[The Enhancement Research of Magnetic Stirring with Laser Irradiation Aqueous Solution on ICP Source Radiation].

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.

[Enhancement effect of laser-processed aqueous solution on ICP source radiation].

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.

[Effect of flat-mirror device on laser-induced plasma radiation characteristics].

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.

Four microRNAs promote prostate cell proliferation with regulation of PTEN and its downstream signals in vitro.

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.

Enhancement effects of flat-mirror reflection on plasma radiation.

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.

[Effect of ultrasonic cavitation on ICP source radiation intensity].

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.

Role and fate of SP100 protein in response to Rep-dependent nonviral integration system.

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.

[The enhancement effect of potassium additives on ICP source radiation].

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.

[Research on radiation intensity of nanosecond pulse laser-induced soil plasma].

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.

[Effects of laser shot frequency on plasma radiation characteristics].

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.

[Enhancement of the radiation of laser-induced stainless steel plasmas by prefabricated keyhole].

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.

Immunogenic comparison for two different recombinant chimeric peptides (CP12 and CP22) containing one or two copies of three linear B cell epitopes from ß-hCG subunit.

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.

[Effect of KCI additive on laser-induced soil plasma radiation].

In order to improve laser-induced breakdown spectroscopy for low-level elements testing capability, the enhancement effects of KCl additive on the emission spectra of soil samples were studied. The laser spectrum analytical system is composed of a high-energy neodymium glass laser ablating samples, a multifunctional and automatic scanning spectrometer, and a CCD data acquisition system recording plasma spectra. The electron temperature and electron density of plasmas were calculated by measuring spectral line intensity and stark broadening respectively. The experimental results showed that with the increase in the KCl additive, the spectral intensity, signal-to-background ratio, the electron temperature and the electron density all went up firstly and then down. When 15% KCl was added, the radiation intensity of plasma reached the maximum value, the spectral lines intensity of element Mn, Fe, and Ti increased by 2.23, 1.13 and 2.04 than that without additive respectively, the spectral signal-to-background ratio increased by 1.33, 0.89 and 0.94 times respectively; while the electron temperature and electron density of plasmas were heightened by 14% and 38% respectively.

[Enhancing effect of sample additive on laser-induced plasma radiation].

In order to improve the radiation characteristic of laser-induced plasma, with the national standard soil taken as the target sample, a laser spectrum analytical system which composed of a high-energy neodymium glass laser, a multifunctional and compact integrated spectrometer, and a CCD detector was used to detect the influence of the NaCl sample additive on the laser plasma radiation intensity. The electron temperature and the electron density of the plasmas were also calculated from the lines intensity and stark broadening of emission spectral line respectively. The experimental results indicated that with the increase in the NaCl additive, the spectral intensity, signal-to-background ratio, the electron temperature, and the electron density all went up firstly and then down. When 15% NaCl was added, the radiation intensity of the plasma reached the maximum value, the spectral lines intensity of element Mn, K, Fe, and Ti increased by 39.2%, 42.5%, 53.9% and 33.8% compared to that without additive respectively, the spectral signal-to-background ratio increased by 64.4%, 84.39, 44.55% and 58.2% respectively, while the electron temperature and the electron density of the plasmas were heightened by 0.17 times and 0.36 times respectively.

Grb10 interacts with Bim L and inhibits apoptosis.

Bim is a proapoptotic member of the Bcl-2 family and is primarily involved in the regulation of the intrinsic apoptotic pathway. However, the detail of regulation of Bim's proapoptotic activity has not been clarified yet. Using Bim L as bait, we screened a human fetal cDNA library for interacting proteins and identified Grb10 as an interactor. This interaction was verified by co-immunoprecipitation and intracellular co-localization studies. The potential segment of Bim L that binds Grb10 was identified via a yeast mating test. Grb10 interacted with the DBD (dynein binding domain) of Bim and inhibited apoptosis triggered by overexpression of DBD containing Bim isoforms. The putative phosphorylation sites on DBD of Bim play a role for the anti-proapoptotic activity of Grb10. Our results suggest that Grb10 interacts with Bim L and inhibits its proapoptotic activity in a phosphorylation-dependant manner.

Development of chimeric gene regulators for cancer-specific gene therapy with both transcriptional and translational targeting.

Cancer gene therapy has been of great challenge in achieving maximal high levels of specificity and more rational efficiency in target cancer cell. We herein developed a novel approach for cancer-specific gene therapy using both transcriptional and translational targeting regulation. We integrated the tumor-specific gene promoter of hTERT, the 5'UTR of bFGF-2, the enhancer of woodchuck hepatitis virus post-transcriptional regulatory element (WRE), and/or the 3'UTR of the human EGFR into two major chimeric gene regulators. We found that chimeric gene regulator I (hTERT_5'UTR...WRE_BGHpolyA) enhanced the specificity of expression in hepatocellular carcinoma (HCC) cells up to 300% in total due to increases at both the transcriptional and translational levels but only 120-200% enhancement at the transcriptional level and 120-180% enhancement at the translational level. In addition, chimeric gene regulator II (hTERT_5'UTR...WRE_3'UTR_BGHpolyA) improved the specificity to 550% and also highly strengthened the stability of the mRNA. In vitro cytotoxicity assays demonstrated that HCC cell growth was inhibited by HSV-1 TK expression under the control of both chimeric regulators, with a relative cell viability of approximately 80% for 2 days and approximately 85% for 4 days after transfection, respectively. These observations represent a new approach for highly tumor-specific gene expression and also provide insights into application to cancer gene therapy.

