MicroRNAs （miRNAs） are endogenous, small, non-coding RNAs, which are capable of silencing gene expression at the post-transcriptional level. In this study, we report that miR-205 is significantly underexpressed in b...MicroRNAs （miRNAs） are endogenous, small, non-coding RNAs, which are capable of silencing gene expression at the post-transcriptional level. In this study, we report that miR-205 is significantly underexpressed in breast tumor compared to the matched normal breast tissue. Similarly, breast cancer cell lines, including MCF-7 and MDA-MB- 231, express a lower level miR-205 than the non-malignant MCF-10A cells. Of interest, ectopic expression of miR-205 significantly inhibits cell proliferation and anchorage independent growth, as well as cell invasion. Furthermore, miR- 205 was shown to suppress lung metastasis in an animal model. Finally, western blot combined with the luciferase reporter assays demonstrate that ErbB3 and vascular endothelial growth factor A （VEGF-A） are direct targets for miR-205, and this miR-205-mediated suppression is likely through the direct interaction with the putative miR-205 binding site in the 3＇-untranslated region （3＇-UTR） of ErbB3 and VEGF-A. Together, these results suggest that miR- 205 is a tumor suppressor in breast cancer.展开更多
POU transcription factor OCT4 not only plays an essential role in maintaining the pluripotent and self-renewing state of embryonic stem （ES） cells but also acts as a cell fate determinant through a gene dosage effec...POU transcription factor OCT4 not only plays an essential role in maintaining the pluripotent and self-renewing state of embryonic stem （ES） cells but also acts as a cell fate determinant through a gene dosage effect. However, the molecular mechanisms that control the intracellular OCT4 protein level remain elusive. Here, we report that human WWP2, an E3 ubiquitin （Ub）-protein ligase, interacts with OCT4 specifically through its WW domain and enhances Ub modification of OCT4 both in vitro and in vivo. We first demonstrated that endogenous OCT4 in hu- man ES cells can be post-translationally modified by Ub. Furthermore, we found that WWP2 promoted degradation of OCT4 through the 26S proteasome in a dosage-dependent manner, and the active site cysteine residue of WWP2 was required for both its enzymatic activity and proteolytic effect on OCT4. Remarkably, our data show that the en- dogenous OCT4 protein level was significantly elevated when WWP2 expression was downregulated by specific RNA interference （RNAi）, suggesting that WWP2 is an important regulator for maintaining a proper OCT4 protein level in human ES cells. Moreover, northern blot analysis showed that the WWP2 transcript was widely present in diverse human tissues/organs and highly expressed in undifferentiated human ES cells. However, its expression level was quickly decreased after human ES cells differentiated, indicating that WWP2 expression might be developmentally regulated. Our findings demonstrate that WWP2 is an important regulator of the OCT4 protein level in human ES cells.展开更多
AIM： To determine the effect of hammerhead ribozyme targeting connective tissue growth factor （CCN2） on human hepatic stellate cell （HSC） function.METHODS： CCN2 hammerhead ribozyme cDNA plus two self-cleaving se...AIM： To determine the effect of hammerhead ribozyme targeting connective tissue growth factor （CCN2） on human hepatic stellate cell （HSC） function.METHODS： CCN2 hammerhead ribozyme cDNA plus two self-cleaving sequences were inserted into pTriEx2 to produce pTriCCN2-Rz. Each vector was individually transfected into cultured LX-2 human HSCs, which were then stimulated by addition of transforming growth factor （TGF）-β1 to the culture medium. Semiquantitative RT-PCR was used to determine mRNA levels for CCN2 or collagen I, while protein levels of each molecule in cell /ysates and conditioned medium were measured by ELISA. Cell-cycle progression of the transfected cells was assessed by flow cytometry.RESULTS： In pTriEx2-transfected LX-2 cells, TGF-β1 treatment caused an increase in the mRNA level for CCN2 or collagen I, and an increase in produced and secreted CCN2 or extracellular collagen I protein levels, pTriCCN2-Rz-transfected LX-2 cells showed decreased basal CCN2 or collagen mRNA levels, as well as produced and secreted CCN2 or collagen I protein. Furthermore, the TGF-β1-induced increase in mRNA or protein for CCN2 or collagen I was inhibited partially in pTriCCN2-Rz-transfected LX-2 cells. Inhibition of CCN2 using hammerhead ribozyme cDNA resulted in fewer of the cells transitioning into S phase.CONCLUSION： Endogenous CCN2 is a mediator of basal or TGF-β1-induced collagen I production in human HSCs and regulates entry of the cells into S phase.展开更多
