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Biomedical subjects

Hong Chen

Publications and source records attributed to Hong Chen.

At least 235 records · Page 13Linked to original sources
PubMed2002

Microchip capillary electrophoresis with electrochemical detection.

A novel microchip capillary electrophoresis system with electrochemical detection, using the replaceable microelectrode, is first reported. This kind of electrode can be fabricated in general laboratories and can be replaced quickly with electrodes of different materials according to the requirements of experiments. The end-column electrochemical detection on microchip CE was achieved by fixing the working electrode (such as carbon fiber, Pt, or Au, etc.) through a guide tube on the end of the separation channel. The experiment results indicate that the alignment of the electrode with the channel outlet can be carried out accurately and reproducibly, and therefore, the detection device has low noise and good reproducibility. The detection limit of dopamine is 2.4 x 10(-7) M, which is the lowest result reported so far. The separation and detection of dopamine, 5-hydroxytryptamine and epinephrine using carbon fiber and Pt microdisk electrodes within 50 s was successfully performed.

Biogenic Amines↗
PubMed2002

ATF4 is a mediator of the nutrient-sensing response pathway that activates the human asparagine synthetase gene.

Transcription from the asparagine synthetase (A.S.) gene is increased in response to either amino acid (amino acid response) or glucose (endoplasmic reticulum stress response) deprivation. These two independent pathways converge on the same set of genomic cis-elements within the A.S. promoter referred to as nutrient-sensing response elements (NSRE) 1 and 2, both of which are necessary for gene activation. The NSRE-1 sequence was used to screen ATF/CREB family members by electrophoresis mobility shift assays and supershift by specific antibodies. The results indicated that ATF4 binds to the NSRE-1 sequence and that the amount of the ATF4 complex was increased when extracts from amino acid-deprived or glucose-deprived cells were tested. Using electrophoresis mobility shift assay experiments and a probe that contained both NSRE-1 and NSRE-2, mutation of the NSRE-1 sequence completely prevented formation of the ATF4-containing complexes, whereas mutation of the NSRE-2 sequence did not. Overexpression of ATF4 increased A.S. promoter-driven transcription, whereas an inhibitory dominant negative ATF4 mutant blocked both basal and starvation-enhanced transcription. Collectively, the results provide both in vitro and in vivo evidence for a role of ATF4 in the transcriptional activation of the A.S. gene in response to nutrient deprivation.

Activating Transcription Factor 4↗
PubMed2002

A single motif responsible for ubiquitin recognition and monoubiquitination in endocytic proteins.

Ubiquitination is a post-translation modification in which ubiquitin chains or single ubiquitin molecules are appended to target proteins, giving rise to poly- or monoubiquitination, respectively. Polyubiquitination targets proteins for destruction by the proteasome. The role of monoubiquitination is less understood, although a function in membrane trafficking is emerging, at least in yeast. Here we report that a short amino-acid stretch at the carboxy-termini of the monoubiquitinated endocytic proteins Eps15 and eps15R is indispensable for their monoubiquitination. A similar sequence, also required for this modification, is found in other cytosolic endocytic proteins, such as epsins and Hrs. These sequences comprise a protein motif, UIM (ref. 6), which has been proposed to bind to ubiquitin. We confirm this for the UIMs of eps15, eps15R, epsins and Hrs. Thus, the same motif in several endocytic proteins is responsible for ubiquitin recognition and monoubiquitination. Our results predict the existence of a UIM:ubiquitin-based intracellular network. Eps15/eps15R, epsins and Hrs may function as adaptors between ubiquitinated membrane cargo and either the clathrin coat or other endocytic scaffolds. In addition, through their own ubiquitination, they may further contribute to the amplification of this network in the endocytic pathway.

Adaptor Proteins, Signal Transducing↗
PubMed2002

Superluminal pulse propagation through one-dimensional photonic crystals with a dispersive defect.

The propagation of a pulse through one-dimensional photonic crystals that contain a dispersive and absorptive defect layer doped with two-level atoms is discussed. The dynamical evolution of the pulse inside the photonic crystal is presented. Superluminal negative group velocity (the peak appears at the exit end before it reaches the input end) is discovered. Although the group velocity is larger than c and even negative, the velocity of energy propagation never exceeds the vacuum light speed. The appearance of the superluminal advance or subluminal delay of the pulse peak inside the photonic crystal or at the exit end is due to the wave interference from Bragg reflections.

