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Biomedical subjects

H Tang

Publications and source records attributed to H Tang.

At least 181 records · Page 10Linked to original sources

Functional interaction of the HTLV-1 transactivator Tax with activating transcription factor-4 (ATF4).

The Tax protein of the Human T-cell Leukemia Virus (HTLV) activates the expression of viral mRNA through a three 21 bp repeat enhancer located within the HTLV-1 LTR. Since Tax does not bind to the 21 bp DNA repeats directly, it has been speculated that Tax interacts with cellular protein(s) which mediate binding to the enhancer. We employed the yeast two hybrid system to identify host proteins that are potentially relevant to Tax transactivation. We identified a Tax binding protein encoded from a cDNA expression library derived from peripheral blood lymphocytes. The corresponding cDNA has sequence identity with a known transcription factor, activating factor-4 (ATF-4). ATF-4 also binds to GST-Tax fusion protein in vitro. Tax mutants that did not transactivate the HTLV-1 LTR also failed to bind ATF-4. The critical domain for Tax binding resides in a 85 amino acid stretch in the C-terminus of ATF-4, which contains the basic domain and leucine zipper. We further demonstrated that both full length and N-terminal truncated ATF-4 were able to enhance Tax transactivation. Thus, ATF-4 may act as an adapter between Tax and the TRE (Tax responsive element), and play an important role in Tax-mediated transactivation.

Activating Transcription Factor 4↗

Promoter analysis in living zebrafish embryos identifies a cis-acting motif required for neuronal expression of GATA-2.

We have used zebrafish embryos to dissect the promoter activity of a gene with a complex expression pattern during embryogenesis. GATA-2 is a transcription factor required for hematopoiesis and is dynamically expressed in hematopoietic tissues and in the central nervous system. Using constructs containing zebrafish GATA-2 genomic flanking sequences and the green fluorescent protein (GFP) reporter gene, we demonstrate that distinct regulatory domains are required for hematopoietic, enveloping layer (EVL), and neuronal expression of GATA-2. During gastrulation, GFP expression is confined to the ventral ectoderm and lateral mesoderm and is lacking in the dorsal shield. Cells derived from the regions expressing GFP give rise to hematopoietic progenitors, EVL cells, and neurons. Deletion analysis of the 7.3-kb GATA-2 promoter region revealed that a 1.1-kb DNA sequence is critical for expression of GATA-2 in neurons. Fine mapping revealed that a 31-bp region is required for neuron enhancer activity, and mutagenesis showed that the DNA motif CCCTCCT is essential for GATA-2 promoter activity in the central nervous system of zebrafish. Our use of zebrafish embryos can be exploited as a whole animal system for the dissection of any developmentally regulated vertebrate promoter.

Animals↗

A cellular cofactor for the constitutive transport element of type D retrovirus.

A human nuclear protein that specifically interacts with the constitutive transport element (CTE) of simian retrovirus was identified as adenosine 5'-triphosphate-dependent RNA helicase A. This protein could bind to functional CTE but not to inactive CTE mutants. The interaction of helicase A with CTE was distinct from previously described helicase activity of this protein. Helicase A shuttled from the nucleus to the cytoplasm in the presence of a transcription inhibitor or in cells transiently overexpressing CTE-containing RNA. In vivo colocalization of helicase A and CTE was observed in experiments that combined in situ hybridization and immunostaining. These results suggest that helicase A plays a role in the nuclear export of CTE-containing RNA.

