The finite element method (FEM) plays a valuable role in computer modeling and is beneficial to the mechanicaldesign of various structural parts. However, the elements produced by conventional FEM are easily inaccurat...The finite element method (FEM) plays a valuable role in computer modeling and is beneficial to the mechanicaldesign of various structural parts. However, the elements produced by conventional FEM are easily inaccurate andunstable when applied. Therefore, developing new elements within the framework of the generalized variationalprinciple is of great significance. In this paper, an 8-node plane hybrid finite element with 15 parameters (PHQ8-15β) is developed for structural mechanics problems based on the Hellinger-Reissner variational principle.According to the design principle of Pian, 15 unknown parameters are adopted in the selection of stress modes toavoid the zero energy modes.Meanwhile, the stress functions within each element satisfy both the equilibrium andthe compatibility relations of plane stress problems. Subsequently, numerical examples are presented to illustrate theeffectiveness and robustness of the proposed finite element. Numerical results show that various common lockingbehaviors of plane elements can be overcome. The PH-Q8-15β element has excellent performance in all benchmarkproblems, especially for structures with varying cross sections. Furthermore, in bending problems, the reasonablemesh shape of the new element for curved edge structures is analyzed in detail, which can be a useful means toimprove numerical accuracy.展开更多
As an effective approach to achieve the“dual-carbon”goal,the grid-connected capacity of renewable energy increases constantly.Photovoltaics are the most widely used renewable energy sources and have been applied on ...As an effective approach to achieve the“dual-carbon”goal,the grid-connected capacity of renewable energy increases constantly.Photovoltaics are the most widely used renewable energy sources and have been applied on various occasions.However,the inherent randomness,intermittency,and weak support of grid-connected equipment not only cause changes in the original flow characteristics of the grid but also result in complex fault characteristics.Traditional overcurrent and differential protection methods cannot respond accurately due to the effects of unknown renewable energy sources.Therefore,a longitudinal protection method based on virtual measurement of current restraint is proposed in this paper.The positive sequence current data and the network parameters are used to calculate the virtual measurement current which compensates for the output current of photovoltaic(PV).The waveform difference between the virtual measured current and the terminal current for internal and external faults is used to construct the protection method.An improved edit distance algorithm is proposed to measure the similarity between virtual measurement current and terminal measurement current.Finally,the feasibility of the protection method is verified through PSCAD simulation.展开更多
A porosity-free and contamination-free surface layer with grain sizes ranging from nanometer to micrometer in Fe samples was obtained by surface mechanical attrition treatment (SMAT) technique. Mechanical and wear pro...A porosity-free and contamination-free surface layer with grain sizes ranging from nanometer to micrometer in Fe samples was obtained by surface mechanical attrition treatment (SMAT) technique. Mechanical and wear properties of the surface layer in the SMATed and annealed Fe samples were measured by means of nanoindentation and nanoscratch tests, respectively. Experimental results showed that the hardness of the surface layer in the SMATed Fe sample increased evidently due to the grain refinement. The elastic moduli of the surface layers in the SMATed and annealed Fe samples were unchanged, independent of grain size in the present grain size regime. Compared with the original Fe sample, the wear resistance enhanced and the coefficient of friction decreased in the surface layer of the SMATed Fe sample.展开更多
AIM: We shall construct the small interfering RNA (siRNA) expression cassette (SEC) targeting activated K-ras gene sequence, identify more effective siRNA sequence against K-ras gene in human pancreatic cancer cell li...AIM: We shall construct the small interfering RNA (siRNA) expression cassette (SEC) targeting activated K-ras gene sequence, identify more effective siRNA sequence against K-ras gene in human pancreatic cancer cell line MiaPaCa-2 by SEC and reveal the anti-cancer effects of RNA interference (RNAi) and its therapeutic possibilities. METHODS: Three different sites of SECs were constructed by PCR. K1/siRNA,K2/siRNA and K3/siRNA are located at sites 194,491 and 327, respectively. They were transfected into MiaPaCa-2 cells by liposome to inhibit the expression of activated K-ras. In the interfering groups of sites 194 and 491, we detected the apoptosis in cells by FACS after they were incubated for 48 h, then we tested the alternation of K-ras gene in MiaPaCa-2 cells by RT-PCR immunofluorescence, respectively. RESULTS: Introduction of the Kl/siRNA and K2/siRNA against K-ras into MiaPaCa-2 cells leads to increased apoptosis, and the number of apoptotic cells is increased compared with control cells. The tests of RT-PCR immunofluorescence show the effects of inhibiting expression of activated K-ras gene by RNA interference in the Kl/siRNA and K2/siRNA groups. We also find that the introduction of K3/siRNA has no effect on MiaPaCa-2 cells. CONCLUSION: Kl/siRNA and K2/siRNA can inhibit the expression of activated K-ras but K3/siRNA has no effect, demonstrating that Kl/siRNA and K2/siRNA are effective sequences against K-ras gene and K3/siRNA are not. We conclude that specific siRNA against K-ras expression may be a powerful tool to be used therapeutically against human pancreatic cancer.展开更多