Journal Article↗
PubMed2002

The ENTH domain.

The epsin NH2-terminal homology (ENTH) domain is a membrane interacting module composed by a superhelix of alpha-helices. It is present at the NH2-terminus of proteins that often contain consensus sequences for binding to clathrin coat components and their accessory factors, and therefore function as endocytic adaptors. ENTH domain containing proteins have additional roles in signaling and actin regulation and may have yet other actions in the nucleus. The ENTH domain is structurally similar to the VHS domain. These domains define two families of adaptor proteins which function in membrane traffic and whose interaction with membranes is regulated, in part, by phosphoinositides.

Amino Acid Sequence↗
PubMed2002

Lipid rafts orchestrate signaling by the platelet receptor glycoprotein VI.

The platelet collagen receptor glycoprotein VI (GPVI) couples to the immune receptor adaptor Fc receptor gamma-chain (FcRgamma) and signals using many of the same intracellular signaling molecules as immune receptors. Studies of immune receptor signaling have revealed a critical role for specialized areas of the cell membrane known as lipid rafts, which are enriched in essential signaling molecules. However, the role of lipid rafts in signaling in nonimmune cells such as platelets remains poorly defined. This study shows that GPVI-FcRgamma does not constitutively associate with rafts, but is recruited to lipid rafts following receptor stimulation in both GPVI-expressing RBL-2H3 cells and human platelets. FcRgamma is required for GPVI association with lipid rafts, as mutant GPVI receptors that do not couple to FcRgamma were unable to associate with lipid rafts after receptor clustering. Following GPVI stimulation in platelets, virtually all phosphorylated FcRgamma was found in lipid rafts, but inhibition of FcRgamma phosphorylation did not block receptor association with lipid rafts. This work demonstrates that lipid rafts orchestrate GPVI receptor signaling in platelets in a manner analogous to immune cell receptors and supports a model of GPVI signaling in which FcRgamma phosphorylation is controlled by ligand-dependent association with lipid rafts.

3T3 Cells↗
PubMed2002

The mechanism of growth-inhibitory effect of DOC-2/DAB2 in prostate cancer. Characterization of a novel GTPase-activating protein associated with N-terminal domain of DOC-2/DAB2.

DOC-2/DAB2 is a member of the disable gene family with tumor-inhibitory activity. Its down-regulation is associated with several neoplasms, and serine phosphorylation of its N terminus modulates DOC-2/DAB2's inhibitory effect on AP-1 transcriptional activity. We describe the cloning of DIP1/2, a novel gene that interacts with the N-terminal domain of DOC-2/DAB2. DIP1/2 is a novel GTPase-activating protein containing a Ras GTPase-activating protein homology domain (N terminus) and two other unique domains (i.e. 10 proline repeats and leucine zipper). Interaction between DOC-2/DAB2 and DIP1/2 is detected in normal tissues such as the brain and prostate. Altered expression of these two proteins is often detected in prostate cancer cells. Indeed, the presence of DIP1/2 effectively blocks mitogen-induced gene expression and inhibits the growth of prostate cancer. Thus, DOC-2/DAB2 and DIP1/2 appear to represent a unique negative regulatory complex that maintains cell homeostasis.

Adaptor Proteins, Signal Transducing↗
PubMed2002

[Correlation between polymorphism of IL-4 and IL-10 gene promoter and childhood asthma and their impact upon cytokine expression].

