Biomedical subjects

C Zhu

Publications and source records attributed to C Zhu.

At least 109 records · Page 6Linked to original sources

PubMed2000

Cell mechanics: mechanical response, cell adhesion, and molecular deformation.

As the basic unit of life, the cell is a biologically complex system, the understanding of which requires a combination of various approaches including biomechanics. With recent progress in cell and molecular biology, the field of cell mechanics has grown rapidly over the last few years. This review synthesizes some of these recent developments to foster new concepts and approaches, and it emphasizes molecular-level understanding. The focuses are on the common themes and interconnections in three related areas: (a) the responses of cells to mechanical forces, (b) the mechanics and kinetics of cell adhesion, and (c) the deformation of biomolecules. Specific examples are also given to illustrate the quantitative modeling used in analyzing biological processes and physiological functions.

Animals↗

PubMed2000

Effect of long-term mild hypothermia therapy in patients with severe traumatic brain injury: 1-year follow-up review of 87 cases.

OBJECT: The goal of this study was to investigate the protective effects of long-term (3-14 days) mild hypothermia therapy (33-35 degrees C) on outcome in 87 patients with severe traumatic brain injury (TBI) (Glasgow Coma Scale score < or = 8). METHODS: In 43 patients assigned to a mild hypothermia group, body temperatures were cooled to 33 to 35 degrees C a mean of 15 hours after injury and kept at 33 to 35 degrees C for 3 to 14 days. Rewarming commenced when the individual patient's intracranial pressure (ICP) returned to the normal level. Body temperatures in 44 patients assigned to a normothermia group were maintained at 37 to 38 degrees C. Each patient's outcome was evaluated 1 year later by using the Glasgow Outcome Scale. One year after TBI, the mortality rate was 25.58% (11 of 43 patients) and the rate of favorable outcome (good recovery or moderate disability) was 46.51% (20 of 43 patients) in the mild hypothermia group. In the normothermia group, the mortality rate was 45.45% (20 of 44 patients) and the rate of favorable outcome was 27.27% (12 of 44 patients) (p < 0.05). Induced mild hypothermia also markedly reduced ICP (p < 0.01) and inhibited hyperglycemia (p < 0.05). The rates of complication were not significantly different between the two groups. CONCLUSIONS: The data produced by this study demonstrate that long-term mild hypothermia therapy significantly improves outcomes in patients with severe TBI.

Adult↗

PubMed2000

Tetramethylpyrazine inhibits phytohemagglutinin-induced upregulation of ICAM-1 and LFA-1 mediated leukocyte adhesion.

The aim of this work was to study the effects of tetramethylpyrazine (TMP) on the interaction between lymphocyte function-associated antigen-1 (LFA-1) and intercellular adhesive molecule-1 (ICAM-1) by micropipette technique. A pair of cells, i.e., a human lymphocyte or a S KW-3 cell (human T cell leukemia), LFA-1 was expressed on which cellular surfaces, and a RBC coupled with ICAM-1, were the carriers for LFA-1 and ICAM-1. The adhesion probabilities of this pair of cells were mediated by specific interactions between ICAM-1 and LFA-1. Phytohemagglutinin (PHA) stimulation resulted in the significant increase in the adhesion probability compared to the resting lymphocyte. TMP treatment can inhibit such increase and even make the adhesion probability lower than the resting state. While the LFA-1 expression has not changed significantly with PHA stimulation or with TMP treatment, which indicated that TMP inhibiting effects was realized by a possible conformation change.

Adult↗

PubMed2000

[Detection of core promoter mutations in chronic hepatitis B].

