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Biomedical subjects

G Cheng

Publications and source records attributed to G Cheng.

At least 127 records · Page 7Linked to original sources
PubMed1998

Characterization of the structural and functional changes of hemoglobin in dimethyl sulfoxide by spectroscopic techniques.

Circular dichroism (CD), fourier transform infrared (FTIR), and fluorescence spectroscopy were used to explore the effect of dimethyl sulfoxide (DMSO) on the structure and function of hemoglobin (Hb). The native tertiary structure was disrupted completely when the concentration of DMSO reached 50% (v/v), which was determined by loss of the characteristic Soret CD spectrum. Loss of the native tertiary structure could be mainly caused by breaking the hydrogen bonds, between the heme propionate groups and nearby surface amino acid residues, and by disorganizing the hydrophobic interior of this protein. Upon exposure of Hb to 52% DMSO for ca. 12 h in a D2O medium no significant change in 1652 cm-1 band of the FTIR spectrum was produced, which demonstrated that alpha-helical structure predominated. When the concentration of DMSO increased to 57%: (1) the band at 1652 cm-1 disappeared with the appearance of two new bands located at 1661 and 1648 cm-1; (2) another new band at 1623 cm-1 was attributed to the formation of intermolecular beta-sheet or aggregation, which was the direct consequence of breaking of the polypeptide chain by the competition of S&z.dbnd6;O groups in DMSO with C&z.dbnd6;O groups in amide bonds. Further increasing the DMSO concentration to 80%, the intensity at 1623 cm-1 increased, and the bands at 1684, 1661 and 1648 cm-1 shifted to 1688, 1664 and 1644 cm-1, respectively. These changes showed that the native secondary structure of Hb was lost and led to further aggregation and increase of the content of 'free' amide C&z.dbnd6;O groups. In pure DMSO solvent, the major band at 1664 cm-1 indicated that almost all of both the intermolecular beta-sheet and any residual secondary structure were completely disrupted. The red shift of the fluorescence emission maxima showed that the tryptophan residues were exposed to a greater hydrophilic environment as the DMSO content increased. CO-binding experiment suggested that the biological function of Hb was disrupted seriously even if the content of DMSO was 20%.

Animals↗
PubMed1998

Development of highly selective SH3 binding peptides for Crk and CRKL which disrupt Crk-complexes with DOCK180, SoS and C3G.

Many Src Homology 3 (SH3) domains function as molecular adhesives in intracellular signal transduction. Based on previous ultrastructural studies, short motifs which bind to the first SH3 domains of the adapters Crk and CRKL were selectively mutagenised to generate Crk/CRKL SH3-binding peptides of very high affinity and selectivity. Affinities were increased up to 20-fold compared to the best wildtype sequences, while the selectivity against a similar SH3 domain [Grb2SH3(N)] was not only retained, but sometimes increased. Blot techniques with GST-fusion peptides and in solution precipitation assays with biotinylated high affinity Crk binding peptides (HACBPs) were subsequently used to analyse the binding of these sequences to a large panel of SH3 domain-containing fusion proteins. Only those proteins which contained the CrkSH3(1) or CRKLSH3(1) domains bound efficiently to the HACBPs. A GST-HACBP fusion protein precipitated Crk and CRKL proteins out of 35S-labelled and unlabelled cell lysates. Very little binding of other cellular proteins to HACBP was detectable, indicative of a great preference for Crk and CRKL when compared to the wide variety of other endogenous cellular proteins. Moreover, HACBP disrupted in vitro preexisting Crk-complexes with DOCK180 and the exchange factors SoS and C3G, which are known targets of Crk adapters, in a concentration dependent manner. HACBP-based molecules should therefore be useful as highly selective inhibitors of intracellular signalling processes involving Crk and CRKL.

Adaptor Proteins, Signal Transducing↗
PubMed1998

Negative regulation of T cell homing by CD43.

We report that the cell surface mucin CD43 acts as an anti-adhesin on T lymphocytes. CD43-deficient murine lymphocytes homed significantly more frequently to secondary lymphoid organs than wild-type cells. Intravital microscopy of peripheral lymph node venules revealed that CD43-deficient lymphocytes were twice as likely to tether, roll, and stick than wild-type cells. This effect was due to CD43 interference with the homing receptor, L-selectin, and was most pronounced in venules with low L-selectin ligand density. In vitro, CD43-deficient cells tethered to L-selectin ligands more efficiently and rolled more slowly than wild-type lymphocytes. Thus, CD43 exerts a negative regulatory effect on T cell trafficking by counterbalancing L-selectin-mediated adhesion.

Animals↗
PubMed1998

IP-10 gene transcription by virus in astrocytes requires cooperation of ISRE with adjacent kappaB site but not IRF-1 or viral transcription.

