Biomedical subjects

Guohua Chen

Publications and source records attributed to Guohua Chen.

At least 19 recordsLinked to original sources

PubMed2006

Overexpression of iron regulatory protein 1 suppresses growth of tumor xenografts.

Iron is essential for proliferation of normal and neoplastic cells. Cellular iron uptake, utilization and storage are regulated by transcriptional and post-transcriptional mechanisms. We hypothesized that the disruption of iron homeostasis may modulate the growth properties of cancer cells. To address this, we employed H1299 lung cancer cells engineered for tetracycline-inducible overexpression of the post-transcriptional regulator iron regulatory protein 1 (IRP1). The induction of IRP1 (wild-type or the constitutive IRP1(C437S) mutant) did not affect the proliferation of the cells in culture, and only modestly reduced their efficiency to form colonies in soft agar. However, IRP1 dramatically impaired the capacity of the cells to form solid tumor xenografts in nude mice. Tumors derived from IRP1-transfectants were <20% in size compared to those from parent cells. IRP1 coordinately controls the expression of transferrin receptor 1 (TfR1) and ferritin by binding to iron-responsive elements (IREs) within their mRNAs. Biochemical analysis revealed high expression of epitope-tagged IRP1 in tumor tissue, which was associated with a profound increase in IRE-binding activity. As expected, this response misregulated iron metabolism by increasing TfR1 levels. Surprisingly, IRP1 failed to suppress ferritin expression and did not affect the levels of the iron transporter ferroportin. Our results show that the overexpression of IRP1 is associated with an apparent tumor suppressor phenotype and provide a direct regulatory link between the IRE/IRP system and cancer.

Aconitate Hydratase↗

PubMed2006

A novel composite nanofiltration (NF) membrane prepared from graft copolymer of trimethylallyl ammonium chloride onto chitosan (GCTACC)/poly(acrylonitrile) (PAN) by epichlorohydrin cross-linking.

A novel composite nanofiltration (NF) membrane was prepared by over-coating the PAN ultrafiltration (UF) membrane with a GCTACC thin layer. The effects of membrane preparation techniques and operating conditions on the performance of the composite membrane were studied. The results indicate that a composite NF membrane from 1.0wt% GCTACC casting solution, vaporized for 2h at 50 degrees C, cross-linked for 20h at 50 degrees C and pH approximately 12 with ethanol/epichlorohydrin (50/0.45 wt/wt) had optimum performance. The resultant GCTACC/PAN composite membrane was positively charged. Scanning electron microscopy showed its asymmetric and composite features. At 25 degrees C and 30L/h of cycling flow, the permeability of pure water through this membrane is 6.3L/hm(2)MPa. At 25 degrees C, 1.2MPa and 30L/h of cycling flow, the rejection of 1000mg/L MgCl(2), CaCl(2), MgSO(4), Na(2)SO(4), and NaCl solutions is 0.976, 0.972, 0.897, 0.65, and 0.407, respectively, with fluxes of 6.8, 6.12, 6.12, 5.57, and 5.51L/hm(2), respectively. The order of rejection of different salts follows the decreasing order of MgCl(2), CaCl(2), MgSO(4), NaCl, KCl, Na(2)SO(4), and K(2)SO(4), which reveals the characteristics of the positively charged NF membrane. In addition, the curve for the streaming potential also illustrates the positively charged characteristics of this membrane, with a pressure osmotic coefficient of 11.7mVMPa(-1).

Acrylonitrile↗

PubMed2006

In vivo tumor growth is inhibited by cytosolic iron deprivation caused by the expression of mitochondrial ferritin.

