Based on the statistical characteristics of remote sensing data, the spatial geometric structure characteristics of spectral data and distribution of background, interference and alteration information in characterist...Based on the statistical characteristics of remote sensing data, the spatial geometric structure characteristics of spectral data and distribution of background, interference and alteration information in characteristic space were researched through the analysis of two-dimensional and three-dimensional scatter diagrams. The results indicate that the hyper-space of remote sensing multi-data aggregation belongs to low-dimensional geometric structure, i.e. hyperplane form, and anomalous point groups including alteration information usually dissociate out of hyperplane. Scatter diagrams of remote sensing data band are mainly presented as two distribution forms of single-ellipse and dual-ellipse. Clarifying the relations of three objects of background, disturbance and alteration information in remote sensing images provides an important technical thought and guidance for accurately detecting and extracting remote sensing alteration information.展开更多
AIM: TO inhibit the expression of vascular endothelial growth factor (VEGF) in colon cancer cell line by RNA interference (RNAi).METHODS: Followed the service of E-RNAi, we designed and constructed two kinds of ...AIM: TO inhibit the expression of vascular endothelial growth factor (VEGF) in colon cancer cell line by RNA interference (RNAi).METHODS: Followed the service of E-RNAi, we designed and constructed two kinds of shRNA expression vectors aiming at the VEGF gene, then transfected them into colon cancer HT29 cells by lipofectamineTM 2000. The level of VEGF mRNA was investigated by RT-PCR and Northern blotting. The protein expression of VEGF was observed by immunofluoresence staining and Western blotting.RESULTS: We got two kinds of VEGF specific shRNA expression vectors which could efficiently inhibit the expression of VEGF in HT29 cells. RT-PCR, Northern blotting, immunofluoresence staining and Western blotting showed that inhibition rate for VEGF expression was up to 42%, 89%, 73% and 82%, respectively.CONCLUSION: The expression of VEGF can be inhibited by RNA interference in HT29 cells.展开更多
AIM: To examine the effects of vascular endothelial growth factor (VEGF)-targeted small interfering RNA (siRNA) on proliferation of gastric cancer cellsin vitro.METHODS: Several siRNAs were transfected into huma...AIM: To examine the effects of vascular endothelial growth factor (VEGF)-targeted small interfering RNA (siRNA) on proliferation of gastric cancer cellsin vitro.METHODS: Several siRNAs were transfected into human gastric cancer cell line SGC-7901 with Lipofectamine 2000. Cells not transfected with LipofectamineTM 2000 or scrambled (SCR) siRNA served as controls. The inhibitory effect of siRNA on the expression of VEGF mRNA and protein was detected by RT-PCR and ELISA. MTT assay was used to examine the inhibition rate of cell growth.The change in cell cycling of siRNA-treated cells was detected by flow cytometry.RESULTS: siRNA targeting human VEGF effectively inhibited the proliferation of gastric cancer cell lineSGC-7901 and the distribution of cell cycle. The percentage of G0/G1 phase was significantly higher in siRNA1- and siRNA2-transfected cells than in control cells.The expression of VEGF mRNA was significantly inhibited in siRNA1- and siRNA2-transfected cells compared with that in control cells. VEGF protein notably decreased in siRNA-transfected cells, but had no effect on SCR siRNA.CONCLUSION: VEGF siRNA inhibits proliferation of gastric cancer cells in vitro.展开更多
Trimaran hydrodynamics have been an important research topic in recent years.Trimarans have even been chosen for naval surface combatants.In this case,investigation of a trimaran with different outrigger positions is ...Trimaran hydrodynamics have been an important research topic in recent years.Trimarans have even been chosen for naval surface combatants.In this case,investigation of a trimaran with different outrigger positions is important and necessary for better hydrodynamic performance.This paper focuses on the numerical investigation of trimaran hydrodynamics.The trimaran model used in this study is a 1/80 scale high-speed displacement frigate-type concept developed by the Center for Innovation in Ship Design(CISD)at Naval Surface Warfare Center,Carderock Division(NSWCCD).The numerical simulations were conducted for different outrigger positions at low and moderate Froude numbers by using commercial CFD software solving URANS equations.A verification and validation study was carried out for the numerical method in one configuration and one ship velocity.The existing experimental results for the trimaran resistance in the literature were used for validation.Five different outrigger positions were analyzed and the form factor of each configuration was calculated by the Prohaska method.The total resistance was decomposed to its components using the form factor.The interference factor was calculated for each configuration in terms of total resistance,residual resistance and wave resistance.Also,wave profiles using the longitudinal wave cuts in different locations were obtained both numerically and experimentally.It was concluded that the outrigger position had different effects on the interference,total resistance and wave profile at different Froude numbers.It was also shown that the CFD results were in good agreement with the experimental data in all configurations.In conclusion,this study presents the results of interference effects for different trimaran configurations in terms of wave resistance in addition to the total resistance and residual resistance.The numerical method was validated not only with the total resistance test data but also the longitudinal wave profiles along the hull.展开更多