AIM: Ischemic preconditioning (IP) is a brief ischemic episode,which confers a state of protection against the subsequent long-term ischemia-reperfusion injuries. However, little is known regarding the use of IP befor...AIM: Ischemic preconditioning (IP) is a brief ischemic episode,which confers a state of protection against the subsequent long-term ischemia-reperfusion injuries. However, little is known regarding the use of IP before the sustained cold storage and liver transplantation. The present study was designed to evaluate the protective effect of IP on the long-term preservation of liver graft and the prolonged anhepatic-phase injury.METHODS: Male Sprague-Dawley rats were used as donors and recipients of orthotopic liver transplantation. All livers underwent 10 min of ischemia followed by 10 min of reperfusion before harvest. Rat liver transplantation was performed with the portal vein clamped for 25 min. Tolerance of transplanted liver to the reperfusion injury and liver damage were investigated. The changes in adenosine concentration in hepatic tissue and those of nitric oxide (NO) and tumor necrosis factor (TNF) in serum were also assessed.RESULTS: Recipients with IP significantly improved their one-week survival rate and liver function, they had increased levels of circulating NO and hepatic adenosine, and a reduced level of serum TNF, as compared to controls. Histological changes indicating hepatic injuries appeared improved in the IP group compared with those in control group. The protective effect of IP was also obtained by administration of adenosine,while blockage of the NO pathway using Nco-nitro-L-arginine methyl ester abolished the protective effect of IP.CONCLUSION: IP appears to have a protective effect on the long-term preservation of liver graft and the prolonged anhepatic-phase injuries. NO may be involved in this process.展开更多
AIM:To identify the role of alpha-fetoprotein (AFP) mRNA expression in peripheral blood one week after surgery as a predictor for recurrence of hepatocellular carcinoma (HCC). METHODS: Published studies fulfilling the...AIM:To identify the role of alpha-fetoprotein (AFP) mRNA expression in peripheral blood one week after surgery as a predictor for recurrence of hepatocellular carcinoma (HCC). METHODS: Published studies fulfilling the selection criteria were identified by searching several databases online. After a methodology assessment using a quality scale designed by European Lung Cancer Working Party, data in each research were aggregated by means of meta-analysis. RESULTS: Altogether 368 cases were included in the 9 selected studies, which fulfilled the selection criteria. The quality scores ranged from 35% to 84% with a median score of 55%. The 'design' subscore had the lowest median value (38%). By aggregating the data, a high x2 value (77.576) was presented. The fail-safe number was 136 and 64 for P= 0.05 and 0.01, respectively. CONCLUSION: AFP mRNA expression in peripheral blood 1 wk after surgery correlated with the recurrence of HCC and was a good predictor for tumor recurrence.展开更多
AIM: To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.METHODS: Total RNA was extracted from colon cancer tissues and amplified by reverse...AIM: To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.METHODS: Total RNA was extracted from colon cancer tissues and amplified by reverse-transcription PCR with two primers, which span the whole coding region of ODC. The synthesized ODC cDNA was cloned into vector pQE-30 at restriction sites BamH I and Sal I which constituted recombinant expression plasmid pQE30-ODC. The sequence of inserted fragment was confirmed by DNA sequencing,the fusion protein including 6His-tag was facilitated for purification by Ni-NTA chromatographic column.RESULTS: ODC expression vector was constructed and confirmed with restriction enzyme digestion and subsequent DNA sequencing. The DNA sequence matching on NCBI Blast showed 99 % affinity. The vector was transformed into E.coli M15 and expressed. The expressed ODC protein was verified with Western blotting.CONCLUSION: The ODC prokaryote expression vector is constructed and thus greatly facilitates to study the role of ODC in colorectal carcinoma.展开更多
AIM: To investigate the role of NF-κB activation and zinc finger protein A20 expression in the regulation of maturation of dendritic cells (DCs) derived from liver allografts undergoing acute rejection.