Adenosine Triphosphate↗

Acid-Base Properties of Aqueous Illite Surfaces

In this paper, the acid-base properties of illite/water suspensions are examined using the constant capacitance surface complexation model. On the basis of results of potentiometric titrations and solubility experiments, we conclude that the proton reactions in the supernatants of illite suspensions can be successfully represented by proton reactions of Al(H2O)63+ and Si(OH)4 in water solutions. For illustrating the acidic characteristics of aqueous illite surfaces, two surface protonation models are proposed: (1) one site-one pKa model, identical withSOH right arrow over left arrow identical withSO- + H+, pKaint = 4.12-4.23; (2) two sites-two pKas model, identical withSIOH right arrow over left arrow identical withSIO- + H+, pKintaI = 4.17-4.44, and identical withSIIOH right arrow over left arrow identical withSIIO- + H+, [dipKintaII = 6.35-7.74. Evaluation of these two models indicates that both of them can give good descriptions of the experimental data of systems with different illite concentrations and ionic strengths and that the one site-one pKa model can be considered as a simplification of the two sites-two pKas model. Since both models assume only deprotonation reactions at the illite surfaces, they suggest that the surface behavior of the illite is similar to that of amorphous SiO2. Model assumptions, experimental procedures, and evaluative criteria are detailed in the paper.

Journal Article↗

Adsorption of Copper at Aqueous Illite Surfaces

In this paper, we conducted potentiometric titrations, batch adsorption experiments and FT-IR analysis to study the uptake of copper in illite/water suspensions and then applied the constant capacitance surface complexation model to interpret the reaction mechanism at the aqueous illite surfaces. Our research shows that the copper adsorption at these surfaces is strongly dependent on pH and that the adsorption causes a deprotonation of surface groups. We propose that the uptake of copper in the carbonate-free illite suspensions can be explained by the formation of mononuclear surface complexes, identical withSOCu+ and identical withSOCuOH, and a multinuclear surface complex, identical withSOCu2(OH)2+, followed by the formation of a bulk precipitate, Cu(OH)2(s), or a surface precipitate, identical withSOCu2(OH)3(sp). For the illite suspensions containing carbonates, we propose that the copper-illite interaction can be depicted by the formation of mononuclear surface complexes, identical withSOCu+ and identical withSOCuOH, followed by the formation of a copper hydroxylcarbonate precipitate, Cu2(OH)2CO3(s), rather than a copper hydroxide precipitate. The existence of Cu2(OH)2CO3(s) in the carbonate-containing illite suspensions was identified by FT-IR analysis.

Journal Article↗

Identification and purification of cellular proteins that specifically interact with the RNA constitutive transport elements from retrovirus D.

Human immunodeficiency virus (HIV) encodes a transacting protein, Rev, which interacts with an RNA element (RRE) to mediate nuclear export of unspliced viral mRNA. Recently, the RNA constitutive transport elements (CTE) from Mason-Pfizer monkey virus (MPMV) and simian retrovirus type I (SRV-1) were shown to render Rev-independent expression of gag, pol, or env genes in subgenomic constructs of HIV-1 and to support replication of HIV-1 mutants lacking RRE and Rev. Since CTEs act in cis, in the absence of any viral regulatory proteins, it is widely believed that they interact directly with the cellular export machinery by means of RNA-protein interaction. In this report, Electrophoretic mobility shift and UV-crosslinking assays were carried out to identify nuclear proteins that interact specifically with CTEs from both MPMV and SRV-1. Two of the four proteins (65 and 40 kDa, respectively) that bound CTE RNA did not interact in the same assays with RNA of a nonfunctional CTE mutant generated by site-directed mutagenesis. Both proteins have been partially purified. The nature of these proteins and their roles in RNA intracellular trafficking are currently under investigation.

Animals↗

Comparison of p53 expression in proximal and distal gastric cancer: histopathologic correlation and prognostic significance.

BACKGROUND: The overexpression of p53 has been found to be correlated with prognosis of some carcinomas, including gastric cancer, but no studies have reported on its relationship to the location of gastric cancer. In the present study, we compared the p53 expression of proximal and distal gastric cancer concerning histopathology and prognosis. METHODS: A total of 170 tumors in the patients with proximal (80 cases) and distal (90 cases) gastric cancer were studied by immunohistochemical methods. RESULTS: p53 immunopositivity was detected in 28.8% of all tumors. The p53-positive expression in proximal gastric cancer was higher than in distal gastric cancer (38.8% vs. 20.0%, p < 0.05). A 5-year survival analysis showed that there is no significant difference between tumors that are p53 positive and p53 negative. No correlation was found between p53 expression and histopathology of gastric cancer. CONCLUSION: p53 nuclear staining is not useful as a prognostic indicator or as a parameter in gastric cancer.

