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Biomedical subjects

D Wu

Publications and source records attributed to D Wu.

At least 163 records · Page 9Linked to original sources

Modulation of transforming growth factor beta to platelet-derived growth factor receptor-alpha of human osteoblasts.

OBJECTIVE: To investigate the mechanism and the significance of TGF beta in modulating the expression of Platelet-Derived Growth Factor Receptor-alpha (PDGFR-alpha) in human osteoblasts. METHODS: The osteoblasts were isolated from human fetal calvaria. The percentage of cell increase (PCI) in every 4 hours was calculated to demonstrate the proliferation of osteoblasts affected by PDGF-AA and TGF beta. The osteoblasts were cultured with TGF beta for 24 hours and with PDGF-AA for another 24 hours, and the cells proliferation was shown by PCI too. The osteoblasts were cultured with TGF beta for 24 h, and the PDGFR-alpha of the cells were measured by immunofluorescent analysis. RESULTS: PCI was increased by 48.2% and 22.4% after PDGF-AA and TGF beta were added into the medium for 24 hours respectively (P < 0.05), and PCI decreased after the removal of the two cytokines. Preincubated with TGF beta for 24 hours and then stimulated with PDGF-AA, PCI grew slowly. TGF beta downregulated the expression of the PDGFR-alpha. CONCLUSION: TGF beta can downregulate the mitogenesis of PDGF-AA by lowering the number of PDGFR-alpha.

Cell Division↗

[Mutation of human O6-alkylguanine-DNA alkyltransferase confers resistance to O6-benzylguanine].

OBJECTIVE: O6-alkylguanine-DNA alkyltransferase (O6-AGT), capable of repairing DNA damage, is responsible for tumor cell resistance to nitrosourea. While O6-benzylguanine as a selective inhibitor of AGT helps reverse drug resistance, it would aggravate myelo-suppression. This investigation is to generate AGT mutant and see if it would confer resistance to O6-benzylguanine-induced inhibition but leave its alkyltransferase activity intact. METHODS: Human O6-methylguanine-DNA methyltransferase (MGMT) cDNA was mutated by site-directed mutagenesis. The mutant cDNA was transferred into E. coli and the protein expressed was purified. The activity of the mutant MGMT was determined in vitro with O6-(3H)-methylguanine-DNA as substrate. RESULTS: Two mutant MGMT proteins were obtained: G156A and P140A, with glycine-to-alanine mutation at position 156 and proline-to-alanine mutation at position 140, respectively. The AGT activity of both mutants was similar to that of the wild type MGMT. However, their resistance to O6-benzylguanine was significantly increased up to 105.8 and 13.5 fold, respectively as compared to that of the wild type MGMT. CONCLUSION: The results suggested that transduction of the mutant MGMT herein reported into hematopoietic progenitor cells may lead to their selective resistance to the combined use of O6-benzylguanine and alkylating agents designed to overcome tumor resistance to nitrosourea treatment.

Amino Acid Sequence↗

[The effects of platelet-derived growth factor-AA on the osteoclasts].

OBJECTIVE: To study the effects of platelet-derived growth factor (PDGF)-AA on the osteoclasts. METHODS: The PDGF receptor alpha (PDGFR-alpha) was detected on the osteoclastic membrane with immunoelectronmicroscopy. Acid phosphatase (ACP) and tartrate-resistant acid phosphatase (TRAP) activities were measured kinetically and the introcellular hydrogen ions were calculated on the confocal laser scanning microscope (CLSM) stained with SNAFL. Stained by toluidine blue, the area and number of the resorption pits were determined with the Leica Quantimet 500 system. RESULTS: The colloidal gold precipitated on the cellular membrane, and the TRAP activity had no difference (P > 0.05). Hydrogen ions rose significantly but their release could not be promoted by PDGF-AA. The changes of the resorption pits were not significant. CONCLUSIONS: The PDGF-AA could promote hydrogen ions yielding but not stimulate osteoclastic bone resorption due to the significant TRAP changes of TRAP activities.

Acid Phosphatase↗

[Mitogenesis of platelet-derived growth factors to human osteoblasts modulated by basic fibroblast growth factor].