METHODS: Sixt...AIM: To investigate the role of NF-κB activation and zinc finger protein A20 expression in the regulation of maturation of dendritic cells (DCs) derived from liver allografts undergoing acute rejection.
METHODS: Sixty donor male SD rats and sixty recipient male LEW rats weighing 220-300 g were randomly divided into whole liver transplantation group and partial liver transplantation group. Allogeneic (SD rat to LEW rat) whole and 50 % partial liver transplantation were performed. DCs from liver grafts 0 hour and 4 days after transplantation were isolated and propagated in the presence of GM-CSF in vitro. Morphological characteristics and phenotypical features of DCs propagated for 10 days were analyzed by electron microscopy and flow cytometry, respectively. NF-κB binding activity, IL-12p70 protein and zinc finger protein A20expression in these DCs were measured by EMSA and Western blotting, respectively. Histological grading of rejection was determined.
RESULTS: Allogeneic whole liver grafts showed no signs of rejection on day 4 after the transplantation. In contrast,allogeneic partial liver grafts demonstrated moderate to severe rejection on day 4 after the transplantation. After propagation for 10 days in the presence of GM-CSF in vitro,DCs from allogeneic whole liver grafts exhibited features of immature DC with absence of CD40 surface expression,these DCs were found to exhibit detectable but very low level of NF-κB activity, IL-12 p70 protein and zinc finger protein A20 expression. Whereas, DCs from allogeneic partial liver graft 4 days after transplantation displayed features of mature DC, with high level of CD40 surface expression, and as a consequence, higher expression of IL-12p70 protein, higher activities of NF-κB and higher expression of zinc finger protein A20 compared with those of DCs from whole liver grafts (P＜0.001).
CONCLUSION: These results suggest that A20expression is up-regulated in response to NF-κB activation in mature DCs derived from allogeneic liver grafts undergoing acute rejection. Given the NF-κB inhibition function of this gene, it is suggested that their expression survives to limit NF-κB activation and maturation of DCs,and consequently inhibits the acute rejection and induces acceptance of liver graft.展开更多
AIM: To investigate the effects of cisapride on intestinal bacterial overgrowth (IBO), bacterial and endotoxin translocation, intestinal transit and permeability in cirrhotic rats.METHODS: All animals were assessed wi...AIM: To investigate the effects of cisapride on intestinal bacterial overgrowth (IBO), bacterial and endotoxin translocation, intestinal transit and permeability in cirrhotic rats.METHODS: All animals were assessed with variables including bacterial and endotoxin translocation, intestinal bacterial overgrowth, intestinal transit and permeability.Bacterial translocation (BT) was assessed by bacterial culture of MLN, liver and spleen, IBO by a jejunal bacterial count of the specific organism, intestinal permeability by determination of the 24-hour urinary 99mTc-DTPA excretion and intestinal transit by measurement of the distribution of 51Cr in the intestine.RESULTS: Bacterial translocation (BT) and IBO was found in 48 % and 80 % cirrhotic rats respectively and none in control rats. Urinary excretion of 99mTc-DTPA in cirrhotic rats with BT (22.2±7.8) was greater than these without BT (10.5±2.9). Intestinal transit (geometric center ratio) was significantly delayed in cirrhotic rats (0.31±0.06) and further more delayed in cirrhotic rats with BT (0.24±0.06) than these without BT (0.38±0.11). Cirrhotic rats with IBO had significantly higher rates of intestinal bacterial and endotoxin translocation, slower intestinal transit time and higher intestinal permeability than those without IBO. It was also found that BT was closely associated with IBO and the injury of intestinal barrier. Compared with the placebo group,cisapride-treated rats had lower rates of bacterial/endotoxin translocation and IBO, which was closely associated with increased intestinal transit and improved intestinal permeability by cisapride.CONCLUSION: These results indicate that endotoxin and bacterial translocation in cirrhotic rats may be attributed to IBO and increased intestinal permeability. Cisapride that accelerates intestinal transit and improve intestinal permeability might be helpful in preventing intestinal bacterial and endotoxin translocation.展开更多