OBJECTIVE: To investigate the correlation between the polymorphism of IL-4 and IL-10 gene promoters and the susceptibility of Chinese children to asthma and to study the impact of such polymorphism upon cytokine production. METHODS: The data of medical history, pulmonary function, hypersensitive test, and serum IgE were recorded among 168 unrelated Chinese children with asthma aged 5 approximately 15. Four polymorphism sites of IL-4 and IL-10 gen promoter regions were determined by PCR/RFLP. The peripheral blood monocyte (PBMC) was cultured and stimulated by lipopolysacchride (LPS). Then the contents of IL-4 and IL-10 gene promoter regions were determined by ELISA. 53 age-matched children without asthma were used as control. RESULTS: (1) The allele frequencies of-598 (C/T) site of IL-4 gene promoter was significantly different between Chinese children and children of Caucasian and Afro-American origin (P = 0.01). The allele frequency of -1082 (G/A) site, -819 (C/T) site, and -592 (A/C) site in IL-10 gene promoter was significantly different between Chinese children and their British counterparts (P = 0.01). (2) The serum total IgE level was significantly different among children with different allelic sites in IL-4 and IL-10 gene promoters. A allele in IL-10 - 1082 (G/A) site and T allele in IL-10 - 819 (C/T) site -1082 were associated with elevated total serum IgE (P < 0.01, OR = 3.16; P < 0.05, OR = 1.84). (3) The IL-10 level in the supernatant of PBMC culture of children with homozygous genotypes of -819 TT and ATA/ATA in IL-10 gene promoter after stimulation of LPS was the lowest in comparison with those in the supernatant of PBMC culture of children with other genotypes (all P < 0.05). (4) No correlation was found between -589 (C/T) site in IL-4 gene promoter and asthma. There was no significant difference between the increase of IL-4 level in the supernatant of PBMC culture of children with the two different homozygous genotypes of IL-4 gene promoter. CONCLUSIONS: (1) There is no significant correlation between the polymorphism sites in IL-4 and IL-10 gene promoters and the susceptibility to asthma among Chinese. (2) The polymorphism of IL-10 gene promoter may be an important candidate gene for severity of asthma. The allele -819T plays an important role in increase of the total serum IgE by reducing the upregulation of IL-10 gene expression.

Adolescent↗
PubMed2002

Profiling the alkaline membrane proteome of Caulobacter crescentus with two-dimensional electrophoresis and mass spectrometry.

Attempts at protein profiling in the alkaline pH region using isoelectric focusing have often proved difficult, greatly limiting the scope of proteome analysis. We investigated several parameters using custom pH 8-11 immobilized pH gradients to separate a Caulobacter crescentus membrane preparation. These included sample application, quenching endoosomotic flow and gel matrix composition. Among these factors, the sample application position was the predominant parameter to affect two-dimensional gel quality. Separated proteins were silver stained and profiled using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The use of a prototype MALDI-Q-Tof mass spectrometer assisted identification of several proteins by providing highly informative peptide fragmentation data from the sample digests. Thirty-two unique alkaline proteins were identified in this study, which complements our previously described C. crescentus membrane proteome. Our experiments point towards new options for proteomic researchers aiming to both extend the scope of analysis, and simplify methods of identifying proteins with high confidence.

2-Propanol↗
PubMed2002

Differential regulation of the human gene DAB2IP in normal and malignant prostatic epithelia: cloning and characterization.

Human DAB2IP (for DAB2 interaction protein) is a novel member of the RasGTPase-activating protein family. It interacts directly with DAB2, which suppresses growth of many cancer types. We demonstrated that DAB2IP is often downregulated in human prostate cancer cell lines. The predicted DAB2IP protein (967 amino acids) shares 94.2% homology with the rat DIP1/2 protein. We mapped the promoter of DAB2IP and studied its regulation in normal and malignant prostate cancer cells. This gene is located at 9q33.1-q33.3 and spans approximately 96 kb with 15 exons and 14 introns. The DAB2IP promoter does not contain any typical TATA box-evidenced by the presence of various RNAs with differential transcription starting sites. We further demonstrated that normal prostatic epithelial cells have elevated DAB2IP mRNA compared with cancer cells, which correlates with increased DAB2IP promoter activity. These data indicate that transcriptional regulation of DAB2IP is responsible for the downregulation of DAB2IP expression in prostate cancer cells.

5' Untranslated Regions↗
PubMed2002

The expression of the plasmid DNA encoding TGF-beta 1 in endothelium after injection into the anterior chamber.