Adult↗

Extreme variation in X-linked agammaglobulinemia phenotype in a three-generation family.

BACKGROUND: X-linked agammaglobulinemia is typically a severe life-threatening disease characterized by the failure of B-cell differentiation and antibody production, which manifests in infancy and early childhood. Recently, we reported a novel mutation (Cys145-->STOP) in Bruton's tyrosine kinase in a 51-year-old man who was referred for evaluation because of chronic nasal congestion, recurrent sinusitis, sporadic pneumonia, and a family history suggestive of an X-linked immunodeficiency disease. He had not been treated with gammaglobulin. OBJECTIVE: This study was performed to investigate the clinical and immunologic phenotypes of this patient's other affected male family members. METHODS: A detailed family history and comprehensive review of medical records was carried out. Genetic mutation analysis of the gene encoding Bruton's tyrosine kinase was carried out in the proband's brother and nephew. RESULTS: Clinically affected male family members exhibit marked phenotypic variation with manifestations ranging from extremely mild to severe recurrent infections. Immunologic evaluation revealed extreme variation in immunoglobulin levels, B-cell numbers, and functional antibody titers. Genetic analysis documented a novel mutation in the gene encoding Bruton's tyrosine kinase in the proband, his brother, and his nephew. CONCLUSIONS: Despite their sharing the same genetic abnormality, extreme variation was noted in the immunologic findings and phenotypic expression of affected family members. This family study is extraordinary in that clinically affected male members who did not receive aggressive medical treatment died of the disease in childhood or survived into late adulthood.

Adult↗

Intravenous administration of deaggregated mouse thyroglobulin suppresses induction of experimental autoimmune thyroiditis and expression of both Th1 and Th2 cytokines.

Experimental autoimmune thyroiditis (EAT) can be induced by transfer of mouse thyroglobulin (MTg) and lipopolysaccharide (LPS)-immunized, MTg-activated spleen cells into syngeneic recipients. Recipients of MTg-activated T cells develop lymphocytic EAT, whereas recipients of cells activated by MTg and anti-IL-2R antibody develop a more severe and histologically distinct granulomatous form of EAT. Intravenous administration of deaggregated MTg (dMTg) 7 days before and 5 days after the first immunization of donor mice with MTg/LPS suppresses the induction of both forms of EAT. Thyroid infiltration is significantly decreased in recipients of effector cells from tolerant donors. MTg-specific T cell proliferation is partially suppressed in tolerant mice. Both IgG1 and IgG2a subclasses of anti-MTg autoantibodies are markedly inhibited in both tolerant donor mice and in recipients of tolerant spleen cells. Expression of T cell cytokine gene transcripts (IL-2, IFN gamma, IL-4, IL-10, tumor necrosis factor-alpha and transforming growth factor-beta) are all decreased in spleen cells of donor mice given dMTg. CD8+ T cells were not required for expression of tolerance since depletion of CD8+ T cells in vivo before tolerance induction or in vitro before MTg re-stimulation did not abrogate tolerance induction. Taken together, these results suggest that i.v. administration of dMTg can induce MTg-specific tolerance in both Th1- and Th2-like EAT effector cell precursors, and therefore prevent induction of adoptively transferred EAT.

Adoptive Transfer↗

Induction of experimental antiphospholipid antibody syndrome in PL/J mice following immunization with beta 2 GPI.

