Biomedical subjects

C Zhu

Publications and source records attributed to C Zhu.

At least 19 recordsLinked to original sources

PubMed2002

Determination of toxicokinetic parameters for bioconcentration of water-soluble fraction of petroleum hydrocarbon associated with no. 0 diesel in Changjiang estuary and Jiaozhou bay: model versus mesocosm experiments.

A method is proposed for determination of toxicokinetic parameters for bioconcentration by phytoplankton of the water-soluble fraction (WSF) of petroleum hydrocarbon (PH) associated with No. 0 diesel, in which WSF-PH concentration in phytoplankton cells, C(A(d)), is estimated by subtracting concentration in water (S-bottle) containing a phytoplankton sample from that in a C-bottle without phytoplankton. It was demonstrated that C(A(d)) agrees well with the concentration found by direct ultrasonication extraction of collected cells, C(A(ind)) ( r = 0.88, p < 0.0001), and its uncertainty was about 17.6%. Mesocosms in 25-m3 ethylene vinyl acetate or 4-m3 polyethylene bags were performed at two sites in China: Changjiang Estuary in spring/summer 1998 and Jiaozhou Bay in autumn 1999 and spring/summer 2000. The experiments were designed to determine toxicokinetic parameters, including specific rates of uptake and elimination, and bioconcentration factor (BCF), for bioconcentration of WSF-PH by phytoplankton. A modified kinetic two-compartment model for bioconcentration of WSF-PH by phytoplankton was developed to estimate the toxicokinetic parameters. In the model, the influence of phytoplankton growth on bioconcentration and WSF-PH decline due to biotic and abiotic processes other than bioconcentration, such as volatilization, microbial degradation, phytolysis, and sorption expressed as an exponential-decay equation, are taken into account. Size-dependent BCF was observed in the laboratory experiment. BCFs were 1.0 x 10(4) in summer in Changjiang Estuary, 1.6 x 10(4) in summer, and 1.1 x 10(4) in autumn in Jiaozhou Bay. The difference in BCF may be interpreted by its size dependence.

Biological Availability↗

PubMed2002

Administration of acetylcholine to the spermatic nerve plexus inhibits testosterone secretion in an in vitro isolated rat testis-nerve plexus system.

Strong evidence indicated that spermatic nerves are involved in the regulation of testosterone secretion. Our previous work showed that the inferior spermatic nerves play a more significant role than the superior ones in the regulation of testosterone secretion. However, it is unknown whether traditional neurotransmitters are involved in this regulation. In order to evaluate this point, the present experiments were carried out in an in vitro system where an isolated testis-spermatic nerve plexus preparation was incubated in two separate containers, one for the testis and the other for the nerve plexus and both interconnected by the inferior spermatic nerves. Both tissues were maintained in the same environmental conditions except for the neurotransmitter treatment, applied only to the nerve plexus. Acetylcholine can significantly inhibit the secretion of testosterone until the end of incubation. The present experiments suggest that the secretion of testosterone could be regulated, at least in part, by acetylcholine through the inferior spermatic nerves.

Acetylcholine↗

PubMed2002

Expression of a blood-brain barrier-specific antigen in the reproductive tract of the male rat.

The endothelial barrier antigen (EBA) is a protein expressed specifically by the endothelial cells of the rat brain barrier vessels. This antigen has been described as a 'barrier protein' and is used as a marker for the competent blood-brain barrier. A blood-testis barrier has also been described. However, unlike the blood-brain barrier, which is formed by endothelial cells, the blood-testis barrier is formed mainly by the Sertoli cells, which provide an isolated environment for spermatogenic cells within the seminiferous tubules. Testicular blood vessels express the erythroid glucose transporter protein and other markers, which are strongly expressed in brain blood vessels, and may contribute to the blood-testis barrier. This study was carried out to determine whether Sertoli cells or testicular blood vessels express EBA. Tissues of other organs were used as controls for EBA expression. EBA was expressed by the endothelial cells in most microvessels of the testis, and in a few vessels of the epididymis, seminal vesicle, prostate gland, vas deferens and bladder-neck region. Furthermore, EBA was strongly and consistently detected in epithelial cells of the rete testis and dorsolateral prostate gland, and in a few epithelial cells of the ventral prostate gland, the seminal vesicle and the coagulating gland. However, Sertoli cells, which are the main site of the blood-testis barrier, were negative for EBA. In conclusion, EBA may have a wider role in rat tissues than has been previously appreciated.

