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Biomedical subjects

H Jin

Publications and source records attributed to H Jin.

At least 73 records · Page 4Linked to original sources
PubMed2000

Application of cDNA microarrays in determining molecular phenotype in cardiac growth, development, and response to injury.

BACKGROUND: Normal myocardial development and the tissue response to cardiac stress are accompanied by marked changes in gene expression; however, the extent of these changes and their significance remain to be fully explored. We used cDNA microarrays for gene expression profiling in rat cardiac tissue samples to study developmental transitions and the response to myocardial infarction (MI). METHODS AND RESULTS: Microarrays with rat cDNAs for 86 known genes and 989 anonymous cDNAs obtained by molecular subtraction (representational difference analysis) of mRNA from sham-operated and 6-week post-MI samples were used in 2-color hybridization experiments. Twelve known genes previously associated with myocardial development were identified together with 10 uncharacterized expressed sequence tags and 36 genes not previously associated with cardiac development. After MI, genes associated with myocardial stress and wound healing exhibited differences in magnitude and expression kinetics, and 14 genes not previously associated with MI were identified. In situ hybridization revealed mRNA localization characteristic of wound healing and vascular and cardiomyocyte reactivity. CONCLUSIONS: Tissue analysis of gene expression with cDNA microarrays provides a measure of transcriptional or posttranscriptional regulation and cellular recruitment. Our results demonstrate the complexity of gene regulation in the developing myocardium and show that cDNA microarrays can be used to monitor the evolution of the cardiac stress-inducible phenotype.

Animals↗
PubMed2000

Granulocyte-macrophage colony-stimulating factor in the innate immune response to Pneumocystis carinii pneumonia in mice.

Innate immunity plays an important role in pulmonary host defense against Pneumocystis carinii, an important pathogen in individuals with impaired cell-mediated immunity. We investigated the role of GM-CSF in host defense in a model of P. carinii pneumonia induced by intratracheal inoculation of CD4-depleted mice. Lung GM-CSF levels increased progressively during the infection and were significantly greater than those in uninfected controls 3, 4, and 5 wk after inoculation. When GM-CSF gene-targeted mice (GM-/-) depleted of CD4+ cells were inoculated with P. carinii, the intensities of infection and inflammation were increased significantly compared with those in CD4-depleted wild-type mice. In contrast, transgenic expression of GM-CSF directed solely in the lungs of GM-/- mice (using the surfactant protein C promoter) dramatically decreased the intensity of infection and inflammation 4 wk after inoculation. The concentrations of surfactant proteins A and D were greater in both uninfected and infected GM-/- mice compared with those in wild-type controls, suggesting that this component of the innate response was preserved in the GM-/- mice. However, alveolar macrophages (AM) from GM-/- mice demonstrated impaired phagocytosis of purified murine P. carinii organisms in vitro compared with AM from wild-type mice. Similarly, AM production of TNF-alpha in response to P. carinii in vitro was totally absent in AM from GM-/- mice, while GM-CSF-replete mice produced abundant TNF in this setting. Thus, GM-CSF plays a critical role in the inflammatory response to P. carinii in the setting of impaired cell-mediated immunity through effects on AM activation.

Animals↗
PubMed2000

A novel method for expression and large-scale production of human brain l-glutamate decarboxylase.

l-Glutamate decarboxylase (GAD; EC 4.1.1.15) is the rate-limiting enzyme involved in the synthesis of gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter in the mammalian brain. Imbalance in the conversion of glutamate to GABA has been implicated in a host of human diseases. Studies on the structure, function, and therapeutic use of GAD have been precluded by insufficient quantities of purified active enzyme. Here we report a novel methodology for the expression and large-scale production of enzymatically active, pure, recombinant human GAD65 and GAD67. This method circumvents the sequestering of expressed protein into insoluble inclusion bodies and reduces production of truncated proteins. The availability of sufficient quantities of purified HGAD65 and HGAD67 has allowed for the production of specific polyclonal antibodies that discriminate between the two isoforms. This methodology, in addition to providing key human brain enzymes, may be generally applicable to other systems.

Brain↗
PubMed2000

Integrating functional genomic information into the Saccharomyces genome database.

