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X X Wu

Publications and source records attributed to X X Wu.

32 records · Page 2Linked to original sources
PubMed1997

Antiphospholipid immunoglobulin G antibodies reduce annexin-V levels on syncytiotrophoblast apical membranes and in culture media of placental villi.

OBJECTIVES: The mechanism by which antiphospholipid antibodies are associated with pregnancy loss and thromboembolism has not been established. We previously showed that annexin-V, a phospholipid-binding protein with potent anticoagulant activity, is present on the apical membranes of the syncytiotrophoblasts that line placental villi and that this protein is reduced, by immunohistochemistry, on placentas of patients with antiphospholipid antibodies. We therefore investigated whether annexin-V in apical membranes of placental villi is quantitatively reduced by antiphospholipid antibody immunoglobulin G. STUDY DESIGN: Placentas were obtained from an index patient with antiphospholipid syndrome with intrauterine growth restriction and from a patient with an uncomplicated pregnancy who were both delivered by cesarean section. Apical villous membranes were isolated and annexin-V levels were measured by enzyme-linked immunosorbent assay. We then studied the effects of antiphospholipid immunoglobulin G on placental villous apical annexin-V in vitro. Antiphospholipid immunoglobulin G was isolated from the sera of five different patients with antiphospholipid antibody syndrome along with five paired control immunoglobulin Gs. Short-term cultures were established from normal placental villi and were exposed to the antibodies, after which isolated apical membranes and culture media were immunoassayed for annexin-V levels. RESULTS: Measurements of apical membrane-associated annexin-V from the antiphospholipid placenta showed significantly less apical membrane-associated annexin-V than did the normal placenta (mean +/- SEM: 4.9 +/- 0.4 micrograms/gm villi for antiphospholipid placenta vs 10.2 +/- 0.6 micrograms/gm villi for control, p < 0.001, n = 4). Exposure of placental villous cultures to five different antiphospholipid immunoglobulin Gs for 24 hours resulted in significant reduction of the levels of apical membrane annexin-V (mean +/- SEM: 3.9 +/- 0.3 micrograms/gm villi) compared with paired controls (5.1 +/- 0.3 micrograms/gm villi, p = 0.02). Villi incubated with the different antiphospholipid immunoglobulin Gs had significantly less annexin-V in conditioned media (mean +/- SEM: 45.1 +/- 4.9 ng/gm villi) compared with the paired normal immunoglobulin G control levels (72.6 +/- 11.4 ng/gm villi, p = 0.03). CONCLUSIONS: Antiphospholipid immunoglobulin G reduces the levels of syncytiotrophoblast apical membrane-associated annexin-V in placental villi and the release of annexin-V into surrounding media. Reduction of this anticoagulant protein at the maternal-fetal interface may account for the pregnancy loss observed in patients with antiphospholipid syndrome. Short-term culture of placental villi may offer an in vitro model to further study the mechanism of this effect of antiphospholipid antibodies.

Adult↗
PubMed1997

Autologous bone marrow mixed with HLA-haploidentical allogeneic marrow transplantation for treatment of patients with malignant blood diseases.

We have previously demonstrated that syngeneic marrow mixed with H-2 haploidentical marrow transplantation could provide not only protection against graft-versus-host disease (GVHD) but also anti-leukemic (GVL) effects in mice. In the present studies, we report clinical observations using autologous marrow mixed with HLA-haploidentical allogeneic marrow transplantation for treatment of patients with malignant blood diseases. Sixteen cases, including 12 with acute leukemia and four with advanced malignant lymphoma, were treated by autologous marrow, which was purged in vitro by hyperthemia (42.5 degrees C for 70 min) following incubation for 5 days with interleukin 2 (IL-2) in liquid culture and mixed with HLA haploidentical marrow cells from their sibling or parent. Acute GVHD was not observed in any patient after transplantation. Hematological rescue in the clinical setting was demonstrated in all cases but one who died early from hepatic veno-occlusive disease (VOD). Five cases who were transplanted at the time of CR2 or CR3 and in advanced phase of lymphoma, relapsed 4 to 7 months after transplantation. The relapse rate was 31.3%. None of eight patients who received allogeneic BMT within 2 h after ABMT relapsed with median follow-up of 12 months and two of them died from procedure-related complications. Seven cases are still alive and disease-free with a median follow-up of 12 months. Mixed chimerism was found in 3/6 cases, who had different sex donors, by analysis of sex chromosomes. These results show that mixed transplantation is a safe, effective and new approach to treating patients with malignant tumors. In order to detect the effects of GVL, studies are now in progress in our clinic.