AEM: To investigate the expression pattern of epithelial cell adhesion molecule (Ep-CAM) on normal and malignant colon tissues to evaluate its diagnostic and therapeutic significance. METHODS: cDNA encoding Ep-CAM ext...AEM: To investigate the expression pattern of epithelial cell adhesion molecule (Ep-CAM) on normal and malignant colon tissues to evaluate its diagnostic and therapeutic significance. METHODS: cDNA encoding Ep-CAM extracellular domain was doned by reverse transcription-polymerase chain reaction (RT-PCR) from excised malignant colon tissues and inserted into a glutathione S-transferase (GST)-tagged vector. Ep-CAM-GST fusion protein was induced by isopropyl-p-D-thiogalactopyranoside (IPTG) and purified with glutathione-sepharose. The Ep-CAM-GST fusion protein was mixed with Freund's adjuvant and Balb/c mice were immunized with it. Sp2/0 myeloma cells were fused with the spleen cells of the immunized mice. After having selected by indirect ELISA, the anti-Ep-CAM monoclonal antibodies (MAbs) were generaled and the corresponding ascites were obtained. Finally, the human colon carcinoma tissue array prepared from seventy individual patients was stained with the anti-Ep-CAM MAbs. RESULTS: The isdated Ep-CAM cDNA sequence was identical to the data in GenBank. The expressed fusion protein was almost soluble and had a molecular weight (MW) of 53 ku. Four MAbs against Ep-CAM were obtained and designated as FMU-Epl, FMU-Ep2, FMU-Ep3 and FMU-Ep4 respectively. Among them, FMU-Ep4 could recognize the natural Ep-CAM on Colo205 and SW480 cells, and all of them could be used for immunohistochemical staining of tissue sections. It was fbund that Ep-CAM was distributed differently in normal and various malignant colon tissues, induding squamous cell carcinoma, signet-ring cell carcinoma and adenocarcinoma. In normal colon gland epithelia, Ep-CAM antigen was mainly distributed on the basolateral membrane and in the region between the basolateral membrane and the cytoplastic part near the nuclei, whereas the expression pattern of colon malignancies was mainly on the whole surface of epithelia and the expression was much higher than the normal colon tissues. The staining pattern of tissue array showed in adenocarcinoma and papillary adenocarcinoma, and the expression of Ep-CAM was increased from grade I to grade Ⅲ. CONCLUSION: MAbs against Ep-CAM might be useful for research on the structure and function of Ep-CAM and may have diagnostic and therapeutic value to various colon carcinomas.展开更多
By taking advantage of silylanization, Al2O3 support was modified by organosilane and supported Pd-Cu-Clx/Al2O3 catalysts were prepared. The effects of hydrophobicity on catalyst stability during CO oxidation were inv...By taking advantage of silylanization, Al2O3 support was modified by organosilane and supported Pd-Cu-Clx/Al2O3 catalysts were prepared. The effects of hydrophobicity on catalyst stability during CO oxidation were investigated. The physicochemical properties and redox potential of the catalyst were characterized by N2 adsorption-desorption, XRD, H2-TPR, and XPS. In order to understand the relationship between the oxidation stability of CO and the presence of water, the CO oxidation mechanism was studied by in situ DRIFT. Support pretreatment markedly promoted catalyst stability during CO oxidation; CO conversion was 78% after 150 h at saturated humidity and freezing point. Modification led to an obvious decrease in chloride ion concentration and enhancement in hydrophobicity. The role of water in CO oxidation was complicated. The presence of water favored CO oxidation over active Pd~＋ species and Pd0 reoxidation by Cu^（2＋） species. Meanwhile, water also inhibited the formation of the active Pd~＋ species and helped to produce carbonate species. Compared with the form of the carbonate species, the inhibition of water to produce active Pd~＋ species played the main detrimental role in catalyst stability.展开更多