BACKGROUND: Previous studies have shown that transforming growth factor-beta 1 (TGF-beta 1) is the most potent means of stimulating liver fibrogenesis by myofibroblast-like cells derived from hepatic stellate cells. T...BACKGROUND: Previous studies have shown that transforming growth factor-beta 1 (TGF-beta 1) is the most potent means of stimulating liver fibrogenesis by myofibroblast-like cells derived from hepatic stellate cells. Thus, TGF-beta 1 could be a target for treating hepatic fibrosis. This study aimed to investigate the inhibitory effects of specific TGF-beta 1 small interference RNA (siRNA) on immune hepatic fibrosis induced by Concanavalin A (Con A) in mice. METHODS: Three short hairpin RNAs targeting different positions of TGF-beta 1 were designed and cloned to the plasmid pGenesil-1 to obtain three recombinant expression vectors (pGenesil-TGF-beta 1-ml, pGenesil-TGF-beta 1-m2 and pGenesil-TGF-beta 1-m3). Thirty male Kunming mice were randomly divided into 6 groups: normal, model, control, and three treatment groups. The immune hepatic fibrosis models were constructed by injecting Con A via the tail vein at 8 mg/kg per week for 6 weeks. At weeks 2, 4 and 6, pGenesil-TGF-beta 1-ml, pGenesil-TGF-beta 1-m2 or pGenesi1-TGF-beta 1-m3 was injected by a hydrodynamics-based transfection method via the tail vein at 0.8 ml/10 g within 24 hours after injection of Con A in each of the three treatment groups. The mice in the control group were injected with control plasmid pGenesil-HK at the same dose. All mice were sacrificed at week 7. The levels of hydroxyproline in liver tissue were determined by biochemistry. Liver histopathology was assessed by Van Gieson staining. The expression levels and localization of TGF-beta 1, Smad3, and Smad7 in liver tissue were detected by immunohistochemistry. The expression of TGF-beta 1, Smad3, Smad7 and alpha-smooth muscle actin (alpha-SMA) mRNAs in the liver were assessed by semi-quantitative RT-PCR. RESULTS: The levels of hydroxyproline in the liver tissue of the treatment groups were lower than those of the model group (P<0.01). Histopathologic assay showed that liver fibrogenesis was clearly improved in the treatment groups compared with the model group. The expression levels of TGF-beta 1 and Smad3 of liver tissue were also markedly lower in the treatment groups than in the model group (P<0.01), while the levels of Smad7 were higher in the treatment groups than in the model group (P<0.01). RT-PCR further showed that the expression of TGF-beta 1, Smad3 and alpha-SMA mRNA was significantly inhibited in the treatment groups compared with the model group, while the levels of Smad7 were increased. There was no difference in the above parameters among the three treatment groups or between the control and model groups (P>0.05), but the inhibitory effect of pGenesil-TGF-beta 1-ml was the highest among the treatment groups. CONCLUSIONS: Specific siRNA targeting of TGF-beta 1 markedly inhibited the fibrogenesis of immune hepatic fibrosis induced by Con A in mice. The anti-fibrosis mechanisms of siRNAs may be associated with the down-regulation of TGF-beta 1, Smad3 and alpha-SMA expression and up-regulation of Smad7 expression in liver tissue, which resulted in suppressing the activation of hepatic stellate cells. (Hepatobiliary Pancreat Dis Int 2009; 8: 300-308)展开更多
BACKGROUND: Human gliomas are more likely to express basic fibroblast growth factor-2 (FGF-2) insulin-like growth factor-1(IGF-1), and IGF-1 receptor (IGF-1R) than normal brain tissue. These factors activate si...BACKGROUND: Human gliomas are more likely to express basic fibroblast growth factor-2 (FGF-2) insulin-like growth factor-1(IGF-1), and IGF-1 receptor (IGF-1R) than normal brain tissue. These factors activate signal transduction systems of Ras/MAPK and PI3K/Akl, which promote glioma growth. OBJECTIVE: To utilize RNA interference (RNAi) technique to down-regulate FGF-2, IGF-1, and IGF-1R gene expression, and to investigate the effects of these genes on rat C6 glioma cells, as well as the feasibility of RNAi for treating glioma. DESIGN, TIME AND SETTING: This neurooncological, randomized, controlled, in vivo and in vitro experiment, which used RNAi methodology, was performed at the Laboratory of Molecular Biology, Institute of Biochemistry, Chinese Academy of Sciences between August 2005 and February 2008. MATERIALS: Rat C6 cell lines were purchased from Shanghai Institute of Cellular Biology Affiliated to Chinese Academy of Sciences. Small interfering RNA (siRNA) was synthesized by Shanghai GenePharma. Anti-IGF-1, anti-IGF-1R, anti-FGF-2, anti-mouse and anti-rabbit IgG G1-HRP antibodies were provided by Santa Cruz Biotechnology, USA. Four to six week-old BALB/c nude mice were purchased from the Laboratory Animal Center, Chinese Academy of Sciences. METHODS: C6 glioma cells were transfected with siRNA, which was chemically synthesized in vitro to correspond to endogenous FGF-2, IGF-1, and IGF-1R genes. The inhibition ratio of targeting mRNA expression was detected by semiquantitative RT-PCR, and protein expression was determined by Western blot analysis. C6 glioma cell proliferation was observed using a growth curve C6 glioma cell apoptosis rate and cell cycle were detected by flow cytometry. C6 glioma cell growth regression was observed by transwell migration assay. In addition, nude mouse subcutaneous tumor models were used in this study. For studying the anti-tumor effects of IGF-1 and IGF-1R siRNA, two blank control groups, with six mice each, were set up: A (2.5 μg siRNA was injected one week after C6 cells were inoculated, Le., when tumor volume reached 8 mm × 8 mm) and B (siRNA was injected at the same time with C6 cells were inoculated. To study the effects of FGF-2 siRNA, the groups consisted of a blank control group, negative control group, 2.6 μg siRNA group, 4 μg siRNA group, and 5.3 μg siRNA group, with six mice each. MAIN OUTCOME MEASURES: mRNA and protein inhibition ratio of FGF-2, IGF-1, and IGF-1 R; C6 glioma cell proliferation, apoptosis, and cycle growth arrest; C6 glioma cell growth regression and subcutaneous tumorigenicity rates. RESULTS: All siRNA constructs proved to be effective. After 48 hours, transfection of 200 nmol/L siRNA resulted in a FGF-2 or IGF-1R gene inhibition ratio 〉 80% and an IGF-1 gene inhibition ratio of approximately 70%. Protein expression levels for FGF-2, IGF-1, and IGF-1R decreased in a dose-dependent manner following siRNA transfection, with an inhibition rate 〉 85%, 60%, and 50%, respectively. C6 glioma cell proliferation and apoptosis rates increased in proportion to siRNA. The apoptosis rate of C6 glioma cells induced by FGF-2, IGF-1, and IGF-1R siRNA was 39.96%, 15.07% and 22.47%, respectively (P 〈 0.01). Transfection of 200 nmol/L IGF or IGF-1R siRNA for 48 hours suppressed C6 glioma cell migration. At 30 days after intratumoral injection of 2.6, 4, and 5.3 tJg FGF-2 siRNA, tumor growth regression rate of FGF-2 siRNA was 56%, 67%, and 86%, respectively. The tumor growth regression rate was 71.88% and 45.71%, respectively, when IGF-1 or IGF-1R siRNA was intratumorally injected 1 week after C6 glioma cell transplantation. When IGF-1 or IGF-1 R siRNA was intratumorally injected during C6 glioma cell transplantation, the tumor growth regression rate was 78.13% and 74.29%, respectively. CONCLUSION: siRNA transfection downregulated gene expression of FGF-2, IGF-1, and IGF-1R In addition, siRNA treatment markedly suppressed glioma cell proliferation, growth, and migration, and concomitantly reduced subcutaneous tumorigenicity.展开更多