Mitochondrial ferritin (MtFt) is a mitochondrial iron-storage protein whose function and regulation is largely unknown. Our previous results have shown that MtFt overexpression markedly affects intracellular iron homeostasis in mammalian cells. Using tumor xenografts, we examined the effects of MtFt overexpression on tumor iron metabolism and growth. The expression of MtFt dramatically reduced implanted tumor growth in nude mice. Mitochondrial iron deposition in MtFt-expressing tumors was directly observed by transmission electron microscopy. A cytosolic iron starvation phenotype in MtFt-expressing tumors was revealed by increased RNA-binding activity of iron regulatory proteins, and concomitantly both an increase in transferrin receptor levels and a decrease in cytosolic ferritin. MtFt overexpression also led to decreases in total cellular heme content and heme oxygenase-1 levels. In addition, elevated MtFt in tumors was also associated with a decrease in total aconitase activity and lower frataxin protein level. In conclusion, our study shows that high MtFt levels can significantly affect tumor iron homeostasis by shunting iron into mitochondria; iron scarcity resulted in partially deficient heme and iron-sulfur cluster synthesis. It is likely that deprivation of iron in the cytosol is the cause for the significant inhibition of xenograft tumor growth.

Animals↗

PubMed2006

Ultrasound assisted supercritical fluid extraction of oil and coixenolide from adlay seed.

Oil and coixenolide are important components of adlay seed (Coix lachrymal-jobi L. var. Adlay) with many beneficial functions to human health. In this work, a novel extraction technique--ultrasound assisted supercritical fluid extraction (USFE)--was studied. Effects of operating conditions on the extraction, including extraction temperature (T), pressure (P), time (t), CO(2) flow rate (F) and ultrasonic power (I) were investigated. There are optimum temperatures which gives the maximum extraction yields (EYs) for the supercritical fluid extractions with and without ultrasound. The effect of pressure on EYs for is similar to that of pressure on CO(2) density. Based on the yield of extraction, the favorable conditions for supercritical fluid extraction (SFE) were: T at 45 degrees C, P at 25 MPa, t at 4.0 h and F at 3.5L/h. While ultrasound was applied as in USFE, the following parameters were preferred: T at 40 degrees C, P at 20 MPa, t at 3.5h and F at 3.0 L/h, respectively. The results show that supercritical fluid extraction with the assistance of ultrasound could reduce the temperature, pressure, CO(2) flow rate, as well as time used in the process. Compared with SFE, USFE could give a 14% increase in the yield for extracting oil and coixenolide from adlay seed with less severe operating conditions.

Chromatography, Supercritical Fluid↗

PubMed2006

Electrochemical degradation of amaranth aqueous solution on ACF.

The degradation of Amaranth, a kind of azo dye, has been studied under galvanostatic model with activated carbon fiber (ACF) electrode in aqueous solution with electrochemical method. The ACF was used as anode and cathode, respectively for the decolorization process. The onset oxidation potential and reduction potential for Amaranth on ACF were respectively ascertained at 0.6 and -0.4 V. During the range of -1.1 to 0.50 mA cm(-2), the decolorization was clarified into three processes as electroreduction, adsorption and electrooxidation. There were little contributions to the color and COD removals for the process of adsorption. The color removal can be up to 99% when the current density was 0.50 mA cm(-2). The maximum COD removal was 52% for the process of electrooxidation. Hundred percent color removal was obtained when the current density of -1.0 mA cm(-2) was applied. The maximum COD removal was 62% for the electroreduction. The COD removal results from the adsorption of products for the decolorization process of electrooxidation or electroreduction.

Amaranth Dye↗

PubMed2006

Effects of photodegradation of dissolved organic matter on the binding of benzo(a)pyrene.

