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Biomedical subjects

Peng Wang

Publications and source records attributed to Peng Wang.

At least 55 records · Page 3Linked to original sources

Molecular cloning and characterization of a novel stem-specific gene from Camptotheca acuminata.

In higher plants, P450s participate in the biosynthesis of many important secondary metabolites. Here we reported for the first time the isolation of a new cytochrome P450 cDNA that expressed in a stem-specific manner from Camptotheca acuminata (designated as CaSS), a native medicinal plant species in China, using RACE-PCR. The full-length cDNA of CaSS was 1735 bp long containing a 1530 bp open reading frame (ORF) encoding a polypeptide of 509 amino acids. Bioinformatic analysis revealed that CASS contained a heme-binding domain PFGXGRRXCX and showed homology to other plant cytochrome P450 monooxygenases and hydroxylases. Southern blotting analysis revealed that there was only one copy of the CaSS present in the genome of Camptotheca acuminata. Northern blotting analysis revealed that CaSS expressed, in a tissuespecific manner, highly in stem and lowly in root, leaf and flower. Our study suggests that CaSS is likely to be involved in the phenylpropanoid pathway.

Amino Acid Sequence↗

Role of death receptor and mitochondrial pathways in conventional chemotherapy drug induction of apoptosis.

The molecular mechanism underlying chemotherapy-induced apoptosis is often debated because of contradicting reports of its signaling pathway. The focus of this ongoing debate is on the requirement of a death receptor and its role in subsequent activation of caspase-8. Understanding the precise mechanism responsible for apoptosis and identifying molecules targeted by chemotherapy will allow us to develop better therapeutic strategies that target the inherent abnormalities of cancer cells. To show conventional chemotherapy drugs can trigger the caspase cascade, including caspase-8, -9, -3 and DNA fragmentation factor, Jurkat T leukemia cells were treated with cisplatin or etoposide in a dose-dependent and a time-dependent manner. Cisplatin and etoposide all induced apoptosis in wild-type Jurkat T leukemia cells. On the other hand, when a pan-caspase inhibitor zVAD-FMK was pretreated, apoptosis did not occur, indicating that these chemotherapy drugs mediated caspase-dependent apoptosis. However, the chemotherapy drug induction of apoptosis was not inhibited by treatment of zIETD-FMK, a caspase-8 inhibitor. There was no difference in cell death between wild-type and caspase-8 or FADD-deficient Jurkat cells after treatment of chemotherapy drug. In addition, cisplatin-induced apoptosis is abrogated by the overexpression of either Bcl-2 or Bcl-x(L), which diminished changes of mitochondrial membrane potential and decreased the amount of cytochrome c released from mitochondria. Again, cisplatin-induced apoptosis was not diminished by c-FLIP-overexpression, whereas the c-FLIP-overexpressing cells were less sensitive to TRAIL-induced apoptosis than the wild type cells. Therefore, these results indicate that conventional chemotherapy drug-triggered apoptosis is indispensable, and its pathway is independent of the death receptor.

Antineoplastic Agents↗

Chloroplastic NAD(P)H dehydrogenase in tobacco leaves functions in alleviation of oxidative damage caused by temperature stress.

In this study, the function of the NAD(P)H dehydrogenase (NDH)-dependent pathway in suppressing the accumulation of reactive oxygen species in chloroplasts was investigated. Hydrogen peroxide accumulated in the leaves of tobacco (Nicotiana tabacum) defective in ndhC-ndhK-ndhJ (DeltandhCKJ) at 42 degrees C and 4 degrees C, and in that of wild-type leaves at 4 degrees C. The maximum quantum efficiency of PSII decreased to a similar extent in both strains at 42 degrees C, while it decreased more evidently in DeltandhCKJ at 4 degrees C. The parameters linked to CO(2) assimilation, such as the photochemical efficiency of PSII, the decrease of nonphotochemical quenching following the initial rise, and the photosynthetic O(2) evolution, were inhibited more significantly in DeltandhCKJ than in wild type at 42 degrees C and were seriously inhibited in both strains at 4 degrees C. While cyclic electron flow around PSI mediated by NDH was remarkably enhanced at 42 degrees C and suppressed at 4 degrees C. The proton gradient across the thylakoid membranes and light-dependent ATP synthesis were higher in wild type than in DeltandhCKJ at either 25 degrees C or 42 degrees C, but were barely formed at 4 degrees C. Based on these results, we suggest that cyclic photophosphorylation via the NDH pathway might play an important role in regulation of CO(2) assimilation under heat-stressed condition but is less important under chilling-stressed condition, thus optimizing the photosynthetic electron transport and reducing the generation of reactive oxygen species.