An improvement for application of Dancoff factor is developed. It combines Stamm'ler's two-term method for resonance integral calculation with neutron current method for Dancoff factor calculation. Stamm'l...An improvement for application of Dancoff factor is developed. It combines Stamm'ler's two-term method for resonance integral calculation with neutron current method for Dancoff factor calculation. Stamm'ler's formulation, which is originally derived for the infinite lattice geometry, can be easily revised to contain the Dancoff factor explicitly, while the neutron current method can easily calculate the Dancoff factor for general irregular assembly geometry. For the resonance interference effects the resonance interference factor table is built in pairs of nuclides, only for the interference between 238 U and other resonance nuclides, spanning over a range of background cross-section and number density ratio of the pairing nuclides. A series of verification calculations have been carried out for problems of infinite lattice and single assembly geometry, with two or multiple resonance absorbers. For these verification calculations, our improvement on Dancoff factor application and resonance interference give good results.展开更多
AIM: To explore the effect of silencing of signal transducer and activator of transcription 3 (STAT3) expression by RNA interference (RNAi) on growth of human hepatocellular carcinoma (HCC) in tumorbearing nude...AIM: To explore the effect of silencing of signal transducer and activator of transcription 3 (STAT3) expression by RNA interference (RNAi) on growth of human hepatocellular carcinoma (HCC) in tumorbearing nude mice in vivo.METHODS: To construct the recombinant plasmid of pSilencer 3.0-H1-STAT3-siRNA-GFP (pSHI-siRNA- STAT3) and establish the tumor-bearing nude mouse model of the HCC cell line SMMC7721, we used intratumoral injection together with electroblotting to transfect the recombinant plasmid pSHI-siRNA- STAT3 into the transplanted tumor. The weight of the nude mice and tumor volumes were recorded. STAT3 gene transcription was detected by semi-quantitative reverse transcription polymerase chain reaction (RT- PCR). Level of protein expression and location of STAT3 were determined by Western blotting and immunohistochemical staining. STAT3-related genes such as survivin, c-myc, VEGF, p53 and caspase3 mRNA and protein expression were detected in tumor tissues at the same time. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect apoptosis of tumor cells.RESULTS: The weight of the treated nude mice increased, and the tumor volume decreased markedly compared with those of the mock-treated and negative control groups (P 〈 0.01). The results of RT-PCR and Western blotting showed that mRNA and protein levels of STAT3 declined markedly in the treated group. The change in STAT3-related gene expression in tumor tissues at the mRNA and protein level also varied, the expression of survivin, VEGF and c-myc were obviously reduced, and expression of p53 and caspase3 increased (P 〈 0.01). Most of the tumor tissue ceils in the treated group developed apoptosis that was detected by TUNEL assay.CONCLUSION: Silencing of STAT3 expression by RNAi significantly inhibits expression of STAT3 mRNA and protein, and suppresses growth of human HCC in tumor-bearing nude mice. The mechanism may be related to down-regulation of survivin, VEGF and c-myc and up-regulation of p53 and caspase3 expression. Accordingly, the STAT3 gene may act as an important and effective target in gene therapy of HCC.展开更多
Hybridoma cells display an increase in antibody productivity following exposure to hypertonic conditions. However, the underlying mechanism is not well understood. In the present study, we hypothesize that the nuclear...Hybridoma cells display an increase in antibody productivity following exposure to hypertonic conditions. However, the underlying mechanism is not well understood. In the present study, we hypothesize that the nuclear factor of activated T cells 5 (NFAT5)/tonicity enhancer binding protein (TonEBP) functions to increase the antibody productivity of hybridoma cells. NFAT5 is an osmosensitive mammalian transcription factor. However, its ubiquitous expression in various organs that are not bathed in hypertonic milieu suggests that NFAT5 may also regulate cell growth and function under isotonic conditions. In this study, we examined the expression of NFAT5 in hybridoma cells by Western blot analysis, and found that it increased significantly in hypertonic medium. To further define the function of NFAT5 in hybridoma cells, RNA interference technique was used to downregulate the expression of NFAT5 in SGB-8 cells (a hybridoma cell line). In isotonic medium, antibody productivity ofhybridoma cells was reduced by downregulation of NFAT5 while cell proliferation was not influenced. The results presented here demonstrate that NFAT5 not only plays an important role in osmotic stress response pathway in hybridoma cells but also is essential for optimal antibody productivity.展开更多
Viruses are extremely heterogeneous entities; the size and the nature of their genetic information, as well as the strategies employed to amplify and propagate their genomes, are highly variable. However, as obligator...Viruses are extremely heterogeneous entities; the size and the nature of their genetic information, as well as the strategies employed to amplify and propagate their genomes, are highly variable. However, as obligatory intracellular parasites, replication of all viruses relies on the host cell. Having co-evolved with their host for several million years, viruses have developed very sophisticated strategies to hijack cellular factors that promote virus uptake, replication, and spread. Identification of host cell factors(HCFs) required for these processes is a major challenge for researchers, but it enables the identification of new, highly selective targets for anti viral therapeutics. To this end, the establishment of platforms enabling genome-wide high-throughput RNA interference(HT-RNAi) screens has led to the identification of several key factors involved in the viral lifecycle. A number of genome-wide HT-RNAi screens have been performed for major human pathogens. These studies enable first inter-viral comparisons related to HCF requirements. Although several cellular functions appear to be uniformly required for the life cycle of most viruses tested(such as the proteasome and the Golgi-mediated secretory pathways), some factors, like the lipid kinase Phosphatidylinositol 4-kinase Ⅲα in the case of hepatitis C virus, are selectively required for individual viruses. However, despite the amount of data available, we are still far away from a comprehensive understanding of the interplay between viruses and host factors. Major limitations towards this goal are the low sensitivity and specificity of such screens, resulting in limited overlap between different screens performed with the same virus. This review focuses on how statistical and bioinformatic analysis methods applied to HTRNAi screens can help overcoming these issues thus increasing the reliability and impact of such studies.展开更多
