Biomedical subjects

Z Zeng

Publications and source records attributed to Z Zeng.

At least 55 records · Page 3Linked to original sources

PubMed2000

Characterization of the human cysteinyl leukotriene 2 receptor.

The contractile and inflammatory actions of the cysteinyl leukotrienes (CysLTs), LTC(4), LTD(4), and LTE(4), are thought to be mediated through at least two distinct but related CysLT G protein-coupled receptors. The human CysLT(1) receptor has been recently cloned and characterized. We describe here the cloning and characterization of the second cysteinyl leukotriene receptor, CysLT(2), a 346-amino acid protein with 38% amino acid identity to the CysLT(1) receptor. The recombinant human CysLT(2) receptor was expressed in Xenopus oocytes and HEK293T cells and shown to couple to elevation of intracellular calcium when activated by LTC(4), LTD(4), or LTE(4). Analyses of radiolabeled LTD(4) binding to the recombinant CysLT(2) receptor demonstrated high affinity binding and a rank order of potency for competition of LTC(4) = LTD(4) LTE(4). In contrast to the dual CysLT(1)/CysLT(2) antagonist, BAY u9773, the CysLT(1) receptor-selective antagonists MK-571, montelukast (Singulair(TM)), zafirlukast (Accolate(TM)), and pranlukast (Onon(TM)) exhibited low potency in competition for LTD(4) binding and as antagonists of CysLT(2) receptor signaling. CysLT(2) receptor mRNA was detected in lung macrophages and airway smooth muscle, cardiac Purkinje cells, adrenal medulla cells, peripheral blood leukocytes, and brain, and the receptor gene was mapped to chromosome 13q14, a region linked to atopic asthma.

Adrenal Medulla↗

PubMed2000

Inducers of gamma-glutamylcysteine synthetase and their effects on glutathione synthetase expression.

Synthesis of GSH occurs via two enzymatic steps, the first is catalyzed by gamma-glutamylcysteine synthetase (GCS) and the second is catalyzed by GSH synthetase (GS). A heavy (HS) and light subunit (LS) make up GCS; regulation of both subunits have been well characterized, whereas regulation of GS is largely unknown. In this study, we examined the effects of treatments known to influence the gene expression of GCS subunits on GS expression. Insulin and hydrocortisone treatment of rat hepatocytes or ethanol-feeding of rats for 9 weeks, which increased the expression of GCS-HS only, had no influence on GS expression. However, two-thirds partial hepatectomy in rats which increased the expression of GCS-HS only, also increased GS expression. Treatment of hepatocytes or rats with diethyl maleate, buthionine sulfoximine, tert-butylhydroquinone, or thioacetamide, which increased the expression of both GCS subunits, increased the expression of GS. The GSH synthesis capacity increased 50-100% by treatments that increased only the GCS-HS expression, whereas it increased 161-200% by treatments that increased both GCS-HS and GS expression. Thioacetamide treatment of Chang cells increased cell GSH and GS expression by 50%, but had minimal influence on GCS subunits. Thus, GS induction can further increase the cell's GSH synthetic capacity and in some cells may be as important as GCS in determining the rate of GSH synthesis.

Animals↗

PubMed2000

Identification of receptors for neuromedin U and its role in feeding.

Neuromedin U (NMU) is a neuropeptide with potent activity on smooth muscle which was isolated first from porcine spinal cord and later from other species. It is widely distributed in the gut and central nervous system. Peripheral activities of NMU include stimulation of smooth muscle, increase of blood pressure, alteration of ion transport in the gut, control of local blood flow and regulation of adrenocortical function. An NMU receptor has not been molecularly identified. Here we show that the previously described orphan G-protein-coupled receptor FM-3 (ref. 15) and a newly discovered one (FM-4) are cognate receptors for NMU. FM-3, designated NMU1R, is abundantly expressed in peripheral tissues whereas FM-4, designated NMU2R, is expressed in specific regions of the brain. NMU is expressed in the ventromedial hypothalamus in the rat brain, and its level is significantly reduced following fasting. Intracerebroventricular administration of NMU markedly suppresses food intake in rats. These findings provide a molecular basis for the biochemical activities of NMU and may indicate that NMU is involved in the central control of feeding.

Amino Acid Sequence↗

PubMed2000

Cloning and functional characterization of the 5'-flanking region of human methionine adenosyltransferase 1A gene.

