Biomedical subjects
Publications and source records attributed to X Wang.
The compound Cs10.5[(V16O40)(Si4.5V1.5O10)].3.5H2O is the first example of an extended structure in which a polyoxometallate anion is linked by an extended tetrahedral unit.
The scaling of the reconnection electric field in a collisionless plasma is determined analytically for a model of forced reconnection. In particular, the dependence of the length of the reconnection layer on the ion skin depth and the boundary conditions is calculated explicitly. Analytical results are tested by Hall magnetohydrodynamics simulations.
The Ras-like Rab GTPases regulate vesicle transport in endocytosis and exocytosis. We found that cardiac Rabs1, 4, and 6 are upregulated in a dilated cardiomyopathy model overexpressing beta(2)-adrenergic receptors. To determine if increased Rab GTPase expression can contribute to cardiomyopathy, we transgenically overexpressed in mouse hearts prototypical Rab1a, the small G protein that regulates vesicle transport from endoplasmic reticulum to and through Golgi. In multiple independent mouse lines, Rab1a overexpression caused cardiac hypertrophy that progressed in a time- and transgene dose-dependent manner to heart failure. Isolated cardiac myocytes were hypertrophied and exhibited contractile depression with impaired calcium reuptake. Ultrastructural analysis revealed enlarged Golgi stacks and increased transitional vesicles in ventricular myocytes, with increased secretory atrial natriuretic peptide granules and degenerative myelin figures in atrial myocytes; immunogold studies localized Rab1a to these abnormal vesicular structures. A survey of hypertrophy signaling molecules revealed increased protein kinase C (PKC) alpha and delta, and confocal microscopy showed abnormal subcellular distribution of PKCalpha in Rab1a transgenics. These results indicate that increased expression of Rab1 GTPase in myocardium distorts subcellular localization of proteins and is sufficient to cause cardiac hypertrophy and failure.
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OBJECTIVE: To construct a kind of engineered cell secreting human GDNF and study its possible effects on gene therapy of Parkinson's disease. METHOD: Human GDNF cDNA with Kozak sequence was cloned by RT-PCR, and then was transfected into SH-SY5Y cell line of human neuroblastoma. These engineered cells were co-cultured with primary mesencephalic cells of rats. Dopaminergic neurons were examined by immunohistochemistry. RESULTS: The number of dopaminergic neurons protected by engineered cells increased at least by 95.4% in comparison with the control cells (P < 0.01). The number of dopaminergic neurons protected by engineered cells against MPP+ toxicity increased 9.5-10.8 times (P < 0.01). CONCLUSION: A kind of engineered SH-SY5Y cells secreting human GDNF has been constructed successfully. These cells obviously protect dopaminergic neurons against degeneration and MPP+ toxication and may play an important role in gene therapy of Parkinson's disease.
Model-free approaches (error-prone PCR to introduce random mutations, DNA shuffling to combine positive mutations, and screening of the resultant mutant libraries) have been used to enhance the catalytic activity and thermostability of alpha-aspartyl dipeptidase from Salmonella typhimurium, which is uniquely able to hydrolyze Asp-X dipeptides (where X is any amino acid) and one tripeptide (Asp-Gly-Gly). Under double selective pressures of activity and thermostability, through two rounds of error-prone PCR and three sequential generations of DNA shuffling, coupled with screening, a mutant pepEM3074 with approximately 47-fold increased enzyme activity compared with its wild-type parent was obtained. Moreover, the stability of pepEM3074 is increased significantly. Three amino acid substitutions (Asn89His, Gln153Glu, and Leu205Arg), two of them are near the active site and substrate binding pocket, were identified by sequencing the genes encoding this evolved enzyme. The mechanism of the enhancement of activity and stability was analyzed in this paper.
We discover that epigallocatechin gallate (EGCG) from green tea is an inhibitor of fatty-acid synthase (FAS) from chicken liver. Its inhibition of FAS is composed of reversible fast-binding inhibition, through which 52 microM EGCG can inhibit 50% of the activity of FAS, and irreversible slow-binding inactivation following saturation kinetics with the dissociation constant of 0.352 mM and limiting rate constant of 0.0168 min(-1). The marked inhibition of ketoacyl reduction shows that the inhibition is related to beta-ketoacyl reductase of FAS. The observable protection of NADPH and competitive inhibition of NADPH for ketoacyl reduction indicate that EGCG may compete with NADPH for the same binding site. The synthetic inhibitor C75 does not show obvious fast-binding inhibition, but does exhibit irreversible slow-binding biphasic inactivation, which is demonstrated to be a second-order reaction. That the inactivation by C75 is protected by malonyl-CoA indicates C75 is similar to cerulenin in being a covalent inactivator of the beta-ketoacyl synthase.
