Biomedical subjects

R Zhou

Publications and source records attributed to R Zhou.

At least 73 records · Page 4Linked to original sources

PubMed2000

Cyclic core dendrimer as a new kind of vector for gene transfer into mammalian cells.

Cyclic core dendritic polymer is a new type of synthetic polymers. The ability of generation 4 of the dendrimer with a core of 1,4,7,10-tetraazacyclododecane to function as an effective gene delivery vector was investigated. Results from fluorescence in situ hybridization (FISH) show that the pCH 110 plasmid DNA was transferred into human small intestine cancer metastatic ascites (HICMA) cells induced by this kind of dendrimer as a vector. The transferred LacZ, GFP and luciferase genes were highly expressed in the transfected HICMA, COS-7 and 293 cells. These studies demonstrate that the dendrimer can transfect mammalian cells in vitro which offers an alternatively efficient method for mammalian gene transfer.

Animals↗

PubMed2000

Role of Drosophila IKK gamma in a toll-independent antibacterial immune response.

We have generated, by ethylmethane sulfonate mutagenesis, loss-of-function mutants in the Drosophila homolog of the mammalian I-kappa B kinase (IKK) complex component IKK gamma (also called NEMO). Our data show that Drosophila IKK gamma is required for the Relish-dependent immune induction of the genes encoding antibacterial peptides and for resistance to infections by Escherichia coli. However, it is not required for the Toll-DIF-dependent antifungal host defense. The results indicate distinct control mechanisms of the Rel-like transactivators DIF and Relish in the Drosophila innate immune response and show that Drosophila Toll does not signal through a IKK gamma-dependent signaling complex. Thus, in contrast to the vertebrate inflammatory response, IKK gamma is required for the activation of only one immune signaling pathway in Drosophila.

Animals↗

PubMed2000

Identification of cadherin-11 down-regulation as a common response of astrocytoma cells to transforming growth factor-alpha.

Transforming growth factor-alpha (TGF-alpha) and its receptor are frequently co-expressed in high-grade astrocytomas, suggesting a role for TGF-alpha autocrine/paracrine loops in the malignant progression of astrocytomas. To identify genes that may be critical in mediating TGF-alpha impact on the malignant progression of astrocytomas, we have used cDNA arrays to investigate TGF-alpha effects on the gene expression profile of U-373 MG glioblastoma cells. We found that in these cells approximately 50% of the TGF-alpha regulated genes code for cell motility/invasion-related proteins. TGF-alpha action on the expression of four of these proteins, alpha-catenin, IQGAP1, RhoA, and cadherin-11, was further investigated by immunoblotting in four astrocytoma cell lines and in normal astrocytes. The results demonstrate that the effects of TGF-alpha on IQGAP1, alpha-catenin, and RhoA expression are cell-line dependent. On the other hand, under TGF-alpha treatment, cadherin-11 expression is consistently decreased in all astrocytoma cell lines tested but is increased in normal astrocytes. In addition, we found that cadherin-11 is consistently down-regulated in astrocytomas versus normal brain tissues. Altogether, these results suggest that the down-regulation of cadherin-11 is a frequent molecular event in the neoplastic transformation of astrocytes and that this down-regulation may be initiated and/or amplified by TGF-alpha autocrine/paracrine loops during tumor progression.

Animals↗

PubMed2000

Inhibition of human rhinovirus-induced cytokine production by AG7088, a human rhinovirus 3C protease inhibitor.

Symptom severity in patients with human rhinovirus (HRV)-induced respiratory illness is associated with elevated levels of the inflammatory cytokines interleukin-6 (IL-6) and IL-8. AG7088 is a novel, irreversible inhibitor of the HRV 3C protease. In this study, AG7088 was tested for its antiviral activity and ability to inhibit the production of IL-6 and IL-8 in a human bronchial epithelial cell line, BEAS-2B. Infection of BEAS-2B cells with HRV 14 resulted in the production of both infectious virus and the cytokines IL-6 and IL-8. Treatment of HRV 14-infected cells with AG7088 resulted in a statistically significant (P, <0.05) dose-dependent reduction in the levels of infectious virus as well as IL-6 and IL-8 released into the cell supernatant compared to the results obtained for compound-free infected cells. AG7088 was also able to inhibit the replication of HRV 2 and 16 in BEAS-2B cells. In time-of-addition studies, AG7088 could be added as late as 14 to 26 h after HRV 14 infection of BEAS-2B cells and still result in a statistically significant (P, <0.05) reduction in the levels of infectious virus, IL-6, and IL-8 compared to the results obtained for compound-free infected cells. These findings have implications for the development of an antirhinovirus agent that may not only block virus replication but also diminish symptoms.