Chloroplasts↗

Enantiomeric separation of chiral pesticides by high-performance liquid chromatography on an amylose tris-(S)-1-phenylethylcarbamate chiral stationary phase.

Amylose tris-(S)-1-phenylethylcarbamate chiral stationary phase (CSP) was prepared. The direct enantiomeric separation of chiral pesticides on this CSP had been studied by HPLC. The mobile phase was n-hexane-isopropanol at a flow rate of 1.0 mL/min. The effects of isopropanol content and column temperature on retention and enantioselectivity were investigated. Thirty-two samples were tested, of which ten interacted enantioselectively with the CSP. Five samples were completely resolved and another five underwent near-baseline or partial resolution. The enantiomers were identified by a circular dichroism detector. Linear van't Hoff plots were established and the thermodynamic parameters were thus calculated.

Amylose↗

The role of chloroplast NAD(P)H dehydrogenase in protection of tobacco plant against heat stress.

After incubation at 42 degrees C for more than 48 h, brown damages occurred on the stems of tobacco (Nicotiana tabacum L.) ndhC-ndhK-ndhJ deletion mutant (deltandhCKJ), followed by wilt of the leaves, while less the phenotype was found in its wild type (WT). Analysis of the kinetics of post-illumination rise in chlorophyll fluorescence indicated that the PSI cyclic electron flow and the chlororespiration mediated by NAD(P)H dehydrogenase (NDH) was significantly enhanced in WT under the high temperature. After leaf disks were treated with methyl viologen (MV), photosynthetic apparatus of deltandhCKJ exhibited more severe photo-oxidative damage, even bleaching of chlorophyll. Analysis of P700 oxidation and reduction showed that the NDH mediated cyclic electron flow probably functioned as an electron competitor with Mehler reaction, to reduce the accumulation of reactive oxygen species (ROS). When leaf disks were heat stressed at 42 degrees C for 6 h, the photochemical activity declined more markedly in deltandhCKJ than in WT, accompanied with more evident decrease in the amount of soluble Rubisco activase. In addition, the slow phase of millisecond-delayed light emission (ms-DLE) of chlorophyll fluorescence indicated that NDH was involved in the building-up of transthylakoid proton gradient (deltapH), while the consumption of deltapH was highly inhibited in deltandhCKJ after heat stress. Based on the results, we supposed that the cyclic electron flow mediated by NDH could be stimulated under the heat stressed conditions, to divert excess electrons via chlororespiration pathway, and sustain CO2 assimilation by providing extra deltapH, thus reducing the photooxidative damage.

Chloroplasts↗

Photoinduced charge transfer and efficient solar energy conversion in a blend of a red polyfluorene copolymer with CdSe nanoparticles.

We present measurements of charge transfer and the photovoltaic effect in a blend of the alternating polyfluorene copolymer poly(2,7-(9,9-dioctyl-fluorene)-alt-5,5-(4',7'-di-2-thienyl-2',1',3'-benzothiadiazole)) with branched CdSe nanoparticles. Quasi-steady-state photoinduced absorption measurements identified a long-lived charged species that formed after photoexcitation at room temperature. Photovoltaic devices based on this blend system showed a spectral response extending to 650 nm and gave a solar power conversion efficiency of 2.4% under Air Mass 1.5 Global (AM1.5G) conditions.

Cadmium Compounds↗

Transcription of the putative tumor suppressor gene HCCS1 requires binding of ETS-2 to its consensus near the transcription start site.

The hepatocellular carcinoma suppressor 1 (HCCS1) gene was identified by both positional cloning from a predominant region of loss of heterozygosity (17p13.3) in liver cancer and by functional screening for genes affecting cell proliferation in large-scale transfection assays. Its overexpression results in inhibition of cell proliferation in cell culture and tumor growth in nude mice. To understand its transcription regulation, the promoter architecture has been dissected in detail. The major start of transcription was mapped by primer extension to a C residue, 177 nucleotides upstream of the ATG codon. By assessing the promoter activity of a set of linker-scanning mutants of the minimal promoter (-60 to +148 region) in a transient transfection assay, we found that the +1 to + 40 region is critical to HCCS1 gene transcription, containing binding sites for transcription factors NF-kappaB (-21 to +7 and +40 to +26), p53 (+29 to +9) and ETS (+4 to +20 and +23 to +39). Biochemical and molecular analyses revealed that the ETS transcription factors ETS-2 and Elf-1 bind to the two ETS sites in situ and contribute significantly to the transcriptionally active state of the HCCS1 gene, while NF-kappaB, p53 and two other members of the ETS family (ETS-1 and NERF2) appear to play little role. Our observations provide insight into the mechanistic aspects of HCCS1 transcription regulation.