An experimental investigation is performed to assess the relation of interference performance on the total resistance of a pentamaran model advancing in calm water. For this motivation, the total drag of the ship is ...An experimental investigation is performed to assess the relation of interference performance on the total resistance of a pentamaran model advancing in calm water. For this motivation, the total drag of the ship is performed for several values of asymmetric outrigger configuration and hull separation, altering the Froude number in the range 0.3–0.9. Our results indicate that remarkable changes in resistance require notable changes in transverse distance values (hull separation) when wave interference may occur. In addition, there is no single configuration that consistently outperforms the other configurations across the entire speed range and the optimum interference factor -0.2 appears at a Froude number of 0.45 in S/L=0.33 with the outrigger outer position: asymmetric outboard for A3 configuration.展开更多
Objective: To investigate the role of peroxisome proliferator-activated receptors δ (PPARδ) in inflammatory reaction and its possible mechanism in adipocyte. Methods:Lentivirus-mediated RNA interference (RNAi)...Objective: To investigate the role of peroxisome proliferator-activated receptors δ (PPARδ) in inflammatory reaction and its possible mechanism in adipocyte. Methods:Lentivirus-mediated RNA interference (RNAi) was used to block the expression of PPARδ in 3T3-L1 cells. In order to induce inflammation in 3T3-L1, cells were stimulated with tumor necrosis factor-α(TNFα, 20 ng/ml) for 4 h. The expression of PPARδ, nuclear factor κB (NFκB) and C reactive protein (CRP) were determined by Western blot analysis. Results:The expression of PPARδ was reduced by 80% after RNAi. Blockage of PPARδ promoted the expression of CRP and NFκB in cells stimulated with TNFα but had no effect on normal cells. Conclusion: PPARδ is involved in inflammatory reaction in adipocyte. Blockage of PPARδ can promote the inflammation mediated by inflammatory factors and increase the expression of NFκB and CRP in 3T3-L1 cells stimulated with TNFα.展开更多
Based on the characteristics of the strong volatility of physical property in vertical direction, high gas content, high resource abundance and large exploitation potentiality of coal reservoir in Bide-Santang basin o...Based on the characteristics of the strong volatility of physical property in vertical direction, high gas content, high resource abundance and large exploitation potentiality of coal reservoir in Bide-Santang basin of Zhina coal field, we study the generation mechanism of interlayer interference, propagation rules of reservoir pressure drop and influencing factors of gas productivity in CBM multi-seam drainage in the paper. On the basis of the actual production data of X-2 well of Zhucang syncline in Bide-Santang basin,by simulating the gas production process of a CBM well under the condition of multiple seam with COMET3 numerical simulation software, we analyze the influencing factors of gas productivity during the process of multi-seam drainage, and illuminate the interlayer interference mechanism of multiseam drainage. The results show that permeability, reservoir pressure gradient, critical desorption pressure and fluid supply capacity of stratum have great influence on gas productivity of multi-seam drainage while coal thickness has little influence on it. Permeability, reservoir pressure gradient and fluid supply capacity of stratum affect the propagation velocity of reservoir pressure drop and thereby affect the final gas productivity. Moreover, the influence of critical desorption pressure on gas productivity of multiseam drainage is reflected in the gas breakthrough time and effective desorption area.展开更多
Objective: Lung cancer has emerged as a leading cause of cancer death in the world. Current therapies are ineffective, thus new approaches are needed to improve the therapeutic ratio. RNA interference (RNAi) has sh...Objective: Lung cancer has emerged as a leading cause of cancer death in the world. Current therapies are ineffective, thus new approaches are needed to improve the therapeutic ratio. RNA interference (RNAi) has shown promise in gene silencing in vitro, the potential of which in developing new methods for the therapy of non-small-cell lung cancer (NSCLC) needs to be further tested in vivo. In this study, chemically synthesized double-stranded RNA (dsRNA) targeting epidermal growth factor receptor (EGFR) was transfected into NSCLC cell line SPC-A1 cells and established the tumor burdened athymic nude mice model to investigate whether dsRNA could induce gene silencing in NSCLC cells in vivo. Methods: SPC-A1 was transfected with EGFR sequence-specific dsRNA formulated with Lipofectamine 2000. SPC-A1 cells (1 × 107/ mL) in 200 pL were injected s.c. into the left flank area of the mice to establish the tumor burdened athymic nude mice model. Calculate the tumor growth inhibition rate by measuring the diameter and the weight of the tumor. Immunohistochemistry and Westem blot were used to monitor the reduction in the production of the EGFR protein. Realtime RT-PCR was used to detect the silencing of the EGFR mRNA level. Results: It displayed that EGFR sequence specific dsRNA (dsRNA-EGFR) significantly inhibited the tumor growth in vivo. The tumor growth inhibition rate was 75.03%. The dsRNA-EGFR sequence specifically silenced EGFR with 53.6% of down-regulation of EGFR protein production and 32.3% of silencing of EGFR mRNA level. Conclusion: DsRNA-EGFR showed a blockbuster effect in downregulation of EGFR mRNA level and protein production, and inhibition of tumor growth in vivo.展开更多
基金Project(2006BAB01A06) supported by the National Science and Technology Pillar Program Project during the 11th Five-Year Plan PeriodProject(1212010761503) supported by Land and Resources Investigation Project
文摘Based on the statistical characteristics of remote sensing data, the spatial geometric structure characteristics of spectral data and distribution of background, interference and alteration information in characteristic space were researched through the analysis of two-dimensional and three-dimensional scatter diagrams. The results indicate that the hyper-space of remote sensing multi-data aggregation belongs to low-dimensional geometric structure, i.e. hyperplane form, and anomalous point groups including alteration information usually dissociate out of hyperplane. Scatter diagrams of remote sensing data band are mainly presented as two distribution forms of single-ellipse and dual-ellipse. Clarifying the relations of three objects of background, disturbance and alteration information in remote sensing images provides an important technical thought and guidance for accurately detecting and extracting remote sensing alteration information.