3C Viral Proteases↗

PubMed2000

Immobilization of antibodies on ultraflat polystyrene surfaces.

BACKGROUND: Functional antibody surfaces were prepared on ultraflat polystyrene surfaces by physical adsorption, and the uniform distribution of monoclonal antibodies against hepatitis B surface antigen (anti-HBs) on such surfaces and the presence of dense hepatitis B surface antigen (HBsAg) particles captured by immobilized antibodies were identified. METHODS: A model polystyrene film was spin-coated directly onto a silicon wafer surface. Atomic force microscopy was used to directly monitor the immobilization of anti-HBs antibodies and their specific molecular interaction with HBsAg. Enzyme immunoassay was also used to characterize functional antibody surfaces. RESULTS: A mean roughness of 2 A for areas of 25 microm(2) was produced. We found a uniform distribution of anti-HBs antibodies on ultraflat polystyrene surfaces and the presence of dense HBsAg particles bound to such anti-HBs surfaces after incubation with HBsAg. CONCLUSIONS: This study confirmed the potential of preparing dense, homogeneous, highly specific, and highly stable antibody surfaces by immobilizing antibodies on polystyrene surfaces with controlled roughness. It is expected that such biofunctional surfaces could be of interest for the development of new solid-phase immunoassay techniques and biosensor techniques.

Adsorption↗

PubMed2000

Biological function of a novel gene overexpressed in human hepatocellular carcinoma.

OBJECTIVE: To clone the full-length of a differentially expressed cDNA fragment, LC27, and study its biological function tentatively. METHODS: Northern blot was used to analyze the expression pattern of LC27 in hepatocellular carcinoma, matched nontumor liver tissues, fetal liver and normal adult liver tissues, as well as BEL-7402 hepatocellular carcinoma cell line ESTs splicing and 5' rapid amplification of cDNA ends (5' RACE) were used to clone the full-length of LC27 cDNA. An antisense oligodeoxynucleotide approach was used to investigate the biological role of the gene in the proliferation of BEL-7402 cells. RESULTS: A 2186 bp novel cDNA with an open reading frame encoding a 283 amino acid protein was cloned. Analysis of the deduced amino acid sequence indicated that it is 38% (88/229) identical to human Golgi 4-transmembrane spanning transporter MTP. The gene and the encoded protein was termed hepatocellular carcinoma overexpressed transmembrane protein (hotp) and HOTP, respectively. Hotp mRNA was almost undetectable in normal adult liver and fetal liver tissues. However, it was significantly up-regulated in hepatocellular carcinoma and some matched nontumor liver tissues, as well as BEL-7402 cells. The proliferation of BEL-7402 cells was suppressed by an antisense oligodeoxynucleotide against hotp mRNA at a concentration of 50 micrograms/ml. CONCLUSION: HOTP may be an integral membrane transporter protein. The overexpression of the gene in hepatocellular carcinoma may play an important role in hepatocarcinogenesis and disease progression.

Adult↗

PubMed2000

The cellular and molecular mechanism of laminin-glycopeptides on anti-metastasis.