Animals↗

Apoptosis of ATII cells in mice induced by phosgene.

Phosgene inhalation can induced pulmonary edema formation. The purpose of this study was to investigate cell of apoptosis in pulmonary edema mice induced by phosgene. Fifty-two BALB/c mice were random divided into a negative group and a positive group with 26 mice in each. Mice were exposed for 5 min to air and phosgene in the negative group and in the positive one, respectively. The dose of phosgene was 539 ppm. After 4 h of exposure, all mice were anesthetized. Lungs were analyzed for lung wet/dry weight ratio and pathological alternation. The method of isolation and culture of alveolar type II cells (ATII cells) was established to observe their apoptosis by electron microscope and flow cytometry. Apoptosis of lung cells was observed by DNA gel electrophoresis and TUNEL. The lung wet/dry weight ratio was significantly higher in the positive group (6.42 +/- 1.00) than in the negative group (4.25 +/- 0.47, p < 0.05). A large amount of fluid effusion was observed in the alveolus of mice induced by phosgene. Alveolar type II cells were identified by tannic acid staining and electron microscope. The apoptotic signs in alveolar type II cells, alveolar type I cells, eosinophils, macrophages, symphocytes, and ciliated cells were viewed under electron microscope in positive group. The ratio of apoptosis cells (40.26 +/- 7.74) in positive was higher than that (1.58 +/- 1.01, p < 0.001) in the negative group by flow cytometry. DNA ladder alternation was seen through DNA gel electrophoresis. Apoptosis of epithelia and vascular endothelia in lung were found by TUNEL. These results indicate that there is success in establishing a model of pulmonary edema and method of isolation and culture of AT II cells in BALB/c mice. Phosgene can induce apoptosis of cells in the lungs of BALB/c mice, and this indicates that pulmonary edema is related to apoptosis of lung cells in mice, induced by phosgene.

Animals↗

Molecular effect of FadD on the regulation and metabolism of fatty acid in Escherichia coli.

Wild-type Escherichia coli can utilize exogenous long-chain fatty acids (C>12) as a sole carbon source for growth, but it cannot grow on medium-chain fatty acids. However, when the fadD gene from E. coli MG1655 was expressed in E. coli DH5alpha, the latter strain was found to utilize medium-chain fatty acids as its sole carbon source. Various E. coli strains thought to carry subtly different FadR phenotypes were all confirmed as using medium-chain fatty acids as their sole carbon source when the additional fadD was expressed. Semi-quantitative reverse transcription PCR analysis showed that the additional expression of fadD in E. coli strongly induced the expression of both fadE and fadBA. Excessive amounts of medium-chain acyl-CoA would probably bind to FadR and remove the inhibition of fatty acid beta-oxidation.

Base Sequence↗

Removal of antibiotic resistance of live vaccine strain Escherichia coli MM-3 and evaluation of the immunogenicity of the new strain.

MM-3 was a live vaccine strain candidate for protecting neonatal piglets from diarrhea. Designed in the 1980s, a high degree of protection from colibacillosis was afforded to piglets in a challenge study and field trials. However MM-3 had a drawback of carrying the antibiotic resistance gene (chloramphenicol acetyltransferase gene, cat). The introduction of a host-plasmid balanced lethal system into the vaccine was a good idea to solve the problem. The lambda-Red recombination system was adopted in this study to realize the replacement of cat by aspartate-semialdehyde dehydrogenase gene (asd) in the plasmid pMM085. The new plasmid named pMMASD was introduced into an Escherichia coli strain chi6097 and Salmonella typhimurium chi4072 where the asd gene had been knocked out in their chromosomes. Cultured in an Erlenmeyer flask, expression levels of two antigens K88ac fimbriae and heat-labile enterotoxin B subunit (LTB) in cell lysate were similar among MM-3, chi4072(pMMASD) and chi6097(pMMASD). However, chi4072(pMMASD) possessed the more effective secretion mechanism to transport LTB enterotoxin into culture liquid. The relatively higher stability of pMMASD in Salmonella typhimurium chi4072 than that of pMM085 in MM-3 was determined both in vitro in the absence of selective pressure, and in vivo following oral inoculation. Oral immunization of BALB/c mice with chi4072(pMMASD) or chi6097(pMMASD) was sufficient to elicit IgA responses in mucosal tissues as well as systemic IgG antibody responses to the K88 fimbriae, while MM-3 failed to elicit specific antibody responses to K88 fimbriae in mucosal tissues. Among three live strains, only chi4072(pMMASD) could develop strong humoral responses against LTB enterotoxin. The results suggest that chi4072(pMMASD) is expected to be a promising live vaccine strain.

