Biomedical subjects
Publications and source records attributed to Wei He.
Substituted divinyl ketones were studied in the Nazarov cyclization. alpha-Carbomethoxy divinyl ketones underwent efficient Nazarov cyclization with catalytic copper triflate (2 mol %) to give a single cyclopentenone regio- and stereoisomer. The efficiency of the cyclizations correlated with the ability of the substituents to favorably polarize the pi-system of the cationic intermediate.
Novel substituted 2-anilino- and 2-cycloalkylaminoquinoxalines have been found to be useful and selective inhibitors of PDGF-R autophosphorylation. Replacement of an anilino-substituent with substituted cyclohexylamino- or norbornylamino substituents led to significant improvements in the pharmacokinetic profile of these analogues.
RPR127963 demonstrates an excellent pharmacokinetic profile in several species and was found to be efficacious in the prevention of restenosis in a Yucatan mini-pig model upon oral administration of 1-5 mg/kg. The in vitro selectivity profile and SAR of the highly optimized PDGF-R tyrosine kinase inhibitor are highlighted.
Bone morphogenetic proteins (BMPs) regulate multiple cellular processes, including cell differentiation and migration. Their signals are transduced by the kinase receptors BMPR-I and BMPR-II, leading to Smad transcription factor activation via BMPR-I. LIM kinase (LIMK) 1 is a key regulator of actin dynamics as it phosphorylates and inactivates cofilin, an actin depolymerizing factor. During a search for LIMK1-interacting proteins, we isolated clones encompassing the tail region of BMPR-II. Although the BMPR-II tail is not involved in BMP signaling via Smad proteins, mutations truncating this domain are present in patients with primary pulmonary hypertension (PPH). Further analysis revealed that the interaction between LIMK1 and BMPR-II inhibited LIMK1's ability to phosphorylate cofilin, which could then be alleviated by addition of BMP4. A BMPR-II mutant containing the smallest COOH-terminal truncation described in PPH failed to bind or inhibit LIMK1. This study identifies the first function of the BMPR-II tail domain and suggests that the deregulation of actin dynamics may contribute to the etiology of PPH.
Nerve growth factor (NGF) is required for the development of sympathetic neurons and subsets of sensory neurons. Our current knowledge on the molecular mechanisms underlying the biological functions of NGF is in part based on the studies with PC12 rat pheochromocytoma cells, which differentiate into sympathetic neuron-like cells upon NGF treatment. Here we report that the expression of leukemia inhibitory factor receptor (LIFR), one of the signaling molecules shared by several neuropoietic cytokines of the interleukin-6 family, is specifically up-regulated in PC12 cells following treatment with NGF. Attenuation of LIFR signaling through stable transfection of antisense- or dominant negative-LIFR constructs enhances NGF-induced neurite extension in PC12 cells. On the contrary, overexpression of LIFR retards the growth of neurites. More importantly, whereas NGF-induced Rac1 activity is enhanced in antisense-LIFR and dominant negative-LIFR expressing PC12 cells, it is reduced in LIFR expressing PC12 cells. Following combined treatment with NGF and ciliary neurotrophic factor, sympathetic neurons exhibit attenuated neurite growth and branching. On the other hand, in sympathetic neurons lacking LIFR, neurite growth and branching is enhanced when compared with wild type controls. Taken together, our findings demonstrate that LIFR expression can be specifically induced by NGF and, besides its known function in cell survival and phenotype development, activated LIFR signaling can exert negative regulatory effects on neurite extension and branching of sympathetic neurons.
RGS (regulator of G protein signaling) proteins containing the G protein gamma-like (GGL) domain (RGS6, RGS7, RGS9, and RGS11) interact with the fifth member of the G protein beta-subunit family, Gbeta5. This interaction is necessary for the stability of both the RGS protein and for Gbeta5. Consistent with this notion, we have found that elevation of RGS9-1 mRNA levels by transgene expression does not increase RGS9-1 protein level in the retina, suggesting that Gbeta5 levels may be limiting. To examine further the interactions of Gbeta5 and the GGL domain-containing RGS proteins, we inactivated the Gbeta5 gene. We found that the levels of GGL domain-containing RGS proteins in retinas and in striatum are eliminated or reduced drastically, whereas the levels of Ggamma2 and RGS4 proteins remain normal in the absence of Gbeta5. The homozygous Gbeta5 knockout (Gbeta5-/-) mice derived from heterozygous knockout mating are runty and exhibit a high preweaning mortality rate. We concluded that complex formation between GGL domain-containing RGS proteins and the Gbeta5 protein is necessary to maintain their mutual stability in vivo. Furthermore, in the absence of Gbeta5 and all four RGS proteins that form protein complexes with Gbeta5, the animals that survive into adulthood are viable and have no gross defects in brain or retinal morphology.