Animals↗

PubMed2001

Ecology. Giant pandas in a changing landscape.

The giant panda has been restricted to several disjunct montane forest populations, and habitat loss and fragmentation are the primary threats to its survival. For pandas to survive, conservation efforts must focus on larger landscapes rather than individual nature reserves. China recently initiated several policies, including the Natural Forest Conservation Program and Grain-to-Green Policy, which provide a historic opportunity to integrate panda conservation into national policies. Simultaneously, China is promoting the Western China Development Program, which calls for substantial infrastructure and hydropower development and economic investments. Integrating panda conservation into these development policies will be a critical challenge.

Animals↗

PubMed2001

Synthesis and characterization of 2D and 3D structures from organic derivatives of polyoxometalate clusters: role of organic moiety, counterion, and solvent.

New organic derivatives of "inverse-Keggin" polyoxometalate (POM) clusters, [Mo(12)O(46)(AsR)(4)](4-) (R = C(6)H(4)-4-COOH, C(6)H(3)-4-OH-3-NO(2), C(6)H(4)-4-OH), have been synthesized. Structures of the corresponding sodium or iron salts were elucidated by single-crystal X-ray diffraction and shown to be 3D structures connected through hydrogen bonds and/or O-Na-O linkages. Parameters which influence the final solid-state architecture, such as the crystallizing solvent, organic moiety, and counterions, have been examined. The hydrogen-bonding ability of the solvent affects the connectivity of the POM clusters through interactions with the organic group and the inorganic core. The organic groups influence the structure through hydrogen bonds to other organic groups, to neighboring clusters, and/or to solvent molecules. Hydrogen bonding between the organic groups and the solvent appears to inhibit some possible connectivity patterns, such as the hydrogen-bonded dimerization of carboxylic acid groups. Na(+) ions were found directly bonded to the cluster oxo ligands and provided linkages between clusters. Larger cations, such as transition metals, did not show this interaction, and other bonding methods dominated.

Journal Article↗

PubMed2001

Bystander effects of nucleoside analogs phosphorylated in the cytosol or mitochondria.

The efficiency of nucleoside kinase suicide gene therapy for cancer is highly dependent on "bystander" cell killing, i.e., the transfer of cytotoxic phosphorylated nucleoside analogs to cells adjacent to those expressing the suicide enzyme. We have recently studied the possible use of mitochondrial nucleoside kinases as suicide genes. In the present study, we investigated if nucleoside analogs phosphorylated in the mitochondrial matrix cause bystander killing. We used deoxycytidine kinase-deficient Chinese hamster ovary cells reconstituted with deoxycytidine kinase targeted to either the cytosol or mitochondria matrix and determined the bystander cell killing when these cells were incubated with the nucleoside analogs 1-beta-D-arabinofuranosylcytosine and 2',2'-difluorodeoxycytidine. A bystander effect occurred when nucleoside analogs were phosphorylated in the cytosol, but not when these compounds were phosphorylated in the mitochondria. These findings suggest that nucleoside kinases targeted to the mitochondrial matrix have limited use in suicide gene therapy when efficient bystander cell killing is required.

Animals↗

PubMed2001

Brain-specific expression of an exogenous gene after i.v. administration.