Transcription of the IP-10 gene requires interferon (IFN)-stimulated response element (ISRE) and kappaB sites to be induced by lipopolysaccharide (LPS), IFN-gamma, virus, and poly(I:C). A requirement for Stat1 binding to ISRE for IFN-gamma and IFN regulatory factor-1 (IRF-1) binding to ISRE for LPS, poly(I:C), and virus has been reported. We investigated whether viral transcription is required for IP-10 induction and how ISRE interacts with IRF-1 and with two kappaB sites. IP-10 mRNA was induced by Newcastle disease virus and Sendai virus in rat astrocytes and the human astrocytoma U251 cell line. IP-10 was also induced by UV-irradiated virus, which is unable to carry out viral transcription. The minimal IP-10 virus response element (VRE) consists of an ISRE and adjacent kappaB site between -236 and -153, to which p50/p65 NF-kappaB proteins and IRF-like proteins bind. Virus induced NF-kappaB binding to an isolated kappaB sequence adjacent to ISRE. However, no protein binding to isolated ISRE was induced by virus. Virus also induced IP-10 in cells expressing a defective IRF-1 gene. Therefore, effective ISRE activity of IP-10 VRE may require an IRF-like protein binding, which is enhanced by an NF-kappaB heterodimer binding to an adjacent KB site. IRF-1 is not required for virus-induced IP-10 gene expression.

Animals↗
PubMed1998

Experiences with "self service" electronic blood banking.

Electronic verification of ABO compatibility (computer crossmatching) has been accepted as the crossmatching procedure for patients without clinically significant alloantibodies. Computer crossmatching offers several advantages over the immediate spin crossmatch and enables the setting up of a self service blood banking system. Self service blood banking saves manpower, improves crossmatch/transfusion(C/T) ratio and makes the maximum surgical blood ordering schedule (MSBOS) redundant. Blood banking service can also be provided at satellite hospitals without stationing blood banking staff there. We have also developed a novel self service system that does not require expensive computer hardware and networking.

Blood Banks↗
PubMed1998

Development of spontaneous synaptic transmission in the rat spinal cord.

Dorsal root afferents form synaptic connections on motoneurons a few days after motoneuron clustering in the rat lumbar spinal cord, but frequent spontaneous synaptic potentials are detected only after birth. To increase our understanding of the mechanisms underlying the differentiation of synaptic transmission, we examined the developmental changes in properties of spontaneous synaptic transmission at early stages of synapse formation. Spontaneous postsynaptic currents (PSCs) and tetrodotoxin (TTX)-resistant miniature PSCs (mPSCs) were measured in spinal motoneurons of embryonic and postnatal rats using whole cell patch-clamp recordings. Spontaneous PSC frequencies were higher than mPSC frequencies in both embryonic and postnatal motoneurons, suggesting that even at embryonic stages, when action-potential firing rate was low, presynaptic action potentials played an important role in triggering spontaneous PSCs. After birth, the twofold increase in spontaneous PSC frequency was attributed to an increase in action-potential-independent quantal release rather than to a higher rate of action-potential firing. In embryonic motoneurons, the fluctuations in peak amplitude of spontaneous PSCs were normally distributed around single peaks with modal values similar to those of mPSCs. These data indicated that early in synapse differentiation spontaneous PSCs were primarily composed of currents generated by quantal release. After birth, mean mPSC amplitude increased by 50% but mean quantal current amplitude did not change. Synchronous, multiquantal release was apparent in postnatal motoneurons only in high-K+ extracellular solution. Comparison of the properties of miniature excitatory and inhibitory postsynaptic currents (mEPSCs and mIPSCs) demonstrated that mean mEPSC frequency was higher than mIPSC frequency, suggesting that either excitatory synapses outnumbered inhibitory synapses or that the probability of excitatory transmitter release was higher than the release of inhibitory neurotransmitters. The finding that mIPSC duration was several-fold longer than mEPSC duration implied that despite their lower frequency, inhibitory currents could modulate motoneuron synaptic integration by shunting incoming excitatory inputs for prolonged time intervals.

2-Amino-5-phosphonovalerate↗
PubMed1998

Suppressive effects of SP-A on ionomycin-induced IL-8 production and release by eosinophils.

Recent studies have demonstrated that pulmonary surfactant protein (SP)-A plays a potential role in modifying inflammation and immune function. To see whether SP-A could modify IL-8 production and release by eosinophils stimulated with ionomycin, SP-A purified from surfactant recovered from patients with alveolar proteinosis was added to eosinophils isolated by the negative-selection method with immunomagnetic beads, and cultured for 24 h. The concentrations of IL-8 in the cell-free supernatants and cell lysates were then measured by ELISA. SP-A attenuated the production of IL-8 by eosinophils in a concentration-dependent manner. SP-A also attenuated the release of IL-8 from the eosinophils. The addition of SP-A antibody (PE10) reversed these effects of SP-A completely. These data suggest that SP-A may have the potential to modify allergic inflammation by inhibiting the release and production of IL-8 by eosinophils.

