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Biomedical subjects

Q Zhang

Publications and source records attributed to Q Zhang.

At least 685 records · Page 38Linked to original sources

[Experimental study on biological characteristics of tenocyte and fibroblast in rabbit].

In order to study the biological characteristics of tenocyte and fibroblast, the former was obtained from rabbit's tendon, and the latter from rabbits's skin. Both cells were cultured according Heuderson's method. The cell morphology, strapping and expanding time, and the type of collagen fiber synthesized in culture were observed. The results showed that the strapping and expanding time of fibroblast was faster than that of tenocyte. The cellular arrangement of fibroblast was irregular, but that in tenocyte was regular. Type I and III collagen of fibers were found in cultured fibroblost while only type I collagen fibers were found in culture of tenocyte. The tenocyte and fibroblast could be identified individually by strapping and expanding time, arrangement of cells and type of collagen fiber synthesized.

Animals↗

[Morphological observation of combined-culture of tendon cell or fibroblast of rabbit with artificial materials in vitro].

In order to investigate the compatibility and growth between the tendon cell or fibroblast of rabbit and artificial materials, the combined-culture of the two cells with the carbon fiber, terylene and chitin was observed respectively. Results showed as following: in vitro, the compatibility of carbon fiber with these two cells was well, cell-adhesion ability was good as well. Few cells grew on terylene. Chitin inhibited the growth of either cells. No matter the tendon cell or the fibroblast, the amount of cells adhering on the carbon fiber was far more than that on terylene or chitin. When the three materials were interlaced together, the collagen fibers produced by the cells were arranged in direction parallel to the carbon fibers. As the time elapsed, the cells on the carbon fiber distributed evenly and enveloped the material in network-like fashion, this suggested that carbon fiber was a good material for producing living artificial tendon and ligament.

Animals↗

[Detection of p53 gene mutations in human gestational trophoblastic neoplasia by polymerase chain reaction-single strand conformational polymorphism techniques].

Using polymerase chain reaction single strand conformational polymorphism (PCR-SSCP) and EB staining technique, paraffin-embeded sections of 20 hydatidiform mole and 4 choriocarcinoma were detected in the exons 5 and 8 of p53 gene. The results showed that mutations of p53 gene were 0/9 in the normal chorionic villi, 6/20 (30%) in hydatidiform mole and 3/4 in choriocarcinoma. This study suggests that mutations of p53 gene may be an important event in human gestational trophoblastic neoplastigenesis and its progression.

Choriocarcinoma↗

Cross-linking of the NH2-terminal region of fibronectin to molecules of large apparent molecular mass. Characterization of fibronectin assembly sites induced by the treatment of fibroblasts with lysophosphatidic acid.

Cell surface molecules on adherent cells that bind 125I-labeled fibronectin or its 70-kDa N-terminal fragment were identified by cross-linking with factor XIIIa and by photoaffinity labeling. Such cross-linking caused the 70-kDa fragment to become associated irreversibly to cell layers and was greater in cells treated with lysophosphatidic acid, an enhancer of fibronectin assembly and strong modulator of cell shape. Cross-linking of the 70-kDa fragment with factor XIIIa was to molecules that migrated in discontinuous sodium dodecyl sulfate-polyacrylamide gels at the top of the 3.3% stacking gel and near the top of the separating gel. Estimated sizes of these large apparent molecular mass molecules (LAMMs) were >>3 MDa and approximately 3 MDa. The label in 70-kDa fragment conjugated with 125I-sulfosuccinimidyl 2-(p-azidosalicylamido)-1, 3'-dithiopropionate was associated with >>3-MDa LAMMs without reduction and with approximately 3-MDa LAMMs after reduction and transfer of the cleavable label. The LAMMs were expressed on monolayer cells shortly after adherence, required both 1% Triton X-100 and 2 M urea for efficient extraction, and were susceptible to digestion with trypsin but not to cathepsin D digestion. Complexes of 125I-70-kDa fragment and LAMMs were also susceptible to limited acid digestion and Glu-C protease digestion but were not cleaved by chondroitin lyase or heparitinase. Neither the uncleaved complexes nor the cleavage products were immunoprecipitated with anti-fibronectin antibodies directed toward epitopes outside the 70-kDa region. Thus, cell surface molecules that are either very large or not dissociated in sodium dodecyl sulfate comprise the labile matrix assembly sites for fibronectin.

Antineoplastic Agents↗

Experiments on the transmission of Babesia major and Babesia bigemina by Haemaphysalis punctata.