The treatment of brain diseases with gene therapy requires the gene to be expressed throughout the central nervous system, and this is possible by using gene targeting technology that delivers the gene across the blood-brain barrier after i.v. administration of a nonviral formulation of the gene. The plasmid DNA is targeted to brain with pegylated immunoliposomes (PILs) using a targeting ligand such as a peptidomimetic mAb, which binds to a transporting receptor on the blood-brain barrier. The present studies adapt the PIL gene targeting technology to the mouse by using the rat 8D3 mAb to the mouse transferrin receptor. Tissue-specific expression in brain and peripheral organs of different exogenous genes (beta-galactosidase, luciferase) is examined at 1-3 days after i.v. injection in adult mice of the exogenous gene packaged in the interior of 8D3-PIL. The expression plasmid is driven either by a broadly expressed promoter, simian virus 40, or by a brain-specific promoter taken from the 5' flanking sequence of the human glial fibrillary acidic protein (GFAP) gene. The transgene is expressed in both brain and peripheral tissues when the simian virus 40 promoter is used, but the expression of the exogenous gene is confined to the brain when the transgene is under the influence of the brain-specific GFAP promoter. Confocal microscopy colocalizes immunoreactive bacterial beta-galactosidase with immunoreactive GFAP in brain astrocytes. These studies indicate that tissue-specific gene expression in brain is possible after the i.v. administration of a nonviral vector with the combined use of gene targeting technology and tissue-specific gene promoters.

Animals↗

PubMed2001

Dimerization of a selectin and its ligand stabilizes cell rolling and enhances tether strength in shear flow.

Selectins mediate rolling of leukocytes by rapid formation and dissociation of selectin-ligand bonds, which are assumed to require high mechanical strength to prevent premature dissociation by the forces applied in shear flow. This assumption is based largely on the observation that increasing wall shear stress increases only modestly the dissociation of transient leukocyte tethers on very low selectin densities. P-selectin binds to the N-terminal region of P-selectin glycoprotein ligand-1 (PSGL-1), a mucin on leukocytes. Both PSGL-1 and P-selectin are extended homodimers. We perfused transfected cells expressing wild-type dimeric PSGL-1 or a chimeric monomeric form of PSGL-1 on immobilized dimeric or monomeric forms of P-selectin. Cells expressing dimeric or monomeric PSGL-1 tethered to P-selectin at equivalent rates. However, cells expressing dimeric PSGL-1 established more stable rolling adhesions, which were more shear resistant and exhibited less fluctuation in rolling velocities. On low densities of dimeric P-selectin, increasing wall shear stress more rapidly increased transient tether dissociation of cells expressing monomeric PSGL-1 than dimeric PSGL-1. Tether dissociation on low densities of monomeric P-selectin was even more shear sensitive. We conclude that dimerization of both PSGL-1 and P-selectin stabilizes tethering and rolling, probably by increasing rebinding within a bond cluster. Because transient tethers may have more than one bond, the mechanical strength of selectin-ligand bonds is likely to be lower than initially estimated. Tether strength may rely more on bond clusters to distribute applied force.

Animals↗

PubMed2001

Dual mechanisms of 9-beta-D-arabinofuranosylguanine resistance in CEM T-lymphoblast leukemia cells.

The guanine nucleoside analog araG is selectively toxic to T-lymphoblasts and has recently shown promise in treatment of lymphoid malignancies of T-cell origin. The molecular mechanism of this tissue-selective cytotoxicity is, however, yet unclear. AraG is phosphorylated, and thereby pharmacologically activated, by the mitochondrial deoxguanosine kinase and the cytosolic/nuclear deoxycytidine kinase. We have recently shown that araG is predominantly incorporated into mitochondrial DNA of cancer cell lines, which suggests a role of mitochondria as its pharmacological target. In the present study, we have generated araG-resistant CEM T-lymphoblast cell lines and show that araG resistance may occur by two separate molecular mechanisms that can occur sequentially. The first mechanism is associated with a decrease of araG incorporation into mitochondrial DNA, and the second event is associated with loss of dCK activity.

Antineoplastic Agents↗

PubMed2001

[Gene diagnosis for hemophilia a using A combination of 4 techniques].