Adult↗
PubMed1997

The significance of subendothelial von Willebrand factor.

von Willebrand factor (vWf) serves to bridge between receptors on the platelet cytoplasmic membrane and the extracellular matrix. In addition to circulating in plasma, vWf is deposited into the extracellular matrix of the subendothelium where it is associated with type VI collagen microfibrils, but not with the elastin-associated microfibrils which are present in the deepest portion of the subendothelium at the zone of the internal elastic lamina. The reaction of platelets to type VI collagen in flow systems is qualitatively different from the shear rate dependent adhesion and aggregation response which is observed with fibrillar type I collagen, exhibiting a response only at low shear rates. The adhesion response to type VI collagen is dependent upon vWf, GP Ib and the GP IIb-IIIa complex. Platelets exposed to purified fibrillin-containing elastin-associated microfibrils adhere and aggregate at low shear rates; this response appears to involve GP IIb-IIIa but not GP Ib. The data are consistent with the hypothesis that type VI collagen is a physiologically relevant binding site for vWf in subendothelium.

Animals↗
PubMed1996

Morphological relationships of von Willebrand factor, type VI collagen, and fibrillin in human vascular subendothelium.

von Willebrand factor (vWF) plays an important role in the process of platelet adhesion after endothelial injury by serving as a bridge between constituents of the vascular subendothelium and platelet membrane receptors. We previously presented evidence that type VI collagen microfibrils serve as a binding site for vWF in human vascular subendothelium. However, others have proposed that vWF is not associated with type VI collagen but rather with the thicker elastin-associated microfibrils, which contain several proteins including fibrillin. We therefore investigated the relationships among vWF, type VI collagen, and fibrillin in human vascular subendothelium by immunoelectron microscopy using single- and double-labeling immunogold localization techniques. In addition, we observed the three-dimensional ultrastructure of vWF-microfibril complexes by stereo paired micrographs and stereo viewer. We found that vWF co-localizes only with the type VI collagen microfibrils in subendothelium but not with fibrillin microfibrils or striated collagen. The vWF is present in subendothelium in the form of electron-dense aggregates having diameters varying between 65 and 80 nm that are closely associated with, and enmesh, the type VI collagen microfibrils and have structural similarities to intracellular Weibel-Palade bodies. The occasional co-localization of type VI collagen and fibrillin adjacent to internal elastic lamina was observed. These results are consistent with the hypothesis that type VI collagen, but not fibrillin-containing microfibrils, serves as a physiologically relevant binding site for vWF in the vascular subendothelium, where the type VI collagen-vWF complex may play an important role modulating the hemostatic response to vascular injury.

Collagen↗
PubMed1994

Reduction of annexin-V (placental anticoagulant protein-I) on placental villi of women with antiphospholipid antibodies and recurrent spontaneous abortion.