Experiments on the transmission by Haemaphysalis punctata of three large Babesia strains were carried out. Three Babesia-free batches of laboratory reared H. punctata ticks were infected with two strains of Babesia major, B. major (Xingjiang strain), isolated with adult ticks of H punctata and B. major (Henan strain), isolated with H. longicornis) and a strain of Babesia bigemina by feeding them on the calves infected by inoculation of blood stabilates. H. punctata was shown to be capable of transmitting the B. major strains transovarially. The larvae, nymphs and adults developed from female ticks engorged on the calf infected with B. major (Xingjiang strain) transmitted the pathogen to splenectomised calves with prepatent periods of 15, 11 and 12 days, respectively. The calves infested with larvae and nymphs died of babesiosis with parasitemias of 400 and 710 per 1000 erythrocytes. The calf infested with adult ticks survived babesiosis, but the number of erythrocytes and the amount of haemoglobin were reduced greatly. H. punctata transmitted B. major (Henan strain) in the same way. The prepatent periods of the calves infested with larvae, nymph and adult ticks were 9, 10 and 12 days, respectively. Calves infested with larvae survived, but those infested with nymphal and adult ticks died of babesiosis with parasitemias of 410 and 100 per 1000 erythrocytes, respectively. H. punctata ticks did not transmit the B. bigemina strain to splenectomised calves. There were no clinical symptoms and no parasites were discovered in the blood films during a 2 month observation period after the calves were infested with larval, nymphal and adult ticks derived from female ticks engorged on calves inoculated with B. bigemina.

Animals↗

The polymorphism of GSTM1, mutagen sensitivity in colon cancer and healthy control.

The aim of this article is to study the relationship between GSTM1 polymorphism and colon cancer and to compare the chromosomal breakage induced by mutagen in a colon cancer group and healthy controls. Using PCR to identify the GSTM1 genotype, we found the frequency of GSTM1- in colon cancer (n = 19) and control group (n = 23) was 36.8% and 26.1%, respectively (p > 0.05, chi 2-test). The bleomycin-induced chromosomal breakage (break/cell) in the patient group was 0.75 +/- 0.29, and in the control group 0.42 +/- (0.24) (p < 0.05, t-test). The percentage of mutagen sensitivity (b/c > 0.8) in the patient group (68%) was 4 times as high as that in the control group (20%). The mutagen hypersensitivity (b/c > 1.0) in the patient group (47%) was 5 times as high as that in the control group (12%). The odds radio was 6.6.

Adult↗

aFGF, bFGF and NGF differentially regulate neuropeptide expression in dorsal root ganglia after axotomy and induce autotomy.

Using immunohistochemistry and in situ hybridization the in vivo effects of acidic and basic fibroblast growth factor (aFGF, bFGF), and of nerve growth factor (NGF) on the expression of galanin, neuropeptide Y (NPY) and substance P in axotomized dorsal root ganglia (DRGs) were examined. Self-mutilation (autotomy), a supposed pain-related behavior, was investigated after growth factor treatment. One microgram of aFGF, bFGF or NGF was applied directly to the transected sciatic nerve via a capsule. In normal rats 3.2%, 0% and 17.5% of the neuron profiles in the DRGs contained galanin-, NPY- and substance P-like immunoreactivity (LI), respectively. Sciatic nerve transection induced a distinct increase in galanin- and NPY-LIs, but a downregulation of substance P-LI. Thus three days after axotomy 23.5%, 26.9% and 9.8% of the DRG neuron profiles showed immunoreactivity for galanin-, NPY- and substance P-LI, respectively. In vivo administration of aFGF counteracted the axotomy-induced increase in galanin and NPY, whereas bFGF only suppressed NPY upregulation. NGF reversed in the injury-induced decrease in substance P-LI, but had no significant effect on galanin- and NPY-LIs. These results were confirmed by monitoring the mRNA levels for these neuropeptides. Moreover, aFGF was found to induce autotomy in 60% of the rats 3 days after axotomy. NGF produced autotomy in about 30% of the rats. Taken together, the present results suggest (1) that aFGF, bFGF and NGF differentially regulate neuropeptide expression in vivo; (2) that FGFs can inhibit neuropeptide upregulation of some peptides after nerve injury; and (3) that aFGF and NGF may induce pain-related behavior.

Animals↗

The study of DNA oxidative damage in benzene-exposed workers.