The Saccharomyces Genome Database (SGD) stores and organizes information about the nearly 6200 genes in the yeast genome. The information is organized around the 'locus page' and directs users to the detailed information they seek. SGD is endeavoring to integrate the existing information about yeast genes with the large volume of data generated by functional analyses that are beginning to appear in the literature and on web sites. New features will include searches of systematic analyses and Gene Summary Paragraphs that succinctly review the literature for each gene. In addition to current information, such as gene product and phenotype descriptions, the new locus page will also describe a gene product's cellular process, function and localization using a controlled vocabulary developed in collaboration with two other model organism databases. We describe these developments in SGD through the newly reorganized locus page. The SGD is accessible via the WWW at http://genome-www.stanford.edu/Saccharomyces/

Database Management Systems↗
PubMed2000

Correlation of high-resolution breast MR imaging with histopathology; validation of a technique.

A high-resolution three-dimensional surface gradient coil set was used to obtain magnetic resonance (MR) images of breast specimens, using a gradient-echo pulse sequence (TR/TE 1000/8 msec, flip angle 75 degrees), with 117 micrometer in-plane resolution and 1 mm slice thickness. Breast tissues were obtained from one autopsy and three surgical specimens. High-resolution breast MR images and histopathology sections (7 micrometer thickness) were acquired in the same anatomical plane. Radiographs were acquired of the sliced specimens (approximately 5 mm thick) so that images from all three methods could be correlated. It was found that in vitro high-resolution breast MRI correlated well with low-resolution microscopic histology, demonstrating normal anatomy (lobules, ducts, connective tissue strands, blood vessels) and pathology (tumor content, margins, and presence of microcalcifications) of the breast more clearly than conventional pre-gadolinium breast MRI. High-resolution breast MRI may improve specificity, when added to a conventional breast MRI protocol.

Biopsy↗
PubMed2000

Expression of porcine endogenous retrovirus in peripheral blood leukocytes from ten different breeds.

The expression of porcine endogenous retroviruses (PERV) was investigated in primary porcine peripheral blood leukocytes (PBL) of ten different pig breeds. The data suggest that PERV exists in all porcine PBL. A new retroviral element, a foamy-like pol-related sequence, was also detected in PBL. Three types of PERV were detected in almost every animal. The breeding of PERV-free pigs is likely to be difficult. Further studies are required to assess the infectious disease risks associated with xenotransplantation.

Animals↗
PubMed2000

Two novel members of the interleukin-1 receptor gene family, one deleted in Xp22.1-Xp21.3 mental retardation.

X-linked mental retardation is estimated to affect approximately 1 in 600 males. Although numerous genes responsible for syndromic mental retardation have been identified, the study of non-syndromic mental retardation suffers from intrinsic issues of genetic heterogeneity. During the investigation of three brothers with a contiguous gene deletion syndrome of Becker muscular dystrophy, glycerol kinase deficiency, congenital adrenal hypoplasia, and mental retardation, we found their dystrophin gene to be fused tail-to-tail with a gene encoding a novel member of the interleukin-1 receptor family, IL1RAPL1. This gene has a close relative in Xq22, which we call IL1RAPL2. Both IL1RAPL1 and IL1RAPL2 have novel C-terminal sequences not present in other related proteins, and are encoded by very large genes. The 1.8-megabase deletion in these patients removes not only the last exon of the dystrophin gene, the entire glycerol kinase and DAX-1 genes, and the MAGE-B gene cluster, but also three exons encoding the intracellular signalling domain of IL1RAPL1. The literature contains multiple reports of patients with non-syndromic mental retardation in association with an Xp22.1-Xp21.3 microdeletion of a marker which lies within the IL1RAPL1 gene. The gene is also wholly or partially deleted in patients with mental retardation as part of a contiguous deletion syndrome. We suggest that IL1RAPL1, and perhaps IL1RAPL2, are strong candidates for X-linked non-syndromic mental retardation loci, and that molecules resembling IL-1 and IL-18 play a role in the development or function of the central nervous system.

5' Untranslated Regions↗
PubMed2000

The secretory pathway mediates localization of the cell polarity regulator Aip3p/Bud6p.

Aip3p/Bud6p is a regulator of cell and cytoskeletal polarity in Saccharomyces cerevisiae that was previously identified as an actin-interacting protein. Actin-interacting protein 3 (Aip3p) localizes at the cell cortex where cytoskeleton assembly must be achieved to execute polarized cell growth, and deletion of AIP3 causes gross defects in cell and cytoskeletal polarity. We have discovered that Aip3p localization is mediated by the secretory pathway. Mutations in early- or late-acting components of the secretory apparatus lead to Aip3p mislocalization. Biochemical data show that a pool of Aip3p is associated with post-Golgi secretory vesicles. An investigation of the sequences within Aip3p necessary for Aip3p localization has identified a sequence within the N terminus of Aip3p that is sufficient for directing Aip3p localization. Replacement of the N terminus of Aip3p with a homologous region from a Schizosaccharomyces pombe protein allows for normal Aip3p localization, indicating that the secretory pathway-mediated Aip3p localization pathway is conserved. Delivery of Aip3p also requires the type V myosin motor Myo2p and its regulatory light-chain calmodulin. These data suggest that one function of calmodulin is to activate Myo2p's activity in the secretory pathway; this function is likely the polarized movement of late secretory vesicles and associated Aip3p on actin cables.