The authors have invented a superior wind power unit, which is composed of tandem wind rotors and double rotational armature type generator without the traditional stator. This unit is called ＂intelligent wind power ...The authors have invented a superior wind power unit, which is composed of tandem wind rotors and double rotational armature type generator without the traditional stator. This unit is called ＂intelligent wind power unit＂. At upwind type unit, the large-sized front wind rotor and the small-sized rear wind rotor drive the inner and the outer rotational armatures respectively, in keeping the rotational torque counter-balance between both wind rotors/armatures. This paper discusses the acoustic noise emitted from the tandem wind rotors. As for tandem wind rotors, the tip vortex shedding from the front wind rotor may make a loud acoustic noise if the vortex attacks the rear wind rotor. Intelligent wind power unit, however, has no chance to increase the acoustic noise level by the tip vortex because the diameter of the front wind rotor is reasonably larger than the diameter of the rear wind rotor. The vorticity generated in the boundary layer on the blade can be observed obviously at wake flow and can be evaluated quantitatively at flow conditions measured by a hot-wire anemometer at a wind tunnel. The flow conditions have shown that the radial and axial components of vorticities contribute to emit the acoustic noise.展开更多
Objective: The purpose of this study was to analyze the relationship between signal transducer and activator of transcription 3 (STAT3) and carcinoembryonic antigen (CEA) in lung adenocarcinoma cell A549, and to explo...Objective: The purpose of this study was to analyze the relationship between signal transducer and activator of transcription 3 (STAT3) and carcinoembryonic antigen (CEA) in lung adenocarcinoma cell A549, and to explore the value of STAT3 on early diagnosis of lung adenocarcinoma. Methods: The expression of CEA, STAT3 mRNA and it's protein in human lung adenocarcinoma cell A549 and normal human lung cells MRC-5 were tested by immunohistochemistry staining (PV) and quantitative real time fluorescent PCR. The correlation between STAT3 and CEA in human lung adenocarcinoma cell A549 was analyzed. Results: The protein and mRNA levels of STAT3, CEA in lung adenocarcinoma cell A549 were apparently higher than those in normal human lung cells MRC-5. The levels of STAT3 mRNA and it's protein were positively correlated with CEA in lung adenocarcinoma cell A549. Conclusion: STAT3 have the same value in diagnosis of lung adenocarcinoma.展开更多
Objective: We aimed to study the transcription level of VEGF-C in human breast cancer tissue, and explore the correlations with the expression of C-erbB-2. Methods: The expression of VEGF-C mRNA in 51 cases of human b...Objective: We aimed to study the transcription level of VEGF-C in human breast cancer tissue, and explore the correlations with the expression of C-erbB-2. Methods: The expression of VEGF-C mRNA in 51 cases of human breast cancer was assessed by hybridization in situ. The expressions of C-erbB-2 was assessed by immunohistochemistry. Results: The positive rate of VEGF-C mRNA was 54.9% in 51 cases of breast cancer. The transcription level had correlation with tumor size and status of lymph nodes(P < 0.05). The expression of VEGF-C mRNA had a positive correlation with the expression of C-erbB-2(P < 0.05). Conclusion: The up-expression of VEGF-C has a significant correlation with the malignancy level and clinical stage of breast cancer. The combined detection of VEGF-C, C-erbB-2 may help to estimate the prognosis of patients with breast cancer and study on thetherapeutic implications.展开更多