PROBLEM: Immunization with beta 2-glycoprotein I (beta 2 GPI) induces antiphospholipid antibodies (aPL) in normal mice and rabbits. Recently we reported early onset of autoimmunity in MRL/(+2) mice following immunization with beta 2 GPI. There is a close association between aPL with thrombosis, recurrent fetal loss, and intrauterine growth retardation. In this study we evaluated the effect of beta 2 GPI-induced aPL on pregnancy outcomes in an inbred strain of mice (PL/J). METHOD: Three groups of seven-week female PL/J mice (12 per group) were studied. Group A was immunized with beta 2 GPI and group B with ovalbumin; group C was not not immunized. After two booster injections, the mice were tested for aPL, anti-DNA by ELISA, and for ANA by indirect immunofluorescence. Platelet count and pregnancy outcomes were studied at the age of 14 weeks. RESULTS: The aPL and anti-DNA levels were higher at 12 and 14 weeks in group A; the optical densities (OD) were 1.72 +/- 0.6 and 0.699 +/- 0.25 for group A, 0.09 +/- 0.040 and 0.230 +/- 0.47 for group B, and 0.0435 +/- 0.003 and 0.119 +/- 0.26 for group C (comparing group A with groups B and C combined, P < 0.001). ANA titers rose in groups A and B by age, but they were significantly higher at 14 weeks in group A. The mean titers were 1/286, 1/90, and 1/16 for A, B, and C, respectively (P < 0.001). The platelet counts were not significantly different among the three groups. The titer size was significantly smaller in group A, as evidenced by the numbers of viable fetuses among the mice that became pregnant in each group: 0.75, 2.45, and 5.5 in groups A, B, and C, respectively. Seven pregnant mice in group A had complete resorption, seven pregnant mice in group B showed focal (partial) resorption areas, by only one mouse in group C had complete resorption of the embryos, as shown by histopathological studies, although the fecundity rate was similar in the three groups. CONCLUSION: Our data suggest a pathogenic role for beta 2 GPI-induced aPL in the development of experimental models of APS in PL/J mice.

Animals↗

Analysis of osteocalcin expression in transgenic mice reveals a species difference in vitamin D regulation of mouse and human osteocalcin genes.

A line of transgenic mice expressing a human osteocalcin genomic fragment (hOClocus) and a murine MC3T3-E1 cell line containing a stably integrated human osteocalcin promoter construct have been developed to characterize the osteogenic and hormonal regulation of human osteocalcin in vivo and in vitro. In this study, we used these models to demonstrate a species difference in the regulation of the mouse and human osteocalcin genes by vitamin D. Repeated administration of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) to mice carrying the hOClocus transgene resulted in striking increases in serum human osteocalcin, whereas serum mouse osteocalcin levels were unchanged after 24 h and only modestly increased 48 h after the second dose of hormone. 1,25(OH)2D3 increased human calvarial mRNA expression by 1.8-fold and slightly decreased mouse osteocalcin mRNA levels by approximately 1.2-fold. Furthermore, treatment of primary calvarial osteoblasts from these mice with 1,25(OH)2D3 increased human osteocalcin production but inhibited mouse osteocalcin protein accumulation. To investigate further the mechanism for the apparent species difference in vitamin D3 induction of mouse and human osteocalcin, we examined the effect of 1,25(OH)2D3 in an MC3T3-E1 cell line (MC4) containing a stably integrated 3900 bp osteocalcin promoter-luciferase construct. Treatment of MC4 cells with ascorbic acid resulted in parallel increases of the endogenous mouse osteocalcin protein and luciferase reporter activity over a 12-day period. Continuous exposure of MC4 cells to 1,25(OH)2D3 resulted in time-and dose-dependent increases in the activity of the phOC3900 luciferase construct. By contrast, the hormone had no effect on mouse osteocalcin protein concentrations and inhibited its induction by ascorbic acid. However, when cells were treated acutely with 1,25(OH)2D3 at later times during growth in ascorbic acid, the induction of mouse osteocalcin protein was only partially inhibited. In conclusion, our results indicate that common osteogenic signals regulate both mouse and human osteocalcin gene expression, but the mouse gene is resistant to induction by vitamin D. This species difference in vitamin D regulation of osteocalcin appears to result from the failure of 1,25(OH)2D3 to transcriptionally activate the mouse osteocalcin gene.