Animals↗

Application of cell penetrating peptide in magnetic resonance imaging of bone marrow mesenchymal stem cells.

Tracking the distribution and differentiation of stem cells by high-resolution imaging techniques would have significant clinical and research implications. In this study, a model cell-penetrating peptide was used to carry gadolinium particles for magnetic resonance imaging (MRI) of mesenchymal stem cells (MSCs). MSCs were isolated from rat bone marrow and identified by osteogenic differentiation in vitro. The cell-penetrating peptide labeled with fluorescein-5-isothiocyanate (FITC) and gadolinium was synthesized by a solid-phase peptide synthesis method. Fluorescein imaging analysis confirmed that this new peptide could internalize into the cytoplasm and nucleus at room temperature, 4 degrees and 37 degrees . Gadolinium were efficiently internalized into mesenchymal stem cells by the peptide in a time or concentration-dependent manner, resulting in intercellular shortening of longitudinal relaxation enhancements, which were obviously detected by 1.5 Tesla Magnetic Resonance Imaging. Cytotoxicity assay and flow cytometric analysis showed that the intercellular contrast medium incorporation did not affect cell viability at the tested concentrations. The in vitro experiment results suggested that the new constructed peptides could be a vector for tracking MSCs.

Animals↗

Gene expression data analysis in subtypes of ovarian cancer using covariance analysis.

Many studies have used microarray technology to identify the molecular signatures of human cancer, yet the critical features of these often unmanageably large set of signatures remain elusive. We have investigated co-expression pattern in four subtypes of ovarian cancer from 104 cancer patients using covariance analysis, treating each subtype of ovarian cancer as a distinct disease entity. We sought gene pairs that were transcriptionally co-expressed in one or multiple subtypes of ovarian cancer, establishing a high confidence network of 87 genes interconnected by significantly high co-expression links that were observed in at least two subtypes of ovarian cancer. We have shown that certain groups of co-expressed gene pairs are cancer subtype specific, through demonstrating significant differences in co-expression patterns of gene pairs between subtypes of ovarian cancer. In addition, we identified a set of 24 genes that classified patients into specific cancer subtypes with a misclassification error rate of less than 5%. Our findings illustrate how large public microarray gene expression datasets could be exploited for identification of cancer subtype specific molecular signatures, and how to classify cancer patients into specific subtypes of cancer using gene expression profiles.

Biomarkers, Tumor↗

[Expression of rabbit neutrophile peptide-1 in nitrate reductase-deficient mutant of Chlorella ellipsoidea].

Application of transgenic Cholrella as bioreactor to express rabbit neutrophile pepetide-1 (NP-1) shows great practical value. In this paper, an NP-1 expression vector containing two selective marker genes NPTII and nitrate reductase gene was constructed. The NP-1 gene was transformed into the nitrate reductase-deficient mutant nrm-4 of Chlorella ellipsoidea via electroporation, and the transgenic alga expressed the active NP-1 were obtained.

Animals↗

Double-wavelength Fano resonance and enhanced coupled-resonator-induced transparency in a double-microcavity resonator system.

We present a theoretical study on resonance control in a double-microcavity resonator system coupled to a waveguide with gain in one microresonator and loss in the other. We demonstrate the variation of the output spectra in the waveguide from initial double-wavelength-wide symmetric resonance to double-wavelength sharper asymmetric Fano profiles when gain is introduced in one of the two microresonators, as well as the inversion of the Fano resonance pattern when gain is introduced in the other microresonator. We also investigate the enhanced coupled-resonator-induced-transparency effect in such a system.

Journal Article↗

[Synthesis and characterization of PEG-segmented polyurethane].

PEG (Mn = 1000)-segmented polyurethanes, with hard segment percentage of 40%, 50% and 55% and named PU-H40, PU-H50 and PU-H55 respectively, were synthesized by bulk polymerization. The structure of PU was characterized by FTIR, DSC, and GPC. Mechanical properties, water contact angles and water vapor transmit rate(WVTR) were also tested. FTIR and DSC showed that the degree of microphase separation increased with the hard content. Mechanical test showed the tensile strength of PU-H50 to be 25 MPa, the highest tensile strength of the PU series. By the use of PEG as soft segment, the surface hydrophilicity of the materials increased dramatically. Owing to its high degree of microphase separation and the mobility of soft segment, the water contact angle of PU-H55 attained to 33 degrees. The WVTRs of PU-H40, PU-H50 and PU-H55 were 789 g/m2/24h, 705 g/m2/24h and 623g/m2/24h respectively. These data suggest that the materials are suitable for fabricating such biomedical articles as surgical gloves, wound dressing and medical protective coating.

Biocompatible Materials↗