In the present research work, the pitch-control is carried out such that the rotor blades are rotated around their longitudinal axis while the rotor continues its normal rotation. It is really a challenge to produce a...In the present research work, the pitch-control is carried out such that the rotor blades are rotated around their longitudinal axis while the rotor continues its normal rotation. It is really a challenge to produce a clever design to pitch the rotor blades by the optimal amount so as to maximize the power output at all wind speeds. The mechanism is implemented to a three-blade, horizontal-axis, home-scale wind turbine. The mechanism is powered by a suitable DC （direct-current） motor. The tests were carried out in the open section of a delivery wind tunnel. The air speed was measured by a suitable anemometer. The corresponding rotational speed （rpm） and output voltage at different wind speeds were measured and recorded for calibration of the control system. The mechanism proved to be successful in controlling the pitch angle over a wide range of wind speeds.展开更多
Cr_2Ge_2Te_6is an intrinsic ferromagnetic semiconductor with van der Waals type layered structure,thus represents a promising material for novel electronic and spintronic devices.Here we combine scanning tunneling mic...Cr_2Ge_2Te_6is an intrinsic ferromagnetic semiconductor with van der Waals type layered structure,thus represents a promising material for novel electronic and spintronic devices.Here we combine scanning tunneling microscopy and first-principles calculations to investigate the electronic structure of Cr_2Ge_2Te_6.Tunneling spectroscopy reveals a surprising large energy level shift and change of energy gap size across the ferromagnetic to paramagnetic phase transition,as well as a peculiar double-peak electronic state on the Cr-site defect.These features can be quantitatively explained by density functional theory calculations,which uncover a close relationship between the electronic structure and magnetic order.These findings shed important new lights on the microscopic electronic structure and origin of magnetic order in Cr_2Ge_2Te_6.展开更多
This paper has attempted to study a mechanism of three-dimensional flow around a horizontal axis wind turbine(HAWT) rotor blade. An experimental study of the flow phenomenon in the vicinity of the wind turbine blade i...This paper has attempted to study a mechanism of three-dimensional flow around a horizontal axis wind turbine(HAWT) rotor blade. An experimental study of the flow phenomenon in the vicinity of the wind turbine blade is a challenging endeavor. In this research, the HAWT model with 2.4 m diameter was tested in the large wind tunnel. The flow around the rotating blade surface was measured simultaneously for three velocity components, and two probes were used for the synchronized measurement of three-dimensional flow components. The local velocity was detected for the single seeding particle measured in the point where three pairs of laser beams intersected. Blade sections of interest in this study are composed of radial positions r/R = 0.3, 0.5 and 0.7. Optimum and low tip speed ratio flow characteristics were also compared. The velocity flow vector, skin friction coefficient and bound circulation were calculated from LDV measurements, and the experimental research showed reasonably and clearly the experimental results.展开更多
文摘MicroRNAs （miRNAs） are endogenous, small, non-coding RNAs, which are capable of silencing gene expression at the post-transcriptional level. In this study, we report that miR-205 is significantly underexpressed in breast tumor compared to the matched normal breast tissue. Similarly, breast cancer cell lines, including MCF-7 and MDA-MB- 231, express a lower level miR-205 than the non-malignant MCF-10A cells. Of interest, ectopic expression of miR-205 significantly inhibits cell proliferation and anchorage independent growth, as well as cell invasion. Furthermore, miR- 205 was shown to suppress lung metastasis in an animal model. Finally, western blot combined with the luciferase reporter assays demonstrate that ErbB3 and vascular endothelial growth factor A （VEGF-A） are direct targets for miR-205, and this miR-205-mediated suppression is likely through the direct interaction with the putative miR-205 binding site in the 3＇-untranslated region （3＇-UTR） of ErbB3 and VEGF-A. Together, these results suggest that miR- 205 is a tumor suppressor in breast cancer.
基金Acknowledgments We are grateful to Dr DA Melton （Harvard University） for shar- ing his human ES cells with us. The study was supported by grants from the National High Technology Research and Development Program of China （2006CB943900）, the National Natural Science Foundation of China （General Program, 30500088）, the Shang- hai Jiao Tong University School of Medicine, and the Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. The study was also supported by the Shanghai Leading Academic Deciline Project （S30201）.