OBJECTIVES: To investigate the mechanism by which basic fibroblast growth factor modulates the mitogenesis of platelet-derived growth factor to human osteoblasts. METHODS: The osteoblasts isolated from human fetal calvaria were incubated with PDGF-AB (100 ng/ml) or bFGF(10 ng/ml) combined with PDGF-AB (100 ng/ml); the growth curve was plotted. The(3)H-TdR incorporation of the osteoblasts was measured after the cells were incubated with different combination of bFGF and PDGF-AA or PDGF-BB. After incubated with bFGF (10 ng/ml) for 24 hours, the number of PDGFR-alpha and PDGFR-beta on the membrane of the osteoblasts was detected by fluoroimmunoassay. RESULTS: Four days after PDGF-AB added into the medium, the population of the osteoblasts was larger than that of the control (P < 0.05). The number of the osteoblasts incubated with PDGF-AB (12.1 x 10(4)) was 1.8 times as large as the control (6.8 x 10(4)) in the 10th day (P < 0.05), and that of the osteoblasts incubated with both bFGF and PDGF-AB increased more quickly than the cells only incubated with PDGF-AB. The incorporation of (3)H-TdR into the osteoblasts cultured with bFGF combined with PDGF-AA (533.6 +/- 13.1) was more than that cultured only with PDGF-AA (435.4 +/- 14.8, P < 0.01), so was the incorporation of (3)H-TdR of those cells cultured with bFGF and then PDGF-AA (633.8 +/- 51.5). bFGF up-regulated PDGFR-alpha and down-regulated PDGFR-beta on the surface of the human osteoblasts. CONCLUSION: bFGF elevates the mitogenesis of PDGF-AA or -AB to human osteoblasts by up-regulating PDGFR-alpha.

Cell Proliferation↗

[The rearrangements of immunoglobulin heavy chain and T cell receptor delta genes in patients with acute nonlymphocytic leukemia].

OBJECTIVE: To explore the clinical significance of rearrangements of immunoglobulin heavy chain (IgH) and T cell receptor (TCR) genes in patients with acute nonlymphocytic leukemias. METHODS: Polymerase chain reaction (PCR) was used to study IgH and TCRdelta genes rearrangements in 30 untreated acute nonlymphocytic leukemia patients. RESULTS: IgH gene rearrangements were found in 9 patients, and TCRdelta gene rearrangements in 5 patients. CONCLUSION: The lineage infidelity does occur in acute nonlymphocytic leukemia, showing not only a high frequency of IgH gene rearrangement, but also of TCRdelta gene rearrangement.

Gene Rearrangement↗

[The toxicity of busulphan and cyclophosphamide as the preparative regimen for allogeneic peripheral blood stem cell transplantation].

OBJECTIVE: To analyse the treatment-related complications of busulphan and cyclophosphamide (BU-CY) as the conditioning regimen for allogeneic peripheral blood stem cell transplantation (allo-PBSCT). METHODS: The clinical data of 40 leukemia patients undergoing allo-PBSCT between June 1997 and May 1999 in our BMT center were retrospectively analysed. RESULTS: Recovery of neutrophil and platelet was achieved at a median of day +13 (9 - 28) and day +12 (7 - 60) respectively. Acute GVHD occurred in 17 of 40 patients (42.5%) with grade II-IV in 10 patients (25%). Chronic GVHD developed in 21 out of 30 evaluable patients (70%). Mild to severe mucositis occurred in 30 patients (75%), and 4 of them had severe esophagitis with bleeding. Haemorrhagic cystitis developed in 8/40 (20%) patients, the median time of its onset was day +100 (+7 to +165). Six of 40 patients (15%) developed interstitial pneumonia (IP), 5 of them were due to cytomegalovirus infection, and the remaining one due to pneumocystis carinii infection. No hepatic veno-occlusive disease was observed and no seizure occurred. During the median follow-up of 480 (300 - 1000) days, 4 (10%) patients relapsed and 8 (20%) patients died of the transplant-related complications. The 3 year leukemia-free survival rate was 70%. CONCLUSION: BU (domestic busulfan)-CY regimen is relatively easy to administer and well tolerated, with low extramedullary toxicities.

Acute Disease↗

[Expression of citrate transporter mRNA in the kidneys of rats with metabolic acidosis].

The purpose of the present study was to examine whether metabolic acidosis affects the expression of rat renal Na(+)/citrate cotransporter. Female Wistar rats were pair-fed with normal rat chow and drinking water (control) or water with 0.28 mol/L NH4Cl (metabolic acidosis). The mRNA of two renal Na(+)/citrate cotransporters, which were respectively expressed on apical and basolateral membrane, were measured by Northern blot with two probes, SDCT1 and SDCT2. Animals were sacrificed on day 1,3 and 7. On the 1st day, the blood plasma HCO(-)3 of acidosis group decreased significantly (P<0.01), but the mRNA abundance did not change. On the 3rd day in the acidosis group, the blood plasma HCO(-)3 increased slightly more than that on the 1st day, but was still significantly lower than that of the control group (P<0.01). Both the probes detected some increase in mRNA of brush border and basolateral Na(+)/citrate cotransporter. On the 7th day, the blood plasma HCO(-)3 of the acidosis group continuously increased and there was no significant difference between the two groups. The abundance of brush border and basolateral Na(+)/citrate cotransporter mRNA increased, but there was no difference between those of the 3rd day and the 7th day. It is concluded that metabolic acidosis can induce increase of Na(+)/citrate cotransporter mRNA, which may be responsible for hypocitraturia.