基金Supported in part by research grants from the National Natural Science Foundation of China,No.30300298the National Natural Science Foundation of China's Joint Research Fund for Overseas Chinese Young Scholars,No.30228026
文摘AIM: TO inhibit the expression of vascular endothelial growth factor (VEGF) in colon cancer cell line by RNA interference (RNAi).METHODS: Followed the service of E-RNAi, we designed and constructed two kinds of shRNA expression vectors aiming at the VEGF gene, then transfected them into colon cancer HT29 cells by lipofectamineTM 2000. The level of VEGF mRNA was investigated by RT-PCR and Northern blotting. The protein expression of VEGF was observed by immunofluoresence staining and Western blotting.RESULTS: We got two kinds of VEGF specific shRNA expression vectors which could efficiently inhibit the expression of VEGF in HT29 cells. RT-PCR, Northern blotting, immunofluoresence staining and Western blotting showed that inhibition rate for VEGF expression was up to 42%, 89%, 73% and 82%, respectively.CONCLUSION: The expression of VEGF can be inhibited by RNA interference in HT29 cells.
基金Supported by Science Project of Qingdao Science and Technology Administration, No. 03-1-YN-14
文摘AIM: To examine the effects of vascular endothelial growth factor (VEGF)-targeted small interfering RNA (siRNA) on proliferation of gastric cancer cellsin vitro.METHODS: Several siRNAs were transfected into human gastric cancer cell line SGC-7901 with Lipofectamine 2000. Cells not transfected with LipofectamineTM 2000 or scrambled (SCR) siRNA served as controls. The inhibitory effect of siRNA on the expression of VEGF mRNA and protein was detected by RT-PCR and ELISA. MTT assay was used to examine the inhibition rate of cell growth.The change in cell cycling of siRNA-treated cells was detected by flow cytometry.RESULTS: siRNA targeting human VEGF effectively inhibited the proliferation of gastric cancer cell lineSGC-7901 and the distribution of cell cycle. The percentage of G0/G1 phase was significantly higher in siRNA1- and siRNA2-transfected cells than in control cells.The expression of VEGF mRNA was significantly inhibited in siRNA1- and siRNA2-transfected cells compared with that in control cells. VEGF protein notably decreased in siRNA-transfected cells, but had no effect on SCR siRNA.CONCLUSION: VEGF siRNA inhibits proliferation of gastric cancer cells in vitro.
基金The second author was supported by the Scientific and Technological Research Council of Turkey(TUBITAK)2219 International Postdoctoral Research Fellowship Program。
文摘Trimaran hydrodynamics have been an important research topic in recent years.Trimarans have even been chosen for naval surface combatants.In this case,investigation of a trimaran with different outrigger positions is important and necessary for better hydrodynamic performance.This paper focuses on the numerical investigation of trimaran hydrodynamics.The trimaran model used in this study is a 1/80 scale high-speed displacement frigate-type concept developed by the Center for Innovation in Ship Design(CISD)at Naval Surface Warfare Center,Carderock Division(NSWCCD).The numerical simulations were conducted for different outrigger positions at low and moderate Froude numbers by using commercial CFD software solving URANS equations.A verification and validation study was carried out for the numerical method in one configuration and one ship velocity.The existing experimental results for the trimaran resistance in the literature were used for validation.Five different outrigger positions were analyzed and the form factor of each configuration was calculated by the Prohaska method.The total resistance was decomposed to its components using the form factor.The interference factor was calculated for each configuration in terms of total resistance,residual resistance and wave resistance.Also,wave profiles using the longitudinal wave cuts in different locations were obtained both numerically and experimentally.It was concluded that the outrigger position had different effects on the interference,total resistance and wave profile at different Froude numbers.It was also shown that the CFD results were in good agreement with the experimental data in all configurations.In conclusion,this study presents the results of interference effects for different trimaran configurations in terms of wave resistance in addition to the total resistance and residual resistance.The numerical method was validated not only with the total resistance test data but also the longitudinal wave profiles along the hull.