Dissolved organic matter (DOM) in natural waters can bind various organic pollutants, and the affinity of this binding is strongly influenced by the chemical characteristics of the DOM and water pH. This study examined the effects of photochemically induced alteration of the DOM's chemical properties and water pH on the binding of benzo(a)pyrene (BaP). Time- and pH-series of solar-simulated irradiations were performed on a natural water sample and aqueous DOM solutions prepared from aquatic and soil humic substances. The binding affinity of BaP, expressed as a partition coefficient of a compound to DOM, decreased substantially after the DOM samples were irradiated over environmentally relevant radiation doses and pH ranges. The lowering of the pH due to the photoproduction of acidic products often partly offsets the reduction of the binding affinity caused by direct photoalteration of the DOM's chemical structure. The decrease of the binding affinity, after correction for the photoinduced pH change, was positively correlated with the decrease in the molecular weight and the aromaticity of the DOM in the course of irradiation. Increasing O(2) abundance accelerated the decrease of the binding affinity as a result of enhanced DOM photodegradation. Visible light played a more important role in reducing the molecular weight and aromaticity of the DOM than in reducing the content of dissolved organic carbon (DOC) via photoremineralization while the reverse was true for UV radiation, indicating that photochemical reduction of the binding affinity may occur in natural waters at depths greater than UV radiation can reach. A decrease of the affinity of DOM for binding BaP will increase the free dissolved fraction of BaP and thus its availability and toxicity to aquatic organisms. The results from this study may have similar implications for organic pollutants other than BaP.

Benzo(a)pyrene↗

PubMed2006

Induction of silencing effect of Swedish mutant amyloid precursor protein by RNA interference.

Over-expression of APP and Swedish mutation could cause some familial early onset AD. In this study, a primary screening was conducted of effective small interference RNAs (siRNAs) targeted wild type APP (APPwt) and Swedish mutant APP (APPswe). One siRNA targeting APPwt and the other siRNA targeting APPswe were designed. All these siRNAs were endogenously expressed by siRNAs expressing plasmids. COS-7 cells were transiently co-transfected with APP-GFP recombinant plasmids and siRNA expression vector. The silencing effect of each siRNA was quantitatively assessed by the level of expression of green fluorescent protein (GFP). It was found that the siRNAs silenced APPwt and APPswe to different degrees. siRNA directed against APPswe was more effective in suppressing the expression of fusion gene of APPswe than that of APPwt. The silencing effect of siRNA directed against APPswe indicating allele-specific silencing property of the siRNAs. Therefore, siRNAs directed against APP play an important role both in the therapeutic study of Alzheimer disease and functional exploration ofAPP gene.

Amyloid beta-Protein Precursor↗

PubMed2006

Sodium nitroprusside promotes IRP2 degradation via an increase in intracellular iron and in the absence of S nitrosylation at C178.

In iron-replete cells the posttranscriptional regulator IRP2 undergoes ubiquitination and proteasomal degradation. A similar response occurs in cells exposed to sodium nitroprusside (SNP), an NO-releasing drug. It has been proposed that nitroprusside ([Fe(CN)5NO]2-) fails to donate iron into cells and that it promotes IRP2 degradation via S nitrosylation at C178. This residue is located within a stretch of 73 amino acids, earlier proposed to define an iron-dependent degradation domain. Surprisingly, we show that IRP2 bearing a C178S mutation or a Delta73 deletion is sensitive to degradation not only by ferric ammonium citrate (FAC) but also by SNP. Moreover, FAC and SNP attenuate the RNA-binding activities of IRP2 and its homologue IRP1 with similar kinetics. Actinomycin D, cycloheximide, succinylacetone, and dimethyl-oxalylglycine antagonize IRP2 degradation in response to both FAC and SNP, suggesting a common mechanistic basis. IRP2 is not only sensitive to fresh, but also to photodegraded SNP and remains unaffected by S-nitrosoglutathione (GSNO), an established nitrosation agent. Importantly, both fresh and photodegraded SNP, but not GSNO, promote a >4-fold increase in the calcein-accessible labile iron pool. Collectively, these results suggest that IRP2 degradation by SNP does not require S nitrosylation but rather represents a response to iron loading.

Cysteine↗

PubMed2006

[Studies on the preparation and characterization of immobilized neutral protease by carboxymethyl chitosan microsphere].