文摘POU transcription factor OCT4 not only plays an essential role in maintaining the pluripotent and self-renewing state of embryonic stem （ES） cells but also acts as a cell fate determinant through a gene dosage effect. However, the molecular mechanisms that control the intracellular OCT4 protein level remain elusive. Here, we report that human WWP2, an E3 ubiquitin （Ub）-protein ligase, interacts with OCT4 specifically through its WW domain and enhances Ub modification of OCT4 both in vitro and in vivo. We first demonstrated that endogenous OCT4 in hu- man ES cells can be post-translationally modified by Ub. Furthermore, we found that WWP2 promoted degradation of OCT4 through the 26S proteasome in a dosage-dependent manner, and the active site cysteine residue of WWP2 was required for both its enzymatic activity and proteolytic effect on OCT4. Remarkably, our data show that the en- dogenous OCT4 protein level was significantly elevated when WWP2 expression was downregulated by specific RNA interference （RNAi）, suggesting that WWP2 is an important regulator for maintaining a proper OCT4 protein level in human ES cells. Moreover, northern blot analysis showed that the WWP2 transcript was widely present in diverse human tissues/organs and highly expressed in undifferentiated human ES cells. However, its expression level was quickly decreased after human ES cells differentiated, indicating that WWP2 expression might be developmentally regulated. Our findings demonstrate that WWP2 is an important regulator of the OCT4 protein level in human ES cells.
基金Supported by National Natural Scientific Foundation No.30872236 to Run-Ping Gao and NIH 5R01AA016003 to David R Brigstock
文摘AIM： To determine the effect of hammerhead ribozyme targeting connective tissue growth factor （CCN2） on human hepatic stellate cell （HSC） function.METHODS： CCN2 hammerhead ribozyme cDNA plus two self-cleaving sequences were inserted into pTriEx2 to produce pTriCCN2-Rz. Each vector was individually transfected into cultured LX-2 human HSCs, which were then stimulated by addition of transforming growth factor （TGF）-β1 to the culture medium. Semiquantitative RT-PCR was used to determine mRNA levels for CCN2 or collagen I, while protein levels of each molecule in cell /ysates and conditioned medium were measured by ELISA. Cell-cycle progression of the transfected cells was assessed by flow cytometry.RESULTS： In pTriEx2-transfected LX-2 cells, TGF-β1 treatment caused an increase in the mRNA level for CCN2 or collagen I, and an increase in produced and secreted CCN2 or extracellular collagen I protein levels, pTriCCN2-Rz-transfected LX-2 cells showed decreased basal CCN2 or collagen mRNA levels, as well as produced and secreted CCN2 or collagen I protein. Furthermore, the TGF-β1-induced increase in mRNA or protein for CCN2 or collagen I was inhibited partially in pTriCCN2-Rz-transfected LX-2 cells. Inhibition of CCN2 using hammerhead ribozyme cDNA resulted in fewer of the cells transitioning into S phase.CONCLUSION： Endogenous CCN2 is a mediator of basal or TGF-β1-induced collagen I production in human HSCs and regulates entry of the cells into S phase.
文摘The authors have invented a superior wind power unit, which is composed of tandem wind rotors and double rotational armature type generator without the traditional stator. This unit is called ＂intelligent wind power unit＂. At upwind type unit, the large-sized front wind rotor and the small-sized rear wind rotor drive the inner and the outer rotational armatures respectively, in keeping the rotational torque counter-balance between both wind rotors/armatures. This paper discusses the acoustic noise emitted from the tandem wind rotors. As for tandem wind rotors, the tip vortex shedding from the front wind rotor may make a loud acoustic noise if the vortex attacks the rear wind rotor. Intelligent wind power unit, however, has no chance to increase the acoustic noise level by the tip vortex because the diameter of the front wind rotor is reasonably larger than the diameter of the rear wind rotor. The vorticity generated in the boundary layer on the blade can be observed obviously at wake flow and can be evaluated quantitatively at flow conditions measured by a hot-wire anemometer at a wind tunnel. The flow conditions have shown that the radial and axial components of vorticities contribute to emit the acoustic noise.
文摘Objective: The purpose of this study was to analyze the relationship between signal transducer and activator of transcription 3 (STAT3) and carcinoembryonic antigen (CEA) in lung adenocarcinoma cell A549, and to explore the value of STAT3 on early diagnosis of lung adenocarcinoma. Methods: The expression of CEA, STAT3 mRNA and it's protein in human lung adenocarcinoma cell A549 and normal human lung cells MRC-5 were tested by immunohistochemistry staining (PV) and quantitative real time fluorescent PCR. The correlation between STAT3 and CEA in human lung adenocarcinoma cell A549 was analyzed. Results: The protein and mRNA levels of STAT3, CEA in lung adenocarcinoma cell A549 were apparently higher than those in normal human lung cells MRC-5. The levels of STAT3 mRNA and it's protein were positively correlated with CEA in lung adenocarcinoma cell A549. Conclusion: STAT3 have the same value in diagnosis of lung adenocarcinoma.