OBJECTIVE: To improve the gene diagnosis,carrier detecting and prenatal diagnosis for hemophilia A families to a maximum. METHODS: Long distance PCR (LD-PCR) was used to detect FVIII gene inversion among 26 patients with severe HA and their female offspring to identify those with FVIII gene inversion and its carriers. Three gene linkage analysis based upon PCR, Bcl I PCR/RFLP, St14 VNTR/PCR analysis and two simple dinucleotides repeats polymorphism analysis were used among 19 non-inversion HA families for indirect diagnosis. RESULTS: Sixteen families with severe HA were detected by LD-PCR among the 26 hemophilia A families. Seven were confirmed as with FVIII gene inversion (43.8%). The diagnostic rates by the above mentioned three indirect diagnostic techniques among nineteen families without gene inversion were 84.2%, 68.4% and 89.5% respectively. Confirmed diagnosis was successfully made among all of the HA families by the four techniques. CONCLUSION: Gene diagnosis and carrier detecting can be confirmed among almost all of the HA families can be diagnosed by the combined use of the four direct and indirect gene diagnostic methods.

Adolescent↗

PubMed2001

[Effects of helicobacter pylori infection on gastric mucosaia: a 5-year cohort study].

OBJECTIVE: To observe the changes in severity of gastritis and intestinal metaplasia by following-up a group of patients for 5 years to explore the effect of Helicobacter pylori (Hp) on the disease process. METHODS: In this prospective cohort study, patients were selected from 7 enterprises by regular endoscopic examination in Zhongshan Hospital from July 1993 to June 1994. Recruited patients were those with upper abdominal discomfort and diagnosed as chronic gastritis endoscopically and pathologically. Hp-positive patients were given either eradication therapy or symptomatic treatment according to their willingness. All the patients were followed-up for 5 years. RESULTS: Among the total 75 patients (30 were Hp positive and 45 were Hp negative), 24 Hp-negative cases were recruited as control group. Among 45 originally Hp-positive patients, except for one drop-out case, 16 cases in whom Hp was eradicated were defined as the eradication group and the remaining 28 patients with persistent Hp infection were assigned into the infection group. In the control group, in only 17% approximately 33% of patients the gastritis was worsened. In the infection group, 29% approximately 42% of patients had no changes in their gastritis severity, however in 43% approximately 57% of patients the disease was worsened. While in the eradication group, in 50% of patients the disease was improved, and in only 25% of patients the disease was aggravated, with an aggravalion rate much lower than that in infection group. For intestinal metaplasia (IM), in the control group, 16% approximately 25% of patients had their IM aggravated, but no changes occurred in 50% approximately 67% of patients. In the infection group, the rate of IM worsening increased to 43%-57%, while no IM worsening could be seen in eradication group, 75.0% of the patients in this group maintained their original status. CONCLUSIONS: Eradication of Hp could not only ameliorate the severity of gastritis, but also inhibit the development of IM.

Adult↗

PubMed2001

Independent regulation of cardiac Kv4.3 potassium channel expression by angiotensin II and phenylephrine.

Hypertrophied cardiac myocytes exhibit prolonged action potentials and decreased transient outward potassium current (I(to)). Because Kv4.3 is a major contributor to I:(to), we studied regulation of its expression in neonatal rat cardiac myocytes in response to the known stimulators of cardiac myocyte hypertrophy, angiotensin II (Ang II) and phenylephrine (PE). RNase protection assays and immunoblots revealed that Ang II and PE each downregulate Kv4.3 mRNA and protein. However, although PE induces a faster and more extensive hypertrophic response than Ang II, the PE effect on Kv4.3 mRNA develops slowly and is sustained, whereas Ang II rapidly and transiently decreases Kv4.3 mRNA expression. Turnover measurements revealed that Kv4.3 mRNA is very stable, with a half-life >20 hours. This suggests that Ang II must destabilize the channel mRNA. In contrast, PE does not affect the rate of Kv4.3 mRNA degradation. To test for transcriptional regulation, the 5' flanking region of the rat Kv4.3 gene was cloned, and Kv4.3 promoter-reporter constructs were expressed in cardiac myocytes. Whereas Ang II was found to have no effect on transcription, PE inhibits Kv4.3 promoter activity. Pharmacological experiments also indicate that PE and Ang II act independently to downregulate Kv4.3 gene expression. Thus, regulation of Kv4.3 gene expression is not a simple secondary response to hypertrophy. Rather, Ang II and PE use different mechanisms to decrease Kv4.3 channel expression in neonatal rat cardiac myocytes.