OBJECTIVE: To study the core promoter mutation in chronic hepatitis B and its effect on viral serology. METHODS: HBV core promoter gene fragments were amplified by using mismatched PCR combined with a restriction fragment length polymorphism assay. The PCR products were digested with Bcl I and subjected to electrophoresis on agarose gels. RESULTS: We investigated the core promoter mutation in 89 patients with HBV DNA positive. The patterns of restriction fragment length polymorphism (RFIP) of the core promoter gene were distinguished and verified by direct sequencing. The combined mutations of nucleotides (nt) 1762 and 1764 in the core promoter from A to T and G to A were detected in 47 individuals, 21 of 43 HBeAg positive patients and 26 of 46 anti- HBe positive cases were found infected with this combined mutant. These two groups showed no significant difference (P > 0.05). The combined mutation was found in 11 of the 15 patients with medium and severe chronic hepatitis. In 5 chronic hepatitis B patients with coexistence of wild-type and mutant a tendency of superior accumulation of mutant was found. CONCLUSIONS: This study suggests that the core promoter mutations commonly exist in Chinese chronic hepatitis B patients. The mutation can only reduce the pre-core mRNA transcribing efficiency, but cannot discontinue the synthesis of HBeAg. The effect on serology in this mutation is different from that in pre-core stop 28 mutation. The superior accumulation of mutations seems relating to the degree of chronic liver disease.

Adolescent↗

PubMed2000

[A 1:2 matched case-control study on the risk factors of uterine myomas].

OBJECTIVE: To investigate the risk factors and their association with Uterine myomas. METHODS: A 1:2 matched case-control study was carried out in Guangzhou including 211 cases and 422 controls. Data were analyzed with conditional logistic regression. RESULTS: Results showed that Pelvic inflammatory, Erosion of Cervix, Menopathy, Adnexitis and family history of Uterine myomas were identified as risk factors, with odds ratios of 3.101, 2.904, 3.942, 7.742 and 2.426 respectively while delayed menstruation was proved to be a preventive factor, with odds ratios of 0.841. CONCLUSION: Uterine myomas seemed to be related to Pelvic inflammatory, Erosion of Cervix, Adnexitis, Menopathy and family history of Uterine myomas.

Adult↗

PubMed2000

[Effect of high-lipid diet on glomerular mesangial matrix in adriamycin-induced nephrotic rats].

OBJECTIVE: To determined the effect of hypercholesterolemia induced by high-lipid diet on glomerulosclerosis. METHODS: Urinary protein excretion and serum cholesterol were assayed, Image analysis and techniques of pathology, immunohistochemistry, molecular biology were used to determine morphological changes in glomeruli and the production of glomerular mesangial matrixes in adriamycin-induced nephrotic syndrome (NS) and non-NS rats fed with standard chow and high-lipid chow. RESULTS: The serum total cholesterol level was significantly higher in rats with high-lipid chow in both non-NS[(2.2 +/- 0.3) g/L vs (0.9 +/- 0.1) g/L, P < 0.01] and NS[(9.5 +/- 0.2) g/L vs (2.3 +/- 0.3) g/L, P < 0.01]. The urinary protein excretion was significantly higher in the high-lipid diet rats than that in standard chow rats [(76.2 +/- 24.2) mg/24 h vs (44.8 +/- 13.6) mg/24 h, P < 0.05] in NS rats. Significant increases in the mesangial matrix and mesangial cells were observed in rats with high-lipid diet in both NS and non-NS group. Especially, more obvious pathological changes were found in NS group, such as lipid deposits and foam cell formation in mesangial areas, and progressing to focal and segmental glomerulosclerosis in some glomeruli. The immunohistochemical assay showed that the production of fibronectin was increased in the rats with high-lipid diet in non-NS group, whereas, the production of three major components (collagen IV, fibronectin, and laminin) was increased in NS group, especially in the rats with high-lipid chow. The increased expression of laminin mRNA was also detected with slot blotting in both NS and non-NS rats with high-lipid chow, and it was more obvious in the rats with NS. CONCLUSION: Our findings indicated that diet-induced hyperlipidemia can lead to over-production of mesangial matrix components, and further aggravate glomerulosclerosis in adriamycin-induced nephrosis.

Animals↗

PubMed1999

Tyrosine replacement in P-selectin glycoprotein ligand-1 affects distinct kinetic and mechanical properties of bonds with P- and L-selectin.

