-Jun N-terminal kinase and MAP kinase pathways. Interestingly, this mutant cell line lacks expression of IKK gamma, a non-catalytic component of the IKK complex. Expression of exogenous IKK gamma in the mutant cells restored NF-kappa B activation by both the T-cell costimulation agents and Tax. These findings provide genetic evidence for the requirement of IKK gamma in NF-kappa B signaling triggered by both T-cell costimulatory signals and HTLV-I Tax protein.

Antibodies, Monoclonal↗

PubMed2000

MAPK pathways activate and phosphorylate the osteoblast-specific transcription factor, Cbfa1.

The bone-specific transcription factor, Cbfa1, regulates expression of the osteocalcin (OCN) gene and is essential for bone formation. However, little is known about the mechanisms regulating Cbfa1 activity. This work examines the role of the MAPK pathway in regulating Cbfa1-dependent transcription. Stimulation of MAPK by transfecting a constitutively active form of MEK1, MEK(SP), into MC3T3-E1 preosteoblast cells increased endogenous OCN mRNA, while a dominant negative mutant, MEK(DN), was inhibitory. MEK(SP) also stimulated activity of a 147-base pair minimal OCN promoter, and this stimulation required an intact copy of OSE2, the DNA binding site for Cbfa1. Effects of MEK(SP) were specific to Cbfa1-positive osteoblast-like cells. A purified His-tagged Cbfa1 fusion protein was directly phosphorylated by activated recombinant MAPK in vitro. Furthermore, (32)P metabolic labeling studies demonstrated that MEK(SP) clearly enhanced phosphorylation of Cbfa1 in intact cells, while MEK(DN) decreased phosphorylation. The specific MEK1/MEK2 inhibitor, PD98059, inhibited extracellular matrix-dependent up-regulation of the OCN promoter, indicating that the MAPK pathway and, presumably, Cbfa1 phosphorylation are also required for responsiveness of osteoblasts to extracellular matrix signals. This study is the first demonstration that Cbfa1 is controlled by MAPKs and suggests that this pathway has an important role in the control of osteoblast-specific gene expression.

Animals↗

PubMed2000

A novel Rb- and p300-binding protein inhibits transactivation by MyoD.

The retinoblastoma protein (Rb) regulates both the cell cycle and tissue-specific transcription, by modulating the activity of factors that associate with its A-B and C pockets. In skeletal muscle, Rb has been reported to regulate irreversible cell cycle exit and muscle-specific transcription. To identify factors interacting with Rb in muscle cells, we utilized the yeast two-hybrid system, using the A-B and C pockets of Rb as bait. A novel protein we have designated E1A-like inhibitor of differentiation 1 (EID-1), was the predominant Rb-binding clone isolated. It is preferentially expressed in adult cardiac and skeletal muscle and encodes a 187-amino-acid protein, with a classic Rb-binding motif (LXCXE) in its C terminus. Overexpression of EID-1 in skeletal muscle inhibited tissue-specific transcription. Repression of skeletal muscle-restricted genes was mediated by a block to transactivation by MyoD independent of G(1) exit and, surprisingly, was potentiated by a mutation that prevents EID-1 binding to Rb. Inhibition of MyoD may be explained by EID-1's ability to bind and inhibit p300's histone acetylase activity, an essential MyoD coactivator. Thus, EID-1 binds both Rb and p300 and is a novel repressor of MyoD function.

Acetyltransferases↗