Peripheral blood lymphocyte 8-hydroxy-2-deoxyguanosine (8-OHdG), were detected in 87 benzene-exposed and 30 control subjects by high performance liquid chromatograph coupled with an electrochemical detector system (HPLC-EC). The air concentration of benzene and its homologes in the workplace, urinary trans,trans-muconic acid (TTMA) as an internal dose of benzene exposure, were examined. The lymphocyte micronuclei (MN) as genotoxic and white blood cell (WBC) count as well as the myelotoxic markers of benzene were examined. Exposure to low, medium and high concentrations of benzene resulted in increased levels of 8-OHdG, which were 4.67, 26.12 and 29.89/10(5) dG, respectively, However, the 8-OHdG level observed in the control group was 3.738/10(5) dG). A good correlation between 8-OHdG formation and the groups exposed to external and internal benzene was observed (r = 0.77, 0.64, respectively). There was also a correlation between 8-OHdG and MN formation (r = 0.50). WBC levels were within normal range in all benzene-exposed subjects. It may be concluded that: benzene induced DNA oxidative damage in occupational exposure workers. The major factors influencing blood the 8-OHdG level were sex and toluene.

8-Hydroxy-2'-Deoxyguanosine↗

Activation of Jak/STAT proteins involved in signal transduction pathway mediated by receptor for interleukin 2 in malignant T lymphocytes derived from cutaneous anaplastic large T-cell lymphoma and Sezary syndrome.

Signaling through the interleukin 2 receptor (IL-2R) involves phosphorylation of several proteins including Jak3, STAT5, and, in preactivated cells, STAT3. In the present study, we examined the functional status of the IL-2R-associated Jak/STAT pathway in malignant T lymphocytes from advanced skin-based lymphomas: anaplastic large T-cell lymphoma (ALCL) and Sezary syndrome (SzS). Proliferation of three ALCL cell lines (PB-1, 2A, and 2B) was partially inhibited by rapamycin, a blocker of some of the signals mediated by IL-2R, but not by cyclosporin A, FK-506, and prednisone, which suppress signals mediated by the T-cell receptor. All the cell lines expressed on their surface the high-affinity IL-2R (alpha, beta, and gamma c chains). They showed basal, constitutive phosphorylation, and coassociation of Jak3, STAT5, and STAT3. Weak basal phosphorylation of IL-2R gamma c was also detected. In regard to SzS, peripheral blood mononuclear cells from 10 of 14 patients showed basal phosphorylation of Jak3, accompanied by phosphorylation of STAT5 in 9 patients, and STAT3 in 4 patients. However, in vitro overnight culture of SzS cells without exogenous cytokines resulted in markedly decreased Jak3 and STAT5 phosphorylation, which could be reversed by stimulation with IL-2. This indicates that the basal phosphorylation of Jak3 and STAT5 in freshly isolated SzS cells is induced rather than constitutive. The basal activation of the Jak/STAT pathway involved in IL-2R signal transduction in ALCL and SzS cells reported here suggests that this pathway may play a role in the pathogenesis of cutaneous T-cell lymphomas, although the mechanism (induced versus constitutive) may vary between different lymphoma types.

Cell Membrane↗

Cooperativity in F-actin: binding of gelsolin at the barbed end affects structure and dynamics of the whole filament.

We have studied the effect of gelsolin, a Ca-dependent actin-binding protein, on the microsecond rotational dynamics of actin filaments, using time-resolved phosphorescence (TPA) and absorption anisotropy (TAA) of erythrosin iodoacetamide attached to Cys374 on actin. Polymerization of actin in the presence of gelsolin resulted in substantial increases in the rate and amplitude of anisotropy decay, indicating increased rotational motion. Analysis indicates that the effect of gelsolin cannot be explained by increased rates of overall (rigid-body) rotations of shortened filaments, but reflects changes in intra-filament structure and dynamics. We conclude that gelsolin induces (1) a 10 degrees change in the orientation of the absorption dipole of the probe relative to the actin filament, indicating a conformational change in actin, and (2) a threefold decrease in torsional rigidity of the filament. This result, which is consistent with complementary electron microscopic observations on the same preparations, directly demonstrates long-range cooperativity in F-actin, where a conformational change induced by the binding of a single gelsolin molecule to the barbed end is propagated along inter-monomer bonds throughout the actin filament.

Actin Cytoskeleton↗

Expression of corticotropin-releasing factor in inflamed tissue is required for intrinsic peripheral opioid analgesia.