Angiotensin II↗

PubMed2001

2'-O-Acyl/alkyl-substituted arabinosyl nucleosides as inhibitors of human mitochondrial thymidine kinase.

Introduction of a bulky lipophilic acyl entity at the 2'-OH position of both 1-beta-D-arabinofuranosylthymine (araT) and (E)-5-(2-bromovinyl)-1-beta-D-arabinofuranosyluracil (BVaraU), consistently resulted in a marked ( approximately 10-fold) increase in the inhibitory activity of these new arabinosyl nucleoside analogues for the mitochondrial thymidine kinase (TK-2)-catalysed conversion of 2 microM [methyl-(3)H]dThd to [methyl-(3)H]dTMP. The most potent derivatives were inhibitory to [methyl-(3)H]dThd phosphorylation by TK-2 within the lower micromolar concentration range. Substitution of the arabinosyl nucleoside derivatives with the acyl groups also dramatically increased the selectivity of these compounds. The inhibitory activity of araT and BVaraU to dThd phosphorylation by other related nucleoside kinases, including herpes simplex virus type 1 TK, varicella-zoster virus TK, and cytosolic TK-1, was completely annihilated upon 2'-O-acyl substitution (IC(50) > or = 1000 microM). Kinetic analysis revealed purely competitive inhibition of 2'-O-acyl-BVaraU against TK-2-catalysed thymidine phosphorylation (K(i)/K(m): 2.3). However, 2'-O-acyl-BVaraU was extremely poorly converted to the corresponding arabinosyl nucleoside 5'-monophosphate by TK-2 as revealed by [gamma-(32)P]phosphate transfer studies from [gamma-(32)P]ATP. Thus, the 2'-O-acyl derivatives of BVaraU did not behave as substrates, but rather as potent and highly selective inhibitors of TK-2. This is the first report on such a highly selective arabinosyl nucleoside inhibitor of mitochondrial TK-2, and opens perspectives for the rational design of selective mitochondrial TK-2 inhibitors.

Adenosine Triphosphate↗

PubMed2001

Characterization of an activation protein-1-binding site in the murine interleukin-12 p40 promoter. Demonstration of novel functional elements by a reductionist approach.

Interleukin (IL)-12 is a heterodimeric cytokine produced by macrophages in response to intracellular pathogens and provides an obligatory signal for the differentiation of T-helper-1 cells. We previously reported an analysis of the IL-12 p40 promoter in RAW264.7 macrophages. Multiple control elements were involved in activation of transcription by bacterial products. A critical control element, located between -96 and -88, interacts with C/EBP family members. In this study, using a strategy to demonstrate functional activity in a minimal promoter context, three novel cis-acting elements are found to have an important role in IL-12 p40 promoter activation by lipopolysaccharide. One of these elements is characterized in detail. Mutations from -79 to -74 in the murine IL-12 p40 promoter significantly reduce lipopolysaccharide-induced promoter activity. Electrophoretic mobility shift assays demonstrate binding of AP-1 family members to this region. Spacing between the C/EBP and AP-1 site is important for promoter activation, suggesting cooperativity between these elements. c-Jun and a mutant c-Jun molecule activate the IL-12 p40 promoter and synergistically activate the promoter when co-expressed with C/EBPbeta. Finally, this region of the promoter is demonstrated to be a target for mitogen-activated protein kinase and toll-like receptor signaling pathways.

Animals↗

PubMed2001

A premature-termination mutation in the Mus musculus cyclin-dependent kinase 3 gene.

Our understanding of the mammalian cell cycle is due in large part to the analysis of cyclin-dependent kinase (CDK) 2 and CDK4/6. These kinases are regulated by E and D type cyclins, respectively, and coordinate the G(1)/S-phase transition. In contrast, little is known about CDK3, a homolog of CDK2 and cell division cycle kinase 2 (CDC2). Previous studies using ectopic expression of human CDK3 suggest a role for this kinase in the G(1)/S-phase transition, but analysis of the endogenous kinase has been stymied by the low levels of protein present in cells and by the absence of an identifiable cyclin partner. Herein we report the presence of a single point mutation in the CDK3 gene from several Mus musculus strains commonly used in the laboratory. This mutation results in the replacement of a conserved tryptophan (Trp-187) within kinase consensus domain IX with a stop codon. The protein predicted to be encoded by this allele is truncated near the T loop, which is involved in activation by CDK-activating kinase. This mutation also deletes motif XI known to be required for kinase function and is, therefore, expected to generate a null allele. In stark contrast, CDK3 from two wild-mice species (Mus spretus and Mus mus castaneus) lack this mutation. These data indicate that CDK3 is not required for M. musculus development and suggest that any functional role played by CDK3 in the G(1)/S-phase transition is likely to be redundant with another CDK.