Strawberry is increasingly used as a model plant for research on fruit growth and development.The transient gene manipulation(TGM)technique is widely used to determine the function of plant genes,including those in st...Strawberry is increasingly used as a model plant for research on fruit growth and development.The transient gene manipulation(TGM)technique is widely used to determine the function of plant genes,including those in strawberry fruits.However,its reliable application for the precise identification of gene function has been difficult owing to the lack of conditional optimization.In this study,we found that successful transient gene manipulation requires optimization,with the vector type,temperature,and fruit developmental stage being three major factors determining success.Notably,we found that transient gene manipulation was feasible only from the large green fruit stage onwards,making it especially suitable for identifying genes involved in strawberry fruit ripening.Furthermore,we established a method called percentage difference of phenotype(PDP),in which the functional effect of a gene could be precisely and efficiently identified in strawberry fruits.This method can be used to estimate the functional effect of a gene as a value from 0 to 100%,such that different genes can be quantitatively compared for their relative abilities to regulate fruit ripening.This study provides a useful tool for accelerating research on the molecular basis of strawberry fruit ripening.展开更多
基金the National Natural Science Foundation of China(No.11572210).
文摘The finite element method (FEM) plays a valuable role in computer modeling and is beneficial to the mechanicaldesign of various structural parts. However, the elements produced by conventional FEM are easily inaccurate andunstable when applied. Therefore, developing new elements within the framework of the generalized variationalprinciple is of great significance. In this paper, an 8-node plane hybrid finite element with 15 parameters (PHQ8-15β) is developed for structural mechanics problems based on the Hellinger-Reissner variational principle.According to the design principle of Pian, 15 unknown parameters are adopted in the selection of stress modes toavoid the zero energy modes.Meanwhile, the stress functions within each element satisfy both the equilibrium andthe compatibility relations of plane stress problems. Subsequently, numerical examples are presented to illustrate theeffectiveness and robustness of the proposed finite element. Numerical results show that various common lockingbehaviors of plane elements can be overcome. The PH-Q8-15β element has excellent performance in all benchmarkproblems, especially for structures with varying cross sections. Furthermore, in bending problems, the reasonablemesh shape of the new element for curved edge structures is analyzed in detail, which can be a useful means toimprove numerical accuracy.
基金funded by State Grid Anhui Electric Power Co.,Ltd.Science and Technology Project(52120021N00L)the National Key Research and Development Program of China(2022YFB2400015).
文摘As an effective approach to achieve the“dual-carbon”goal,the grid-connected capacity of renewable energy increases constantly.Photovoltaics are the most widely used renewable energy sources and have been applied on various occasions.However,the inherent randomness,intermittency,and weak support of grid-connected equipment not only cause changes in the original flow characteristics of the grid but also result in complex fault characteristics.Traditional overcurrent and differential protection methods cannot respond accurately due to the effects of unknown renewable energy sources.Therefore,a longitudinal protection method based on virtual measurement of current restraint is proposed in this paper.The positive sequence current data and the network parameters are used to calculate the virtual measurement current which compensates for the output current of photovoltaic(PV).The waveform difference between the virtual measured current and the terminal current for internal and external faults is used to construct the protection method.An improved edit distance algorithm is proposed to measure the similarity between virtual measurement current and terminal measurement current.Finally,the feasibility of the protection method is verified through PSCAD simulation.
基金Financial support from the National Natural Science Foundation of China,the Ministry of Science and Technology of China(Grant G1999064505)NEDO International Joint Research Grant Program(01MB5)Ministry of Research of France(Grant 2001882,CPER EN2040)is acknowledged.
