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1.
Guang Pu Xue Yu Guang Pu Fen Xi ; 36(9): 2966-70, 2016 Sep.
Article in Chinese | MEDLINE | ID: mdl-30085488

ABSTRACT

At present, the way to introduce the sample into the inductively coupled plasma atomic emission spectrometry (ICP-AES) light source is still in the form of solution. In order to improve the treatment effect of the aqueous solution and change its physical properties, the surface tension and viscosity under different experimental conditions were measured with magnetic stirring combined with laser irradiation. . The treated samples were introduced into the inductively coupled plasma (ICP) to measure the spectral line intensity, signal-to-background ratio, excitation temperature and electron density emitted by the ICP source. The experimental results showed that: when the magnetic stirrer rotate speed was 1 197 r·min-1, the laser power density was 0.227 6 W·cm-2 and irradiation for 15 min, the surface tension and viscosity of the solution were decreased by 27.85% and 8.66% respectively than those of the untreated solution. As to the element spectral lines of As 188.980 nm, Cd 214.439 nm, Cr 267.716 nm, Cu 324.754 nm, Hg 253.652 nm and Pb 220.353nm: the intensity was enhanced 32.07%, 65.36%, 18.27%, 32.29%, 19.38% and 54.28%; the signal-to-background ratio increased by 25.13%, 60.97%, 18.18%, 27.69%, 21.11% and 48.93%, respectively. The enhancement of the plasma radiation was explained to a certain extent by measuring the excitation temperature and electron density of the plasma. The processing method of the aqueous solution can effectively improve the spectral intensity and signal-to-background ratio of the ICP. Compared with the laser irradiation aqueous solution separately, this method significantly shortened the processing time, improve the efficiency. This method is simple, with no secondary pollution in the treatment of the sample solution, convenient popularization and use.

2.
Guang Pu Xue Yu Guang Pu Fen Xi ; 35(1): 203-7, 2015 Jan.
Article in Chinese | MEDLINE | ID: mdl-25993849

ABSTRACT

In order to change the physical properties of aqueous solution and improve the radiation intensity of the ICP emission spectrum, the effects of different laser power density and irradiation time on the surface tension and viscosity of aqueous solution were investigated by using near infrared laser at 976 nm and CO2 laser at 10. 6 µm to irradiate aqueous solution orthogonally, then the enhancement of ICP spectral intensity with processed solution was discussed. The results showed that the surface tension and viscosity of aqueous solution reduced by 42. 13% and 14. 03% compared with the untreated, and the atomization efficiency increased by 51.26% at the laser power density 0. 265 7 W . cm-2 of 976 nm and 0. 206 9 W . cm-2 of CO2 laser with 40 min irradiation time. With the optimized aqueous solution introduced into the ICP source, the spectral line intensity of sample elements As, Cd, Cr, Hg and Pb was enhanced by 46.29%, 94. 65%, 30. 76%, 33.07% and 94. 58% compared to the untreated aqueous solution, while the signal-to-background ratio increased by 43. 84%, 85. 35%, 28. 71%, 34. 37% and 90. 91%, respectively. Plasma temperature and electron density also increased by 5. 94% and 1. 18% respectively. It is obvious that the method of double-beam laser orthogonal irradiation on solution can reduce the surface tension and viscosity of aqueous solution significantly, and raise the radiationintensity of ICP source, and will provide a better condition for detecting the trace heavy metal elements in water samples.

