Biomedical subjects

Xiaoming Yang

Publications and source records attributed to Xiaoming Yang.

At least 37 records · Page 2Linked to original sources

PubMed2004

Hepatocyte growth factor-induced proliferation of hepatic stem-like cells depends on activation of NF-kappaB.

BACKGROUND/AIMS: Hepatocyte growth factor (HGF) regulates proliferation of hepatic stem cells. Transcription factor nuclear factor kappa B (NF-kappaB) has been demonstrated as a key mediator for cell growth regulation. We investigated the role of NF-kappaB in HGF-mediated cellular proliferation responses in a rat liver-derived hepatic stem-like cell line WB-F344. METHODS: Cell proliferation was determined by incorporation of [3H]thymidine. Phosphorylation of ERK1/2, p38 MAPK, Akt and IkappaBalpha by HGF stimulation was detected by Western blotting. NF-kappaB activation was determined by electrophoretic mobility shift assay and NF-kappaB-mediated SEAP reporter assay. NF-kappaB activation was inhibited by treatment with an IkappaBalpha dominant-negative vector or inhibitor BAY-11-7082. RESULTS: We found that stimulation of WB-F344 cells with HGF promoted cell proliferation and effectively protected WB-F344 cells from apoptosis induced by TNF-alpha. We also observed activation of ERK1/2, p38 MAPK, Akt and NF-kappaB signaling pathways by HGF in WB-F344 cells. HGF-induced cell proliferation was partly blocked by pre-treatment of the cells with inhibitors against MEK1 or p38 MAPK, and completely blocked using an inhibitor for NF-kappaB activity. Furthermore, it was demonstrated that IkappaB mutant that suppressed NF-kappaB activity completely blocked HGF-induced cell proliferation. CONCLUSIONS: NF-kappaB activity is required for HGF-induced proliferation in hepatic stem-like cell line WB-F344, and this activity requires ERK1/2 and p38 MAPK pathways.

Animals↗

PubMed2004

SU5416 is a potent inhibitor of hepatocyte growth factor receptor (c-Met) and blocks HGF-induced invasiveness of human HepG2 hepatoma cells.

BACKGROUND/AIMS: SU5416 is a potent inhibitor of receptor tyrosine kinases, including those of the vascular endothelial growth factor receptor, stem cell factor receptor, and platelet-derived growth factor receptor. Because of the overwhelming evidence favoring the role of aberrant hepatocyte growth factor (HGF)/Met signaling in the pathogenesis of various human cancers, various inhibitor strategies have been employed to therapeutically target this receptor. METHODS: Cell proliferation was determined by incorporation of [(3)H] thymidine. Invasiveness was assayed in Boyden Chambers with 8 microm Matrigel coated filters. Phosphorylation of ERK1/2, Akt by HGF stimulation was detected by Western blotting. RESULTS: We found that SU5416 inhibited motility scattering and the invasive activity of a hepatocellular carcinoma cell line HepG2 in vitro and growth in primary cultured hepatocytes induced by HGF. Consequently, tyrosine autophosphorylation of the c-met induced by HGF was inhibited in these cells by SU5416 in a dose-dependent manner. Furthermore, ERK1/2 and Akt phosphorylation, the signaling events down-stream of c-met activation were reduced. Moreover, SU5416 caused reversion in NIH3T3 fibroblasts transformed by the oncogenic form of the receptor, Tpr-Met. CONCLUSIONS: Inhibition of various solid tumors growth and metastasis by SU5416 may be partially attributed to blocking activation of the hepatocyte growth factor receptor.

3T3 Cells↗

PubMed2004

[Study on antitumor activities of ginkgolic acids from Ginkgo sarcotestas in vitro].

OBJECTIVE: To study the influence of ginkgolic acids on human tumor cells and normal cells. METHODS: Ginkgolic acids (total concentration 90%) was prepared from ginkgo sarcotestas. The inhibitive effect of ginkgolic acids on human tumor cells and normal cells lines was examined by MTT assay. RESULTS: When the concentration was 5.0 micrograms/ml, ginkgolic acids obviously inhibited the growth of tumor cells and didn't influence the normal cells. Inhibitive rate of ginkgolic acids on LTEP-a-2 was 59.1%. High-concentration ginkgolic acids had inhibitive effect on the growth of tumor cells and normal cells. CONCLUSION: Ginkgolic acids had a obvious inhibitive effect on tumor cells in vitro. When the concentration was under 5.0 micrograms/ml, ginkgolic acids didn't influence the growth of normal cells.