OBJECTIVE: The mechanism by which antiphospholipid antibodies are associated with pregnancy loss and thromboembolic conditions has yet to be elucidated. Annexin-V, an anticoagulant phospholipid-binding protein, is normally present in syncytiotrophoblasts lining the placental villi, where it may play a role in the maintenance of intervillous blood fluidity. We therefore investigated the distribution of annexin-V in placentas of patients with antiphospholipid antibodies in situ and then used short-term villous cultures to study the direct effect of antiphospholipid antibodies on the immunolocation of annexin-V. STUDY DESIGN: We performed a blinded study by means of computerized morphometric analysis of placental tissues that were stained for annexin-V with affinity-purified polyclonal antibody in an avidin-biotin peroxidase system. The distribution of villous surface annexin-V on cross sections of placentas of patients with antiphospholipid antibodies was compared with that of placentas from patients with uncomplicated pregnancies, elective abortions, and pregnancy losses not associated with antiphospholipid antibodies (n = 8 for each group). We quantitated villous surface annexin-V in cultured placental villi that were incubated with antiphospholipid antibodies immunoglobulin G compared with normal immunoglobulin G and measured annexin-V levels by enzyme-linked immunosorbent assay in conditioned media and in the villi. RESULTS: The mean villous surface annexin-V of the group with antiphospholipid antibodies was 26.2% +/- 17% (SD) versus 93.9% +/- 5.7% in the normal control group (p < 0.0001). Villi from patients undergoing elective abortions and with pregnancy losses that were not attributed to antiphospholipid antibodies also showed higher mean villous surface annexin-V levels (86.9% +/- 10.6% and 83.5% +/- 11.3%, respectively, p < 0.0001). Organ culture of normal placental villi with affinity-purified immunoglobulin G from patients with antiphospholipid antibodies showed a dose-dependent decrease of villous surface annexin-V over a concentration range of 1.5 micrograms/ml to 1.5 mg/ml. Annexin-V concentrations in conditioned media were significantly lower in the presence of antiphospholipid antibodies immunoglobulin G compared with normal immunoglobulin G (49.4 +/- 8.9 ng/gm wet weight vs 57.2 +/- 11.5 ng/gm, respectively, p < 0.05). In contrast, the mean level of annexin-V in placental villi incubated with antiphospholipid antibodies immunoglobulin G was greater than in villi incubated with normal immunoglobulin G, 1328 +/- 130 ng/gm wet weight versus 1183 +/- 165 ng/gm (p < 0.02). CONCLUSIONS: Patients with antiphospholipid antibodies and a history of previous pregnancy losses have a significant reduction in annexin-V immunostaining on placental villous surfaces, and antiphospholipid antibodies immunoglobulin G can directly decrease levels of villous surface annexin-V on cultured placental villi. Assays of annexin-V in the conditioned media and cell pellets of cultured placental villi suggest that the mechanism for antiphospholipid antibodies-mediated reduced annexin-V surface staining is an inhibition of annexin-V transport to the villous surface rather than displacement by antiphospholipid antibodies from the surface. This antiphospholipid antibodies-induced deficiency of placental surface annexin-V may contribute to the placental thrombosis observed in these patients.

Abortion, Habitual↗
PubMed1994

The expression of the placental anticoagulant protein, annexin V, by villous trophoblasts: immunolocalization and in vitro regulation.

We evaluated the histological and ultrastructural localization of the potent anticoagulant protein, annexin V, at the light and electron microscopic levels, using immunohistochemistry and an immunogold method. Annexin V was found to localize to the microvillar surface of the villous syncytiotrophoblasts. Isolated villous-derived trophoblasts were then utilized to evaluate the expression of annexin 1 protein mRNA in response to syncytialization in vitro, as well as to exposure to adenylate cyclase and protein kinase C agonists. Levels of immunoreactive annexin V released into the conditioned media and associated with cell protein were assessed by ELISA while levels of annexin V mRNA were evaluated by Northern analysis. No significant change in either media or cell-associated annexin V concentrations were detected over time in culture or in response to 1.5 mM 8-bromo-cyclic-adenosine-monophosphate (8-b-cAMP) or 0.15 nM phorbol ester myristic acid (PMA). These results indicate that annexin V is ideally positioned to inhibit intervillous thrombosis and maintain the fluidity of the intervillous circulation. Moreover, the absence of trophoblast annexin V regulation by intracellular second messenger regulators suggests that this crucial placental anticoagulant factor is constitutively produced.

Annexin A5↗
PubMed1993

[Dig-DNA probe for etiologic diagnosis in tuberculosis patients].

We have detected the sputa DNA with nonradioactive probe which is labelled to the specific sequence DNA, 31bp-188bp, by dig-11-dUTP. It showed that both the Sensitivity and specificity are satisfactory. The positive rate of the 80 specimens are 56.25%, and the 30 positive specimens of sputum culture are all positive when detected by the Dig-DNA probe.

Base Sequence↗
PubMed1993

Prolonged disease-free survival after ablative chemoradiotherapy and autologous bone marrow transplantation in adult malignant lymphoma.

From June 1983 to December 1991, 21 adult patients with intermediate or high-grade malignant lymphoma (ML) were treated by ablative chemoradiotherapy, including vincristine, cytosine arabinoside, BCNU and cyclophosphamide plus total lymphoid or body irradiation with boost irradiation over bulky and original tumor areas (Hd-VCCA+TL(B) I) together with autologous bone marrow transplantation (ABMT). Five patients were in advanced stage, 2 in drug-resistant relapse, 6 in drug sensitive relapse, 6 in first complete remission (CR1) and 2 in CR2. One with marrow involvement at ABMT. The 8-year disease-free survival after ABMT in patients with Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) in 89% and 63%, respectively, with a median follow-up up to 34 months. This study demonstrated that our Hd-VCCA+TL(B) I regimen and ABMT performed early in CR or drug-sensitive relapse of adult poor prognosis lymphoma, may potentially cure more than 70% of them. The toxicity of the present treatment is tolerable. The results confirm the value of ABMT in the treatment of adult ML, and suggest that it is necessary to purge the residual tumor cells in the bone marrow at ABMT in patients with marrow infiltration, or lymphoblastic lymphoma.