Amino Acid Sequence↗
PubMed2000

Antiviral properties of a series of 1,6-naphthyridine and 7, 8-dihydroisoquinoline derivatives exhibiting potent activity against human cytomegalovirus.

A series of 1,6-naphthyridine (L. Chan, H. Jin, T. Stefanac, J. F. Lavallee, G. Falardeau, W. Wang, J. Bedard, S. May, and L. Yuen, J. Med. Chem. 42:3023-3025, 1999) and isoquinoline (L. Chan, H. Jin, T. Stefanac, W. Wang, J. F. Lavallee, J. Bedard, and S. May, Bioorg. Med. Chem. Lett. 9:2583-2586, 1999) analogues exhibiting a high level of anti-human cytomegalovirus (HCMV) activity were investigated in a series of studies aimed at better understanding the mechanism of action of some representatives of this class of compounds. In vitro antiviral profiling revealed that these compounds were active against a narrow spectrum of viruses, essentially the human herpesviruses and type 2 rhinovirus. In HCMV assays, a 39- to 223-fold lower 50% inhibitory concentration was obtained for compound A1 than for ganciclovir against strains AD 169 and Towne. In addition, ganciclovir, foscarnet, cidofovir, and BDCRB (2-bromo-5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole)-resistant HCMV strains remained susceptible to 1,6-naphthyridines and 7, 8-dihydroisoquinolines tested in this study, supporting the view that a novel mechanism of action could be involved. Drug combination studies showed a small but significant synergistic antiviral effect between compound B2 and ganciclovir. Cytotoxicity profiling of representative compounds under various cell growth conditions indicated a generally similar cytotoxic effect, relative to ganciclovir, in log-phase growing cells. However, in stationary cells, a relatively higher level of toxicity was observed than that for control compound. Effect of time of drug addition showed that the anti-HCMV activity of compound A1, ganciclovir, and cidofovir was lost at approximately the same time (72 h postinfection), indicating that the compound was affecting events at the early and late stage of virus replication. This interpretation is also supported by reduction of de novo synthesis of pp65 tegument protein and lack of any effect of the compound on viral adsorption. A reduction of the HCMV enhancer-promoter-directed luciferase expression was also observed in a stably transfected cell line when compound A1 was present at relatively high concentrations.

Antiviral Agents↗
PubMed2000

Respiratory syncytial virus that lacks open reading frame 2 of the M2 gene (M2-2) has altered growth characteristics and is attenuated in rodents.

The M2 gene of respiratory syncytial virus (RSV) encodes two putative proteins: M2-1 and M2-2; both are believed to be involved in the RNA transcription or replication process. To understand the function of the M2-2 protein in virus replication, we deleted the majority of the M2-2 open reading frame from an infectious cDNA clone derived from the human RSV A2 strain. Transfection of HEp-2 cells with the cDNA clone containing the M2-2 deletion, together with plasmids that encoded the RSV N, P, and L proteins, produced a recombinant RSV that lacked the M2-2 protein (rA2DeltaM2-2). Recombinant virus rA2DeltaM2-2 was recovered and characterized. The levels of viral mRNA expression for 10 RSV genes examined were unchanged in cells infected with rA2DeltaM2-2, except that a shorter M2 mRNA was detected. However, the ratio of viral genomic or antigenomic RNA to mRNA was reduced in rA2DeltaM2-2-infected cells. By use of an antibody directed against the bacterially expressed M2-2 protein, the putative M2-2 protein was detected in cells infected with wild-type RSV but not in cells infected with rA2DeltaM2-2. rA2DeltaM2-2 displayed a small-plaque morphology and grew much more slowly than wild-type RSV in HEp-2 cells. In infected Vero cells, rA2DeltaM2-2 exhibited very large syncytium formation compared to that of wild-type recombinant RSV. rA2DeltaM2-2 appeared to be a host range mutant, since it replicated poorly in HEp-2, HeLa, and MRC5 cells but replicated efficiently in Vero and LLC-MK2 cells. Replication of rA2DeltaM2-2 in the upper and lower respiratory tracts of mice and cotton rats was highly restricted. Despite its attenuated replication in rodents, rA2DeltaM2-2 was able to provide protection against challenge with wild-type RSV A2. The genotype and phenotype of the M2-2 deletion mutant were stably maintained after extensive in vitro passages. The attenuated phenotype of rA2DeltaM2-2 suggested that rA2DeltaM2-2 may be a potential candidate for use as a live attenuated vaccine.