Acidosis↗

[An experiment on disinfection using high power microwave].

This experimental disinfection was intended to examine the effect of high power microwave on bacteria, and the putative nonthermal effect. The bacteria of the experimental groups were irradiated by high power microwave in different intensities and time and were compared with the controls--bacteria heated only. The principles of disinfection by means of high power microwave were studied. The results showed that the temperature did not change after the dry bacteria were irradiated by high power microwave in different intensities and time, but the number of bacteria alive decreased significantly(P < 0.05). The high power microwave's effect of disinfection was greater in the wet bacteria group than in the dry bacteria group and the bacteria heated group. These data indicate that the disinfection effect of high power microwave is higher than that of heating. High power microwave can kill dry bacteria, and it has the nonthermal effect in addition to its thermal effect.

Disinfection↗

[Effects of 1 alpha(OH)D3 on trabecular and cortical bone in ovariectomized rats].

OBJECTIVE: To study the pharmaceutical effects of 1 alpha(OH)D3 on trabecular and cortical bone in ovariectomized (OVX) rats. METHODS: 41 female Wistar rats of six months old were randomly divided into 5 groups: (1) Baseline control; (2) Sham control; (3) 6 weeks after OVX (OVXb); (4) 14 weeks after OVX (OVXe); (5) OVX + 0.1 microgram/(kg.d) 1 alpha(OH)D3 (O + VD), treatment started 6 weeks after OVX and lasted 8 weeks. Histomorphometry analysis of tibia, peripheral quantitative computed tomography (pQCT) scanning of femur, bone biomechanical test and serum and urinary biochemical parameters were determined. RESULTS: The levels of bone turnover indexes increased in OVX rats, OVX also resulted in reducing of trabecular, bone mass and biomechanical properties. The ratio of urinary deoxypyridinoline crosslink/creatinine was decreased by 67.0% in O + VD group compared with OVXe group 67.0% [(43.50 +/- 11.20) nmol.L-1/mmol.L-1 vs(131.80 +/- 14.90) nmol.L-1/mmol.L-1, P < 0.01]. Percent trabecular area (Cn-BV/TV) was increased by 89.8% in O + VD group compared with OVXe group (11.03 +/- 0.73 vs 5.81 +/- 1.29, P < 0.05). Trabecular bone mineral content and density were increased by 77.3% and 91.3% compared with OVXe respectively (P < 0.05). Although cancellous maximal load and stiffness increased in O + VD group, but no statistical significance. 1 alpha(OH)D3 also enhanced polar moment of inertia (PMI) and maximal load of cortical bone in femur compared with OVXe (23.70 +/- 1.63 vs 18.23 +/- 1.41, P < 0.01 and 171.69 +/- 9.92 vs 147.58 +/- 11.29, P < 0.05 respectively). CONCLUSION: 1 alpha(OH)D3 inhibited the higher bone turnover induced by OVX, increased trabecular bone in proximal tibia and bone mass in distal femur. 1 alpha(OH)D3 also improved the mechanical properties of cortical bone in femur.

Animals↗

[A quantum chemical study of pi-back-donation bond and Raman intensity of 1 sigma + electronic state of Pt-CO molecule].

The Raman spectroscopic properties of Pt-CO molecule have been investigated based on the electronic state 1 sigma + determined by the HF and B3LYP methods. The result shows that the calculated stretching vibrational frequencies of the Pt-C and C-O bonds depend on the method and the basis sets used. It indicates that it is important to adopt an appropriate method to describe pi-donation and pi-back-donation bond. The result of the differential Raman scattering cross section for the stretching vibrations of the Pt-C and C-O bond shows that the latter is significantly larger value compared to the former.

Adsorption↗

[Photoluminescent properties of organic film in flat optical microcavity].

The microcavity is sandwiched between a quarterwavelength distributed Bragg reflector(DBR) and a metal Ag reflective mirror. A single layer of a Tris(8-quinolinolato) aluminum (Alq) film was used as the light-emitting layer. The photoluminescent properties of the optical microcavity and that of the Alq film were studied at the same excitation condition. Compared with the Alq film, the significantly narrowed spectral emission linewidth from 90 nm to 10 nm was observed, the PL emission intensity of the microcavity at the resonant mode is enhanced by the order of 1. The spectral narrowing and intensity enhancement of the microcavity is attributed to the microcavity effect.