Antibodies, Monoclonal↗
PubMed1998

[Experimental study of homograft of repeated freezing-thawing treated and ultra-low-temperature treated tendon].

In order to compare the immunogenecity and biological properties of homologous tendon grafts after treatment from different methods of freezing, tendons from chickens received repeated freezing-thawing treatment or ultra-low-temperature treatment, and then, the post-treatment tendons were preserved in liquid nitrogen for 3 months before transplantation. The autogenous tendon transplantation was served as the control. It was found that in the group of repeated freezing-thawing treated tendons, the tendon cells all died and while in the ultra-low temperature treated tendons the active rate of tendon cells was 92.5% +/- 3.4%, and the histological observation showed that transplantation of frozen tendons would result in extensive infiltration of inflammatory cells in the grafted tendons and the peritendinous adhesion was serious than that of the autografts. The active flexion function, hydroxyproline levels and the biomechanical analysis showed no significant differences between the repeated freezing-thawing treated homografts and the ultra-low-temperature treated homografts, and that the autografts was definitely superior to the homografts. The conclusions were: (1) Transplantation of the homologous tendons from the two different methods of freezing could receive considerable success and there was no significant difference between them; (2) Transplantation of frozen homologous tendon graft might give successful result which was probably due to the preservation of the cellular activity of the tendon cells following freezing treatment and elimination of the antigen presenting cells in the tendon as well, and (3) Although the cellular components of the tendon were damaged and the antigenicity of the tendon was lowered, it did not necessarily mean that homologous tendon graft would always be successful in transplantation.

Animals↗
PubMed1998

[Association of fragile histidine triad gene (FHIT) with susceptibility to esophageal cancer. A preliminary study].

OBJECTIVE: To identify loss of heterozygosity(LOH) and homozygous deletion on loci of fragile histidine triad gene(FHIT). METHODS: Gene regions have been detected in 73 pairs of esophageal cancer samples using microsatellite DNA markers. Deletions of FHIT cDNA on 14 fresh esophageal cancer(EC) samples and 2 EC cell lines were screened by RT-PCR and DNA sequencing. RESULTS: High frequency of LOH was detected on loci D3S1480, D3S1481, D3S1234. Of 14 esophageal cancer samples 9(64.3%) had deletions of the FHIT gene, 7 of them were deletions in exon 1-4, and 5 of them were deletions in exon 5-9, the coding region. The whole FHIT gene was found deleted in an EC cell line, EC8733. CONCLUSION: It is suggested that the FHIT gene may play an important role in the genesis of EC. However, further study is needed to ascertain whether FHIT gene is indeed an EC susceptibility gene.

Acid Anhydride Hydrolases↗
PubMed1998

[Effects of the glucoprotein component of musk on functions of rat polymorphonuclear leukocytes activated by LTB4 in vitro].

To investigate the effects of musk-1, a glucoprotein component isolated from the water extract of musk, on some functions of rat polymorphonuclear leukocytes activated by LTB4, an in vitro incubation system with rat polymorphonuclear leukocytes was used. The superoxide anion production was determined by cytochrome C reduction, and the beta-glucuronidase and lysozyme release was quantitated by enzyme reactions in which phenolphthaleinglucuronic acid and micrococcus lysodeikticus were used as the substrates. In comparison with the control, musk-1 at final concentrations of 1 microgram/ml-100 micrograms/ml can increase the superoxide anion production by 28.7%-202.1% and decrease the beta-glucuronidase and lysozyme release by 3%-46% and 6%-32% respectively in rat polymorphonuclear leukocytes. It is concluded that musk-1 can significantly affect the functions of rat polymorphonuclear leukocytes activated by LTB4. One of the mechanisms of this anti-inflammatory action of musk may consist in the inhibition of lysosomal enzyme release.

Animals↗
PubMed1998

Lack of structural rearrangement in c-kit and stem cell factor genes in Hong Kong Chinese patients with myelodysplastic syndromes or acute myeloid leukaemia.

Stem cell factor is a haemopoietic growth factor that interacts with the c-kit-encoded transmembrane tyrosine kinase receptor during signal transduction in haemopoietic progenitor stem cells. We have screened 127 Chinese patients with myelodysplastic syndromes or acute myeloid leukaemia for structural rearrangements in the stem cell factor and c-kit genes using Southern blot analysis. No structural rearrangements were detected in any of the bone marrow samples that were tested. It seems that structural rearrangements in the stem cell factor and c-kit genes are rare in Hong Kong patients who have a haematological malignancy.