Selectins are adhesion molecules that initiate tethering and rolling of leukocytes on the vessel wall. Rolling requires rapid formation and breakage of selectin-ligand bonds that must have mechanical strength to resist premature dissociation by the forces applied in shear flow. P- and L-selectin bind to the N-terminal region of P-selectin glycoprotein ligand-1 (PSGL-1), a mucin on leukocytes. To define determinants on PSGL-1 that contribute to the kinetic and mechanical properties of bonds with selectins, we compared rolling of transfected preB cells expressing P- or L-selectin on transfected cell monolayers expressing wild-type PSGL-1 or PSGL-1 constructs with substitutions in targeted N-terminal residues. Rolling through P- or L-selectin required a Thr or Ser at a specific position on PSGL-1, the attachment site for an essential O-glycan, but required only one of three nearby Tyr residues, which are sites for Tyr-SO(3) formation. The adhesive strengths and numbers of cells rolling through P- or L-selectin were similar on wild-type PSGL-1 and on each of the three PSGL-1 constructs containing only a single Tyr. However, the cells rolled more irregularly on the single-Tyr forms of PSGL-1. Analysis of the lifetimes of transient tethers on limiting densities of PSGL-1 revealed that L-selectin dissociated faster from single-Tyr than wild-type PSGL-1 at all shears examined. In sharp contrast, P-selectin dissociated faster from single-Tyr than wild-type PSGL-1 at higher shear but not at lower shear. Thus, tyrosine replacements in PSGL-1 affect distinct kinetic and mechanical properties of bonds with P- and L-selectin.

Amino Acid Sequence↗

PubMed1999

Cloning of mouse mitochondrial thymidine kinase 2 cDNA.

Phosphorylation of anti-viral nucleoside analogs by mitochondrial thymidine kinase 2 (TK2) has been implicated as a mechanism for the mitochondrial toxicity caused by several of these compounds. We have cloned the cDNA of mouse TK2 and shown that the enzyme is targeted to the mitochondria when expressed in fusion with the green fluorescent protein. This is the first report on the cloning of a mitochondrial TK2 and will contribute to elucidate the role of TK2 in the pharmacological activation of nucleoside analogs.

Amino Acid Sequence↗

PubMed1999

A family of mammalian F-box proteins.

Ubiquitin-mediated destruction of regulatory proteins is a frequent means of controlling progression through signaling pathways [1]. F-box proteins [2] are components of modular E3 ubiquitin protein ligases called SCFs, which function in phosphorylation-dependent ubiquitination ([3] [4] [5], reviewed in [6] [7]). F-box proteins contain a carboxy-terminal domain that interacts with substrates and a 42-48 amino-acid F-box motif which binds to the protein Skp1 [2] [3] [4]. Skp1 binding links the F-box protein with a core ubiquitin ligase composed of the proteins Cdc53/Cul1, Rbx1 (also called Hrt1 and Roc1) and the E2 ubiquitin-conjugating enzyme Cdc34 [8] [9] [10] [11]. The genomes of the budding yeast Saccharomyces cerevisiae and the nematode worm Caenorhabditis elegans contain, respectively, 16 and more than 60 F-box proteins [2] [7]; in S. cerevisiae, the F-box proteins Cdc4, Grr1 and Met30 target cyclin-dependent kinase inhibitors, G1 cyclins and transcriptional regulators for ubiquitination ([3] [4] [5] [8] [10], reviewed in [6] [7]). Only four mammalian F-box proteins (Cyclin F, Skp1, beta-TRCP and NFB42) have been identified so far [2] [12]. Here, we report the identification of a family of 33 novel mammalian F-box proteins. The large number of these proteins in mammals suggests that the SCF system controls a correspondingly large number of regulatory pathways in vertebrates. Four of these proteins contain a novel conserved motif, the F-box-associated (FBA) domain, which may represent a new protein-protein interaction motif. The identification of these genes will help uncover pathways controlled by ubiquitin-mediated proteolysis in mammals.

Amino Acid Sequence↗

PubMed1999

Antizyme2 is a negative regulator of ornithine decarboxylase and polyamine transport.

The antizyme family consists of closely homologous proteins believed to regulate cellular polyamine pools. Antizyme1, the first described, negatively regulates ornithine decarboxylase, the initial enzyme in the biosynthetic pathway for polyamines. Antizyme1 targets ornithine decarboxylase for degradation and inhibits polyamine transport into cells, thereby diminishing polyamine pools. A polyamine-stimulated ribosomal frameshift is required for decoding antizyme1 mRNA. Recently, additional novel conserved members of the antizyme family have been described. We report here the properties of one of these, antizyme2. Antizyme2, like antizyme1, binds to ornithine decarboxylase and inhibits polyamine transport. Using a baculovirus expression system in cultured Sf21 insect cells, both antizymes were found to accelerate ornithine decarboxylase degradation. Expression of either antizyme1 or 2 in Sf21 cells also diminished their uptake of the polyamine spermidine. Both forms of antizyme can therefore function as negative regulators of polyamine production and transport. However, in contrast to antizyme1, antizyme2 has negligible ability to stimulate degradation of ornithine decarboxylase in a rabbit reticulocyte lysate.