The method of gene transfer into corneal endothelium was investigated to provide a foundation for the study of TGF-beta 1 gene transfer to inhibit corneal graft rejection. Two days after direct injection of pMAM TGF-beta 1 mediated by liposome into the anterior chamber of rabbits, one half of corneas were made into paraffin slides and the endothelial layer was carefully torn from the other half to make a single layer slide of endothelia. By means of immunohistochemical technique, the plasmid pMAM TGF-beta 1 expression product TGF-beta 1 in the endothelia was detected. Specific TGF-beta 1 expression was positive in the endothelia on both the paraffin slide and the single layer slide. The results showed that by direct injection into the anterior chamber, foreign plasmid DNA could be transferred into the endothelia and its expression was obtained. This may provide a foundation for further study on TGF-beta 1 participating in local induction of corneal immune tolerance.

Animals↗
PubMed2002

Experimental study of plasmid TGF-beta 1 DNA gene transfer with lipofectamine into rabbit corneal epithelial cells in vitro.

To investigate whether the TGF-beta 1 plasmid DNA carried by lipofectamine could be introduced into cultured rabbit corneal epithelial cells, specific expression of the plasmid pMAM TGF-beta 1 in the cultured corneal epithelial cells was studied. Two days after 12 h of transfection of pMAMT-GF-beta 1 mediated by lipofectamine into the cultured corneal epithelial cells, the TGF-beta 1 protein expression specific for pMAMTGF-beta 1 in the cells was detected by means of immunohistochemical staining and the positive rate was 23.37%. The results suggested that foreign plasmid DNA could be effectively delivered into cultured rabbit corneal epithelial cells by means of lipofectamine, and this will provide a promising method of studying TGF-beta 1 on the mechanism of physiology and pathology concerned with corneal epithelial cells.

Animals↗
PubMed2002

Catheter ablation of atrial flutter in a heart transplant recipient.

In the transplanted heart with biatrial anastomosis, atrial flutter is common and is amenable to catheter ablation. Although this arrhythmia is isthmus dependent, the unique atrial architecture with a suture line through the inferior vena cava-tricuspid annulus isthmus makes the substrate atypical. A cardiac transplant recipient with atrial flutter underwent successful catheter ablation. Five weeks after the procedure, the patient died of a myocardial infarction. The autopsy and histological findings are described and correlated with the electroanatomic map obtained during the ablation. Due to the atrial suture lines, atrial flutter following cardiac transplantation is an isthmus dependent arrhythmia with a different arrhythmogenic substrate. The electrical isthmus (atrial tissue from the tricuspid annulus to the suture line) in these hearts is smaller than the anatomic isthmus.

Atrial Flutter↗
PubMed2002

The mechanism for transcriptional activation of the human ATA2 transporter gene by amino acid deprivation is different than that for asparagine synthetase.

After amino acid deprivation, the mRNA content for both asparagine synthetase (AS) and the system A transporter ATA2 is increased. The purpose of the reported experiments was to characterize the molecular mechanism for the ATA2 gene and to contrast the ATA2 regulatory characteristics with those of AS. Amino acid limitation was initiated by incubation of HepG2 human hepatoma cells in either amino acid-free Krebs-Ringer bicarbonate buffer or culture medium lacking the single amino acid histidine. For ATA2, like AS, the elevated mRNA content was due to increased transcription. However, there were fundamental differences between the mechanisms for nutrient regulation of the AS and ATA2 genes. When cells were deprived of amino acids, there was a lag period of approximately 4 h before an increase in AS mRNA occurred, whereas the elevation of ATA2 mRNA was readily detectable at 2-4 h. Consistent with these observations, de novo protein synthesis was absolutely required for the activation of the AS gene, but the increase in ATA2 mRNA was largely independent of protein synthesis. Furthermore, in contrast to AS, transcription from the ATA2 gene was not increased by glucose deprivation. Given this lack of ATA2 transcriptional activation by glucose starvation and that the induction of the AS gene by glucose or amino acid starvation is mediated by common genomic elements, it is likely that the ATA2 gene does not contain the same genomic amino acid-responsive cis-elements as the AS gene.

Amino Acid Transport System A↗
PubMed2002

Characterization and regulation of a cloned ovine gastrointestinal peptide transporter (oPepT1) expressed in a mammalian cell line.