The functional receptor complex of ciliary neurotrophic factor (CNTF), a member of the gp130 family of cytokines, is composed of CNTF, the CNTF receptor alpha (CNTFR), gp130, and the leukemia inhibitory factor receptor (LIFR). However, the nature of the receptor-mediated interactions in this complex has not yet been resolved. To address this issue we have determined the solution structure of the C-terminal or BC domain of CNTFR and studied the interactions of CNTFR with LIFR and gp130. We reported previously that the membrane distal cytokine-binding domain (CBD1) of LIFR could interact in vitro with soluble CNTFR (sCNTFR) in the absence of CNTF. Here we show that the CBD of human gp130 can also bind in vitro to sCNTFR in the absence of CNTF. In addition, the gp130 CBD could compete with the LIFR CBD1 for the binding of sCNTFR. Substitution of residues in the gp130 CBD, the LIFR CBD1, and the CNTFR BC domain that are expected to be involved in receptor-receptor interactions significantly reduced their interactions. An NMR chemical shift perturbation study of the interaction between the BC domains of CNTFR and gp130 further mapped the interaction surface. These data suggest that both the gp130 CBD and the LIFR CBD1 interact with CNTFR in a similar way and provide insights into the nature of the CNTF receptor complex.
The genetic variation and spatial distribution of the ectomycorrhizal fungus Tricholoma terreum was studied in an artificial Larix principis-rupprechtii stand in China. The 33 sporophores studied belonged to distinct genotypes, based on analysis of random amplified polymorphic DNA (RAPD) markers. The genets of T. terreum were small and no larger than 0.5 m. Two major phenetic groups, i.e. eight individuals in group 1 and 25 in group 2, were identified by principal component analysis and the unweighted pair group method with arithmetic means of simple matching coefficients, respectively. The genetic diversity as determined by the Shannon-Weaver diversity index was 4.52 for these 33 sporophores. Furthermore, there was a slightly lower diversity index in group 1 (4.28) than in group 2 (4.46). It is suggested that sexual spore propagation is very important in the life history of T. terreum in this larch forest.
Memory impairment progressing to dementia is the main clinical symptom of Alzheimer's disease (AD). Deposition of the amyloid-beta peptide (Abeta) in brain, particularly its 42-amino acid isoform (Abeta42), has been shown to play a primary and crucial role in the pathogenesis of AD. In this study we have developed a recombinant adeno-associated virus (AAV) vaccine against AD. This vaccine could express CB-Abeta42 (cholera toxin B subunit and Abeta42 fusion protein) in vivo. A single administration of the AAV-CB-Abeta42 vaccine induced a prolonged, strong production of Abeta-specific serum IgG in transgenic mice that overexpressed the London mutant of amyloid precursor protein (APP/V717I), and resulted in improved ability of memory and cognition, decreased Abeta deposition in the brain, and a resultant decrease in plaque-associated astrocytosis. Our results extended the immunological approaches for the treatment and prevention of AD to an oral, intranasal, or intramuscular route that might be better tolerated in human patients than repetitive parental immunizations in the presence of adjuvant. AAV has attracted tremendous interest as a promising vector for gene delivery. Our results raised the possibility that AAV-CB-Abeta42 vector immunization may provide the basis of a novel and promising Alzheimer's disease vaccination program.
CD28 is the requisite co-stimulatory molecule in the activation of T cells and in the generation of immune responses. But expression of CD28 declined and oxidants accumulated in the elderly. Although accumulation of reactive oxygen species (ROS) during senescence has been reported extensively, the effect of oxidants on CD28-expression remains totally unknown. In this study, we tried to address the molecular mechanism underlying the decrease in CD28-expression of Jurkat T cells cultured in H2O2. Our results indicate that H2O2 could partially block the expression of CD28. This correlates well with a change of nuclear protein binding activity to the motif of site alpha of the CD28 gene, while the site beta-binding activity remained unaltered. On the other hand, since caspase-3 is activated by H2O2, inhibitors of caspase-3 should increase the expression of CD28. What is more interesting is the fact that the site alpha-binding activity was mostly restored after caspase-3 inhibitors had being added. However, caspase-3 is not activated by caspase-8. Maybe it is activated by caspase-9, which is triggered by cytochrome c. We believe that the procaspase-3 is activated by ROS, and the active caspase-3 can induce the change of the site alpha-binding activity, causing a decrease in CD28 expression.
Ligation of CD28, which is present on the majority of CD4(+)T cells, promotes proliferation and immune responses. However, expression of CD28 declines with aging, and apoptosis may contribute to this decline. We have investigated the molecular mechanism underlying the decrease in CD28 expression in Jurkat T cells cultured with FasL. FasL blocks expression of CD28 at the transcriptional level. This correlates with activation of caspase cascades: active caspase-3 can be detected and inhibitors of caspase-3 and caspase-8 increase CD28 promoter activity and CD28 expression. These findings are consistent with the hypothesis that apoptosis plays a key role in the age-related decline of CD28 expression and hence in immunosenescence.