Amino Acid Sequence↗

PubMed1999

[Detection of ABO genotypes by simultaneous PCR-RFLP method]

OBJECTIVE: To study human ABO genotyping by means of multiplex PCR and restriction fragment length polymorphism. METHODS: Two specific fragments of ABO gene were simultaneously amplified in a single tube, and then the double restriction digestion with RE Kpn I and Alu I was performed in the same tube. The amplified products were analyzed by PAGE and silver staining. A total of 125 Han unrelated individuals living in Wuhan were genotyped. RESULTS: Six genotypes of ABO were detected and the distribution was in good agreement with Hardy-Weinberg equilibrium. CONCLUSION: The results demonstrate that PCR-RFLP based approach is convenient, reliable and should be of value in forensic application.

Journal Article↗

PubMed1999

The mixed lineage kinase DLK utilizes MKK7 and not MKK4 as substrate.

Mixed lineage kinases DLK (dual leucine zipper-bearing kinase) and MLK3 have been proposed to function as mitogen-activated protein kinase kinase kinases in pathways leading to stress-activated protein kinase/c-Jun NH2-terminal kinase activation. Differences in primary protein structure place these MLK (mixed lineage kinase) enzymes in separate subfamilies and suggest that they perform distinct functional roles. Both DLK and MLK3 associated with, phosphorylated, and activated MKK7 in vitro. Unlike MLK3, however, DLK did not phosphorylate or activate recombinant MKK4 in vitro. In confirmatory experiments performed in vivo, DLK both associated with and activated MKK7. The relative localization of endogenous DLK, MLK3, MKK4, and MKK7 was determined in cells of the nervous system. Distinct from MLK3, which was identified in non-neuronal cells, DLK and MKK7 were detected predominantly in neurons in sections of adult rat cortex by immunocytochemistry. Subcellular fractionation experiments of cerebral cortex identified DLK and MKK7 in similar nuclear and extranuclear subcellular compartments. Concordant with biochemical experiments, however, MKK4 occupied compartments distinct from that of DLK and MKK7. That DLK and MKK7 occupied subcellular compartments distinct from MKK4 was confirmed by immunocytochemistry in primary neuronal culture. The dissimilar cellular specificity of DLK and MLK3 and the specific substrate utilization and subcellular compartmentation of DLK suggest that specific mixed lineage kinases participate in unique signal transduction events.

Animals↗

PubMed1999

Specific association of the gene product of PKD2 with the TRPC1 channel.

The function(s) of the genes (PKD1 and PKD2) responsible for the majority of cases of autosomal dominant polycystic kidney disease is unknown. While PKD1 encodes a large integral membrane protein containing several structural motifs found in known proteins involved in cell-cell or cell-matrix interactions, PKD2 has homology to PKD1 and the major subunit of the voltage-activated Ca2+ channels. We now describe sequence homology between PKD2 and various members of the mammalian transient receptor potential channel (TRPC) proteins, thought to be activated by G protein-coupled receptor activation and/or depletion of internal Ca2+ stores. We show that PKD2 can directly associate with TRPC1 but not TRPC3 in transfected cells and in vitro. This association is mediated by two distinct domains in PKD2. One domain involves a minimal region of 73 amino acids in the C-terminal cytoplasmic tail of PKD2 shown previously to constitute an interacting domain with PKD1. However, distinct residues within this region mediate specific interactions with TRPC1 or PKD1. The C-terminal domain is sufficient but not necessary for the PKD2-TRPC1 association. A more N-terminal domain located within transmembrane segments S2 and S5, including a putative pore helical region between S5 and S6, is also responsible for the association. Given the ability of the TRPC to form functional homo- and heteromultimeric complexes, these data provide evidence that PKD2 may be functionally related to TRPC proteins and suggest a possible role of PKD2 in modulating Ca2+ entry in response to G protein-coupled receptor activation and/or store depletion.