3T3 Cells↗

[Detection of p53 gene mutations in hepatocellular carcinoma].

This study screened 32 cases of hepatocellular carcinoma (HCC) from Chengdu detecting HBV DNA. HBsAg and p53 mutations by using Southern blot hybridization, immunohistochemistry and polymerase chain reaction/restriction enzyme digest methods, respectively. The results revealed all the cases had been infected by HBV; the frequency of HBV DNA integration into HCC cell was 72%, and the positive staining for HBsAg 96.3%(26/27). Of the 32 cases, 8 showed nuclear staining of p53 protein (25%), no mutation of p53 gene at all in 27 nontumorours liver tissues was identified. The resluts suggest the inactivation of p53 function may play a significant role in the genesis of HBV-associated HCC; however, the largely negative p53 mutation results in the current study conversely indicate that hepatocarcinogenesis may even involve other comprehensive mechanisms. Further studies are worth doing to evaluate the possible contribution of HBV to the p53 mutation in HCC.

Adult↗

[The bred technique of Zaocys dhumnades].

This paper reports the observational data on the laying egg, hatching, life habit, relationship between capacity for eating and growth speed of Zaocys dhumnades bred in enclosure-like garden of snake, summarizes an available breeding technique.

Animal Feed↗

[Determination of glutamine in intestinal mucosa by pre-column derivatization/reversed-phase high performance liquid chromatography with fluorescence detection].

A high performance liquid chromatographic pre-column derivatization method with fluorescence detection for the analysis of glutamine (Gln) in rat intestinal mucosa is presented. Gln was derivatized with o-phthalaldehyde and 3-mercaptopropionic acid under an alkaline condition and separated by reversed-phase liquid chromatography on a Lichrosorb RP18 column (150 mm x 4.6 mm i.d., 5 microm). The mobile phase was consisted of 50 mmol/L phosphate buffer (pH 7.0)-acetonitrile (94:6, V/V) with a flow rate of 2.0 mL/min. The excited and emitted wavelength were selected at 230 nm and 389 nm respectively. The volume ratio of samples and derivatization reagent solution was 4:1 (V/V). The detection limit of Gln was 25 micromol/L (S/N= 3.5) and the regression equation was A = 16.9405C + 179.9339, r = 0.9996 at the linear range of 50-3200 micromol/L. The day-to-day deviation was 6.97% (n = 3) and the retention time of Gln was 3.158 min. This method is rapid, simple and highly sensitive, and has been applied to the determination of Gln in intestinal mucosa.

Animals↗

Correlation between retinal fluorescein angiography and blood viscosity and other factors in patients with primary open angle glaucoma.

OBJECTIVE: To investigate the correlation among retinal fluorescein angiography, blood viscosity, and other factors in patients with open angle glaucoma (POAG). METHODS: Multiple step regression analysis was made to investigate the correlation between each of the following blood vessel filling times: the arm-choroid (A-CT), arm-retinal artery (A-AT), and retinal artery-venous (A-VT) of the fundus fluorescein angiography (FFA) in 122 eyes with POAG and each of the following related factors in hemorrheology: whole blood apparent viscosity at low, medium and high shear rates, plasma viscosity and hematocrit. Also, the same analysis was applied to investigate the correlation between each of the A-AT, A-VT of the FFA in 70 eyes with POAG and the following factors: systolic blood pressure, diastolic blood pressure, age and whole blood apparent viscosity at low shear rate. RESULTS: The whole blood apparent viscosity at low shear rate was closely related to A-CT and A-AT, while hematocrit was closely related to A-VT of the FFA. The whole blood apparent viscosity at low shear rate and age, especially the whole blood apparent viscosity, was closely related to A-AT, A-VT of the FFA. CONCLUSION: Blood viscosity can affect the filling times of the FFA in POAG.

Blood Viscosity↗