OBJECTIVE: To further study the anti-metastasis mechanism of laminin-glycopeptides on carcinoma cell proliferation, apoptosis and the secretion of matrix metalloproteinases. METHODS: Human hepatocellular carcinoma cells in serum free medium were incubated on laminin-coated substrate with or without laminin-glycopeptides at a final concentration of 50 micrograms/ml. The total number of surviving cells after incubating for the indicated time was assayed by MTT assay. DNA synthesis of the incubated cells was detected by 3H-TdR incorporation. Cell cycle was analysed by FACS. The mitotic index of Giemsa stained cells was assessed. Cell apoptosis was detected by both FACS and an acridine orange staining method. Matrix metalloproteinase secretion was analysed by gelatin zymography. RESULTS: The total number of surviving cells incubated on laminin in the absence of laminin-glycopeptides was significantly larger than that in the presence of laminin-glycopeptides. Laminin promoted 3H-TdR incorporation of carcinoma cells, decreased the percentage of cells in G1 phase and increased the percentage of cells in S phase. In contrast, laminin-glycopeptides could inhibit the effect of laminin as shown by 3H-TdR incorporation and cell cycle analysis. The percentage of cells in G2 + M phase and the mitotic index among various groups showed no significant difference. Matrix metalloproteinases secretion from cells treated by laminin-glycopeptides was much less compared to that without the treatment by laminin-glycopeptides. CONCLUSION: Laminin may stimulate cell proliferation, while laminin-glycopeptides could significantly inhibit the effect of laminin by inhibiting DNA synthesis and arresting the carcinoma cell cycle from G1 to S phase. These effects may inhibit not only tumor growth of the primary carcinoma, but also the establishment of metastases at ectopic tissues. Laminin-glycopeptides could also inhibit the secretion of matrix metalloproteinases from carcinoma cells and this may contribute to their decreased invasive and metastatic phenotype. This study further revealed the cellular and molecular mechanism of laminin-glycopeptides on anti-metastasis.

Apoptosis↗

PubMed2000

[Mechanism of enhanced invasiveness of human hepatocellular carcinoma by integrin alpha 6 beta 1].

OBJECTIVE: To study the mechanism of enhanced invasiveness of human hepatocellular carcinoma (HCC) by laminin (LN) receptor integrin alpha 6 beta 1. METHODS: A stable transfectant of HCC Bel 7402 cell line, the alpha 6 beta 1 receptors of which were replaced by non-functional LN receptor alpha 6 beta 4-TR using dominant negative strategy, was used. Motility of the non-transfected, mock transfected and transfected tumor cells was assessed by the number of cells migrated to an area scratched out of cells on tumor cell monolayer. Tumor cell invasion was examined by cell penetration through a Matrigel layer in Boyden chamber. Matrix metalloproteinases (MMPs) secretion was detected by gelatin zymography. RESULTS: The migration of HCC Bel 7402 cells expressing dominant negative alpha 6 beta 1 was significantly decreased. Their invasive capability was decreased by 46.7%. The secretion of MMP-9 was almost totally inhibited and the activated MMP-2 was decreased by 43.9%. CONCLUSION: The decrease in cell motility, invasiveness and MMPs secretion of HCC cells expressing dominant negative alpha 6 beta 4 TR implies that LN-integrin interaction plays important role in HCC progression.

Carcinoma, Hepatocellular↗

PubMed2000

[Sampling survey for iodine content of iodized salt in household of CFNSS in 1998].

The sampling survey for iodine content of iodized salt was conducted in 36 food and nutrition surveillance sites of CFNSS in 1998 in China, which covered 26 provinces, 362 portions of salt samples. The average compliance rate was 76.0%, 83.2% in Urban area and 71.9% in Rural. Iodine content of salt less then 20 mg/kg at the rural households was 20.4%, and 9.9% in urban.

Female↗

PubMed2000

[The electron microscopic study of AchR and ultrastructure of neuromuscular junction in recurrent laryngeal nerve reinnervation].

OBJECTIVE: The purpose of this study was to observe the distribution of Acetylcholine receptor (AchR) and ultrastructural changes of the neuromuscular junction after laryngeal reinnervation. METHODS: alpha-bungarotoxin-horseradish peroxidase (alpha-BT-HRP) conjugate electron microscopic immunocytochemical methods and computer analysis of intensity of immunocytochemical reaction were applied to observe the distribution and relative quantity of junctional N-AchR. RESULTS: The results showed that the synaptic nerve terminals and presynaptic membrane were segmentally disrupted and almost disappeared completely and were replaced by Schwann's cells at 3 weeks after neurorrhaphy. At 6 weeks, some of the regenerating axons grew to the neuromuscular junctions and all the muscle fibers were reinnervated at 12 weeks. But the original postsynaptic membranes were only partially in contact with the regenerated nerve terminals. These conditions persisted until 18 weeks after neurorrhaphy, when remyelination was found in the vicinity of the neuromuscular junctions. The N-AchR reaction product on the postsynaptic membrane were increased greatly about 1.28 times more than the normal at 3 weeks after neurorrhaphy. The reactive products decreased greatly which was only 0.47% of that in the normal at 6 weeks. It remained low until 12 weeks after neurorrhaphy, which was 0.72% of the normal side. It increased obviously to 0.83 times of normal sides at 18 weeks and didn't changed there after. CONCLUSION: The state of reinnervation determines the quantity of neuromuscular junctional N-AchR.