3.
Guang Pu Xue Yu Guang Pu Fen Xi ; 34(9): 2337-41, 2014 Sep.
Article in Chinese | MEDLINE | ID: mdl-25532321

ABSTRACT

In order to reduce the effect of the spectral line self-absorption on the analysis result in the laser induced plasma and enhance the qualities of spectrum, the spectral information was recorded by the spectral analysis system consisting of a modular multifunctional grating spectrometer and a CCD detector etc., and the electron temperature and electron density of the plasma were measured with the spectroscopic methods. A plane mirror device was used to constraint the laser plasma, and a reasonable explanation was got through comparing the linear evolution under different experimental conditions and measuring the temperature, electronic density and sample evaporation. The result shows that when an appropriate plane mirror device was used to constraint the laser plasma, the axial temperature of the plasma increased and the radial distribution of the plasma becomes uniform; the electron density increased dramatically; however, obviously sample evaporation decreased, which may be the reasons for being able to effectively reduce the level of self-absorption spectral lines. Therefore, the plane mirror device could reduce the self-absorption effect in the laser-induced plasma. This makes it possible to choose a sensitive line that acts as analysis line in the quantitative analysis of the major elements. In other words, this promotes the measurement precision in the laser-induced break-down spectroscopy.

4.
Guang Pu Xue Yu Guang Pu Fen Xi ; 34(6): 1667-70, 2014 Jun.
Article in Chinese | MEDLINE | ID: mdl-25358184

ABSTRACT

To enhance the intensity of inductively coupled plasma-atomic emission spectrum and improve the detection level of trace heavy metal elements, the surface tension and viscosity of the aqueous solution processed by near-infrared laser at wave-length of 976 nm were studied in the present paper. The influences of the treated solution on the spectral line intensity and signal-to-background ratio of the ICP source were observed. The results showed that when the laser irradiation time was 60 min and the power density was 0.329 6 W x cm(-2), the surface tension and viscosity of the solution decreased by 36.73% and 9.73% respectively compared to the untreated solution. Under the optimum conditions, the aqueous solution treated by the laser irradiation was introduced into the ICP source. By measuring the intensity of emission spectrum of the sample elements, the spectral line intensity of Cd, Cr, Cu, Hg, and Pb was enhanced by about 73.52%, 22.97%, 33.86%, 24.44% and 65.59% compared to the untreated solution, while the signal-to-background ratio increased by 76.03%, 21.74%, 32.17%, 22.68% and 65.32%, respectively. Spectral line intensity and signal-to-background ratio of the ICP source were significantly improved so that the foundation was established for reducing the analysis detection limits. Further more, the surface tension and viscosity of the processed aqueous solution remain the same within 30 minutes standing time with the stable physical properties. This simple and easy method of laser-processed aqueous solution helps improve the detection capabilities of ICP spectrometry.

5.
Appl Opt ; 52(25): 6295-9, 2013 Sep 01.
Article in English | MEDLINE | ID: mdl-24085090

ABSTRACT

Laser-induced breakdown spectroscopy quality can be improved by using a nanosecond Nd:YAG laser pulse to excite soil samples. To investigate how flat-mirror reflection affects the radiation characteristics of laser-induced plasma, emission spectra of sample elements were recorded using a grating spectrometer and photoelectric detection system. Placing a planar mirror vertically on the sample surface (10 mm mirror to plasma-center axis distance) for flat-mirror reflection increased spectral line intensities of Mg, Al, Fe, and Ba by 93.06%, 159.63%, 93.43%, and 94.61%, respectively. Signal-to-noise ratio increased by 17.56%, 40.21%, 31.29%, and 30%. The radiation enhancement mechanism was clarified using measured plasma parameters.