English Abstract↗

[EPR spectroscopy studies on surface reactivity of quartz].

With the technique of Electron Paramagnetic Resonance Spectroscopy (EPR), we studied the surface reactivity of quartz on adsorbing Cu(II) ions from aqueous solution over the pH range from 2 to 11. The surface sorption mass of Cu(II), gamma Cu, is as a function of pH. The Cu(II) surface coverage for powered quartz samples changes from 0 to 10.23%. The EPR spectra can be demonstrated with the coexistence of several binding forms of Cu(II) ions on the surface, that is, (SO)2Cu at pH = 4.7, SOCu2+ at pH = 6.5, SOCu (OH)(4)3- at pH = 7, and SOH...Cu(OH)2(s) at pH = 10. Therefore, the sorption tendency that the Cu(II) ions are either adsorbed at isolate sites or form multinuclear complexes versus three-dimensional precipitates onto the surface of quartz is strongly dependent on the natures of quartz surface and aqueous solution medium.

Copper↗

Caffeine-sensitive Ca2+ stores in carp retinal bipolar cells.

High K+- or caffeine-induced Ca2+ signal was studied in freshly dissociated carp retinal ON-type bipolar cells using a confocal laser-scanning microscope. In response to 35 mM K+ exposure, a rise in [Ca2+]i appeared in both the terminal and soma, but was absent after removal of external Ca2+ or in the presence of 100 microM nifedipine. It is indicated that, for high K+-induced increase of [Ca2+]i, Ca2+ influx through voltage-gated L-type Ca2+ channels is essential and Ca2+ entry through reversed Na+/Ca2+ exchange may be negligible. Interestingly, caffeine-induced elevation of [Ca2+]i was restricted to the soma, and could be abolished by 50 microM ryanodine, suggesting that caffeine-sensitive Ca2+ stores gated by ryanodine receptors were present in the soma but not in the terminal of bipolar cells. After treatment with 50 microM ryanodine for 20 min, the peak of the Ca2+ transients evoked by 35 mM K+ in the soma decreased to 48.2+/-5.7% of the control. The results suggest that depolarization-evoked Ca2+ influx can cause Ca2+ release from caffeine-sensitive Ca2+ stores, and in turn amplify Ca2+ signal in the soma of retinal bipolar cells.

Animals↗

Thermal Conversion of a Helical Coil into a Three-Dimensional Chiral Framework.

Thermal dehydration results in the solid-state supramolecular conversion of the helical coordination polymer [{[Cu(sala)](2)(H(2)O)}(n)] into the chiral three-dimensional covalent open network [{Cu(sala)}(n)] (shown schematically). X-ray crystallography reveals that hydrogen bonding plays a key role in this process. H(2)sala=N-(2-hydroxybenzyl)-L-alanine

Journal Article↗

Clinical and angiographic outcomes are similar with half, single, or multiple contiguous Palmaz-Schatz stent implantations for a single coronary stenosis.

We compared the immediate and 6-month clinical and angiographic outcomes in patients undergoing a half, a single, or multiple contiguous stent implantations for a single coronary stenosis. Four hundred forty-three consecutive patients, who underwent elective Palmaz-Schatz stent implantations for 542 stenoses between November 1995 and July 1998, were analyzed. Sixty-three patients with 78 stenoses received a half stent (group A), 346 patients with 395 stenoses received a single stent (group B), and 68 patients with 69 stenoses received multiple overlapping stents (group C) for a single coronary stenosis. Seventy-eight half stents were implanted in 78 stenoses in group A, 395 stents in 395 stenoses in group B, and 141 stents in 69 stenoses in group C. The baseline characteristics were similar in the 3 groups. There were no deaths, no subacute thrombosis, and no vascular complications. Forty-nine patients with 57 stenoses in group A, 280 patients with 326 stenoses in group B, and 59 patients with 60 stenosis in group C underwent 6-month follow-up coronary angiography; the restenotic rate per patient was 10% in group A, 20% in group B, and 24% in group C (NS); the restenotic rate per stenosis was 9% in group A, 18% in group B, and 23% in group C (NS). Follow-up of 18 +/- 3 months revealed no differences in mortality, reinfarction, recurrent angina, target narrowing angioplasty, and elective coronary artery bypass surgery among the 3 groups. The overall cardiac event-free survival was 90%, 82%, and 83% in groups A, B, and C, respectively (p = 0.275). Thus, the procedural success rate, the in-hospital morbidity, and the long-term outcome are similar with coronary stenting using a half, a single, or multiple overlapping Palmaz-Schatz stents for a single stenosis.