Adolescent↗
PubMed1993

[Screening of anti-simian immunodeficiency virus (SIV) drugs from Chinese medicinal herbs].

The inhibitory activities of more than 40 species of Chinese medicinal herbs or their single chemical components against simian immunodeficiency virus (SIV) have been studied. The study revealed that four species of the medicinal herbs and a single chemical component had more than 50% inhibition of SIV antigen expression and five other herbs' inhibitory rate of SIV antigen expression was between 30-50%. The results showed that Chinese medicinal herbs could inhibit the SIV activity.

Antiviral Agents↗
PubMed1993

Co-localization of von Willebrand factor and type VI collagen in human vascular subendothelium.

The binding of von Willebrand factor (vWF) to subendothelium constitutes an important initial step in the process of platelet adhesion to exposed subendothelium following blood vessel injury. We previously demonstrated that vWF is present in human vascular subendothelium and recently found that a 150 kd vWF-binding protein, which we extracted from subendothelium, is type VI collagen. Although we have established that vWF and type VI collagen bind in vitro, it is not known whether these two proteins are associated in the vascular subendothelium in situ. We, therefore, 1) investigated the morphological effects of our biochemical extraction procedure on human umbilical veins by scanning and transmission electron microscopy, 2) analyzed the subendothelial extract by immunofluorescence for the presence of vWF and collagens and by electron microscopy for morphological characteristics, and 3) localized vWF and type VI collagen in subendothelium by immunofluorescence and by single- and double-label immunoelectron microscopic studies with protein A-conjugated gold particles. We found that the surface exposed following de-endothelialization is composed of microfibrils and contains very little fibrillar collagen. The subendothelium is stripped after sodium dodecyl sulfate-urea extraction, and the extract itself contains immunoreactive vWF and type VI collagen but no immunoreactive type I or III fibrillar collagens. Immunofluorescence and immunoelectron microscopic studies showed that vWF and type VI collagen are both present in subendothelium, where both co-localized to microfibrils. In conclusion, vWF that binds to type VI collagen in vitro, also co-localizes with type VI collagen in subendothelium, where both are associated with microfibrils. Type VI collagen, therefore appears to serve as a biologically significant binding site for vWF in vivo and may thereby play a role in mediating platelet adhesion to exposed subendothelium following vascular injury.

Collagen↗
PubMedDate not supplied

Chemoimmunosensitization of the T24 human bladder cancer line to Fas-mediated cytotoxicity and apoptosis by cisplatin and 5-fluorouracil.

Previous studies have demonstrated that cisplatin (CDDP) sensitizes bladder cancer cells to Fas-mediated cytotoxicity and that CDDP also enhances the cytotoxic effect of 5-fluorouracil (5-FU). These agents mediate apoptosis and may share common intracellular pathways leading to cell killing. We reasoned that combination treatment with CDDP, 5-FU and anti-Fas monoclonal antibody (mAb) might overcome the drug-resistance. We investigated whether CDDP, 5-FU and anti-Fas mAb synergize in cytotoxicity and apoptosis against the T24 bladder cancer line. Cytotoxicity was monitored by a one day microculture tetrazolium dye assay. Treatment of T24 cells with anti-Fas mAb in combination with CDDP or 5-FU resulted in a synergistic cytotoxic effect. In addition, combination treatment of T24 cells with CDDP, 5-FU and anti-Fas mAb further enhanced the synergistic cytotoxicity achieved by two agents. The synergy achieved in cytotoxicity with CDDP, 5-FU and anti-Fas mAb was also achieved in apoptosis. The current study demonstrates that combination treatment of bladder cancer cells with CDDP, 5-FU and anti-Fas mAb overcomes their resistance. In addition, the sensitization required low concentrations of CDDP and 5-FU, and thus supporting the potential in vivo application of combination of CDDP, 5-FU and immunotherapy in the treatment of drug-resistant bladder cancer.

Antibodies, Monoclonal↗