文摘A porosity-free and contamination-free surface layer with grain sizes ranging from nanometer to micrometer in Fe samples was obtained by surface mechanical attrition treatment (SMAT) technique. Mechanical and wear properties of the surface layer in the SMATed and annealed Fe samples were measured by means of nanoindentation and nanoscratch tests, respectively. Experimental results showed that the hardness of the surface layer in the SMATed Fe sample increased evidently due to the grain refinement. The elastic moduli of the surface layers in the SMATed and annealed Fe samples were unchanged, independent of grain size in the present grain size regime. Compared with the original Fe sample, the wear resistance enhanced and the coefficient of friction decreased in the surface layer of the SMATed Fe sample.
文摘AIM: We shall construct the small interfering RNA (siRNA) expression cassette (SEC) targeting activated K-ras gene sequence, identify more effective siRNA sequence against K-ras gene in human pancreatic cancer cell line MiaPaCa-2 by SEC and reveal the anti-cancer effects of RNA interference (RNAi) and its therapeutic possibilities. METHODS: Three different sites of SECs were constructed by PCR. K1/siRNA,K2/siRNA and K3/siRNA are located at sites 194,491 and 327, respectively. They were transfected into MiaPaCa-2 cells by liposome to inhibit the expression of activated K-ras. In the interfering groups of sites 194 and 491, we detected the apoptosis in cells by FACS after they were incubated for 48 h, then we tested the alternation of K-ras gene in MiaPaCa-2 cells by RT-PCR immunofluorescence, respectively. RESULTS: Introduction of the Kl/siRNA and K2/siRNA against K-ras into MiaPaCa-2 cells leads to increased apoptosis, and the number of apoptotic cells is increased compared with control cells. The tests of RT-PCR immunofluorescence show the effects of inhibiting expression of activated K-ras gene by RNA interference in the Kl/siRNA and K2/siRNA groups. We also find that the introduction of K3/siRNA has no effect on MiaPaCa-2 cells. CONCLUSION: Kl/siRNA and K2/siRNA can inhibit the expression of activated K-ras but K3/siRNA has no effect, demonstrating that Kl/siRNA and K2/siRNA are effective sequences against K-ras gene and K3/siRNA are not. We conclude that specific siRNA against K-ras expression may be a powerful tool to be used therapeutically against human pancreatic cancer.
基金Supported by the National Natural Science Foundation of China,No.30200278 China Postdoctoral Science Foundation,No.2001-5
文摘AIM: Ischemic preconditioning (IP) is a brief ischemic episode,which confers a state of protection against the subsequent long-term ischemia-reperfusion injuries. However, little is known regarding the use of IP before the sustained cold storage and liver transplantation. The present study was designed to evaluate the protective effect of IP on the long-term preservation of liver graft and the prolonged anhepatic-phase injury.METHODS: Male Sprague-Dawley rats were used as donors and recipients of orthotopic liver transplantation. All livers underwent 10 min of ischemia followed by 10 min of reperfusion before harvest. Rat liver transplantation was performed with the portal vein clamped for 25 min. Tolerance of transplanted liver to the reperfusion injury and liver damage were investigated. The changes in adenosine concentration in hepatic tissue and those of nitric oxide (NO) and tumor necrosis factor (TNF) in serum were also assessed.RESULTS: Recipients with IP significantly improved their one-week survival rate and liver function, they had increased levels of circulating NO and hepatic adenosine, and a reduced level of serum TNF, as compared to controls. Histological changes indicating hepatic injuries appeared improved in the IP group compared with those in control group. The protective effect of IP was also obtained by administration of adenosine,while blockage of the NO pathway using Nco-nitro-L-arginine methyl ester abolished the protective effect of IP.CONCLUSION: IP appears to have a protective effect on the long-term preservation of liver graft and the prolonged anhepatic-phase injuries. NO may be involved in this process.
基金Supported by the National Key Research and Development Program,No. 2001BA703BO4
文摘AIM:To identify the role of alpha-fetoprotein (AFP) mRNA expression in peripheral blood one week after surgery as a predictor for recurrence of hepatocellular carcinoma (HCC). METHODS: Published studies fulfilling the selection criteria were identified by searching several databases online. After a methodology assessment using a quality scale designed by European Lung Cancer Working Party, data in each research were aggregated by means of meta-analysis. RESULTS: Altogether 368 cases were included in the 9 selected studies, which fulfilled the selection criteria. The quality scores ranged from 35% to 84% with a median score of 55%. The 'design' subscore had the lowest median value (38%). By aggregating the data, a high x2 value (77.576) was presented. The fail-safe number was 136 and 64 for P= 0.05 and 0.01, respectively. CONCLUSION: AFP mRNA expression in peripheral blood 1 wk after surgery correlated with the recurrence of HCC and was a good predictor for tumor recurrence.