We hypothesize that the decline of male gonadal hormones may play a role in age-related decrease of calcitonin gene-related peptide (CGRP) synthesis and release. Orchidectomized rats were raised with or without testosterone replacement and CGRP levels in serum and some tissues as well as the perfusate from the isolated mesenteric arterial bed (MAB) were measured at 1, 2 and 4 months after orchidectomy. CGRP levels of serum and tissues, and CGRP release from MAB were significantly elevated after 1 month and decreased after 4 months in orchidectomized rats. The changes were restored by testosterone replacement. Our results indicate that the age-related decline of testosterone might contribute to the age-related decrease of CGRP synthesis and release.
Calcitonin gene-related peptide (CGRP), is produced in dorsal root ganglia (DRG) neurons and released from primary afferent neurons to mediate hemodynamic effects and neurogenic inflammation. In this work, we determined whether lipopolysaccharide (LPS), an inflammatory stimulator, could trigger CGRP release from cultured DRG neurons and if so, which cellular signaling pathway was involved in this response. Cytoplasmic concentration of calcium ([Ca(2+)](i)) plays a key role in neurotransmitter release, therefore [Ca(2+)](i) was also determined in cultured DRG cells using fluo-3/AM. The results showed that LPS (0.1-10 microg/ml) evoked CGRP release in a time- and concentration-dependent manner from DRG neurons. LPS also increased [Ca(2+)](i) in a concentration-dependent manner. The protein kinase C (PKC) inhibitors, calphostin C 0.5 microM or RO-31-8220 0.1 microM, and the cAMP-dependent protein kinase (PKA) specific inhibitor RP-CAMPS 30 microM or nonspecific inhibitor H8 1 microM inhibited 1 microg/ml LPS-evoked CGRP release and [Ca(2+)](i) increase from DRG neurons. The cGMP-dependent protein kinase (PKG) inhibitor Rp-8-pCPT-cGMPS 30 microM did not block the LPS response. These data suggest that LPS may stimulate CGRP release and [Ca(2+)](i) elevation through PKC and PKA, but not PKG signaling pathway in DRG neurons of neonatal rats.
The authors conducted a simulation study to evaluate whether exclusion of the early mortality (deaths occurring during a prespecified period immediately after baseline) reduces confounding of the exposure-mortality relation by preexisting disease. The simulation specified an exposure that decreased mortality risk in the absence of confounding and then introduced confounding by preexisting disease that biased the "true" protective effect of exposure towards greater risk. In 2,000 cohorts, exclusion of the early mortality (deaths occurring during the first 25 months of a 60-month follow-up period) did not alter the mean hazard ratio for exposure under conditions of confounding by preexisting disease that produced a constant, threefold increase in mortality risk during follow-up (the mean hazard ratio was 1.72 for all subjects and 1.72 after exclusion of the early mortality). However, when the authors specified confounding by preexisting disease which produced a threefold increase in mortality risk that attenuated over time, exclusion of the early mortality consistently identified the "true" protective effect of exposure (the mean hazard ratio was 1.07 for all subjects and 0.31 after exclusion of the early mortality). Thus, under conditions of confounding by preexisting disease which produces an increase in mortality risk that attenuates over time--an effect that does have empirical support-the early mortality exclusion can be very effective in revealing the "true" exposure-mortality relation.