Methionine adenosyltransferase (MAT) is an essential cellular enzyme which catalyses the formation of S-adenosylmethionine, the principal methyl donor and precursor for polyamines. In mammals, two different genes, MAT1A and MAT2A, encode for liver-specific and non-liver-specific MAT respectively. We previously described a switch in the MAT expression from MAT1A to MAT2A in human liver cancer, which offered the cancerous cell a growth advantage. Loss of MAT1A expression was due to lack of gene transcription. To study regulation of the MAT1A gene, we have cloned and characterized a 1.9 kb 5'-flanking region of the human MAT1A gene. One transcriptional start site, located 25 nt downstream from a consensus TATA box, was identified by primer extension and RNase protection assays. The promoter contains several consensus binding sites for CAAT enhancer binding protein (C/EBP) and hepatocyte-enriched nuclear factor (HNF), transcriptional factors important in liver-specific gene expression. The human MAT1A promoter was able to efficiently drive luciferase expression in Chang cells, a human liver cell line, but not in HeLa cells. Sequential deletion analysis of the promoter revealed two DNA regions upstream of the translational start site, -705 to -839 bp and -1111 to -1483 bp, which are involved in positive and negative gene regulation, respectively. Specific protein binding to these regions was confirmed by electrophoretic-mobility-shift and DNase I footprinting assays. Similar to the situation with the rat MAT1A, glucocorticoid treatment also increased human MAT1A expression and promoter activity in a dose- and time-dependent manner.

5' Untranslated Regions↗

PubMed2000

Siglec-8. A novel eosinophil-specific member of the immunoglobulin superfamily.

We describe the characterization of siglec-8, a novel sialic acid-binding immunoglobulin-like lectin that is expressed specifically by eosinophils. A full-length cDNA encoding siglec-8 was isolated from a human eosinophil cDNA library. Siglec-8 is predicted to contain three extracellular immunoglobulin-like domains, a transmembrane region, and a cytoplasmic tail of 47 amino acids. The siglec-8 gene mapped on chromosome 19q13.33-41, closely linked to genes encoding CD33 (siglec-3), siglec-5, siglec-6, and siglec-7. When siglec-8 was expressed on COS cells or as a recombinant protein fused to the Fc region of human IgG(1), it was able to mediate sialic acid-dependent binding to human erythrocytes and to soluble sialoglycoconjugates. Using specific monoclonal antibodies, siglec-8 could be detected only on eosinophils and hence appears to be the first example of an eosinophil-specific transmembrane receptor.

Amino Acid Sequence↗

PubMed2000

Aspartoacylase gene transfer to the mammalian central nervous system with therapeutic implications for Canavan disease.

With the ultimate goal of developing safe and effective in vivo gene therapy for the treatment of Canavan disease and other neurological disorders, we developed a non-viral lipid-entrapped, polycation-condensed delivery system (LPD) for central nervous system gene transfer, in conjunction with adeno-associated virus (AAV)-based plasmids containing recombinant aspartoacylase (ASPA). The gene delivery system was tested in healthy rodents and primates, before proceeding to preliminary studies in 2 children with Canavan disease. Toxicity and expression testing was first carried out in human 293 cells, which demonstrated effective transduction of cells and high levels of functional ASPA activity. We performed in vivo toxicity and expression testing of LPD/pAAVaspa and LPD/pAAVlac in rodents, which demonstrated widespread gene expression for more than 10 months after intraventricular delivery, and local expression in deep brain nuclei and white matter tracts for more than 6 months after intraparenchymal injections, with no significant adverse effects. We also performed intraventricular delivery of LPD/pAAVaspa to 2 cynomologous monkeys, with 2 additional monkeys receiving LPD and saline controls. None of the monkeys demonstrated significant adverse effects, and at 1 month the 2 LPD/pAAVaspa monkeys were positive for human ASPA transcript by reverse transcriptase polymerase chain reaction of brain tissue punches. Finally, we performed the first in vivo gene transfer study for a human neurodegenerative disease in 2 children with Canavan disease to assess the in vivo toxicity and efficacy of ASPA gene delivery. Our results suggest that LPD/pAAVaspa is well tolerated in human subjects and is associated with biochemical, radiological, and clinical changes.

Amidohydrolases↗

PubMed2000

[A new potential promoter region of the human connexin 26 gene].