Amino Acid Sequence↗

PubMed1999

Coexistence of immune-neuro-endocrine substances in the rat central neurons.

To investigate the expression of interleukin-2 (IL-2), metabotropic glutamate receptor subunit 1 (mGluR1) and estrogen receptor (ER) in neurons of the rat central nervous system (CNS) and identify the coexistence possibility of these immune-neuro-endocrine substances in the central neurons, the tri-labeling immunocytochemical technique with different species-specific primary antibodies (goat anti-IL-2 antibody, rabbit anti-mGluR1 antibody and mouse anti-ER antibody) were used to incubate two serial neighbor sections (one for demonstrating IL-2, another for mGluR1 and ER) of the cerebral cortex, medulla oblongata and spinal cord. There were IL-2-, mGluR1- and ER-immunoreactivity (IR)-positive labeled neurons in the above-mentioned central areas. The IL-2-IR production showed brown color, located in the cytoplasm; In the neighbor serial section, the mGluR1-IR, production showed blue-black color, located on the cell membrane; the ER-IR production also showed brown color, located in the cytoplasm and nuclei. There were mGluR1/ER double-labeled cells in the same section, which accounted for about 50%-60% of the total single and double labeled neurons. It was identified by projection check of serial neighbor sections that had mGluR1/ER/IL-2 tri-labeled cells, which accounted for about 30% of total mGluR1/ER double-labeled neurons. The results indicate that mGluR1, ER and Il-2 can coexist in the same rat central neurons, therefore, providing morphological basis for the theory about immune-neuro-endocrine network at the cellular level for the first time.

Animals↗

PubMed1999

Antisense oligodeoxynucleotide inhibits expression of recombinant porcine follicle-stimulating hormone receptor.

To assess the role of follicle-stimulating hormone receptor (FSHR) gene expression in regulating expression of FSHR protein in the plasma membrane, the effects of a porcine FSHR cDNA antisense oligodeoxynucleotide (ODN) on FSHR mRNA levels and 125I-FSH binding were determined in Chinese hamster ovary cells expression recombinant porcine FSHR (pFSHR-CHO cells). An 18-mer phosphorothioate-endcapped antisense ODN that corresponded to the region surrounding the translation initiation codon of the porcine FSHR cDNA was synthesized. An 18-mer nonsense sequence of identical nucleotide composition, which had little homology to known DNA sequences, was synthesized for use as a control. pFSHR-CHO cells were cultured in 24-well plates (10(5) cells/well) in the absence or presence of 1-20 mumol/L antisense or nonsense ODN for 24 h and then assayed for porcine FSHR mRNA, using quantitative reverse transcription and competitive polymerase chain reaction, and for 125I-FSH binding activity. Treatment with 10 mumol/L antisense ODN caused a paradoxical increase in porcine FSHR mRNA from 0.89 +/- 0.06 to 1.64 +/- 0.08 ng/mg total RNA (P < 0.05). Transfection with lipofectamine and 0.33 mumol/L antisense ODN caused an increase in porcine mRNA from 0.95 +/- 0.08 to 1.53 +/- 0.07 ng/mg total RAN. This was probably due to upregulation of mRNA synthesis resulting from inhibition of porcine FSHR protein translation. The nonsense ODN had no effect on porcine FSHR mRNA. Antisense, but not nonsense, ODN (10 mumol/L) inhibited membrane binding of 125I-FSH by 13.6 +/- 0.8% (P < 0.05) in 24 h. Treatment of cells with antisense ODN (10 mumol/L) for 48 h resulted in a 76 +/- 1.5% (P < 0.05) inhibition of 125I-FSH binding. In contrast, transfection with lipofectamine and 0.33 mumol/L antisense ODN at 0 h caused a 76.1 +/- 1.3% (P < 0.05) reduction in binding within 24 h. Binding had returned to 52.3 +/- 2.3% (P < 0.05) of normal by 48 h. These results indicate that an antisense ODN corresponding to the region of the translation start site of the porcine FSHR cDNA is an effective specific inhibitor of porcine FSHR synthesis and that inhibition of receptor synthesis causes a decrease in functional membrane-bound FSHR.