Animals↗

PubMed2004

[Study on anti-bacterium activity of ginkgolic acids and their momomers].

OBJECTIVE: Ginkgolic acids and their three monomers were separated from ginkgo sarcotestas. The anti-bacterium activity of ginkgolic acids were tested. The relation between the anti-bacterium activity and side chain of ginkgolic acid were studied. METHOD: The MIC of ginkgolic acids and their three monomers and salicylic acid were tested. RESULTS: Ginkgolic acid has strong inhibitive effect on G+-bacterium. Salicylic acid has no side chain, so no anti-bacterial activity. When the length of gingkolic acid side chain is C13:0, it has the strongest anti-bacterial activity in three monomers. CONCLUSION: The side chain of ginkgolic acid is the key functional group that possessed anti-bacterial activity. The length of Ginkgolic acid was the main effective factor of anti-bacterial activity.

Anti-Bacterial Agents↗

PubMed2003

Imaging of vascular gene therapy.

Gene therapy is an exciting frontier in medicine today. Many genes have been shown to be useful for treatment of various vascular diseases, including chronic cardiac and limb ischemia syndromes, vasculoproliferative disorder, hypercholesterolemia, atherosclerosis, thrombosis, and hypertension. Precise delivery of genes into target vessels, efficient transfer of genes into vascular cells of the target, and prompt assessment of gene expression over time are three challenging tasks for successful vascular gene therapy. Thus, in vivo imaging methods that can be used to monitor gene delivery and localize gene expression are needed. Modern imaging techniques provide an opportunity to monitor and direct vascular gene therapy. Radiologists play a key role not only in developing and mastering endovascular genetic interventions but also in assessing the success of vascular gene therapy and directing further refinement of vascular gene therapy technology. This article provides an overview of the current status of imaging of vascular gene therapy.

Diagnostic Imaging↗

PubMed2003

MRI-guided coronary catheterization and PTCA: A feasibility study on a dog model.

The aim of this work was to demonstrate the feasibility of MRI-guided coronary artery catheterization and intervention in a dog model. Experiments were performed on 10 healthy dogs. A 9F introducer sheath was placed through a right carotid artery cutdown. A prototype 0.014-inch coronary MRI guidewire, a prototype 7 French MRI-guiding catheter, and two flexible surface coils were connected to a GE 1.5 T CV/i scanner for simultaneous visualization of the guidewire, guiding catheter, and chest anatomy. Images were displayed in real time on an in-room monitor. A nongated, single-slice fast gradient-echo sequence was used to obtain real-time images of the catheters and background anatomy during the intervention. Fifteen selective catheterizations were attempted in the coronary arteries, and all were successful. Selective injection of diluted gadolinium into the MRI-guiding catheter provided dynamic 2D projection coronary angiography in all cases, confirming successful catheterization. Percutaneous transluminal coronary angioplasty (PTCA) was attempted after two catheterizations, and all attempts were successful. Inflation of the balloon angioplasty catheter was performed successfully in the left anterior and circumflex arteries. Our results indicate that coronary artery catheterization and intracoronary balloon angioplasty are feasible with MRI guidance only. MRI guidance may be used as an alternative to X-ray guidance in coronary artery interventions in the future.

Angioplasty, Balloon, Coronary↗

PubMed2003

Phased-array MRI of canine prostate using endorectal and endourethral coils.