OBJECTIVE: To explain regulatory mechanism of the human connexin 26(Cx26) gene. METHODS: A DNase-1 hypersensitive 1.6 kb fragment upstream of the 5'-terminus of the Cx26 gene was sequenced and assayed with CAT reporter system. RESULTS: The Cx26-1.6 kb sequence serves as a powerful promoter. CONCLUSION: The Cx26-1.6 kb fragment contains two GT boxes (centering at-6158 and -6213 bp), and a TATA-less TTAAAA box (-6237/-6232 bp) which is another promoter region of the human Cx26.

Base Sequence↗

PubMed2000

Detailed allelic loss mapping on 7q32 in nasopharyngeal carcinoma.

OBJECTIVE: The present research group had found high frequency of loss of heterozygosity(LOH) of D7S500-D7S495 on chromosome band 7q32 in nasopharyngeal carcinoma, this study was conducted to further find the deletion region. METHODS: Higher density of loci on 7q32 in 30 tumors was studied by using microsatellite analysis. RESULTS: The frequency of LOH was 63.3%. The highest frequency of LOH was identified at the loci of D7S500-D7S509-D7S495, among which a common deletion of D7S509 was found. CONCLUSION: A putative tumor suppressor gene may be present around D7S509.

Chromosomes, Human, Pair 7↗

PubMed2000

[Clinical analysis of 224 cases of small intestine disease].

OBJECTIVE: To increase the knowledge of small intestine disease, improve the diagnostic rate and discusses the examination methods. METHODS: 224 cases of small intestine disease were studied and their major symptoms, etiology, diagnostic methods were analyzed. RESULTS: The major symptoms of small intestine disease were abdominal pain (58%), abdominal mass (18%), fever (17%) and hematochezia (12%). The most common disease of small intestine was malignant tumor (32%), others were diverticulum (29%), leiomyomas (7%), hemorrhagic necrotizing enteritis (6%), Crohn's disease (4%). Double-contrast enteroclysis and laparotomy were still the major diagnostic methods. CONCLUSION: Malignant tumor was the most common disease of small intestine. More attention should be paid to the diagnosis of small intestine disease.

Adolescent↗

PubMed2000

[Prevalence of cag A and vac A subtypes of Helicobacter pylori in Guangzhou].

OBJECTIVE: To investigate the prevalence of cytotoxin associated gene (cag A) and vacuolating cytotoxin gene (vac A) subtypes of Helicobacter pylori (Hp) from patients with upper gastrointestinal diseases in Guangzhou, and to assess the relationship between cag A, vac A subtypes and Hp related gastrointestinal diseases. METHODS: 191 Hp strains were isolated from patients with different gastrointestinal diseases in Guangzhou. Bacterial DNA from all these strains was extracted. cag A and vac A alleles were typed using PCR with specific primers. RESULTS: In Guangzhou, Hp cag A positive rate was 85.3% (163/191). cag A positive rate was significantly higher in gastric cancer and peptic ulcer than that in chronic gastritis (P < 0.05). There were six vac A mosaicisms, including 168 for s1a/m2 (88.0%), 14 for s1a/m1b (7.3%), 6 for s1b/m2 (3.1%), 1 for s1b/m1b (0.5%), 1 for s2/m2 (0.5%) and 1 for s1a/m1b-m2 (0.5%). The rate of vac A s1a/m2 subtype in chronic gastritis, peptic ulcer and gastric carcinomas was 87.5% (70/80), 87.5% (84/96) and 93.3% (14/15) respectively. There was no significant relationship between cag A and vac A subtypes (P > 0.05). CONCLUSION: cag A(+) and vac A s1a/m2 subtype of Hp is predominant in patients with upper gastrointestinal diseases in Guangzhou, cag A(+) strains are further more common in patients with gastric cancer and peptic ulcer.

Adolescent↗

PubMed2000

[Cloning and expression analysis of human dystonia/deafness peptide like gene].

OBJECTIVE: To clone a new deafness associated gene. METHODS: Molecular database search, RACE and cDNA library screen were applied. RESULTS: A new gene, named dystonia/deafness peptide like (DDPL) (GeneBank Accession Number: DDPL2 AF165967) and mapped to 11q22.1-22.2, was cloned. DDPL had two different mRNA splicing types and accordingly encoded 83 and 51 amino acids. DDPL transcripts were detected in a range of adult and fetal tissues. A pseudogene of DDPL, named DDPLphi which was shown lying within Xq25-q26, showed 96.9% sequence identity with DDPL1 cDNA sequence across 382 bp. CONCLUSION: DDPL lies within the 5.2 Mb interval of 11q22.1-q22.2 between D11S939 and D11S1347, and has a high identity with the DFN-1/MTS associated gene-DDP.