Journal Article↗
PubMed1998

[Supercritical fluid extraction and gas chromatographic analysis of four organochlorine pesticides in vegetable].

Supercritical fluid extraction(SFE) of pesticide residues of chlorothalonil, aldrin, dieldrin, and endrin from chives was evaluated. A mixture of vegetable sample: anhydrous magnesium sulfate(2:3) was used for extraction. The extract was analyzed by GC/ECD. The optimum SFE conditions for the pesticides in spiked samples were studied by orthogonal experiment. The chosen conditions were 30.40 MPa, 40 degrees C, 1 min static time, dynamic extraction with 15 mL of CO2, and collection in 3 mL of ethyl acetate. All the pesticides were extracted satisfactorily from their matrix.

Chive↗
PubMed1998

[Effects of isorhapotigenin and resveratrol on function of peripheral blood polymorphonuclear leukocytes from rabbits].

In effects of isorhapotigenin and resveratrol on chemotaxis and release of beta-glucuronidase from rabbit peripheral blood polymorphonuclear leukocytes stimulated with formylmethionyl-leucyl-phenyl-phenylalanine (fMLPP) were studied. The optimal concentration of fMLPP for the induction of chemotaxis of polymorphonuclear leukocytes was approximately at 5 x 10(-10) mol.L-1. fMLPP, at the concentration range of 10(-10)-10(-6) mol.L-1, dose-dependently increased the release of beta-glucuronidase from polymorphonuclear leukocytes. Isorhapotigenin and resveratrol both inhibited the chemotaxis of polymorphonuclear leukocytes stimulated with fMLPP(5 x 10(-10) mol.L-1). Isorhapotigenin, at the concentration range of 10(-5)-10(-4) mol.L-1, dose-dependently inhibited the release of beta-glucuronidase from polymorphonuclear leukocytes stimulated with fMLPP(10(-8) mol.L-1). Resveratrol, at the concentrations of 1 x 10(-5) and 3 x 10(-5) mol.L-1, was shown to dose-dependently inhibit beta-glucuronidase release from polymorphonuclear leukocytes stimulated with fMLPP(10(-8) mol.L-1).

Animals↗
PubMed1998

[Effects of leukotrienes on production of interleukin 6 from mouse peritoneal macrophages].

In the present study, effects of leukotrienes on IL-6 production by mouse resident peritoneal macrophages were explored with a bioassay method involving IL-6 dependent murine hybridoma B9 cell line. The results showed that LTB4, LTC4 and LTD4 can enhance the production of IL-6 from cultured peritoneal macrophages of mice. The optimal concentractions for LTB4, LTC4 and LTD4 were found to be 1.45 x 10(-6), 6.9 x 10(-8) and 8.05 x 10(-8) mol.L-1, respectively. The results suggest that peptide leukotrienes might play important role on IL-6 production in the local milieu of inflammation.

Animals↗
PubMed1997

Low-frequency stimulation of afferent Adelta-fibers induces long-term depression at primary afferent synapses with substantia gelatinosa neurons in the rat.

Impulses in primary afferent nerve fibers may produce short- or long-lasting modifications in spinal nociception. Here we have identified a robust long-term depression (LTD) of synaptic transmission in substantia gelatinosa neurons that can be induced by low-frequency stimulation of primary afferent Adelta-fibers. Synaptic transmission between dorsal root afferents and neurons in the substantia gelatinosa of the spinal cord dorsal horn was examined by intracellular recording in a transverse slice dorsal root preparation of rat spinal cord. Conditioning stimulation of dorsal roots with 900 pulses given at 1 Hz (10 V, 0.1 msec) produced LTD of EPSP amplitudes in substantia gelatinosa neurons to 41 +/- 10% of control that lasted for at least 2 hr. When A- and C-fibers were recruited, conditioning stimulation was as effective as A-fiber stimulation alone. After LTD, synaptic strength could be increased to its original level by applying a second, high-frequency tetanic stimulus to the dorsal root, indicating that LTD is reversible and not attributable to damage of individual synapses. Bath application of the GABAA receptor antagonist bicuculline and glycine receptor antagonist strychnine did not affect LTD. When NMDA receptors were blocked by bath application of D-2-amino-5-phosphonovaleric acid, LTD was abolished or strongly reduced. Loading substantia gelatinosa neurons with Ca2+ chelator BAPTA also blocked or reduced LTD. After incubation of slices with calyculin A, a selective and membrane permeable inhibitor of protein phosphatases 1 and 2A, LTD was not attenuated. We propose that this form of LTD may be relevant for long-lasting segmental antinociception after afferent stimulation.

2-Amino-5-phosphonovalerate↗