文摘BACKGROUND: Previous studies have shown that transforming growth factor-beta 1 (TGF-beta 1) is the most potent means of stimulating liver fibrogenesis by myofibroblast-like cells derived from hepatic stellate cells. Thus, TGF-beta 1 could be a target for treating hepatic fibrosis. This study aimed to investigate the inhibitory effects of specific TGF-beta 1 small interference RNA (siRNA) on immune hepatic fibrosis induced by Concanavalin A (Con A) in mice. METHODS: Three short hairpin RNAs targeting different positions of TGF-beta 1 were designed and cloned to the plasmid pGenesil-1 to obtain three recombinant expression vectors (pGenesil-TGF-beta 1-ml, pGenesil-TGF-beta 1-m2 and pGenesil-TGF-beta 1-m3). Thirty male Kunming mice were randomly divided into 6 groups: normal, model, control, and three treatment groups. The immune hepatic fibrosis models were constructed by injecting Con A via the tail vein at 8 mg/kg per week for 6 weeks. At weeks 2, 4 and 6, pGenesil-TGF-beta 1-ml, pGenesil-TGF-beta 1-m2 or pGenesi1-TGF-beta 1-m3 was injected by a hydrodynamics-based transfection method via the tail vein at 0.8 ml/10 g within 24 hours after injection of Con A in each of the three treatment groups. The mice in the control group were injected with control plasmid pGenesil-HK at the same dose. All mice were sacrificed at week 7. The levels of hydroxyproline in liver tissue were determined by biochemistry. Liver histopathology was assessed by Van Gieson staining. The expression levels and localization of TGF-beta 1, Smad3, and Smad7 in liver tissue were detected by immunohistochemistry. The expression of TGF-beta 1, Smad3, Smad7 and alpha-smooth muscle actin (alpha-SMA) mRNAs in the liver were assessed by semi-quantitative RT-PCR. RESULTS: The levels of hydroxyproline in the liver tissue of the treatment groups were lower than those of the model group (P<0.01). Histopathologic assay showed that liver fibrogenesis was clearly improved in the treatment groups compared with the model group. The expression levels of TGF-beta 1 and Smad3 of liver tissue were also markedly lower in the treatment groups than in the model group (P<0.01), while the levels of Smad7 were higher in the treatment groups than in the model group (P<0.01). RT-PCR further showed that the expression of TGF-beta 1, Smad3 and alpha-SMA mRNA was significantly inhibited in the treatment groups compared with the model group, while the levels of Smad7 were increased. There was no difference in the above parameters among the three treatment groups or between the control and model groups (P>0.05), but the inhibitory effect of pGenesil-TGF-beta 1-ml was the highest among the treatment groups. CONCLUSIONS: Specific siRNA targeting of TGF-beta 1 markedly inhibited the fibrogenesis of immune hepatic fibrosis induced by Con A in mice. The anti-fibrosis mechanisms of siRNAs may be associated with the down-regulation of TGF-beta 1, Smad3 and alpha-SMA expression and up-regulation of Smad7 expression in liver tissue, which resulted in suppressing the activation of hepatic stellate cells. (Hepatobiliary Pancreat Dis Int 2009; 8: 300-308)
基金the National Natural Science Foundation of China,No.30371459Science and Technology Development Fund of Shanghai,No.034047
文摘BACKGROUND: Human gliomas are more likely to express basic fibroblast growth factor-2 (FGF-2) insulin-like growth factor-1(IGF-1), and IGF-1 receptor (IGF-1R) than normal brain tissue. These factors activate signal transduction systems of Ras/MAPK and PI3K/Akl, which promote glioma growth. OBJECTIVE: To utilize RNA interference (RNAi) technique to down-regulate FGF-2, IGF-1, and IGF-1R gene expression, and to investigate the effects of these genes on rat C6 glioma cells, as well as the feasibility of RNAi for treating glioma. DESIGN, TIME AND SETTING: This neurooncological, randomized, controlled, in vivo and in vitro experiment, which used RNAi methodology, was performed at the Laboratory of Molecular Biology, Institute of Biochemistry, Chinese Academy of Sciences between August 2005 and February 2008. MATERIALS: Rat C6 cell lines were purchased from Shanghai Institute of Cellular Biology Affiliated to Chinese Academy of Sciences. Small interfering RNA (siRNA) was synthesized by Shanghai GenePharma. Anti-IGF-1, anti-IGF-1R, anti-FGF-2, anti-mouse and anti-rabbit IgG G1-HRP antibodies were provided by Santa Cruz Biotechnology, USA. Four to six week-old BALB/c nude mice were purchased from the Laboratory Animal Center, Chinese Academy of Sciences. METHODS: C6 glioma cells were transfected with siRNA, which was chemically synthesized in vitro to correspond to endogenous FGF-2, IGF-1, and IGF-1R genes. The inhibition ratio of targeting mRNA expression was detected by semiquantitative RT-PCR, and protein expression was determined by Western blot analysis. C6 glioma cell proliferation was observed using a growth curve C6 glioma cell apoptosis rate and cell cycle were detected by flow cytometry. C6 glioma cell growth regression was observed by transwell migration assay. In addition, nude mouse subcutaneous tumor models were used in this study. For studying the anti-tumor effects of IGF-1 and IGF-1R siRNA, two blank control groups, with six mice each, were set up: A (2.5 μg siRNA was injected one week after C6 cells were inoculated, Le., when tumor volume reached 8 mm × 8 mm) and B (siRNA was injected at the same time with C6 cells were inoculated. To study the effects of FGF-2 siRNA, the groups consisted of a blank control group, negative control group, 2.6 μg siRNA group, 4 μg siRNA group, and 5.3 μg siRNA group, with six mice each. MAIN OUTCOME MEASURES: mRNA and protein inhibition ratio of FGF-2, IGF-1, and IGF-1 R; C6 glioma cell proliferation, apoptosis, and cycle growth arrest; C6 glioma cell growth regression and subcutaneous tumorigenicity rates. RESULTS: All siRNA constructs proved to be effective. After 48 hours, transfection of 200 nmol/L siRNA resulted in a FGF-2 or IGF-1R gene inhibition ratio 〉 80% and an IGF-1 gene inhibition ratio of approximately 70%. Protein expression levels for FGF-2, IGF-1, and IGF-1R decreased in a dose-dependent manner following siRNA transfection, with an inhibition rate 〉 85%, 60%, and 50%, respectively. C6 glioma cell proliferation and apoptosis rates increased in proportion to siRNA. The apoptosis rate of C6 glioma cells induced by FGF-2, IGF-1, and IGF-1R siRNA was 39.96%, 15.07% and 22.47%, respectively (P 〈 0.01). Transfection of 200 nmol/L IGF or IGF-1R siRNA for 48 hours suppressed C6 glioma cell migration. At 30 days after intratumoral injection of 2.6, 4, and 5.3 tJg FGF-2 siRNA, tumor growth regression rate of FGF-2 siRNA was 56%, 67%, and 86%, respectively. The tumor growth regression rate was 71.88% and 45.71%, respectively, when IGF-1 or IGF-1R siRNA was intratumorally injected 1 week after C6 glioma cell transplantation. When IGF-1 or IGF-1 R siRNA was intratumorally injected during C6 glioma cell transplantation, the tumor growth regression rate was 78.13% and 74.29%, respectively. CONCLUSION: siRNA transfection downregulated gene expression of FGF-2, IGF-1, and IGF-1R In addition, siRNA treatment markedly suppressed glioma cell proliferation, growth, and migration, and concomitantly reduced subcutaneous tumorigenicity.