基金Supported by grants from the Liaoning Province Science&Technology Development Funds(No.2012225019)Sub-topics of Major Drug Discovery Platform in the Twelfth-Five Year Research Program of China(No.2012ZX09303016-002)
文摘Objective: We aimed to study the transcription level of VEGF-C in human breast cancer tissue, and explore the correlations with the expression of C-erbB-2. Methods: The expression of VEGF-C mRNA in 51 cases of human breast cancer was assessed by hybridization in situ. The expressions of C-erbB-2 was assessed by immunohistochemistry. Results: The positive rate of VEGF-C mRNA was 54.9% in 51 cases of breast cancer. The transcription level had correlation with tumor size and status of lymph nodes(P < 0.05). The expression of VEGF-C mRNA had a positive correlation with the expression of C-erbB-2(P < 0.05). Conclusion: The up-expression of VEGF-C has a significant correlation with the malignancy level and clinical stage of breast cancer. The combined detection of VEGF-C, C-erbB-2 may help to estimate the prognosis of patients with breast cancer and study on thetherapeutic implications.
文摘In the present research work, the pitch-control is carried out such that the rotor blades are rotated around their longitudinal axis while the rotor continues its normal rotation. It is really a challenge to produce a clever design to pitch the rotor blades by the optimal amount so as to maximize the power output at all wind speeds. The mechanism is implemented to a three-blade, horizontal-axis, home-scale wind turbine. The mechanism is powered by a suitable DC （direct-current） motor. The tests were carried out in the open section of a delivery wind tunnel. The air speed was measured by a suitable anemometer. The corresponding rotational speed （rpm） and output voltage at different wind speeds were measured and recorded for calibration of the control system. The mechanism proved to be successful in controlling the pitch angle over a wide range of wind speeds.
基金supported by the Basic Science Center Project of NSFC(51788104)the MOST of China(2015CB921000)+6 种基金the support from Tsinghua University Initiative Scientific Research Program and NSFC(11774196)S.H.Z.is supported by the National Postdoctoral Program for Innovative Talents of China(BX201600091)the China Postdoctoral Science Foundation(2017M610858)the support of the National Key Research and Development Program(2016YFA0300404)NSFC Grant(11674326)the Joint Funds of NSFC and the Chinese Academy of Sciences’Large-Scale Scientific Facility(U1432139)supported in part by the Beijing Advanced Innovation Center for Future Chip(ICFC)
文摘Cr_2Ge_2Te_6is an intrinsic ferromagnetic semiconductor with van der Waals type layered structure,thus represents a promising material for novel electronic and spintronic devices.Here we combine scanning tunneling microscopy and first-principles calculations to investigate the electronic structure of Cr_2Ge_2Te_6.Tunneling spectroscopy reveals a surprising large energy level shift and change of energy gap size across the ferromagnetic to paramagnetic phase transition,as well as a peculiar double-peak electronic state on the Cr-site defect.These features can be quantitatively explained by density functional theory calculations,which uncover a close relationship between the electronic structure and magnetic order.These findings shed important new lights on the microscopic electronic structure and origin of magnetic order in Cr_2Ge_2Te_6.
文摘This paper has attempted to study a mechanism of three-dimensional flow around a horizontal axis wind turbine(HAWT) rotor blade. An experimental study of the flow phenomenon in the vicinity of the wind turbine blade is a challenging endeavor. In this research, the HAWT model with 2.4 m diameter was tested in the large wind tunnel. The flow around the rotating blade surface was measured simultaneously for three velocity components, and two probes were used for the synchronized measurement of three-dimensional flow components. The local velocity was detected for the single seeding particle measured in the point where three pairs of laser beams intersected. Blade sections of interest in this study are composed of radial positions r/R = 0.3, 0.5 and 0.7. Optimum and low tip speed ratio flow characteristics were also compared. The velocity flow vector, skin friction coefficient and bound circulation were calculated from LDV measurements, and the experimental research showed reasonably and clearly the experimental results.