AS1. 398 neutral protease was immobilized onto carboxymethyl chitosan with glutaraldehyde as cross-linking agent. The effects of pH, time of cross-linking, amount of cross-linking agent and the ratio of enzyme to carrier on the activity of the immobilized enzyme were demonstrated, and the optimum immobilization condition of AS1. 398 neutral protease was established. Also studied was the characterization of immobilized enzyme,including pH, temperature, Km and the stability of storage.

Bacterial Proteins↗

PubMed2006

[Study on preparation of the pH sensitive hydroxyethyl chitin/poly (acrylic acid) hydrogel and its drug release property].

Hydroxyethyl chitin (HECH) is a water soluble chitin derivative made by etherification of chitin, ethylene chlorohydrin was used as etherification reagent in this reaction. A novel interpenetrating polymer network (IPN) composed of HECH/PAA was prepared. The IR spectra confirmed that HECH/PAA was formed through chemical bond interaction. The sensitivity of this hydrogel to temperature and pH was studied. The swelling ratio of this hydrogel in artificial intestinal juice is much greater than that in artificial gastric juice. The IPN hydrogel exhibited a typical pH-sensitivity, and its degree of swelling ratio increased with the increase of temperature. The sustained-release drug system of Dichlofenac potassium was prepared by using HECH/PAA as the drug carrier. The release experiment showed a perfect release behavior in artificial intestinal juice. This IPN is expected to be used as a good drug delivery system of enteric medicine.

Acrylates↗

PubMed2005

Inhibition of transferrin receptor 1 transcription by a cell density response element.

TfR1 (transferrin receptor 1) mediates the uptake of transferrin-bound iron and thereby plays a critical role in cellular iron metabolism. Its expression is coupled to cell proliferation/differentiation and controlled in response to iron levels and other signals by transcriptional and post-transcriptional mechanisms. It is well established that TfR1 levels decline when cultured cells reach a high density and in the present study we have investigated the underlying mechanisms. Consistent with previous findings, we demonstrate that TfR1 expression is attenuated in a cell-density-dependent manner in human lung cancer H1299 cells and in murine B6 fibroblasts as the result of a marked decrease in mRNA content. This response is not associated with alterations in the RNA-binding activity of iron regulatory proteins that are indicative of a transcriptional mechanism. Reporter assays reveal that the human TfR1 promoters contains sequences mediating cell-density-dependent transcriptional inhibition. Mapping of the human and mouse TfR1 promoters identified a conserved hexa-nucleotide 5'-GAGGGC-3' motif with notable sequence similarity to a previously described element within the IGF-2 (insulin-like growth factor-2) promoter. We show that this motif is necessary for the formation of specific complexes with nuclear extracts and for cell-density-dependent regulation in reporter gene assays. Thus the TfR1 promoter contains a functional 'cell density response element' (CDRE).

Animals↗

PubMed2005

Fast removal and recovery of Cr(VI) using surface-modified jacobsite (MnFe2O4) nanoparticles.

In this work, the effectiveness of surface-modified jacobsite (MnFe2O4) nanoparticles was investigated for the removal and recovery of Cr(VI) from synthetic wastewater. Ten nanometer modified MnFe2O4 nanoparticles were produced to be a new adsorbent using a co-precipitation method followed by a surface redox reaction. The equilibrium time for Cr(VI) adsorption onto modified MnFe2O4 nanoparticles was as short as 5 min, and the adsorption data fit the Langmuir model well. The maximum uptake of 31.5 mg of Cr(VI)/g of modified MnFe2O4 was obtained at pH 2, which was comparable with other common adsorbents such as activated carbon and sawdust. The effects of ligands (EDTA, SO4(2-), NH4+) and ionic strength were studied in a pH range of 2-10. EDTA and SO4(2-) inhibited the adsorption of Cr(VI) over the entire pH range studied, whereas NH4+ enhanced the uptake of Cr(VI) at pH greater than 6.5. The mechanisms leading to Cr(VI) adsorption by modified MnFe2O4 nanoparticles were determined by X-ray diffraction and X-ray photoelectron spectroscopy to be a combination of electrostatic interaction and ion exchange. Regeneration studies indicated the potential reuse of the modified MnFe2O4 nanoparticles without sacrificing adsorption capacity and the possible recycling of Cr(VI) without changing the valence.