Functional differentiation between Rep-mediated site-specific integration and transcriptional repression of the adeno-associated viral p5 promoter.

The adeno-associated virus (AAV) p5 promoter controls expression of Rep68 and Rep78, which are responsible for specific integration of the viral genome into the AAVS1 site of the human genome. The p5 promoter contains a Rep-binding element (RBE) sequence that acts as a substrate of the Rep proteins for both site-specific integration of p5 itself and transcriptional suppression of the p5 promoter. To differentiate these two Rep-mediated functions, we dissected the p5 core structure TATA/RBE/YY1+1 through a series of mutations. Mutations in the TATA box or YY1+1 region of p5IEE significantly reduced Rep-mediated site-specific integration (RMSSI) and p5 promoter transcriptional activity, but only the TATA box is involved in Rep-mediated transcriptional suppression (RMTS). Point mutations at nucleotides 266, 267, 268, 270, and 273 of the GAGTGAGC motif in p5 RBE significantly reduced RMSSI efficiency. However, only p5G270T lost the affinity of Rep binding and had significant reduction of RMTS. It appears that RMTS is determined by the affinity of p5RBE for Rep whereas RMSSI requires more stringent conditions. Thus, RMTS and RMSSI can be differentiated by point mutations in the p5 promoter, which is useful in gene therapy in a helper vector to drive Rep expression, as the mutant promoters seldom integrate themselves but remain the RMTS feature for reduced cytotoxicity caused by a high level of Rep protein.

PhiC31 integrase interacts with TTRAP and inhibits NFkappaB activation.

Phage PhiC31 integrase-mediated gene delivery is believed to be safer than using retroviral vectors since the protein confines its insertion of the target gene to a limited number of sites in mammalian genomes. To evaluate its safety in human cells, it is important to understand the interactions between this integrase and cellular proteins. Here we show that PhiC31 integrase interacts with TTRAP as presented by yeast two-hybrid and co-immunoprecipitation assays. Reducing the expression of endogenous TTRAP can increase the efficiency of PhiC31 integrase-mediated integration. A possible effect of interaction between PhiC31 integrase and TTRAP was highlighted by the fact that PhiC31 integrase inhibited the NFkappaB activation mediated by IL-1 in a dose-dependent manner. Because low dose of PhiC31 integrase can mediate considerable recombination events, we suggest that low dose of PhiC31 integrase be used when this integrase is applied in human cells.

[Study of enhancement effect of laser-induced crater on plasma radiation].

Single pulses exported from high-energy neodymium glass laser were used to act on the same position of soil sample surface repeatedly, and the plasma emission spectra generated from sequential laser pulse action were collected by spectral recording system. The experimental results show that the laser-induced soil plasma radiation was enhanced continuously under the confinement effect of the crater walls, and the line intensities and signal-to-background ratios both had different improvements along with increasing the number of acting pulses. The photographs of the plasma image and crater appearance were taken to study the plasma shape, laser-induced crater appearance, and the mass of the ablated sample. The internal mechanism behind that laser-induced crater enhanced plasma radiation was researched. Under the sequential laser pulse action, the forming plasma as a result enlarges gradually first, leading to distortion at the trail of plasma plume, and then, its volume diminishes slowly. And also, the color of the plasma changes from buff to white gradually, which implies that the temperature increases constantly. The laser-induced crater had a regular shape, that is, the diameter increased from its bottom to top gradually, thus forming a taper. The mass of the laser-ablated substance descends along with increasing the amount of action pulse. Atomization degree of vaporized substance was improved in virtue of the crater confinement effect, Fresnel absorption produced from the crater walls reflection, and the inverse bremsstrahlung, and the plasma radiation intensity was enhanced as a result.

[Preliminary study of the association between human thrombospondin and gastric cancer].

OBJECTIVE: To study the possible role of human thrombospondin (hPWTSR) in gastric cancer and explore its potential to serve as the target for gastric cancer diagnosis and intervention. METHODS: Using pLexA-hPWTSR as the bait, a premade pB42AD-based fetal brain cDNA library was constructed to identify the interacting proteins. The expression pattern of hPWTSR in gastric cancer tissues and a gastric cancer cell line was observed to investigate the correlation between hPWTSR expression and the biological behaviors of the tumor. The possibility of hPWTSR as a potential gastric cancer marker was evaluated. RESULTS: Fifty-seven independent clones were isolated from 107 clones screened. Sequence analysis indicated that the 57 positive clones represented the products of 12 genes. A RT-PCR-based expression pattern revealed that the expression of hPWTSR in gastric cancer tissues and a gastric cancer cell line was lower than that in the corresponding normal tissues, but no mutations were identified by the subsequent sequence analysis. CONCLUSIONS: hPWTSR interacts with adhesion-related proteins and tumor-related genes, and its expression is lowered in gastric cancer tissues and gastric cancer cell line. hPWTSR might play a role in gastric cancer development, especially in metastasis and might be used as a potential gastric cancer marker. The exact functions of hPWTSR and its potential clinical value still await further study.