Immune cell-derived opioid peptides can activate opioid receptors on peripheral sensory nerves to inhibit inflammatory pain. The intrinsic mechanisms triggering this neuroimmune interaction are unknown. This study investigates the involvement of endogenous corticotropin-releasing factor (CRF) and interleukin-1beta (IL-1). A specific stress paradigm, cold water swim (CWS), produces potent opioid receptor-specific antinociception in inflamed paws of rats. This effect is dose-dependently attenuated by intraplantar but not by intravenous alpha-helical CRF. IL-1 receptor antagonist is ineffective. Similarly, local injection of antiserum against CRF, but not to IL-1, dose-dependently reverses this effect. Intravenous anti-CRF is only inhibitory at 10(4)-fold higher concentrations and intravenous CRF does not produce analgesia. Pretreatment of inflamed paws with an 18-mer 3'-3'-end inverted CRF-antisense oligodeoxynucleotide abolishes CWS-induced antinociception. The same treatment significantly reduces the amount of CRF extracted from inflamed paws and the number of CRF-immunostained cells without affecting gross inflammatory signs. A mismatch oligodeoxynucleotide alters neither the CWS effect nor CRF immunoreactivity. These findings identify locally expressed CRF as the predominant agent to trigger opioid release within inflamed tissue. Endogenous IL-1, circulating CRF or antiinflammatory effects, are not involved. Thus, an intact immune system plays an essential role in pain control, which is important for the understanding of pain in immunosuppressed patients with cancer or AIDS.

Analgesics, Opioid↗

Nuclear localization of type II cAMP-dependent protein kinase during limb cartilage differentiation is associated with a novel developmentally regulated A-kinase anchoring protein.

Differentiation of chicken limb cartilage is accompanied by a rise in intracellular cyclic AMP, an inducer of cartilage-specific gene expression. A basic approximately 35-kDa protein, designated p35, is the major nuclear substrate for cAMP-dependent protein kinase (PKA) during this process. Here we show that whereas both precartilage and cartilage nuclei contain p35, only precartilage nuclei contain PKA. The phosphorylation of p35 in isolated cell fractions was used as an index of changes in the cellular compartmentalization of components of PKA during chondrogenesis. Both the catalytic subunit and type II regulatory subunit (RII) of PKA were present in the precartilage nuclear fraction, but were undetectable or present in only trace amounts in the cartilage nuclear fraction. Furthermore, a novel approximately 150-kDa A-kinase anchoring protein (AKAP), which binds to RII, was detected in the nuclear matrix of precartilage nuclei but, like RII, was virtually absent in the nuclei of fully differentiated cartilage cells. In limb mesenchymal cells undergoing chondrogenesis in culture a corresponding set of changes occurred: cartilage differentiation was accompanied by a marked reduction in the amounts of both nuclear RII and nAKAP150. These observations indicate that type II PKA holoenzyme is imported into the mesenchymal cell nucleus prior to chondrogenesis, an event that appears to depend on the activity of the developmentally regulated nAKAP150.

Affinity Labels↗

Ca2+/calmodulin-dependent protein kinase type IV in dorsal root ganglion: colocalization with peptides, axonal transport and effect of axotomy.

Using the indirect immunofluorescence technique, the distribution of Ca2+/calmodulin-dependent protein kinase IV (CaM kinase IV) was studied in dorsal root ganglia (DRGs) and the sciatic nerve under normal circumstances and after axotomy and nerve ligation. CaM kinase IV-like immunoreactivity (-LI) was observed mainly in small DRG neurons but also in some large ones with the immunoreactivity mainly confined to the cell nuclei and with varying levels in the cytoplasm. CaM kinase IV-LI was present in around 1/4 of all CGRP-positive neurons and in the vast majority of the somatostatin-positive neurons. The enzyme levels decreased markedly after axotomy. The enzyme was also observed in axons in the sciatic nerve and accumulated both proximal and distal to a ligation. The present results suggest that CaM kinase is not of direct importance for upregulation of neuropeptides in DRG neurons after nerve injury. In addition to a nuclear function it may also play a role in the peripheral processes of DRG neurons.

Animals↗

Microsecond rotational dynamics of actin: spectroscopic detection and theoretical simulation.

We have investigated the microsecond rotational dynamics of F-actin with transient phosphorescence anisotropy (TPA) spectroscopy, and analyzed the data to determine the relative contributions from rigid-body rotations and from intrafilament bending and twisting, using a theoretical model developed for DNA dynamics by Schurr and co-workers. The fits of the data to the model were constrained by independently determining the orientation of the dye's absorption dipole (by transient absorption anisotropy, TAA) and the actin filament length distribution (by electron microscopy). We conclude that (1) the Schurr theory enables calculation of the torsional flexibility of actin independent of any contribution from rigid body rotations of the whole filament, (2) the TPA decays cannot be explained by rigid-body or bending rotations, but reflect primary twisting motions within actin filaments, and (3) the dynamic properties of actin filaments are best ascribed to a continuous elasticity. This analysis establishes a firm methodological foundation for future studies of the effects or perturbations of the dynamics of actin on its functional properties.