Journal Article↗

PubMed2005

Removal and recovery of Cr(VI) from wastewater by maghemite nanoparticles.

Hexavalent chromium existing in the effluent is a major concern for the metal-processing plant. In this study, a new method combining nanoparticle adsorption and magnetic separation was developed for the removal and recovery of Cr(VI) from wastewater. The nanoscale maghemite was synthesized, characterized, and evaluated as adsorbents of Cr(VI). Various factors influencing the adsorption of Cr(VI), e.g., pH, temperature, initial concentration, and coexisting common ions were studied. Adsorption reached equilibrium within 15 min and was independent of initial Cr concentration. The maximum adsorption occurred at pH 2.5. The adsorption data were analyzed and fitted well by Freundlich isotherm. Cr(VI) adsorption capacity of maghemite nanoparticles was compared favorably with other adsorbents like activated carbon and clay. Competition from common coexisting ions such as Na+, Ca2+, Mg2+, Cu2+, Ni2+, NO3-, and Cl- was ignorable, which illustrated the selective adsorption of Cr(VI) from wastewater. Regeneration studies verified that the maghemite nanoparticles, which underwent six successive adsorption-desorption processes, still retained the original metal removal capacity. In addition, the adsorption mechanisms were investigated using X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and Raman spectroscopic techniques.

Adsorption↗

PubMed2005

Nitric oxide inhibits the degradation of IRP2.

Iron-regulatory protein 2 (IRP2), a posttranscriptional regulator of iron metabolism, undergoes proteasomal degradation in iron-replete cells, while it is stabilized in iron deficiency or hypoxia. IRP2 also responds to nitric oxide (NO), as shown in various cell types exposed to pharmacological NO donors and in gamma interferon/lipopolysaccharide-stimulated macrophages. However, the diverse experimental systems have yielded conflicting results on whether NO activates or inhibits IRP2. We show here that a treatment of mouse B6 fibroblasts or human H1299 lung cancer cells with the NO-releasing drug S-nitroso-N-acetyl-penicillamine (SNAP) activates IRP2 expression. Moreover, the exposure of H1299 cells to SNAP leads to stabilization of hemagglutinin (HA)-tagged IRP2, with kinetics analogous to those elicited by the iron chelator desferrioxamine. Similar results were obtained with IRP2(Delta)(73), a mutant lacking a conserved, IRP2-specific proline- and cysteine-rich domain. Importantly, SNAP fails to stabilize HA-tagged p53, suggesting that under the above experimental conditions, NO does not impair the capacity of the proteasome for protein degradation. Finally, by employing a coculture system of B6 and H1299 cells expressing NO synthase II or IRP2-HA cDNAs, respectively, we demonstrate that NO generated in B6 cells stabilizes IRP2-HA in target H1299 cells by passive diffusion. Thus, biologically synthesized NO promotes IRP2 stabilization without compromising the overall proteasomal activity. These results are consistent with the idea that NO may negatively affect the labile iron pool and thereby trigger responses to iron deficiency.

Animals↗

PubMed2005

[Rapid propagation of Smilax glabra in vitro].

In vitro propagation of Smilax glabra Roxb. was studied. The results indicated that sterilized explants cultured on MS + 6-BA 1.0 mg/L + NAA 0.1 mg/L could lead to budding fastly, the medium MS + 6-BA 1.0 mg/L + NAA 0.1 mg/L + 15% CM suitable for the proliferation, 3/2MS + 6-BA 0.05 mg/L + 4% Sugar suitable for buds strong. Treatment on the medium H (modified) + NAA 0.5 mg/L was best for rooting. The survival rate of the test-tube plantlets was 95% in transplanting on media contained turf-sand (1:1).