6.
PLoS One ; 8(9): e75885, 2013.
Article in English | MEDLINE | ID: mdl-24098737

ABSTRACT

BACKGROUND: Phosphatase and tensin homologue (PTEN), as a tumor suppressor, plays vital roles in tumorigenesis and progression of prostate cancer. However, the mechanisms of PTEN regulation still need further investigation. We here report that a combination of four microRNAs (miR-19b, miR-23b, miR-26a and miR-92a) promotes prostate cell proliferation by regulating PTEN and its downstream signals in vitro. METHODOLOGY/PRINCIPAL FINDINGS: We found that the four microRNAs (miRNAs) could effectively suppress PTEN expression by directly interacting with its 3' UTR in prostate epithelial and cancer cells. Under-expression of the four miRNAs by antisense neutralization up-regulates PTEN expression, while overexpression of the four miRNAs accelerates epithelial and prostate cancer cell proliferation. Furthermore, the expression of the four miRNAs could, singly or jointly, alter the expression of the key components in the phosphoinositide 3-kinase (PI3K)/Akt pathway, including PIK3CA, PIK3CD, PIK3R1 and Akt, along with their downstream signal, cyclin D1. CONCLUSIONS: These results suggested that the four miRNAs could promote prostate cancer cell proliferation by co-regulating the expression of PTEN, PI3K/Akt pathway and cyclin D1 in vitro. These findings increase understanding of the molecular mechanisms of prostate carcinogenesis and progression, even provide valuable insights into the diagnosis, prognosis, and rational design of novel therapeutics for prostate cancer.


Subject(s)
Cell Proliferation , Gene Expression Regulation, Neoplastic/physiology , MicroRNAs/physiology , PTEN Phosphohydrolase/metabolism , Prostate/cytology , Prostatic Neoplasms/physiopathology , Blotting, Western , DNA Primers/genetics , Gene Expression Regulation, Neoplastic/genetics , Humans , Luciferases , Male , MicroRNAs/genetics , Plasmids/genetics , Reverse Transcriptase Polymerase Chain Reaction
7.
Guang Pu Xue Yu Guang Pu Fen Xi ; 33(8): 2039-42, 2013 Aug.
Article in Chinese | MEDLINE | ID: mdl-24159841

ABSTRACT

To improve the quality of laser-induced breakdown spectroscopy, flat-mirror device was proposed. The effects of flat-mirror device on the radiation characteristics of laser-induced plasma were studied. The experimental results showed that when the device consisted of three flat-mirrors placed around the plasma, the spectral line intensity of Mg, Fe, Ba, Ti and Al increases by about 116.2%, 96.43%, 90.93%, 102.1% and 98.57% than that without flat-mirror device, and the signal-to-noise raises by around 39.17%, 32.48%, 38.07%, 39.95% and 21.30%,respectively. By measuring the plasma parameters, the mechanism of the radiation enhancement obtained with the device consisting of three flat-mirrors was explained. This method was an effective way to improve the detection capacity of LIBS.

8.
Guang Pu Xue Yu Guang Pu Fen Xi ; 33(5): 1338-41, 2013 May.
Article in Chinese | MEDLINE | ID: mdl-23905347

ABSTRACT

In order to increase the intensity of inductively coupled plasma radiation and reduce the detection limit of analysis, the experiment studied on the change of surface tension and viscosity of the water samples which were processed by the ultrasonic cavitation, meanwhile the influence of cavitation effect to samples' spectral intensity and signal-to-background ratio was researched. The experimental results showed that the surface tension and viscosity of sample solution initially decreased and then increased as the ultrasonic power and cavitation time monotonously increased, and the minimum value could be achieved at the ultrasonic power of 50W and the cavitation time of 15 minutes. Under the best experiment condition (the ultrasonic power of 50W and the cavitation time of 15 min), the results revealed that the spectral lines intensity of element Al, Cd) Mn, Ni, Pb and Zn were increased around 56.73%, 57.23%, 44.57%, 43.20%, 39.04% and 40.19% than that without cavitation treatment, spectral signal-background ratio increased about 61.54%, 64.86%, 40.95%, 52.27%, 37.84% and 40.84%, respectively. Thus it can be seen that cavitation-processed water solution can improve the quality of Inductively Coupled Plasma-atomic emission spectrum.