Adult↗

PubMed2000

[The effects of lipoproteins on human vascular endothelial gap junctional intercellular communication].

The effects of HDL, LDL and ascorbic acid on cultured human vascular endothelial gap junctional intercellular communication (GJIC) were observed using fluorescence recovery after photobleaching assay under confocal laser scanning microscopy in this study. The results showed that in culture, human vascular endothelial cells could produce functional gap junctions. The fluorescence recovery rates of the LDL group were 6.691 +/- 2.634, 4.153 +/- 2.125 and 2.441 +/- 0.720 respectively; when these rates were compared with the control and with each other, significant differences were found(P < 0.05). When the fluorescence recovery rates of the HDL group and Vitamin C group were compared with the control, no significant differences were found(P > 0.05). The fluorescence recovery rates of the LDL + HDL group were 3.500 +/- 0.890, 4.339 +/- 1.126 and 5.243 +/- 1.278(n = 15) respectively, when these rates were compared with the control and with each other, significant differences were found (P < 0.05). These suggested that LDL should be able to inhibit GJIC in human vascular endothelial cells, and the inhibition could be diminished by HDL or ascorbic acid. Inhibition of vascular endothelial GJIC may be one of the atherosclerogenic mechanisms of LDL, whereas HDL and ascorbic acid may be conducive to the improvement of endothelial GJIC. But it may also be only a compensatory behavior of endothelial cells to injuries.

Arteriosclerosis↗

PubMed2000

[Transthoracic three-dimensional echocardiography for determination of left ventricular mass in patients with dilated cardiomyopathy].

This study was performed to compare left ventricular mass (LVM) measurement by one- (1D), two- (2D) and 3D echocardiography with magnetic resonance imaging (MRI) in patients with dilated cardiomyopathy (DCM). 36 patients with DCM underwent imaging by conventional 1D and 2D echocardiography as well as transthoracic 3D echocardiographic data acquisition. Also patients were imaged with cardiac MRI. LVM was determined. There was no significant correlation (r = 0.42, P > 0.05) for measuring LVM between 1D echocardiography and MRI in patients with DCM. A significant correlation was obtained between 2D echocardiography and MRI (r = 0.64, P < 0.01) and between 3D and MRI (r = 0.78, P < 0.01) in determination of LVM. Compared with 1D and 2D echocardiography, the 3D analysis achieved a significantly higher agreement with the MRI results (1D; 399.2 g, 2D: 285.9 g, 3D: 172.6 g versus MRI: 199.1 g). Interobserver variability was 5.1% for measuring LVM by 3D echocardiography. These results suggest that LVM of patients with DCM can be accurately determined by transthoracic 3D echocardiography and this method could be used in clinical practice.

Adolescent↗

PubMed2000

[Partial laryngectomy plus radiotherapy versus partial laryngectomy alone for laryngeal carcinoma].

OBJECTIVE: To investigate the effect of radiotherapy in the combined treatment of laryngeal cancer with partial laryngectomy. METHODS: One hundred and seventy-one cases of laryngeal cancer were randomly separated into three groups of partial laryngectomy, partial laryngectomy with preoperative radiotherapy (dose 40-50Gy) and partial laryngectomy with postoperative radiotherapy(dose 51-70 Gy). The surgical procedures included cordectomy, vertical partial laryngectomy, horizontal partial laryngectomy and near total laryngectomy. RESULTS: The overall 5-year survival rate was 82% (123/150). The 5-year survival rates for surgery alone group (56 cases), surgery with preoperative radiotherapy group(65 cases) and surgery with postoperative radiotherapy group(50 cases) were 85.7%, 80.7% and 79.5% respectively. There was no significant difference among the groups(chi 2 = 0.703, P = 0.704). Analysis of survival rates revealed a significant difference among the different stages(chi 2 = 12.248, P = 0.007). All 171 cases except 3 that had near total laryngectomy achieved satisfactory phonation. CONCLUSION: Neither preoperative radiotherapy nor postoperative radiotherapy can improve the 5-year survival rate of laryngeal cancer with partial laryngectomy. All patients with partial laryngectomy got satisfactory phonation.