A four-channel phased array consisting of one surface coil, two endorectal coils, and one flexible endourethral loop coil was designed for MRI of the canine prostate. The endorectal coils provide high signal in the posterior region of the prostate, while the endourethral and surface coils are sensitive to the central and anterior regions of the prostate. Gel phantom experiments indicate that the proposed phased-array configuration generates 15 times more signal-to-noise ratio (SNR) than a combination of two surface coils and one endorectal coil within the posterior region of the prostate; the performance of the two configurations is comparable near the anterior prostate surface. Ultimate intrinsic SNR (UISNR) analysis was used to compare the proposed phased array's performance to the best possible SNR for external coils. This analysis showed that the proposed phased array outperforms the best-case external coil within the posterior and central regions of the prostate by up to 20 times. In canine experiments in vivo, high-resolution fast spin-echo (FSE) images of the prostate were obtained with a pixel size of 230 microm obtained in 3 min 12 s. The proposed phased-array design potentially can be used to increase the accuracy of prostate cancer staging and the feasibility of MR-guided prostate interventions.

Animals↗

PubMed2003

Lentiviral vectors with two independent internal promoters transfer high-level expression of multiple transgenes to human hematopoietic stem-progenitor cells.

Lentiviral vectors (LVs) offer several advantages over traditional oncoretroviral vectors. LVs efficiently transduce slowly dividing cells, including hematopoietic stem-progenitor cells (HSCs), resulting in stable gene transfer and expression. Additionally, recently developed self-inactivating (SIN) LVs allow promoter-specific transgene expression. For many gene transfer applications, transduction of more than one gene is needed. We obtained inconsistent results in our attempts to coexpress two transgenes linked by an internal ribosomal entry site (IRES) element in a single bicistronic LV transcript. In more than six bicistronic LVs we constructed containing a gene of interest followed by an IRES and the GFP reporter gene, GFP fluorescence was undetectable in transduced cells. We therefore investigated how to achieve consistent and efficient coexpression of two transgenes by LVs. In a SIN LV containing the elongation factor 1alpha promoter, we included a second promoter from cytomegalovirus, the phosphoglycerate kinase gene, or the HLA-DRalpha gene. Using a single LV containing two constitutive promoters, we achieved strong and sustained expression of both transgenes in transduced engrafting CD34(+) HSCs and their progeny, as well as in other human cell types. Thus, such dual-promoter LVs can coexpress multiple transgenes efficiently in a single target cell and will enable many gene transfer applications.

Animals↗

PubMed2003

In vivo intravascular MR imaging: transvenous technique for arterial wall imaging.

PURPOSE: To determine, in vivo, the potential for transvenous magnetic resonance (MR) imaging of the arterial wall and to assess appropriate MR pulse sequences for this method. MATERIALS AND METHODS: MR imaging was performed on 19 vessels (right renal artery, N = 9; left renal artery N = 2; external iliac artery, N = 4; abdominal aorta, N = 4) in nine swine. The animals were either low-density lipoprotein receptor knockout (N = 5) or Yucatan mini-pigs fed an atherogenic diet for 6 to 11 weeks (N = 4). The intravascular MR coil/guide wire (IVMRG) (Surgi-Vision, Gaithersburg, MD) was introduced via the external iliac vein into the inferior vena cava (IVC). The following electrocardiograph-gated MR pulse sequences were obtained: T1-weighted precontrast with and without fat saturation and T1-weighted postcontrast with fat saturation. Two observers scored wall signal and conspicuity and classified the vessel as normal, abnormal, or stented. Images were compared with histopathologic findings. RESULTS: The T1-weighted precontrast without fat saturation, T1-weighted precontrast with fat saturation, and T1-weighted postcontrast images correlated with histopathologic findings in 12 of 15 vessels, eight of 10 vessels, and 14 of 16 vessels, respectively. Abnormal histopathologic findings included: arterial wall thickening (N = 3), arterial dissection (N = 2), focal fibrous plaque (N = 2), adherent thrombus (N = 1). The T1-weighted postcontrast images were not compromised by artifacts and had the highest score for vessel wall signal and conspicuity. T1-weighted precontrast images were compromised by chemical shift artifact and poor blood suppression. Negligible artifacts were created by the platinum stent. CONCLUSION: The T1-weighted fat saturated postcontrast pulse sequence was superior to other sequences for transvenous MR imaging of the arterial wall.

Animals↗

PubMed2003

Temperature-sensitive growth and decreased thermotolerance associated with relA mutations in Escherichia coli.