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OBJECTIVE: To observe the effect of NPS R-467, a calcimimetic, on PTH secretion of cultured parathyroid cells from severe secondary hyperparathyroidism (SHPT) patient. METHODS: The parathyroid tissue from a severe SHPT patient (iPTH: 2000 pg/ml) who had underwent parathyroidectomy was cultured in medium containing 1.05 mmol/L Ca and 0.8 mmo/L Mg. After 90 minutes' culture, NPS R-467 or NPS S-467, as negative control, with the final concentrations of 25, 50, and 100 nmol/L was added. The supernatant of medium was aspirated after 15, 30, and 60 minutes and iPTH therein was measured by radioimmunoassay. RESULTS: There was no difference between the PTH concentrations in NPS R-467 group and NPS S-467 15 minutes after the reagents were added (1.8 ng/10(5) cells). In the NPS S-467 group, the PTH secretion was elevated along with the culture time, at 60 minutes up to 2.3 ng/10(5) cells. The PTH concentration of NPS R-467 was 80% of that of control group 30 minutes later and 60% of that in control group 60 minutes later. No difference was found among the effects of NPS R-467 at different concentrations. CONCLUSION: The calcimimetic NPS R-467 significantly suppresses the PTH secretion, even in severe uremic hyperparathyroidism patient. It is an effective medicine for hyperparathyroidism.
Efflux of glutamate during cerebral ischemia is known to contribute to brain cell death via processes of excitotoxicity. However, gamma-aminobutyric acid (GABA) is also released during ischemia, and may be protective. In this study, we used in vivo microdialysis to map the efflux of glutamate and GABA from central core and peripheral zones of focal ischemia in mouse brain. We show that the temporal profiles of glutamate and GABA efflux are significantly different in core versus peripheral zones. Calculation of glutamate/GABA ratios demonstrate that, in the core, there is a significant increase above baseline ratios during the first 30 mm of ischemia, which then rapidly renormalizes. In contrast, no significant changes in glutamate/GABA ratios were seen in the ischemic periphery. These data suggest that imbalances in glutamate versus GABA efflux may be an initial trigger of excitotoxic brain damage in the core but not the peripheral zones of focal cerebral ischemia.
BACKGROUND: Following cleavage by caspase 8, the C-terminus of Bid translocates from the cytosol to the mitochondria that is dependent upon structures formed by the mitochondrial-specific lipid cardiolipin. Once associated with mitochondria, truncated Bid (tBid) causes the potent release of cytochrome c, endonuclease G, and smac. RESULTS: We investigated whether tBid localizes specifically to the contact sites of mitochondria purported to be rich in cardiolipin. A point mutation changing the glycine at position 94 to glutamic acid in the BH3 domain of tBid (tBidG94E) was principally used because mitochondria treated with this mutant tBid displayed better preservation of the outer membrane than those treated with wild type tBid. Additionally, tBidG94E lowers the cytochrome c releasing activity of tBid without affecting its targeting to mitochondria. Electron microscope tomography coupled with immunogold labeling was used as a new hybrid technique to investigate the three-dimensional distributions of tBid and tBidG94E around the mitochondrial periphery. The statistics of spatial point patterns was used to analyze the association of these proteins with contact sites. CONCLUSIONS: Immunoelectron tomography with statistical analysis confirmed the preferential association of tBid with mitochondrial contact sites. These findings link these sites with cardiolipin in tBid targeting and suggest a role for Bcl-2 family members in regulating the activity of contact sites in relation to apoptosis. We propose a mechanism whereby Bcl-2 proteins alter mitochondrial function by disrupting cardiolipin containing contact site membranes.
Membrane-type matrix metalloproteinase 1 (MT1-MMP) plays a critical role in extracellular matrix remodeling under both physiological and pathological conditions. However, the mechanisms controlling its activity on the cell surface remain poorly understood. In this study, we demonstrate that MT1-MMP is regulated by endocytosis. First, we determined that Con A induces proMMP-2 activation in HT1080 cells by shifting endogenous MT1-MMP from intracellular compartments to cell surface. This phenotype was mimicked by the cytoplasmic truncation mutant MT1 Delta C with more robust pro-MMP-2 activation and cell surface expression than wild-type MT1-MMP in transfected cells. MT1 Delta C was subsequently shown to be resistant to Con A treatment whereas MT1-MMP remains competent, suggesting that Con A regulates MT1-MMP activity through cytoplasmic domain-dependent trafficking. Indeed, MT1-MMP was colocalized with clathrin on the plasma membrane and with endosomal antigen 1 in endosomes. Internalization experiments revealed that MT1-MMP is internalized rapidly in clathrin-coated vesicles whereas MT1 Delta C remains on cell surface. Coexpression of a dominant negative mutant of dynamin, K44A, resulted in elevation of MT1-MMP activity by interfering with the endocytic process. Thus, MT1-MMP is regulated by dynamin-dependent endocytosis in clathrin-coated pits through its cytoplasmic domain.