Activities of vascular smooth muscle cells (SMCs) such as proliferation, migration, and matrix production contribute to restenosis following clinical interventions of angioplasty and stent placement. Because activation of platelet-derived growth factor (PDGF)-receptor tyrosine kinase (PDGFr-TK) influences these processes and promotes restenosis, TKI963, an inhibitor of the PDGFr-TK was discovered, and its efficacy was evaluated in blocking stent-induced restenosis as analyzed by intravascular ultrasound (IVUS). TKI963, a low-molecular-weight compound, inhibited the cell-free PDGFbetar-TK with a K(i) value of 56 +/- 14 nM. TKI963 also inhibited PDGF-dependent events in human aortic SMCs (e.g., in situ PDGFr autophosphorylation, mitogenesis, chemotaxis, and collagen production with median inhibitory concentration values of approximately 300 nM) without affecting the activity of a series of membrane receptor tyrosine kinases and intracellular serine/threonine kinases. In vivo, stent-induced restenosis in the swine coronary artery was reduced by oral administration of TKI963 (1.25, 2.5, and 5 mg/kg BID, for 28 days). Late lumen cross-sectional area (CSA) loss, plaque CSA growth, and plaque volume in the stent determined by IVUS were dose-relatedly decreased (33-62% at 1.25 mg/kg BID to 66-92% at 5 mg/kg BID, depending on the parameter) compared with controls. TKI963 treatment of </=1 week following stent placement had no effect on the prevention of restenosis. TKI963, a selective, orally bioavailable inhibitor of the PDGFr-TK, dose-relatedly reduced stent-induced restenosis and did so by inhibiting PDGF-dependent activities that occur as late events following stent placement.
OBJECTIVE: The objective of this study was to investigate the telomerase activity of phytohemagglutinin-activated lymphocytes from patients with Alzheimer disease. BACKGROUND: There is impairment of immune function in patients with Alzheimer disease, and the perturbation of immune system is involved the pathogenesis of Alzheimer disease. However, the mechanism of the impairment is unclear so far. METHODS: We analyzed telomerase activities of phytohemagglutinin-activated lymphocytes from 187 cases of patients with Alzheimer disease, 53 cases of patients with vascular dementia, and 80 cases of age-matched healthy controls, respectively. Telomerase activity was measured using the telomeric repeat amplification protocol-based telomerase polymerase chain reaction enzyme-linked immunosorbent assay. We also detected the proliferation activity of peripheral blood mononuclear cells from 10 cases of patients with Alzheimer disease or 10 cases of age-matched healthy controls by [3H] thymidine incorporation. RESULTS: Telomerase activity of phytohemagglutinin-activated lymphocytes in Alzheimer disease patients was significantly elevated compared with healthy controls (P < 0.01) and vascular dementia patients (P < 0.01), respectively. There was significant statistical correlation between the telomerase activities of lymphocytes and the degree of dementia in Alzheimer disease patients. The proliferation activity of peripheral blood mononuclear cells to phytohemagglutinin was significantly decreased in Alzheimer disease patients compared with age-matched healthy controls (P < 0.01). CONCLUSIONS: Our data suggest that there could be an accelerated telomere dysfunction in lymphocytes of Alzheimer disease patients, which induces the increase of telomerase activity and the decrease of proliferation activity of lymphocytes, and subsequently results in the impairment of immune function in Alzheimer disease patients.
The scaling properties of the fluctuations of EEG time series are used in an investigation of acute stroke in humans. We use detrended fluctuation analysis to characterize the fluctuations in 10-second time series in terms of two dimensionless scaling exponents. The statistics of these scaling exponents across 129 scalp sites define measures which may be used to distinguish normal subjects from those with acute cerebral ischemia. By their nature, these statistics emphasize the global properties of EEG dynamics. Simulation of a focal anomaly which accurately reproduces the mean scaling exponents for stroke subjects contradicts the data for the variances, which we take as evidence that the effect of stroke on EEG is global.