Animals↗

PubMed1999

Selective inhibition of spinal cord neurite outgrowth and cell survival by the Eph family ligand ephrin-A5.

The Eph family tyrosine kinase receptors and their ligands, the ephrins, have been shown to play critical roles in cell migration, tissue morphogenesis, and axonal guidance in many different systems. However, their function in the spinal cord has not been examined carefully. We showed in this study that several Eph receptors, including EphA3, Eph A4, and Eph A5, are expressed in the ventral spinal cord in partially overlapping patterns, with EphA5 exhibiting the most widespread transcription in the entire ventral spinal cord during early development. Complementary to the receptor expression, a ligand of these receptors, ephrin-A5, is transcribed in the dorsal half of the spinal cord. Consistent with the spatial location of receptor expression, the ligand selectively inhibits neurite outgrowth and induces cell death of the ventral, but not the dorsal, spinal cord neurons. These observations suggest that interactions between the Eph family receptors and ligands exerts negative influences on ventral spinal cord neurons and thus may play important roles in regulating morphogenesis and axon guidance in the spinal cord.

Animals↗

PubMed1999

A role for the Eph ligand ephrin-A3 in entorhino-hippocampal axon targeting.

Neurons of layers II and III of the entorhinal cortex constitute the major afferent connection of the hippocampus. The molecular mechanisms that target the entorhinal axons to specific layers in the hippocampus are not known. EphA5, a member of the Eph receptor family, which has been shown to play critical roles in axon guidance, is expressed in the entorhinal cortex, the origin of the perforant pathway. In addition, ligands that interact with EphA5 are expressed in distinct hippocampal regions during development of the entorhino-hippocampal projection. Of these ligands, ephrin-A3 mRNA is localized both in the granular cell layer of the dentate gyrus and in the pyramidal cell layer of the cornu ammonis, whereas ephrin-A5 mRNA is only expressed in the pyramidal cell layer of the cornu ammonis. In the dentate gyrus, the ligand protein is not present in the termination zone of the entorhinal efferents (the outer molecular layer of the dentate gyrus) but is concentrated in the inner molecular layer into which entorhinal efferents do not grow. We used outgrowth and stripe assays to test the effects of ephrin-A3 and ephrin-A5 on the outgrowth behavior of entorhinal axons. This functional analysis revealed that entorhinal neurites were repelled by ephrin-A3 but not by ephrin-A5. These observations suggest that ephrin-A3 plays an important role in the layer-specific termination of the perforant pathway and that this ligand may interact with the EphA5 receptor to restrict entorhinal axon terminals in the outer molecular layer of the dentate gyrus.

3T3 Cells↗

PubMed1999

Structure-assisted design of mechanism-based irreversible inhibitors of human rhinovirus 3C protease with potent antiviral activity against multiple rhinovirus serotypes.

Human rhinoviruses, the most important etiologic agents of the common cold, are messenger-active single-stranded monocistronic RNA viruses that have evolved a highly complex cascade of proteolytic processing events to control viral gene expression and replication. Most maturation cleavages within the precursor polyprotein are mediated by rhinovirus 3C protease (or its immediate precursor, 3CD), a cysteine protease with a trypsin-like polypeptide fold. High-resolution crystal structures of the enzyme from three viral serotypes have been used for the design and elaboration of 3C protease inhibitors representing different structural and chemical classes. Inhibitors having alpha,beta-unsaturated carbonyl groups combined with peptidyl-binding elements specific for 3C protease undergo a Michael reaction mediated by nucleophilic addition of the enzyme's catalytic Cys-147, resulting in covalent-bond formation and irreversible inactivation of the viral protease. Direct inhibition of 3C proteolytic activity in virally infected cells treated with these compounds can be inferred from dose-dependent accumulations of viral precursor polyproteins as determined by SDS/PAGE analysis of radiolabeled proteins. Cocrystal-structure-assisted optimization of 3C-protease-directed Michael acceptors has yielded molecules having extremely rapid in vitro inactivation of the viral protease, potent antiviral activity against multiple rhinovirus serotypes and low cellular toxicity. Recently, one compound in this series, AG7088, has entered clinical trials.