BACKGROUND: Thalidomide has been shown to have antiangiogenic effects in preclinical models as well as a significant antitumor effect in hematologic tumors such as multiple myeloma. The authors performed this Phase II study to determine the activity, toxicity profile, and antiangiogenic effect of thalidomide in patients with locoregionally recurrent or metastatic squamous cell carcinoma of the head and neck. METHODS: Twenty-one patients with recurrent or metastatic squamous cell carcinoma of the head and neck were treated with single-agent thalidomide. All patients had received radiation therapy, and most had undergone surgery (95%) and/or chemotherapy (90%). Thalidomide was initiated at 200 mg;3>daily and increased to a target dose of 1000 mg daily. Patients continued treatment until disease progression, unacceptable toxicity, or death occurred. RESULTS: All 21 patients eventually developed progressive disease. Median time to progression was 50 days (95% confidence interval, 28-70), with median overall survival time of 194 days (95% lower confidence boundary, 151), similar to the progression and survival times reported for this patient group with other agents. Thalidomide was generally well tolerated, with few patients experiencing Grades 3 to 4 toxicities. Serum vascular endothelial growth factor and basic fibroblast growth factor levels increased in six of seven patients, for whom paired serum samples were available and all of whom had progressive disease. CONCLUSIONS: In this heavily pretreated population of patients with advanced squamous cell carcinoma of the head and neck, thalidomide does not appear to have single-agent antitumor activity. Further evaluation of the mechanism of action of thalidomide is indicated. Potentially, future evaluations of thalidomide may be performed in combination with other antiangiogenic or cytotoxic agents in patients with earlier stage disease or in patients with minimal residual disease.
OBJECTIVES: The study evaluated the relationship between plasma cardiotrophin-1 (CT-1) concentration and left ventricular (LV) mass in dilated cardiomyopathy (DCM) patients with congestive heart failure (CHF). BACKGROUND: Cardiotrophin-1 is a newly identified member of the interleukin-6 (IL-6) family of cytokines and one of the endogenous ligands for gp130 signaling pathways in the heart, and it has potent hypertrophic and survival effects on cardiac myocytes. However, the clinical significance of CT-1 is poorly understood. METHODS: We measured the plasma CT-1 level in 51 consecutive patients with DCM. Patients were classified into two groups: small LV mass index group and large LV mass index group, based on the median level of LV mass index. RESULTS: The plasma CT-1 level was increased in DCM patients with the severity of CHF and was significantly higher in the large LV mass group than in the small LV mass group, despite the absence of a difference in LV ejection fraction between the two groups. In addition, there was a significant positive correlation between the plasma CT-1 level and the LV mass index (r = 0.627, p < 0.0001). According to stepwise multivariate analyses among hemodynamic and neurohumoral factors, a high plasma CT-1 level showed an independent and significant positive relationship with a large LV mass index in patients with DCM. CONCLUSIONS: These results indicate that the plasma CT-1 level is increased in patients with DCM and is significantly correlated with the LV mass index, suggesting that CT-1 plays an important role in structural LV remodeling in patients with DCM.
OBJECTIVE: To investigate and analyse HLA A/B local haplotype gene frequency (HGF) in the females of Han nationality in Hunan. METHODS: HLA-A, B local antigen polymorphisms were investigated via microcytotoxicity assay. Haplotype was inferred by means of linkage disequilibrium parameter. RESULTS: Among the 46 haplotypes, HGF high were: A11-B60 (HGF = 0.1381), A2-B60 (HGF = 0.0861), A24-B48 (HGF = 0.0709). Three haplotype's linkage disequilibrium parameter have significant difference, they are A2-B52, A11-B13, A24-B52. CONCLUSION: The female of Han Nationality of Hunan HLA-A, B especially B local antigen frequency exists difference sompare with Han Nationality in Hunan and other areas in China of mix public of both sexes. When study relativity of the female disease of Han Nationality of Hunan with HAL, should notice HLA distributing particularity, be assure data veracity. Among the 46 haplotypes, A2-B52, A11-B13, A24-B52 three haplotypes exist significant linkage disequilibrium, probably they are own haplotypes at the female of Han Nationality in Hunan.