Animals↗

PubMed1999

Analysis of competition binding between soluble and membrane-bound ligands for cell surface receptors.

Binding of the Fc portion of IgG coated on targets to Fcgamma receptors (e.g., CD16) expressed on leukocytes (i.e., 2D binding) is an initiating step for immune responses such as phagocytosis or antibody-dependent cellular cytotoxicity. In vivo, circulating leukocytes are exposed to plasma IgG. The competition from soluble IgG (i.e., 3D binding) may affect the 2D binding. Many cell surface receptors, CD16 included, have soluble counterparts. While their physiological significance is not clear, receptor-based competitive inhibition therapy, in which soluble receptors, ligands, and their analogs are employed to compete with surface-bound receptors and ligands to prevent unwanted adhesion, is widely used to treat various diseases. To provide a quantitative basis for design of these therapeutic approaches, we developed a mathematical model for 2D and 3D competition binding. The model relates cell-surface adhesion (in the presence and absence of dislodging forces) to the concentration of the soluble competitor, the densities of the surface-bound receptors and ligands, as well as the binding affinities of the 2D and 3D interactions. Binding of CD16-expressing cells to an IgG-coated surface in the presence of a soluble competitor (IgG or anti-CD16 antibody) was quantified by a centrifugation assay. The agreement between experiment and theory supports the validity of the model, which could be useful in predicting the efficacy of the competitor.

Animals↗

PubMed1999

Probabilistic modeling of shear-induced formation and breakage of doublets cross-linked by receptor-ligand bonds.

A model was constructed to describe previously published experiments of shear-induced formation and breakage of doublets of red cells and of latexes cross-linked by receptor-ligand bonds (. Biophys. J. 65:1318-1334; Tees and Goldsmith. 1996. Biophys. J. 71:1102-1114;. Biophys. J. 71:1115-1122). The model, based on McQuarrie's master equations (1963. J. Phys. Chem. 38:433-436), provides unifying treatments for three distinctive time periods in the experiments of particles in a Couette flow in which a doublet undergoes 1) formation upon two-body collision between singlets; 2) evolution of bonds at low shear rate; and 3) break-up at high shear rate. Neglecting the applied force at low shear rate, the probability of forming a doublet per collision as well as the evolution of probability distribution of bonds in a preformed doublet were solved analytically and found to be in quite good agreement with measurements. At high shear rate with significant force acting to accelerate bond dissociation, the predictions for break-up of doublets were obtained numerically and compared well with data in both individual and population studies. These comparisons enabled bond kinetic parameters for three types of particles cross-linked by two receptor-ligand systems to be calculated, which agreed well with those computed from Monte Carlo simulations. This work can be extended to analyze kinetics of receptor-ligand binding in cell aggregates, such as those of neutrophils and platelets in the circulation.

Cell Aggregation↗

PubMed1999

Intra-arterial blood pressure reference ranges, death and morbidity in very low birthweight infants during the first seven days of life.

OBJECTIVES: We aimed to: (1) assess the association of average, low, high and variable mean blood pressure (mbp) on death and the common morbidities of very low birthweight infants, and in doing so, (2) to derive representative reference ranges for mbp in very low birthweight infants. STUDY DESIGN: This five year retrospective study assessed 1 min computer recordings of intra-arterial mbp in 232 very low birthweight infants over the first 7 days of life in a tertiary NICU. Four measures of mbp were assessed: average, variability, maximum (per time period), and percentage of time with a mean blood pressure less than the infant's gestation. Correlation was made with death and the development of intraventricular haemorrhage (IVH), periventricular leukomalacia (PVL) and retinopathy of prematurity (ROP). RESULTS: The mbp increased with increasing birthweight and postnatal age (though with a slight decrease on days 6 and 7). Birthweight, gestation and colloid support (adjusted for birthweight and gestation) were the only factors significantly associated with mbp. IVH was predominantly associated with a low and variable mbp on the day IVH was noted or the day before. PVL and ROP were not associated with blood pressure. CONCLUSIONS: These reference ranges include more infants and data than previously published and relate mbp in this cohort to morbidity and mortality. They could assist clinicians in judging appropriate mbp for birthweight.

Aging↗