Culture Media↗

PubMed2004

Surface and aggregate properties of an amphiphilic derivative of carboxymethylchitosan.

A new kind of amphiphilic derivative of carboxymethylchitosan, a group of (2-hydroxyl-3-butoxyl)propylcarboxymethylchitosans (HBP-CMCHS), has been synthesized, and the surface and aggregate properties have been studied by means of surface tension, surface pressure and fluorescence measurements. HBP-CMCHS can adsorb on the surface to decrease the surface tension of the solution. The adsorption film was quite stable, which can make the relative compressed pressure increase dramatically with the decrease of the surface area. In solution, hydrophobic aggregations were identified by the decrease in the ratio of the fluorescence emission intensity of the first and third pyrene vibronic peaks ( I(1)/ I(3)). Results showed that the aggregation began to form at a concentration similar to that of the polymer transfer to the air-water interface. Aggregate formation of the polymers is a gradually compact process with hydrophobic associations. Increase of DS and addition of NaCl to the HBP-CMCHS solution can make the surface tension decrease, make the aggregation occur at lower concentration, and make the aggregation more hydrophobic.

Adsorption↗

PubMed2004

Iron-mediated degradation of IRP2, an unexpected pathway involving a 2-oxoglutarate-dependent oxygenase activity.

Iron regulatory protein 2 (IRP2), a central posttranscriptional regulator of cellular and systemic iron metabolism, undergoes proteasomal degradation in iron-replete cells. The prevailing model postulates that the mechanism involves site-specific oxidation of 3 cysteine residues (C168, C174, and C178) within a 73-amino-acid (73-aa) degradation domain. By expressing wild-type and mutated versions of IRP2 in H1299 cells, we find that a C168S C174S C178S triple mutant, or a deletion mutant lacking the entire "73-aa domain," is sensitive to iron-mediated degradation, like wild-type IRP2. The antioxidants N-acetylcysteine, ascorbate, and alpha-tocopherol not only fail to stabilize IRP2 but, furthermore, promote its proteasomal degradation. The pathway for IRP2 degradation is saturable, which may explain earlier data supporting the "cysteine oxidation model," and shows remarkable similarities with the degradation of the hypoxia-inducible factor 1 alpha (HIF-1 alpha): dimethyl-oxalylglycine, a specific inhibitor of 2-oxoglutarate-dependent oxygenases, stabilizes IRP2 following the administration of iron to iron-deficient cells. Our results challenge the current model for IRP2 regulation and provide direct pharmacological evidence for the involvement of 2-oxoglutarate-dependent oxygenases in a pathway for IRP2 degradation.

Animals↗

PubMed2003

High-performance Ti/BDD electrodes for pollutant oxidation.

Deposition of stable boron-doped diamond (BDD) films on Ti substrates is believed to be very difficult. In the present study, the stability of Ti/BDD electrodes has been significantly improved by using an organic additive, CH2(OCH3)2. The improved electrodes had service lives of 175-264 h under accelerated life test conditions, which are 2.3-3.0 times longer than the service lives of electrodes prepared with the conventional H2 + CH4 mixture. Raman spectroscopy, X-ray diffraction (XRD), and scanning electron microscopy (SEM) examinations demonstrated thatthe films had well-defined diamond features. The current efficiency (CE) obtained on Ti/BDD was 46.9-78.5% in oxidizing acetic acid, maleic acid, phenol, and dyes, which is 1.6-4.3-fold higher than that obtained on the typical Ti/ Sb2O5-SnO2 electrode. We used a Ti/BDD electrode prepared with H2 + CH4 + CH2(OCH3)2 for over 300 h; its activity remained superior. The successful development of stable and active Ti/BDD electrodes significantly increases the feasibility of industrial applications of anodic oxidation in wastewater treatment.

Boron↗