Fibrin deposition and thrombosis within the microvasculature is now appreciated to play a pivotal role in the hepatocellular injury observed in experimental and human viral hepatitis. Importantly, the pathways by which fibrin generation is elicited in viral hepatitis may be mechanistically distinct from the classical pathways of coagulation induced by mechanical trauma or bacterial lipopolysaccharide (LPS). In the setting of murine hepatitis virus strain-3 (MHV-3) infection, a member of the Coronaviridae, activated endothelial cells and macrophages express distinct cell-surface procoagulants, including a novel prothrombinase, Fgl2/fibroleukin, which are important for both the initiation and localization of fibrin deposition. To assess the role of Fgl2/fibroleukin in murine viral hepatitis we generated a Fgl2/fibroleukin-deficient mouse. Peritoneal macrophages isolated from Fgl2/fibroleukin-/- mice did not generate a procoagulant response when infected with MHV-3. Fibrin deposition and liver necrosis were markedly reduced, and survival was increased in mice infected with MHV-3. To address the relevance of Fgl2/fibroleukin in human chronic viral hepatitis we studied patients with minimal and marked chronic hepatitis B. We detected robust expression of Fgl2/fibroleukin mRNA transcripts and protein in liver tissue isolated from patients with marked chronic hepatitis B. Fibrin deposition was strongly associated with Fgl2/fibroleukin expression. Collectively, these data indicate a critical role for Fgl2/fibroleukin in the pathophysiology of experimental and human viral hepatitis.
OBJECTIVE: To evaluate the value of incremental dynamic CT in the diagnosis of pulmonary sclerosing hemangioma (sclerotic hemangioma). METHODS: Thin-section CT at 2.0 mm thickness and 2.0 mm interval was performed before and after administration of contrast material in 20 cases of pulmonary sclerosing hemangioma. RESULTS: Pulmonary sclerosing hemangiomas were shown to be round or oval masses, with smooth margins, homogeneous parenchymal density, and occasional calcification. Homogeneous enhancement was evident in all cases, with maximum CT values ranging from 90 to 110 HU. CONCLUSION: Spiral dynamic CT is useful in the diagnosing of sclerosing hemangioma in the lungs.
OBJECTIVE: gamma delta T cells from the peripheral blood of nasopharyngeal carcinoma(NPC) patients were expanded in vitro and their cytotoxic activity to NPC lines was analysed. A role of gamma delta T cell of immune mechanism in development of NPC was discussed. METHOD: The solid-phase monoclonal anti-TCR gamma delta McAb was used to expand the gamma delta T cells from peripheral blood of seven patients with NPC and six healthy donors. Then we detected the cytotoxicities of these gamma delta T cell against tumour cell lines including Daudi Burkitt's lymphoma and the cell lines of NPC, CNE1 and CNE2, at different effector:target (E:T) ratio by MTT method. The immunophenotyping of gamma delta T cells was detected by flow cytometry. RESULT: gamma delta T cell were obtained with a high purity (> 82%) from healthy individuals and NPC patients in which 90% of expanded cells were V gamma 9/V delta 2 subtype. The results of cytotoxicity assays revealed that activated gamma delta T cells from both healthy donors and patients lysed Daudi, CNE1 and CNE2 cells at similar level(P > 0.05) when E:T ratios increased, the level of cytotoxicity to each of the targets of CNE1 and CNE2 increased. These gamma delta T cells exhibited highest level of cytotoxicity to the Daudi cells in same ration of effector to target cell. CONCLUSION: gamma delta T cell of NPC could significantly kill NPC in vitro.
UNLABELLED: Gated SPECT allows combined assessment of regional myocardial perfusion and left ventricular function. The aim of this study was to address the prognostic value of gated SPECT performed during dobutamine stress testing and during rest on patients with acute myocardial infarction treated with thrombolysis. METHODS: Eighty-eight consecutive patients with uncomplicated acute myocardial infarction who underwent predischarge (3-7 d after admission) dobutamine (5-40 microg/kg of body weight per minute in 3-min dose increments) and rest gated (99m)Tc-sestamibi SPECT were followed for a mean of 48 mo (range, 4-64 mo). RESULTS: Eighteen cardiac events (8 cardiac deaths and 10 nonfatal myocardial infarctions) occurred. Ischemia at dobutamine SPECT imaging (summed difference score or=>or= 1) was present in 60% of the patients. In patients without ischemia, there was a lower event rate (11%), compared with patients with mild ischemia (18%) and moderate-to-severe ischemia (40%) (P < 0.05). Patients with events showed also a higher summed difference score, compared with patients without events (2.3 +/- 1.6 vs. 1.3 +/- 1.6, P < 0.05). Independent predictors of events were the number of segments with preserved (99m)Tc-sestamibi uptake at rest and the number of akinetic or dyskinetic segments with preserved (99m)Tc-sestamibi uptake and preserved wall thickening (global chi(2) of the model, 13.6; P < 0.01). The assessment of the incremental prognostic value of variables added sequentially showed that the addition of the summed difference score added information to perfusion status at rest (P < 0.05). Combined assessment of regional myocardial perfusion and left ventricular function at rest further improved the model (P < 0.05). CONCLUSION: The present study indicated that predischarge (99m)Tc-sestamibi gated SPECT gives prognostic information on patients recovering from acute myocardial infarction. Patients with preserved systolic wall thickening should be regarded as a high-risk subgroup, requiring closer follow-up for appropriate treatment.