Amino Acid Sequence↗
PubMed2000

Effects of exercise training on cardiac function, gene expression, and apoptosis in rats.

This study determined the effects of exercise training on cardiac function, gene expression, and apoptosis. Rats exposed to a regimen of treadmill exercise for 13 wk had a significant increase in cardiac index and stroke volume index and a concomitant decrease in systemic vascular resistance compared with both age-matched and body weight-matched sedentary controls in the conscious state at rest. In exercise-trained animals, there was no change in the expression of several marker genes known to be associated with pathological cardiac adaptation, including atrial natriuretic factor, beta-myosin heavy chain, alpha-skeletal and smooth muscle actins, and collagens I and III. Exercise training, however, produced a significant induction of alpha-myosin heavy chain, which was not observed in rats with myocardial infarction. No histological features of cardiac apoptosis were observed in the treadmill-trained rats. In contrast, apoptotic myocytes were detected in animals with myocardial infarction. In summary, exercise training improves cardiac function without evidence of cardiac apoptosis and produces a pattern of cardiac gene expression distinct from pathological cardiac adaptation.

Actins↗
PubMed2000

Effects of homocysteine on endothelial nitric oxide production.

Hyperhomocysteinemia (HHCy) is an independent and graded cardiovascular risk factor. HHCy is prevalent in patients with chronic renal failure, contributing to the increased mortality rate. Controversy exists as to the effects of HHCy on nitric oxide (NO) production: it has been shown that HHCy both increases and suppresses it. We addressed this problem by using amperometric electrochemical NO detection with a porphyrinic microelectrode to study responses of endothelial cells incubated with homocysteine (Hcy) to the stimulation with bradykinin, calcium ionophore, or L-arginine. Twenty-four-hour preincubation with Hcy (10, 20, and 50 microM) resulted in a gradual decline in responsiveness of endothelial cells to the above stimuli. Hcy did not affect the expression of endothelial nitric oxide synthase (eNOS), but it stimulated formation of superoxide anions, as judged by fluorescence of dichlorofluorescein, and peroxynitrite, as detected by using immunoprecipitation and immunoblotting of proteins modified by tyrosine nitration. Hcy did not directly affect the ability of recombinant eNOS to generate NO, but oxidation of sulfhydryl groups in eNOS reduced its NO-generating activity. Addition of 5-methyltetrahydrofolate restored NO responses to all agonists tested but affected neither the expression of the enzyme nor formation of nitrotyrosine-modified proteins. In addition, a scavenger of peroxynitrite or a cell-permeant superoxide dismutase mimetic reversed the Hcy-induced suppression of NO production by endothelial cells. In conclusion, electrochemical detection of NO release from cultured endothelial cells demonstrated that concentrations of Hcy >20 microM produce a significant indirect suppression of eNOS activity without any discernible effects on its expression. Folates, superoxide ions, and peroxynitrite scavengers restore the NO-generating activity to eNOS, collectively suggesting that cellular redox state plays an important role in HCy-suppressed NO-generating function of this enzyme.

Animals↗
PubMed2000

Sequence analysis and tissue specific expression of human HOXA7.

A 3071-bp fragment containing the human HOXA7 gene was sequenced. It contained two exons, one intron, and two polyadenylation signals (AATAAA) at positions 1844 and 2923. The exon encoded 230 aa residues, while the hexapeptide, homeodomain, and C-terminal acidic domains were detected. When the total sequences were compared with those of murine Hoxa-7, the 5' untranslated region (UTR), exon I, intron, exon II, and 3' UTR exhibited 99, 92, 65, 85, and 72% homology, respectively. Through Northern analysis, about a 1.9-kb transcript was detected in the fetal kidney. Minor transcripts of 1.5 and 1.1-kb were also detected in the fetal liver as well as in the kidney. In the case of adult tissues, most of the tissues tested (lung, liver, skeletal muscle, kidney, pancreas, and placenta), except brain tissue, expressed a 5.3 kb transcript with various intensities. Our results here suggested that not only Abd-B type Hox genes, but the ones in paralogous groups I-VIII could generate multiple transcripts. The characterization of these tissue-specific and stage-specific alternative transcripts would help to understand the regulatory function of the HOXA7 gene during development, and possibly the pathology of human disease caused by Hox genes.