9.
Guang Pu Xue Yu Guang Pu Fen Xi ; 33(3): 595-9, 2013 Mar.
Article in Chinese | MEDLINE | ID: mdl-23705415

ABSTRACT

Abstract A plasma analysis system comprised of Omni-X300 series grating spectrometer, CCD data acquisition system and optical fiber transmission system was utilized in the present paper to realize the real-time acquisition of plasma emission spectra during the process of radio frequency (RF) magnetron sputtering. The plasma emission spectra produced by NiTa, TiAl ceramic targets and NiA1, TiA1 alloy targets were monitored respectively, in addition, the behavior of analysis lines of Ta I 333.991 nm, Ni I 362.473 nm, Al I 396.153 nm and Ti I 398.176 nm with time was obtained, according to which the time of pre-sputtering of the four kinds of target materials was fixed. At the same time, for the TiAl alloy target as the research object, the influence of different powers and pressures on the time of pre-sputtering was studied.

10.
Appl Microbiol Biotechnol ; 97(3): 1141-7, 2013 Feb.
Article in English | MEDLINE | ID: mdl-22419217

ABSTRACT

Previously, we studied an AAVS1 site-specific non-viral integration system with a Rep-donor plasmid and a plasmid containing adeno-associated virus integration element. Our earlier study focused on the plasmid vector itself, but the cellular response to the system was still unknown. SP100 is a member of the promyelocytic leukemia nuclear bodies. It is involved in many cellular processes such as transcriptional regulation and the cellular intrinsic immune response against viral infection. In this study, we revealed that SP100 inhibited the Rep-dependent nonviral integration. Conversely, transient expression of Rep78 increased the degradation of SP100. This degradation was inhibited by treatment with MG132, an inhibitor of the ubiquitin proteasome. SP100 and Rep78 are both located in the nucleolus, which provides the spatial possibility for their interaction. Rep78 was coimmunoprecipitated with the enhanced green fluorescent protein (EGFP)-SP100 fusion protein but not EGFP, which verified the interaction between Rep78 and SP100. These results have enriched our knowledge about the cellular protein SP100 and Rep-dependent nonviral integration. It may lead to an improvement in the application of Rep-related transgene integration method and in the selection of target cells.


Subject(s)
Antigens, Nuclear/metabolism , Autoantigens/metabolism , Dependovirus/physiology , Virus Integration , Cell Line , DNA-Binding Proteins/metabolism , Dependovirus/genetics , Humans , Plasmids , Protein Binding , Viral Proteins/metabolism
11.
Guang Pu Xue Yu Guang Pu Fen Xi ; 32(6): 1654-7, 2012 Jun.
Article in Chinese | MEDLINE | ID: mdl-22870659

ABSTRACT

In order to improve the quality of inductively coupled plasma-atomic emission spectrum and reduce the detection limit of analysis, the influence of potassium additives into water samples on water samples' spectral intensity and signal-to-background ratio was studied. The excitation temperature and electron density of plasma were measured through multi-line slope and the Stark broadening method. The results demonstrated that the plasma spectral intensity intensity increases to a various degree after adding potassium additives into the sample solution. When the content of the potassium is 1.0 g x L(-1), the spectral lines intensity of element Al, Cr, Cu, Mn, Ni and Zn was increased by 8.62%, 32.29%, 108.45%, 6.06%, 64.98% and 54.99% respectively, the spectral signal-background ratio increased by about 7.90%, 30.95%, 104.60%, 5.21%, 66.00% and 52.82%, respectively. Under the conditions of the content of potassium is 1.0 g x L(-1) in the sample, the plasma excitation temperature increased by about 239.69 K than that without additive, and the electron density increased by about 4.99 x 10(11) cm(-3). It is thus clear that potassium additives can improve the quality of inductively coupled plasma-atomic emission spectrum.