The relA gene of Escherichia coli encodes guanosine 3',5'-bispyrophosphate (ppGpp) synthetase I, a ribosome-associated enzyme that is activated during amino acid starvation. The stringent response is thought to be mediated by ppGpp. Mutations in relA are known to result in pleiotropic phenotypes. We now report that three different relA mutant alleles, relA1, relA2, and relA251::kan, conferred temperature-sensitive phenotypes, as demonstrated by reduced plating efficiencies on nutrient agar (Difco) or on Davis minimal agar (Difco) at temperatures above 41 degrees C. The relA-mediated temperature sensitivity was osmoremedial and could be completely suppressed, for example, by the addition of NaCl to the medium at a concentration of 0.3 M. The temperature sensitivities of the relA mutants were associated with decreased thermotolerance; e.g., relA mutants lost viability at 42 degrees C, a temperature that is normally nonlethal. The spoT gene encodes a bifunctional enzyme possessing ppGpp synthetase and ppGpp pyrophosphohydrolase activities. The introduction of the spoT207::cat allele into a strain bearing the relA251::kan mutation completely abolished ppGpp synthesis. This ppGpp null mutant was even more temperature sensitive than the strain carrying the relA251::kan mutation alone. The relA-mediated thermosensitivity was suppressed by certain mutant alleles of rpoB (encoding the beta subunit of RNA polymerase) and spoT that have been previously reported to suppress other phenotypic characteristics conferred by relA mutations. Collectively, these results suggest that ppGpp may be required in some way for the expression of genes involved in thermotolerance.

Culture Media↗

PubMed2003

In vivo US monitoring of catheter-based vascular delivery of gene microspheres in pigs: feasibility.

In this study, the authors tested the feasibility of using ultrasonography (US) to monitor catheter-based vascular gene microsphere delivery. Polymeric biodegradable microspheres (mean diameter, 5 microm) were prepared by using a double-emulsion technique to encapsulate DNA-plasmid-encoding green fluorescent protein (GFP) genes. With use of gene-delivery catheters, GFP microspheres were locally delivered into the left femoral arterial walls of six pigs; the contralateral arteries were not infused with microspheres and thus served as negative control vessels. The delivery procedures were monitored with high-frequency (8-15-MHz) transducer US. The effectiveness of monitoring with US was compared with the effectiveness of monitoring with immunohistochemical anti-GFP staining. A highly echogenic "star burst" sign around the entire vessel wall was seen at US and correlated with immunohistochemical findings that showed the destination of the gene microspheres. Study results demonstrate the potential of US for monitoring catheter-based vascular gene microsphere delivery in vivo.

Animals↗

PubMed2003

[Antitumor effect of alcohol extract from Sesamum indicum flower on S180 and H22 experimental tumor].

OBJECTIVE: To investigate the antineoplasm effect of extract from Sesamum indicum L. flower. METHOD: Observing the effects of alcohol extract from Sesamum indicum flower on tumor growth in sarcoma 180 (S180) and Heps 22 (H22) tumorigenic mouse, and on weight of immune organs. RESULT: 6, 3, 1.5 g/kg extract showed inhibiting effect on tumor growth obviously, and had not distinct effect on weight of thymus and spleen in mice sarcoma 180 and Heps 22. CONCLUSION: The alcohol extract from Sesamum indicum flower showed obvious antitumor effect.

Animals↗

PubMed2002

The essential role of Cited2, a negative regulator for HIF-1alpha, in heart development and neurulation.

Cited2 is a cAMP-responsive element-binding protein (CBP)/p300 interacting transcriptional modulator and a proposed negative regulator for hypoxia-inducible factor (HIF)-1alpha through its competitive binding with HIF-1alpha to CBP/p300. Disruption of the gene encoding Cited2 is embryonic lethal because of defects in the development of heart and neural tube. Morphological and Doppler echocardiographic analyses of Cited2(-/-) embryos reveal severe cardiovascular abnormalities, including pulmonic arterial stenosis and ventricular septal defects accompanied by high peak outflow velocities, features of the human congenital cardiac defect termed tetralogy of Fallot. The mRNA levels of several HIF-1alpha-responsive genes, such as vascular endothelial growth factor (VEGF), Glut1, and phosphoglycerate kinase 1, increased in the Cited2(-/-) hearts. The increase of VEGF levels is significant, because defects in the Cited2(-/-) embryos closely resemble the major defects observed in the VEGF transgenic embryos. Finally, compared with wild-type, cultured fibroblasts from Cited2(-/-) embryos demonstrate an enhanced expression of HIF-1alpha-responsive genes under hypoxic conditions. These observations suggest that functional loss of Cited2 is responsible for defects in heart and neural tube development, in part because of the modulation of HIF-1 transcriptional activities in the absence of Cited2. These findings demonstrate that Cited2 is an indispensable regulatory gene during prenatal development.