Amino Acid Sequence↗

PubMed2001

Quantifying the impact of membrane microtopology on effective two-dimensional affinity.

Just as interactions of soluble proteins are affected by the solvent, membrane protein binding is influenced by the surface environment. This is particularly true for adhesion receptors because their function requires tightly apposed membranes. We sought to demonstrate, and further, to quantify the possible scale of this phenomenon by comparing the effective affinity and kinetic rates of an adhesion receptor (CD16b) placed in three distinct environments: red blood cells (RBCs), detached Chinese hamster ovary (CHO) cells, and K562 cells. Effective affinity reflects both the intrinsic receptor-ligand kinetics and the effectiveness of their presentation by the host membranes. Expression of CD16b, a low affinity Fcgamma receptor, was established by either transfection or spontaneous insertion via its glycosylphosphatidylinositol anchor. Binding to IgG-coated RBCs, measured using a micropipette method, indicated a 50-fold increase in effective affinity for receptors on RBCs over CHO and K562 cells, whereas the off rates were similar for all three. Electron microscopy confirmed that specific tight contacts were broad in RBC-RBC conjugates but sparse in CHO-RBC conjugates. We suggest that through modulation of surface roughness the cytoskeleton can greatly impact the effectiveness of adhesion molecules, even those with no cytoplasmic structures. Implications for locomotion and static adhesion are discussed.

Animals↗

PubMed2001

Structure-activity studies of 5-substituted pyridopyrimidines as adenosine kinase inhibitors.

The synthesis and SAR of a novel series of non-nucleoside pyridopyrimidine inhibitors of the enzyme adenosine kinase (AK) are described. It was found that pyridopyrimidines with a broad range of medium and large non-polar substituents at the 5-position potently inhibited AK activity. A narrower range of analogues was capable of potently inhibiting adenosine phosphorylation in intact cells indicating an enhanced ability of these analogues to penetrate cell membranes. Potent AK inhibitors were found to effectively reduce nociception in animal models of thermal hyperalgesia and persistent pain.

Adenosine Kinase↗

PubMed2001

Clostridium perfringens prototoxin-induced alteration of endothelial barrier antigen (EBA) immunoreactivity at the blood-brain barrier (BBB).

It has been reported that the severe cerebral edema produced in experimental animals by Clostridium perfringens (Cl p) type D epsilon toxin can be prevented by prior treatment with its precursor prototoxin due to competitive binding to endothelial cells (ECs) at the blood-brain barrier (BBB). In this study we investigate the effects of the prototoxin on the BBB, without added toxin. The integrity of the BBB was assessed by its ability to prevent leakage of endogenous albumin. ECs at the BBB were studied by immunocytochemistry for any alteration in the endothelial barrier antigen (EBA), a molecular marker for the intact BBB. Immunocytochemistry showed rapid but mild opening of the BBB to endogenous albumin. Light and electron immunocytochemistry showed qualitative and quantitative reduction in EBA immunoreactivity, with a spectrum of changes at time intervals from 1 h to 14 days post-prototoxin injection. Some vessels with ultrastructural changes and widening of the perivascular space retained EBA immunoreactivity. Many vessels showed partial or complete loss of EBA staining with minimal widening of the perivascular space and edema. Recovery of EBA expression was still incomplete at 14 days postinjection. This is the first report to show endothelial cell damage, mild reversible cerebral edema, and alteration in BBB markers following administration of Cl p prototoxin. This model of mild brain edema may be useful for BBB studies.

Animals↗