文摘An improvement for application of Dancoff factor is developed. It combines Stamm'ler's two-term method for resonance integral calculation with neutron current method for Dancoff factor calculation. Stamm'ler's formulation, which is originally derived for the infinite lattice geometry, can be easily revised to contain the Dancoff factor explicitly, while the neutron current method can easily calculate the Dancoff factor for general irregular assembly geometry. For the resonance interference effects the resonance interference factor table is built in pairs of nuclides, only for the interference between 238 U and other resonance nuclides, spanning over a range of background cross-section and number density ratio of the pairing nuclides. A series of verification calculations have been carried out for problems of infinite lattice and single assembly geometry, with two or multiple resonance absorbers. For these verification calculations, our improvement on Dancoff factor application and resonance interference give good results.
基金Supported by The Science and Technology Fund of Jilin Province,No. 200505219
文摘AIM: To explore the effect of silencing of signal transducer and activator of transcription 3 (STAT3) expression by RNA interference (RNAi) on growth of human hepatocellular carcinoma (HCC) in tumorbearing nude mice in vivo.METHODS: To construct the recombinant plasmid of pSilencer 3.0-H1-STAT3-siRNA-GFP (pSHI-siRNA- STAT3) and establish the tumor-bearing nude mouse model of the HCC cell line SMMC7721, we used intratumoral injection together with electroblotting to transfect the recombinant plasmid pSHI-siRNA- STAT3 into the transplanted tumor. The weight of the nude mice and tumor volumes were recorded. STAT3 gene transcription was detected by semi-quantitative reverse transcription polymerase chain reaction (RT- PCR). Level of protein expression and location of STAT3 were determined by Western blotting and immunohistochemical staining. STAT3-related genes such as survivin, c-myc, VEGF, p53 and caspase3 mRNA and protein expression were detected in tumor tissues at the same time. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect apoptosis of tumor cells.RESULTS: The weight of the treated nude mice increased, and the tumor volume decreased markedly compared with those of the mock-treated and negative control groups (P 〈 0.01). The results of RT-PCR and Western blotting showed that mRNA and protein levels of STAT3 declined markedly in the treated group. The change in STAT3-related gene expression in tumor tissues at the mRNA and protein level also varied, the expression of survivin, VEGF and c-myc were obviously reduced, and expression of p53 and caspase3 increased (P 〈 0.01). Most of the tumor tissue ceils in the treated group developed apoptosis that was detected by TUNEL assay.CONCLUSION: Silencing of STAT3 expression by RNAi significantly inhibits expression of STAT3 mRNA and protein, and suppresses growth of human HCC in tumor-bearing nude mice. The mechanism may be related to down-regulation of survivin, VEGF and c-myc and up-regulation of p53 and caspase3 expression. Accordingly, the STAT3 gene may act as an important and effective target in gene therapy of HCC.
文摘Hybridoma cells display an increase in antibody productivity following exposure to hypertonic conditions. However, the underlying mechanism is not well understood. In the present study, we hypothesize that the nuclear factor of activated T cells 5 (NFAT5)/tonicity enhancer binding protein (TonEBP) functions to increase the antibody productivity of hybridoma cells. NFAT5 is an osmosensitive mammalian transcription factor. However, its ubiquitous expression in various organs that are not bathed in hypertonic milieu suggests that NFAT5 may also regulate cell growth and function under isotonic conditions. In this study, we examined the expression of NFAT5 in hybridoma cells by Western blot analysis, and found that it increased significantly in hypertonic medium. To further define the function of NFAT5 in hybridoma cells, RNA interference technique was used to downregulate the expression of NFAT5 in SGB-8 cells (a hybridoma cell line). In isotonic medium, antibody productivity ofhybridoma cells was reduced by downregulation of NFAT5 while cell proliferation was not influenced. The results presented here demonstrate that NFAT5 not only plays an important role in osmotic stress response pathway in hybridoma cells but also is essential for optimal antibody productivity.