To investigate the kinetics of peptide transport by the peptide transporter, PepT1, Chinese hamster ovary cells were transfected with an expression vector containing our cloned ovine PepT1 cDNA. Transport was assessed by uptake studies using the radiolabeled dipeptide, [(3)H]-Gly-Sar. Expression of oPepT1 was detected at 8-24 h post-transfection with an optimal time of 16-24 h. Uptake of Gly-Sar by oPepT1 was pH-dependent with an optimal pH of 5.5-6.0, concentration-dependent and saturable with an apparent K(m) value of 1.0 +/- 0.1 mmol/L and a maximum velocity of 14.3 +/- 0.4 nmol/(mg protein x 40 min). Competition studies with nonradiolabeled peptides and [(3)H]-Gly-Sar showed that all di- and tripeptides inhibited uptake of [(3)H]-Gly-Sar. In addition, three tetrapeptides (Met-Gly-Met-Met, Pro-Phe-Gly-Lys, and Val-Gly-Ser-Glu) also inhibited [(3)H]-Gly-Sar uptake. There was no inhibition of [(3)H]-Gly-Sar uptake detected in the presence of nonradiolabeled free amino acids. Treatment of the cells with staurosporine, an inhibitor of protein kinase C (PKC) significantly increased the transport system. This increase was specific and could be blocked if treatment was done in the presence of phorbol 12-myristate-13-acetate (PMA), an activator of PKC. The staurosporine- and PMA-induced changes in peptide transport activity were not affected by cotreatment with cycloheximide. These data demonstrate that the transport of peptide substrates by oPepT1 in transfected mammalian cells is similar to that in microinjected Xenopus oocytes and that PKC phosphorylation plays a regulatory role in oPepT1 function.

Animals↗
PubMed2002

Molecular cloning and functional expression of a chicken intestinal peptide transporter (cPepT1) in Xenopus oocytes and Chinese hamster ovary cells.

To study peptide absorption in chickens, an intestinal peptide transporter cDNA (cPepT1) was isolated from a chicken duodenal cDNA library. The cDNA was 2914 bp long and encoded a protein of 714 amino acid residues with an estimated molecular size of 79.3 kDa and an isoelectric point of 7.48. cPepT1 protein is similar60% identical to PepT1 from rabbits, humans, mice, rats and sheep. Sixteen dipeptides, three tripeptides and four tetrapeptides that contained the essential amino acids Met, Lys and(or) Trp were used for functional analysis of cPepT1 in Xenopus oocytes and Chinese hamster ovary cells. For most di- and tripeptides tested, the substrate affinities were in the micromolar range, indicating that cPepT1 has high affinity for these peptides. Lys-Lys and Lys-Trp-Lys were exceptions, with substrate affinities in the millimolar range. Neither free amino acids nor tetrapeptides were transported by cPepT1. Northern blot analysis using a full-length cPepT1 cDNA as the probe demonstrated that cPepT1 is expressed strongly in the duodenum, jejunum and ileum, and at lower levels in kidney and ceca. The present study demonstrated for the first time the presence and functional characteristics of a peptide transport system from an avian species.

Amino Acid Sequence↗
PubMed2002

Smoking in chronic schizophrenic inpatients in taiwan.

OBJECTIVE: This study investigated the prevalence of smoking and its association with the clinical characteristics of Chinese inpatients with chronic schizophrenia. METHOD: Schizophrenic patients hospitalized in chronic wards were assessed using Brief Psychiatric Rating Scale (BPRS), Abnormal Involuntary Movements Scale (AIMS) and Folstein Mini-Mental Status Examination (MMSE) testing. RESULTS: Of 257 patients, 105 smoked and 4 had ceased. Males exhibited a higher prevalence of smoking than females (p < 0.001). Smoking was not significantly associated with age at onset (AAO), chlorpromazine equivalents, MMSE, AIMS, BPRS positive symptom subscale, BPRS negative symptom subscale or total BPRS scores. Smokers had higher BPRS general subscales. CONCLUSION: Compared to the general population, smoking prevalence was slightly higher in schizophrenic males, double in schizophrenic females, but no difference in refractory schizophrenic clozapine users. Smoking did not affect patient AAO or daily antipsychotic dose. Patients with a higher BPRS general subscale may smoke to relieve affective symptoms.

Adult↗