Angioplasty, Balloon, Coronary↗

Suppression of glycogen synthase kinase activity is not sufficient for leukemia enhancer factor-1 activation.

Glycogen synthase kinase-3 (GSK) can be regulated by different signaling pathways including those mediated by protein kinase Akt and Wnt proteins. Wnt proteins are believed to activate a transcription factor leukemia enhancer factor-1 (LEF-1) by inhibiting GSK, and Akt was shown to phosphorylate GSK and inhibit its kinase activity. We investigated the effect of an activated Akt on the accumulation of cytosolic beta-catenin and LEF-1-dependent transcription. Although the activated Akt, mAkt, clearly inhibited the kinase activity of GSK, mAkt alone did not induce accumulation of cytosolic beta-catenin or activate LEF-1-dependent transcription. On the contrary, coexpressed Wnt-1 and Frat activated LEF-1 but did not show significant inhibition of GSK-mediated phosphorylation of a peptide substrate. However, mAkt could act synergistically with Wnt-1 or Frat to activate LEF-1. In addition, the interaction of GSK for Axin appeared to decrease in the presence of mAkt, whereas the interaction for Frat remained unchanged. Consistently, a GSK mutant with substitution of a Phe residue for residue Tyr-216, which showed one-fifth of kinase activity of the wild-type GSK, exhibited a reduced association for Axin than the wild-type GSK. These results suggest that inhibition of GSK kinase activity is not sufficient for activation of LEF-1 but may facilitate the activation by reducing the interaction of GSK for Axin. The additional mechanism for LEF-1 activation may require dissociation of GSK from Axin as Frat facilitates the dissociation of GSK from Axin.

3T3 Cells↗

Characterization of stromelysin 1 (MMP-3), matrilysin (MMP-7), and membrane type 1 matrix metalloproteinase (MT1-MMP) derived fibrin(ogen) fragments D-dimer and D-like monomer: NH2-terminal sequences of late-stage digest fragments.

Matrix metalloproteinases (MMPs) participate in physiological remodeling of the extracellular matrix. Recently we determined that both fibrinogen (Fg) and cross-linked fibrin (XL-Fb) are substrates for selected MMPs. Specifically, XL-Fb clots were solubilized by MMP-3 (stromelysin 1) by cleavage at gamma Gly 404-Ala 405, resulting in a D-like monomer fragment. Similarly, MMP-7 (matrilysin) and MT1-MMP (membrane type 1 matrix metalloproteinase) solubilized XL-Fb clots. However, the molecular mass of fragment D-dimer, obtained after MMP-7 and MT1-MMP degradation of XL-Fb, is similar to that of fragment D-dimer from plasmin degradation ( approximately 186 kDa). In contrast, fragment D-like monomer, from MMP-3 degradation of both fibrinogen (Fg) and XL-Fb, is similar to fragment D from plasmin degradation of Fg ( approximately 94 kDa). Reduced chains from MMP-3, MMP-7, and MT1-MMP digests of Fg and XL-Fb were subjected to direct sequence analyses and D/D-dimer alpha-chain showed cleavage at both alpha Asp 97-Phe 98 and alpha Asn 102-Asn 103. Degradation of the beta-chain resulted in microheterogeneity of cleavage sites at beta Asp 123-Leu 124, beta Asn 137-Val 138, and beta Glu 141-Tyr 142, whereas all three enzymes cleaved the gamma-chain at gamma Thr 83-Leu 84. In both Fg and XL-Fb, several cleavage sites obtained by proteolysis with MMP-3, MMP-7, and MT1-MMP were found to be in very close proximity to those obtained by plasmin on these same substrates. That does not occur with other MMPs such as MMP-1, -2, and -9 and MT2-MMP. The degradation of XL-Fb by MMPs suggests both plasmin-dependent and independent mechanisms of fibrinolysis that might be relevant in inflammation, angiogenesis, arthritis, and atherosclerosis.

Binding Sites↗

[New method for the isolation of astroglial cells and oligodendrocytes].

Based on the different properties of cellular adhesions, developmental time-courses and growth pattern of astroglial cells and oligodendrocytes, a method of isolating glial cells in vitro was developed. The observations under inverted light microscope and scan electronic microscope identified that the morphology of isolated cells were consistent with the previous description. Immunocytochemical examination showed the expressions of markers, the glial fibrillary acidic protein (GFAP) of astroglia cells and the myelin basic protein (MBP) of oligodendrocytes were consistent with the isolated cells. All parameters showed that the isolated glial cells were pure, and the new methods were easy and reliable.

Animals↗