Actins↗

Partial purification of an IGF-II receptor/binding protein from the erythroleukemia cell line K562.

We previously reported that insulin-like growth factor II (IGF-II) stimulated clonal growth of an erythroleukemia cell line, K562, in semi-solid agar, an effect not mimicked by insulin-like growth factor I (IGF-I), as IGF-I receptors are generally not expressed in this cell line. Affinity crosslinking of intact K562 cells with 125I-IGF-II revealed that the labeled hormone predominantly bound to a protein with a molecular weight of approximately 75 K. We report here the partial purification of the 75 K IGF-II binding protein from K562 cells. Triton X-100-solubilized K562 cells were subjected to Sephacryl-400, followed by Sephacryl-200 chromatography. Fractions of interest were collected and applied to a Sepharose-IGF-II column or an immuno-affinity column. The immuno-affinity column was prepared using an antiserum against placental membrane-derived material eluted from the Sephacryl-400 column in the elution volume, corresponding to the IGF-II binding protein from K562 cells. An affi-gel 10 affinity column, prepared with a protein A purified IgG fraction of this antiserum (antibody-29), retarded proteins showing binding specificity for IGF-II, with apparent molecular weights of 76 K, 87 K, and 70 K under reducing conditions. These protein bands were similar to the proteins retarded in the IGF-II affinity column, when evaluated by affinity crosslinking and SDS-PAGE. Fractionation of the purified material from the antibody-29 affinity column on Superose 12 revealed 6 protein peaks. Affinity crosslinking of the peak fractions from FPLC resulted in single bands with a molecular weight of 75 K under reducing conditions with variable specificity for IGF-II.

Animals↗

Urinary homovanillic acid and vanillylmandelic acid in workers exposed to carbon disulfide.

Homovanillic acid (HVA) and vanillylmandelic acid (VMA), two end products of dopamine metabolism, were measured in 60 workers exposed to carbon disulfide (CS(2)) in a rayon factory and in 48 unexposed workers. The airborne CS(2) concentrations in eight major exposure zones of the plant were measured monthly over a period of 4 years, from 1990 to 1994. In addition, the exposure concentrations and exposure history of each worker were integrated to estimate the overall lifetime exposure. Industrial hygiene data showed that the geometric mean concentrations of CS(2) in the plant ranged from 2.68 to 20.19 ppm, and more than 15% of the studied population had been repeatedly exposed to CS(2) at concentrations exceeding the ACGIH recommended time-weighted average of 10 ppm. The results showed that there was a significantly lower level and a higher proportion of CS(2) workers with decreased HVA and VMA excretion. However, there were no statistical correlations between the two dopaminergic metabolites and the mean CS(2) concentration, and years of employment. In contrast, significant dose-effect relationships were observed between these two metabolites and the integrated cumulative exposure (ICE) variable. The correlation coefficients for ICE and HVA, and ICE and VMA were -0.35 (p < 0.01) and -0.20 (p <0.05), respectively. These data suggest that chronic exposure to CS(2) was associated with measurable reduction in catecholamine metabolite concentrations. This finding is compatible with the earlier observations in laboratory animals that CS(2) exposures interfere with neurochemical metabolism.

Adult↗

Clinical and genetic analysis of a large pedigree with juvenile myoclonic epilepsy.

Juvenile myoclonic epilepsy is a common type of idiopathic generalized epilepsy characterized by myoclonic, generalized tonic-clonic, and in 30% of patients, absence seizures. We studied a three-generation pedigree of 33 members, 10 of whom were clinically affected with juvenile myoclonic epilepsy or presented with subclinical electroencephalographic (EEG) 3.5- to 6.0-Hz diffuse polyspike-wave or spike-wave complexes. Juvenile myoclonic epilepsy and the EEG trait segregated as an autosomal dominant trait with 70% penetrance. Linkage analysis using this model showed significant linkage to four microsatellite markers centromeric to human leukocyte antigen (HLA) in chromosome 6p. Maximum lod scores of 3.43 at theta(m=f)=0.00 for D6S272, D6S466, D6S257, and D6S402 were obtained. Recombinant events in 2 affected members defined the gene region to a 43-cM interval flanked by D6S258 (HLA region) and D6S313 (centromere). Our results in this large family provide evidence that a gene responsible for juvenile myoclonic epilepsy and the subclinical, 3.5- to 6.0-Hz, polyspike-wave or spike-wave EEG pattern is located in chromosome 6p.

Adolescent↗