文摘Viruses are extremely heterogeneous entities; the size and the nature of their genetic information, as well as the strategies employed to amplify and propagate their genomes, are highly variable. However, as obligatory intracellular parasites, replication of all viruses relies on the host cell. Having co-evolved with their host for several million years, viruses have developed very sophisticated strategies to hijack cellular factors that promote virus uptake, replication, and spread. Identification of host cell factors(HCFs) required for these processes is a major challenge for researchers, but it enables the identification of new, highly selective targets for anti viral therapeutics. To this end, the establishment of platforms enabling genome-wide high-throughput RNA interference(HT-RNAi) screens has led to the identification of several key factors involved in the viral lifecycle. A number of genome-wide HT-RNAi screens have been performed for major human pathogens. These studies enable first inter-viral comparisons related to HCF requirements. Although several cellular functions appear to be uniformly required for the life cycle of most viruses tested(such as the proteasome and the Golgi-mediated secretory pathways), some factors, like the lipid kinase Phosphatidylinositol 4-kinase Ⅲα in the case of hepatitis C virus, are selectively required for individual viruses. However, despite the amount of data available, we are still far away from a comprehensive understanding of the interplay between viruses and host factors. Major limitations towards this goal are the low sensitivity and specificity of such screens, resulting in limited overlap between different screens performed with the same virus. This review focuses on how statistical and bioinformatic analysis methods applied to HTRNAi screens can help overcoming these issues thus increasing the reliability and impact of such studies.
文摘An experimental investigation is performed to assess the relation of interference performance on the total resistance of a pentamaran model advancing in calm water. For this motivation, the total drag of the ship is performed for several values of asymmetric outrigger configuration and hull separation, altering the Froude number in the range 0.3–0.9. Our results indicate that remarkable changes in resistance require notable changes in transverse distance values (hull separation) when wave interference may occur. In addition, there is no single configuration that consistently outperforms the other configurations across the entire speed range and the optimum interference factor -0.2 appears at a Froude number of 0.45 in S/L=0.33 with the outrigger outer position: asymmetric outboard for A3 configuration.
文摘Objective: To investigate the role of peroxisome proliferator-activated receptors δ (PPARδ) in inflammatory reaction and its possible mechanism in adipocyte. Methods:Lentivirus-mediated RNA interference (RNAi) was used to block the expression of PPARδ in 3T3-L1 cells. In order to induce inflammation in 3T3-L1, cells were stimulated with tumor necrosis factor-α(TNFα, 20 ng/ml) for 4 h. The expression of PPARδ, nuclear factor κB (NFκB) and C reactive protein (CRP) were determined by Western blot analysis. Results:The expression of PPARδ was reduced by 80% after RNAi. Blockage of PPARδ promoted the expression of CRP and NFκB in cells stimulated with TNFα but had no effect on normal cells. Conclusion: PPARδ is involved in inflammatory reaction in adipocyte. Blockage of PPARδ can promote the inflammation mediated by inflammatory factors and increase the expression of NFκB and CRP in 3T3-L1 cells stimulated with TNFα.
基金supported by the National Major Special Project of Science and Technology of China (No.2011ZX05034)the Innovation Projects of University Graduates in Jiangsu Province (No.CXLX13_948)+1 种基金the National Natural Science Foundation of China (No.41272178)the Natural Science Foundation of Hunan Province (No.2016JJ4031)
文摘Based on the characteristics of the strong volatility of physical property in vertical direction, high gas content, high resource abundance and large exploitation potentiality of coal reservoir in Bide-Santang basin of Zhina coal field, we study the generation mechanism of interlayer interference, propagation rules of reservoir pressure drop and influencing factors of gas productivity in CBM multi-seam drainage in the paper. On the basis of the actual production data of X-2 well of Zhucang syncline in Bide-Santang basin,by simulating the gas production process of a CBM well under the condition of multiple seam with COMET3 numerical simulation software, we analyze the influencing factors of gas productivity during the process of multi-seam drainage, and illuminate the interlayer interference mechanism of multiseam drainage. The results show that permeability, reservoir pressure gradient, critical desorption pressure and fluid supply capacity of stratum have great influence on gas productivity of multi-seam drainage while coal thickness has little influence on it. Permeability, reservoir pressure gradient and fluid supply capacity of stratum affect the propagation velocity of reservoir pressure drop and thereby affect the final gas productivity. Moreover, the influence of critical desorption pressure on gas productivity of multiseam drainage is reflected in the gas breakthrough time and effective desorption area.
基金Supported by a grant from the Natural Science Foundation of Shanghai (No. 03ZR14004).
文摘Objective: Lung cancer has emerged as a leading cause of cancer death in the world. Current therapies are ineffective, thus new approaches are needed to improve the therapeutic ratio. RNA interference (RNAi) has shown promise in gene silencing in vitro, the potential of which in developing new methods for the therapy of non-small-cell lung cancer (NSCLC) needs to be further tested in vivo. In this study, chemically synthesized double-stranded RNA (dsRNA) targeting epidermal growth factor receptor (EGFR) was transfected into NSCLC cell line SPC-A1 cells and established the tumor burdened athymic nude mice model to investigate whether dsRNA could induce gene silencing in NSCLC cells in vivo. Methods: SPC-A1 was transfected with EGFR sequence-specific dsRNA formulated with Lipofectamine 2000. SPC-A1 cells (1 × 107/ mL) in 200 pL were injected s.c. into the left flank area of the mice to establish the tumor burdened athymic nude mice model. Calculate the tumor growth inhibition rate by measuring the diameter and the weight of the tumor. Immunohistochemistry and Westem blot were used to monitor the reduction in the production of the EGFR protein. Realtime RT-PCR was used to detect the silencing of the EGFR mRNA level. Results: It displayed that EGFR sequence specific dsRNA (dsRNA-EGFR) significantly inhibited the tumor growth in vivo. The tumor growth inhibition rate was 75.03%. The dsRNA-EGFR sequence specifically silenced EGFR with 53.6% of down-regulation of EGFR protein production and 32.3% of silencing of EGFR mRNA level. Conclusion: DsRNA-EGFR showed a blockbuster effect in downregulation of EGFR mRNA level and protein production, and inhibition of tumor growth in vivo.