12.
Chin Med J (Engl) ; 125(12): 2200-4, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22884153

ABSTRACT

BACKGROUND: Vascular smooth muscle cell proliferation is an important process in the development of atherosclerosis and is associated with other cellular processes in atherogenesis. Telmisartan is reported to have partial peroxisome proliferator-activated receptor (PPAR)-γ activating properties and has been referred to as selective PPAR modulators, but valsartan just blocks angiotensin II (AngII) type 1 (AT1) receptors. This study aimed to compare the different effects of telmisartan and valsartan on human aortic smooth muscle cells (HASMCs) proliferation. METHODS: Ability of telmisartan and valsartan to inhibit proliferation of HASMCs was evaluated by the Cell Counting Kit-8 (CCK-8) in continuous cell culture. Whether the antiproliferative effects of telmisartan and valsartan depend on their effects on AngII receptors or activating the peroxisome PPAR-γ was also investigated in this study. RESULTS: Telmisartan inhibited proliferation of HASMCs by 52.4% (P < 0.01) at the concentration of 25 µmol/L and the effect depended on the dose of telmisartan, but valsartan had little effect on HASMCs proliferation (P > 0.05) and no dose response. When tested in cells stimulated with AngII, telmisartan had the same inhibition of HASMCs by 59.2% (P < 0.05) and valsartan also inhibited it by 41.6% (P < 0.05). Telmisartan and valsartan had the same effect on down-regulating AT1 receptor expression and telmisartan was superior to valsartan up-regulating AngII type 2 (AT2) receptor expression. Antiproliferative effects of telmisartan were observed when HASMCs were treated with the PPAR-γ antagonist GW9662 but antiproliferative effects of the PPAR-γ activator pioglitazone were not observed. CONCLUSIONS: Telmisartan, but not valsartan, inhibits HASMCs proliferation and has dose-dependent response without stimulation of AngII. AT2 receptor up-regulation of telmisartan contributes to its greater antiproliferative effects than valsartan. Its PPAR-γ activation does not play a critical role in inhibiting HASMCs proliferation.


Subject(s)
Benzimidazoles/pharmacology , Benzoates/pharmacology , Cell Proliferation/drug effects , Muscle, Smooth, Vascular/cytology , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/drug effects , Tetrazoles/pharmacology , Valine/analogs & derivatives , Humans , Muscle, Smooth, Vascular/metabolism , PPAR gamma/metabolism , Receptor, Angiotensin, Type 1/metabolism , Receptor, Angiotensin, Type 2/metabolism , Telmisartan , Valine/pharmacology , Valsartan
13.
Guang Pu Xue Yu Guang Pu Fen Xi ; 32(1): 37-40, 2012 Jan.
Article in Chinese | MEDLINE | ID: mdl-22497122

ABSTRACT

To improve the quality of laser-induced breakdown spectroscopy, nanosecond pulse laser generated by Nd : YAG laser was used to excite soil sample. The laser-induced plasma spectrum was observed using a grating spectrometer and a photoelectric detection system. The influence of laser output energy ranging from 100 to 500 mJ on the radiation intensity of plasma was studied. The results show that both the line intensity and signal-to-background ratio can be enhanced under the optimized condition that the laser energy is 200 mJ. The quality of spectrum was further improved after the laser beam used to excite the sample was defocused properly. When the defocusing position is + 6 mm, the spectral lines intensity of element Mg, Al, K and Fe increased about 46%, 63%, 59% and 45% compared to that without defocusing respectively. The spectral signal-to-background ratio increased about 11%, 31%, 35% and 38% respectively. This lays a foundation for detection of trace impurity element in soil.