Animals↗

PubMed2002

[Effect of recombinant human augmenter of liver regeneration on gene expression of tissue inhibitor of metalloproteinase-1 in rat with experimental liver fibrosis].

OBJECTIVE: To investigate the influence of recombinant human augmenter of liver regeneration (hALR) on gene expression of tissue inhibitor of metalloproteinase-1 (TIMP1) in rat with experimental liver fibrosis. METHODS: Carbon tetrachloride was injected into the peritoneal cavities of 150 rats twice a week for 8 weeks. Albumin sensitization and caudal vein attack were conducted to another 136 rats for the duration of 3.5 months, so as to establish two kinds of animal model of experimental liver fibrosis. Recombinant hALR at different doses was given during the process of model making. Specimens of liver were taken at different time-points, then the total RNA of liver tissues was isolated and TIMP1 gene expression levels were measured by reverse transcription polymerase chain reaction. RESULTS: In both rat models, TIMP(1) gene expression levels increased gradually during the process of model making. The TIMP(1) gene expression level in CCl4 model treated with low dose hALR was 0.86 +/- 0.13, 1.77 +/- 0.23, 2.78 +/- 0.36, and 3.76 +/- 0.50 respectively 2, 4, 6, and 8 weeks after the beginning of model making, all lower than those in the model group, however, without significant difference. The TIMP(1) gene expression level in CCl4 model treated with high dose hALR was 0.63 +/- 0.10, 1.18 +/- 0.20, 1.89 +/- 0.30, 2.63 +/- 0.33 respectively 2, 4, 6, and 8 weeks after the beginning of model making, significantly lower than those in CCl4 model treated with low dose hALR. In the albumin model group the TIMP(1) gene expression level showed no change during albumin sensitization, significantly increased during caudal vein attack, and reached the peak after medel making. The TIMP(1) gene expression level was lower in albumin model treated with low dose hALR during and after caudal vein attack than in the model group and negative control group, however, without significant difference (both P > 0.05). The TIMP(1) gene expression level in albumin model treated with high dose hALR was 2.33 +/- 0.36 and 4.02 +/- 0.53 during and after caudal vein attack respectively, significantly lower than those in CCl4 model group (0.99 +/- 0.14, 2.03 +/- 0.30, 2.99 +/- 0.43, and 4.13 +/- 0.44 respectively 2, 4, 6, and 8 weeks after beginning of model making respectively) and those in albumin model without hALR treatment and that with low dose hALR (4.13 +/- 0.60 and 5.99 +/- 0.83, and 3.70 +/- 0.82 and 5.63 +/- 0.89 during and after caudal vein attack respectively), and negative control group. CONCLUSION: High dose recombinant human augmenter of liver regeneration is effective in inhibiting gene expression of TIMP(1) in experimental liver fibrosis.

Animals↗

PubMed2002

The COP9 signalosome inhibits p27(kip1) degradation and impedes G1-S phase progression via deneddylation of SCF Cul1.

The COP9 signalosome (CSN) is a conserved protein complex with homologies to the lid subcomplex of the 26S proteasome. It promotes cleavage of the Nedd8 conjugate (deneddylation) from the cullin component of SCF ubiquitin ligases. We provide evidence that cullin neddylation and deneddylation is highly dynamic, that its equilibrium can be effectively modulated by CSN, and that neddylation allows Cul1 to form larger protein complexes. CSN2 integrates into the CSN complex via its C-terminal region and its N-terminal half region is necessary for direct interaction with Cul1. The polyclonal antibodies against CSN2 but not other CSN subunits cause accumulation of neddylated Cul1/Cul2 in HeLa cell extract, indicating that CSN2 is essential in cullin deneddylation. Further, CSN inhibits ubiquitination and degradation of the cyclin-dependent kinase inhibitor p27(kip1) in vitro. Microinjection of the CSN complex impeded the G1 cells from entering the S phase. Moreover, anti-CSN2 antibodies negate the CSN-dependent p27 stabilization and the G1/S blockage, suggesting that these functions require the deneddylation activity. We conclude that CSN inhibits SCF ubiquitin ligase activity in targeting p27 proteolysis and negatively regulates cell cycle at the G1 phase by promoting deneddylation of Cul1.