3C Viral Proteases↗

PubMed1999

Structure-based design of irreversible, tripeptidyl human rhinovirus 3C protease inhibitors containing N-methyl amino acids.

Tripeptide-derived molecules incorporating N-methyl amino acid residues and C-terminal Michael acceptor moieties were evaluated as irreversible inhibitors of the cysteine-containing human rhinovirus 3C protease (3CP). Such compounds displayed good 3CP inhibition activity (k(obs)/[I] up to 610,000 M(-1) s(-1)) and potent in vitro antiviral properties (EC50 approaching 0.03 microM) when tested against HRV serotype-14.

3C Viral Proteases↗

PubMed1999

Identification of a novel gene encoding a p53-associated protein.

p53 exerts important physiological functions in cell-cycle control, gene regulation, cell differentiation, apoptosis and tumor suppression by interacting with many cellular proteins. Using the yeast two-hybrid system, we screened a HeLa cDNA library and identified a novel gene encoding a p53-binding protein (p53BP3). The full-length cDNA of p53BP3 was isolated from a HeLalambdagt10 cDNA library. This predicted protein was composed of 815 amino acids. Sequence analysis indicated that p53BP3 contained two bipartite nuclear localization signals and was confirmed to be a nuclear protein. FISH mapping results showed that this novel gene was located at human chromosome 12, region p11.2-p12.1. Northern blot analysis suggested that p53BP3 was broadly expressed in human tissues. A further study showed that p53BP3 had a homologue in mouse.

Amino Acid Sequence↗

PubMed1999

Structure-based design, synthesis, and biological evaluation of irreversible human rhinovirus 3C protease inhibitors. 3. Structure-activity studies of ketomethylene-containing peptidomimetics.

The structure-based design, chemical synthesis, and biological evaluation of various ketomethylene-containing human rhinovirus (HRV) 3C protease (3CP) inhibitors are described. These compounds are comprised of a peptidomimetic binding determinant and an ethyl propenoate Michael acceptor moiety which forms an irreversible covalent adduct with the active site cysteine residue of the 3C enzyme. The ketomethylene-containing inhibitors typically display slightly reduced 3CP inhibition activity relative to the corresponding peptide-derived molecules, but they also exhibit significantly improved antiviral properties. Optimization of the ketomethylene-containing compounds is shown to provide several highly active 3C protease inhibitors which function as potent antirhinoviral agents (EC90 = <1 microM) against multiple virus serotypes in cell culture.

3C Viral Proteases↗

PubMed1999

Structure-based design, synthesis, and biological evaluation of irreversible human rhinovirus 3C protease inhibitors. 4. Incorporation of P1 lactam moieties as L-glutamine replacements.

The structure-based design, chemical synthesis, and biological evaluation of various human rhinovirus (HRV) 3C protease (3CP) inhibitors which incorporate P1 lactam moieties in lieu of an L-glutamine residue are described. These compounds are comprised of a tripeptidyl or peptidomimetic binding determinant and an ethyl propenoate Michael acceptor moiety which forms an irreversible covalent adduct with the active site cysteine residue of the 3C enzyme. The P1-lactam-containing inhibitors display significantly increased 3CP inhibition activity along with improved antirhinoviral properties relative to corresponding L-glutamine-derived molecules. In addition, several lactam-containing compounds exhibit excellent selectivity for HRV 3CP over several other serine and cysteine proteases and are not appreciably degraded by a variety of biological agents. One of the most potent inhibitors (AG7088, mean antirhinoviral EC90 approximately 0.10 microM, n = 46 serotypes) is shown to warrant additional preclinical development to explore its potential for use as an antirhinoviral agent.

3C Viral Proteases↗

PubMed1999

Ephrin-dependent growth and pruning of hippocampal axons.

Neuronal connections are arranged topographically such that the spatial organization of neurons is preserved by their termini in the targets. During the development of topographic projections, axons initially explore areas much wider than the final targets, and mistargeted axons are pruned later. The molecules regulating these processes are not known. We report here that the ligands of the Eph family tyrosine kinase receptors may regulate both the initial outgrowth and the subsequent pruning of axons. In the presence of ephrins, the outgrowth and branching of the receptor-positive hippocampal axons are enhanced. However, these axons are induced later to degenerate. These observations suggest that the ephrins and their receptors may regulate topographic map formation by stimulating axonal arborization and by pruning mistargeted axons.

3T3 Cells↗