14.
Mol Biol Rep ; 39(1): 343-9, 2012 Jan.
Article in English | MEDLINE | ID: mdl-21556766

ABSTRACT

HIV-1 integrase (HIV-1 IN), a key element of HIV-1-derived lentiviral vectors, is crucial for the stable maintenance of the vector gene by inserting them into host genome. HIV-1 IN has been found to have functions other than integration, such as involving in virion morphology, viral DNA synthesis and viral DNA nuclear import. In our study, the yeast two-hybrid assay identified a tetrapeptide 156KELK159 in HIV-1 IN that was crucial for HIV-1 IN and Daxx interaction. To investigate the functions of the tetrapeptide 156KELK159 of the HIV-1 IN, both the wild type HIV-1 IN and a mutant without 156KELK159 were used to package the EGFP reporter gene contained lentivirus. p24 based titer assay revealed that deleting the tetrapeptide did not affect virus packaging. The result was verified by quantitative real time PCR with viral specific primers. But the 156KELK159 was crucial for lentiviral gene integration. Deleting the tetrapeptide made the percentage of cells expressing the reporter gene significantly decreased and did not affect the level of DNA entered into the cells or nucleus. Real time reverse transcription PCR and FACS were used to detect the lentiviral report gene expression in infection maintaining cells and revealed 156KELK159 did not affect lentiviral vector gene expression. Our results may shed light on the regulatory mechanism of gene integration of lentivirus.


Subject(s)
HIV Integrase/genetics , HIV-1/genetics , Oligopeptides/physiology , Transduction, Genetic/methods , Adaptor Proteins, Signal Transducing/metabolism , Cell Line , Co-Repressor Proteins , DNA Primers/genetics , Genetic Vectors , Green Fluorescent Proteins/metabolism , Humans , Molecular Chaperones , Nuclear Proteins/metabolism , Oligopeptides/genetics , Plasmids/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Two-Hybrid System Techniques
15.
Guang Pu Xue Yu Guang Pu Fen Xi ; 32(11): 2916-9, 2012 Nov.
Article in Chinese | MEDLINE | ID: mdl-23387149

ABSTRACT

To improve the quality of laser-induced breakdown spectroscopy, nanosecond pulse laser generated by Nd:YAG laser was used to excite soil sample. The intensity and signal-to-background ratio of A1 I 394.401 nm, Ba I 455.403 nm, Fe I 430.791 nm and Ti I 498.173 nm were observed using a grating spectrometer and a photoelectric detection system. The effects of laser shot frequency (5, 10 and 15 Hz)on the radiation characteristics of laser-induced plasma was studied. The experimental results show that as compared with the laser shot frequency of 5 Hz, the spectral line intensity of A1, Ba, Fe and Ti increased by about 50.94%, 112.7%, 107.46%, and 99.38% at 15 Hz respectively under the same laser energy, while the spectral signal-to-background ratio increased by about 15.16%, 24.08%, 40.26% and 72.06% respectively. The effects mechanism of the laser shot frequency on radiation characteristics of plasma is explained by measuring plasma parameters.

16.
Biochem Biophys Res Commun ; 415(4): 702-6, 2011 Dec 02.
Article in English | MEDLINE | ID: mdl-22086178

ABSTRACT

Cdc20 is a co-activator of the anaphase-promoting complex/cyclosome (APC/C complex), which recruits substrates at particular phases of the cell cycle and mediates their degradation. Sp100 is a PML-NB scaffold protein, which localizes to nuclear particles during interphase and disperses from them during mitosis, participates in viral resistance, transcriptional regulation, and apoptosis. However, its metabolism during the cell cycle has not yet been fully characterized. We found a putative D-box in Sp100 using the Eukaryotic Linear Motif (ELM) predictor database. The putative D-box of Sp100 was verified by mutational analysis. Overexpression of Cdc20 resulted in decreased levels of both endogenous Sp100 protein and overexpressed Sp100 mRNA in HEK 293 cells. Only an overexpressed D-box deletion mutant of Sp100 accumulated in HEK293 cells that also overexpressed Cdc20. Cdc20 knockdown by cdc20 specific siRNA resulted in increased Sp100 protein levels in cells. Furthermore, we discovered that the Cdc20 mediated degradation of Sp100 is diminished by the proteasome inhibitor MG132, which suggests that the ubiquitination pathway is involved in this process. However, unlike the other Cdc20 substrates, which display oscillating protein levels, the level of Sp100 protein remains constant throughout the cell cycle. Additionally, both overexpression and knockdown of endogenous Sp100 had no effect on the cell cycle. Our results suggested that sp100 is a novel substrate of Cdc20 and it is degraded by the ubiquitination pathway. The intact D-box of Sp100 was necessary for this process. These findings expand our knowledge of both Sp100 and Cdc20 as well as their role in ubiquitination.