Animals↗

PubMed2002

Development of an intravascular heating source using an MR imaging guidewire.

PURPOSE: To develop a novel endovascular heating source using a magnetic resonance (MR) imaging guidewire (MRIG) to deliver controlled microwave energy into the target vessel for thermal enhancement of vascular gene transfection. MATERIALS AND METHODS: A 0.032-inch MRIG was connected to a 2.45-GHz microwave generator. We 1) calculated the microwave power loss along the MRIG, 2) simulated the power distribution around the MRIG, 3) measured the temperature increase vs. input power with the MRIG, and 4) evaluated the thermal effect on the balloon-compressed/microwave-heated aorta of six living rabbits. In addition, during balloon inflation, we also simultaneously generated high-resolution MR images of the aortic wall. RESULTS: The power loss was calculated to be 3.9 dB along the MRIG. The simulation-predicted power distribution pattern was cylindrically symmetric, analogous to the geometry of vessels. Under balloon compression, the vessel wall could be locally heated at 41 degrees C with no thermal damage apparent on histology. CONCLUSION: This study demonstrates the possibility of using the MRIG as a multifunctional device, not only as a receiver antenna to generate intravascular high-resolution MR images of atherosclerotic plaques and as a conventional guidewire to guide endovascular interventions during MR imaging, but also as a potential intravascular heating source to produce local heat for thermal enhancement of vascular gene transfection.

Animals↗

PubMed2002

CAND1 binds to unneddylated CUL1 and regulates the formation of SCF ubiquitin E3 ligase complex.

The SCF ubiquitin E3 ligase regulates ubiquitin-dependent proteolysis of many regulatory proteins such as p27(Kip1), IkappaB, and beta-catenin. We report the isolation of a CUL1 binding protein, p120(CAND1). We found the majority of CUL1 is in a complex with CAND1 and ROC1 independent of SKP1 and F box protein SKP2. Both in vivo and in vitro, CAND1 prevents the binding of SKP1 and SKP2 to CUL1 while dissociation of CAND1 from CUL1 promotes the reverse reaction. Neddylation of CUL1 or the presence of SKP1 and ATP causes CAND1 dissociation. Our data suggest that CAND1 regulates the formation of the SCF complex, and its dissociation from CUL1 is coupled with the incorporation of F box proteins into the SCF complex, causing their destabilization.

Adenosine Triphosphate↗

PubMed2002

Thermal effect of intravascular MR imaging using an MR imaging-guidewire: an in vivo laboratory and histopathological evaluation.

BACKGROUND: Intravascular magnetic resonance (MR) imaging to guide interventional procedures is a rapidly growing field. A primary concern with these new techniques is their thermal safety. The purpose of this study was to evaluate, in vivo, the thermal effect of an MR imaging-guidewire (MRIG) for intravascular MR imaging (IVMRI). MATERIAL/METHODS: Two indications of potentially adverse local heating were investigated: blood coagulation disorders and pathologic changes in target vessels. Experiments were performed on ten rabbits with a 1.5 T MR scanner. Using a 0.64-mm MRIG as the RF receiver, we imaged the target aorta using a fast spin-echo pulse sequence with an average specific absorption rate (SAR) of 0.6 W/kg. The total MR imaging time was approximately 70 minutes. RESULTS: There were no abnormal value changes of the coagulation factors between pre- and post-IVMRI, no clinical manifestations of blood coagulation disorders, and, histopathologically, no thermal damage in target vessels. CONCLUSIONS: This study demonstrates, from a pathophysiological point of view, the potential safe use of the MR imaging-guidewire for intravascular MR imaging. Further study is required to precisely define the boundaries of these safe operating parameters.

Animals↗