3' Untranslated Regions↗
PubMed2000

Combined AFLP and RFLP mapping in two hexaploid oat recombinant inbred populations.

A combined RFLP and AFLP map was constructed for hexaploid oat (Avena spp.). The segregation of AFLP markers was scored in two hexaploid oat recombinant inbred line (RIL) populations, the 'Kanota' x 'Ogle' RFLP population, and a population derived from 'Clintland64' and 'IL86-5698', barley yellow dwarf virus (BYDV)-sensitive and BYDV-tolerant lines, respectively. More than 300 AFLP markers were scored in each population, of which 97 could be scored in both populations. AFLP markers were linked to RFLP markers in 32 of 36 'Kanota' x 'Ogle' RFLP linkage groups. The addition of the AFLP markers to the 'Kanota' x 'Ogle' RFLP data set combined markers from four pairs of linkage groups and increased the size of the map from 1402 cM to 2351 cM. Thirty linkage groups were observed in the 'Clintland64' x 'IL86-5698' population, two of which could be consolidated by comparing the maps from both populations. The AFLP and RFLP markers showed very similar distributions in the 'Kanota' x 'Ogle' population with a tendency of each type of marker to cluster with markers of the same type. The placement of a set of AFLP markers on the 'Kanota' x 'Ogle' linkage map will enrich the RFLP map and allow others to relate AFLP markers for agronomically important genes to the reference 'Kanota' x 'Ogle' linkage map.

Avena↗
PubMed2000

[Progress in researches on anti-tumor agent L-asparaginase for treatment of leukemia].

L-asparaginase is a very effective anti-tumor agent. To take the most advantage of enzyme treatment method, the natural L-asparaginase could be chemically modified, entrapped or immobilized into/onto other carriers. The extracorporeal shunt system with immobilized enzyme reactor deserves to receive special attention. It provides an entirely new idea for leukemia treatment and is proved to be very prospective in the future leukemia cure.

Antineoplastic Agents↗
PubMed2000

Changes in coagulation and tissue plasminogen activator after the treatment of cerebral infarction with lumbrokinase.

This paper aimed to investigate the effect of lumbrokinase on the anticoagulation and fibrinolysis in treating cerebral infarction. Lumbrokinase was used in patients with cerebral infarction. Patients were randomly divided into treatment group (n = 31) and control group (n = 20). Single blind method was used in this investigation. The Chinese stroke score was used to evaluate the results of treatment before and after administration of lumbrokinase. Kaolin partial thromboplastin time (KPTT), prothrombin time (PT), fibrinogen content, vWF content were analyzed, and tissue plasminogen activator (t-PA) activity, plasminogen activator inhibitor (PAI) activity, D-dimer level were assayed. In both groups, the stroke score decreased after administration, but in the treatment group, it was more obvious. In the treatment group, KPTT was prolonged, t-PA activity and D-dimer level increased, while the content of fibrinogen decreased significantly. There were no significant changes of PT and PAI activity in both groups. It is concluded that lumbrokinase is beneficial to the treatment of cerebral infarction. The effect of lumbrokinase is related to the inhibition of intrinsic coagulation pathway and the activation of fibrinolysis via an increase of t-PA activity.

Aged↗
PubMed2000

Impaired gp-91phox gene expression and dysfunction of peripheral blood neutrophils in patients maintaining hemodialysis.

OBJECTIVE: To investigate expressions of gp-91phox and IL-8 gene and the function of neutrophils in hemodialysis (HD) patients, and to understand the relationship between uremia and immune. METHODS: Gene expression was studied by means of reversal transcription polymerase chain reaction (RT/PCR) from the untreated and treated with lipopolysaccharide (LPS) neutrophils in controls and HD patients. Phagocytosis and bactericidal assays were measured as a decrease in the number of viable intracellular and extracellular bacteria by colony counts. RESULTS: Freshly isolated neutrophils express gp-91phox mRNA in controls while no gp-91phox mRNA was detected in HD patients. When challenged with LPS, gp-91phox mRNA exhibited decreased expression. IL-8 mRNA in HD patients could be spontaneously expressed. Phagocytosis and intracellular killing of neutrophils, when exposed to Staphylococcus aureus, were decreased obviously compared with controls. CONCLUSIONS: The impaired gp-91phox gene expression of neutrophils indicates the impaired NADPH-oxidase system and the untreated neutrophils spontaneously express IL-8 mRNA in the HD patients, which is consistent with suppressed phagocytosis and intracellular killing capacity.

Cell Division↗