Subject(s)
Antigens, Nuclear/metabolism , Autoantigens/metabolism , Cell Cycle Proteins/metabolism , Ubiquitin-Protein Ligase Complexes/metabolism , Ubiquitination , Anaphase-Promoting Complex-Cyclosome , Cdc20 Proteins , Cell Cycle Proteins/genetics , Gene Knockdown Techniques , Gene Silencing , HEK293 Cells , Humans , RNA, Small Interfering/genetics , Substrate Specificity
17.
Guang Pu Xue Yu Guang Pu Fen Xi ; 31(9): 2361-5, 2011 Sep.
Article in Chinese | MEDLINE | ID: mdl-22097827

ABSTRACT

The prefabricated keyhole effects on the radiation characteristic of laser-induced stainless steel plasma were investigated. A high-energy neodymium glass pulse laser was used to ablate stainless steel sample in air at atmospheric pressure. Combined-type multi-function grating spectroscope and CCD spectral acquainting and processing system were used to record plasma spectrum. The electron temperature and the full width at half maximum of spectral line, respectively. The study results showed that the spectral intensity and signal-to-background ratio of laser plasma increase in the range of 71.5%-125.8% and 7.6%-18.5% respectively when a laser beam (-5 J) acted on the stainless steel sample on which prefabricated keyholes (d = 1.5 mm, h = 0.8 mm) were placed. The plasma temperature and electron density increased by about 1 200 K and 1.21 x 10(16) cm(-3), respectively. This proved that prefabricated keyhole had a significant enhancement effect on the radiation of laser-induced stainless steel plasma.

18.
Fa Yi Xue Za Zhi ; 27(4): 260-4, 2011 Aug.
Article in Chinese | MEDLINE | ID: mdl-21913554

ABSTRACT

OBJECTIVE: To evaluate the potential usefulness of DNA methylation in individual discrimination of monozygotic twins by investigating the differences of DNA methylation profiles in monozygotic twins' blood samples. METHODS: Blood samples from 22 pairs of monozygotic twins were obtained with informed consent. Genomic DNA extracts were bisulfite treated followed by detection with Infinium HumanMethylation27 BeadChip Assays(Illumina, USA). Epigenetic distances between each pair of monozygotic twins and each pair of unrelated individuals of same gender were calculated with Euclidean distance algorithms. Distribution of epigenetic distance in monozygotic twin group was statistically compared with that in unrelated individuals. RESULTS: Difference of epigenetic distance between male and female pairs was not statistically significant in unrelated individual group or in monozygotic twin group (P = 0.0695 and 0.4825, respectively). Epigenetic distance of monozygotic twins was significantly lower than that of unrelated individual pair of same gender (Median: 6.02 vs 7.20, P = 0.0002). However, all the epigenetic distance in monozygotic twin group or in unrelated individuals were significantly higher than 4.00 (P < 0.000 1). CONCLUSION: DNA methylation profiles of monozygotic twin's blood samples were significantly different with each other, which was similar to that in unrelated individuals of same gender. These results indicated that DNA methylation was a useful biomarker in individual discrimination of monozygotic twins.