Adolescent↗

PubMed2000

[The measurement of temperature with Raman scattering spectra of polycrystal (SrNO3)2].

We have developed a new method for the determination of temperature with Raman scattering spectra. The strontium nitrate was chosen as an experiment substance. It has two bands of Raman scattering, and their wavenumbers are 738 and 1,057 cm-1 in 600-1,700 cm-1. We have obtained the intensities of their Stokes and Anti-Stokes scattering, then we have got two temperatures of strontium nitrate according to the formula. The result from the 738 cm-1 scattering band has larger error than that from the 1,057 cm-1. This may account for the noise of the photocounter. This no touch method is fit to measure the temperature of the molecular.

Crystallization↗

PubMed1999

Identification of urotensin II as the endogenous ligand for the orphan G-protein-coupled receptor GPR14.

Urotensin II (UII) is a neuropeptide with potent cardiovascular effects. Its sequence is strongly conserved among different species and has structural similarity to somatostatin. No receptor for UII has been molecularly identified from any species so far. GPR14 was cloned as an orphan G protein-coupled receptor with similarity to members of the somatostatin/opioid receptor family. We have now demonstrated that GPR14 is a high affinity receptor for UII and designate it UII-R1a. HEK293 cells and COS-7 cells transfected with rat GPR14 showed strong, dose-dependent calcium mobilization in response to fish, frog, and human UII. Radioligand binding analysis showed high affinity binding of UII to membrane preparations isolated from HEK293 cells stably expressing rat GPR14. In situ hybridization analysis showed that GPR14 was expressed in motor neurons of the spinal cord, smooth muscle cells of the bladder, and muscle cells of the heart. The identification of the first receptor for UII will allow better understanding of the physiological and pharmacological roles of UII.

Amino Acid Sequence↗

PubMed1999

Characterization of the human cysteinyl leukotriene CysLT1 receptor.

The cysteinyl leukotrienes-leukotriene C4(LTC4), leukotriene D4(LTD4) and leukotriene E4(LTE4)-are important mediators of human bronchial asthma. Pharmacological studies have determined that cysteinyl leukotrienes activate at least two receptors, designated CysLT1 and CysLT2. The CysLT1-selective antagonists, such as montelukast (Singulair), zafirlukast (Accolate) and pranlukast (Onon), are important in the treatment of asthma. Previous biochemical characterization of CysLT1 antagonists and the CysLT1 receptor has been in membrane preparations from tissues enriched for this receptor. Here we report the molecular and pharmacological characterization of the cloned human CysLT1 receptor. We describe the functional activation (calcium mobilization) of this receptor by LTD4 and LTC4, and competition for radiolabelled LTD4 binding to this receptor by the cysteinyl leukotrienes and three structurally distinct classes of CysLT1-receptor antagonists. We detected CysLT1-receptor messenger RNA in spleen, peripheral blood leukocytes and lung. In normal human lung, expression of the CysLT1-receptor mRNA was confined to smooth muscle cells and tissue macrophages. Finally, we mapped the human CysLT1-receptor gene to the X chromosome.

Amino Acid Sequence↗

PubMed1999

Mitochondrial DNA deletions are associated with ischemia and aging in Balb/c mouse brain.

Deletions in the mitochondrial DNA (mtDNA) of Balb/c mouse cerebrums, resembling deletions found in elderly humans or in patients with certain disorders, were detected by PCR. Analysis was carried out on mice of various ages and on mice in which the bilateral common carotid arteries had been incompletely ligated to reconstruct cerebral ischemia. A 3,867 bp mtDNA deletion was present only in old or ischemic mouse groups. Among the non-ischemic groups, it was found in 0 of 12 weaning, 0 of 12 young, and four of eight old mice. Among the ischemic groups, it was found in 12 of 17 young and 11 of 11 old mice. Moreover, the percentage of total mtDNA containing deletions was 22% for the old non-ischemic group, 37% for the young ischemic group, and 69% for the old ischemic group. In addition, PCR analysis detected two other deletions of 3,726 bp and 4,236 bp in 4 of the 11 old ischemic cerebrums. The results indicate that mtDNA deletions are associated with aging, that ischemia increases the incidence of mtDNA deletions, and that mtDNA deletions resulting from ischemia are more likely to occur in old mice than in young mice.

Age Factors↗