Subject(s)
DNA Methylation , Epigenomics , Genome, Human/genetics , Twins, Monozygotic/genetics , Adolescent , Adult , Aged , Chromosomes, Human, X/genetics , CpG Islands , Female , Genetic Markers , Genetic Variation , Humans , Male , Middle Aged , Oligonucleotide Array Sequence Analysis/methods , Sex Factors , Young Adult
19.
Acta Biochim Pol ; 58(1): 67-73, 2011.
Article in English | MEDLINE | ID: mdl-21383994

ABSTRACT

Phage ΦC31 integrase is a potential vector for the insertion of therapeutic genes into specific sites in the human genome. To understand the mechanism involved in ΦC31 integrase-mediated recombination, it is important to understand the interaction between the integrase and cellular proteins. Using a yeast two-hybrid system with pLexA-ΦC31 integrase as bait, we screened a pB42AD human fetal brain cDNA library for potential interacting cellular proteins. From the 106 independent clones that were screened, 11 potential interacting clones were isolated, of which one encoded C-terminal fragment of Sp100. The interaction between Sp100 and ΦC31 integrase was further confirmed by yeast mating and co-immunoprecipitation assays. The hybridization between a ΦC31 integrase peptide array and an HEK293 cell extract revealed that residues 81RILN84 in the N-terminus of ΦC31 integrase are responsible for the interaction with Sp100. Knocking down endogenous Sp100 with Sp100-specific siRNA increased ΦC31 integrase-mediated recombination but did not impact reporter gene expression. Therefore, endogenous Sp100 may interact with ΦC31 integrase and inhibit the efficiency of ΦC31 integrase-mediated recombination.


Subject(s)
Antigens, Nuclear/metabolism , Autoantigens/metabolism , Bacteriophages/enzymology , Brain/metabolism , Integrases/metabolism , Antigens, Nuclear/genetics , Autoantigens/genetics , Brain/embryology , Gene Library , HEK293 Cells , Humans , Immunoprecipitation , Protein Binding , RNA, Small Interfering , Two-Hybrid System Techniques
20.
J Biotechnol ; 151(1): 15-21, 2011 Jan 10.
Article in English | MEDLINE | ID: mdl-21084058

ABSTRACT

To develop a superior chimeric peptide (CP) vaccine of human chorionic gonadotropin (hCG), two CP antigens (named CP12 and CP22) encoding one or two copies of three linear B cell epitopes from the ß-hCG subunit and six foreign T cell epitopes, including two promiscuous TCEs from hepatitis B surface antigen and tetanus toxoid, were constructed and biosynthesized. The hCG CP12 and CP22 of 21 or 23 kDa, respectively, were expressed in Escherichia coli at the level of ~1% of total cell proteins when inserted into thermo-inducible pBV221 expression vector. The purified CP12 and CP22 proteins with >95% relative homogeneity are immunogenic, and elicited antibodies against the ß5, ß9 and ß8 BCEs of ß-hCG in both rabbits and three different inbred strains of mice. A mouse uterine weight study in Balb/c mice demonstrated that the CP12 and CP22 antigens with an additional ß5 neutralizing epitope enhanced the in vivo bio-neutralization capacity of the induced antibodies compared to the C-terminal immunogen of ß-hCG. We propose that the biosynthesized CP22, possessing with two copies of three BCEs, represents a novel candidate antigen for an hCG contraceptive or tumor therapeutic vaccine.


Subject(s)
Chorionic Gonadotropin, beta Subunit, Human/immunology , Epitopes, B-Lymphocyte/immunology , Vaccines, Subunit/immunology , Animals , Blotting, Western , Chorionic Gonadotropin, beta Subunit, Human/chemistry , Chorionic Gonadotropin, beta Subunit, Human/metabolism , Electrophoresis, Polyacrylamide Gel , Epitopes, B-Lymphocyte/chemistry , Epitopes, B-Lymphocyte/metabolism , Escherichia coli , Female , Mice , Organ Size , Rabbits , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology , Recombinant Fusion Proteins/metabolism , Sensitivity and Specificity , Stereoisomerism , Uterus/metabolism , Vaccines, Subunit/chemistry , Vaccines, Subunit/genetics , Vaccines, Subunit/metabolism , Vaccines, Synthetic/chemistry , Vaccines, Synthetic/genetics , Vaccines, Synthetic/immunology , Vaccines, Synthetic/metabolism
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