基金Scientific Research Fund of national Ministry of Health,No.98-1-173
文摘AIM: To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.METHODS: Total RNA was extracted from colon cancer tissues and amplified by reverse-transcription PCR with two primers, which span the whole coding region of ODC. The synthesized ODC cDNA was cloned into vector pQE-30 at restriction sites BamH I and Sal I which constituted recombinant expression plasmid pQE30-ODC. The sequence of inserted fragment was confirmed by DNA sequencing,the fusion protein including 6His-tag was facilitated for purification by Ni-NTA chromatographic column.RESULTS: ODC expression vector was constructed and confirmed with restriction enzyme digestion and subsequent DNA sequencing. The DNA sequence matching on NCBI Blast showed 99 % affinity. The vector was transformed into E.coli M15 and expressed. The expressed ODC protein was verified with Western blotting.CONCLUSION: The ODC prokaryote expression vector is constructed and thus greatly facilitates to study the role of ODC in colorectal carcinoma.
文摘AIM: To investigate the role of NF-κB activation and zinc finger protein A20 expression in the regulation of maturation of dendritic cells (DCs) derived from liver allografts undergoing acute rejection.
METHODS: Sixty donor male SD rats and sixty recipient male LEW rats weighing 220-300 g were randomly divided into whole liver transplantation group and partial liver transplantation group. Allogeneic (SD rat to LEW rat) whole and 50 % partial liver transplantation were performed. DCs from liver grafts 0 hour and 4 days after transplantation were isolated and propagated in the presence of GM-CSF in vitro. Morphological characteristics and phenotypical features of DCs propagated for 10 days were analyzed by electron microscopy and flow cytometry, respectively. NF-κB binding activity, IL-12p70 protein and zinc finger protein A20expression in these DCs were measured by EMSA and Western blotting, respectively. Histological grading of rejection was determined.
RESULTS: Allogeneic whole liver grafts showed no signs of rejection on day 4 after the transplantation. In contrast,allogeneic partial liver grafts demonstrated moderate to severe rejection on day 4 after the transplantation. After propagation for 10 days in the presence of GM-CSF in vitro,DCs from allogeneic whole liver grafts exhibited features of immature DC with absence of CD40 surface expression,these DCs were found to exhibit detectable but very low level of NF-κB activity, IL-12 p70 protein and zinc finger protein A20 expression. Whereas, DCs from allogeneic partial liver graft 4 days after transplantation displayed features of mature DC, with high level of CD40 surface expression, and as a consequence, higher expression of IL-12p70 protein, higher activities of NF-κB and higher expression of zinc finger protein A20 compared with those of DCs from whole liver grafts (P＜0.001).
CONCLUSION: These results suggest that A20expression is up-regulated in response to NF-κB activation in mature DCs derived from allogeneic liver grafts undergoing acute rejection. Given the NF-κB inhibition function of this gene, it is suggested that their expression survives to limit NF-κB activation and maturation of DCs,and consequently inhibits the acute rejection and induces acceptance of liver graft.
基金the National Natural Science Foundation No.30070340
文摘AIM: To investigate the effects of cisapride on intestinal bacterial overgrowth (IBO), bacterial and endotoxin translocation, intestinal transit and permeability in cirrhotic rats.METHODS: All animals were assessed with variables including bacterial and endotoxin translocation, intestinal bacterial overgrowth, intestinal transit and permeability.Bacterial translocation (BT) was assessed by bacterial culture of MLN, liver and spleen, IBO by a jejunal bacterial count of the specific organism, intestinal permeability by determination of the 24-hour urinary 99mTc-DTPA excretion and intestinal transit by measurement of the distribution of 51Cr in the intestine.RESULTS: Bacterial translocation (BT) and IBO was found in 48 % and 80 % cirrhotic rats respectively and none in control rats. Urinary excretion of 99mTc-DTPA in cirrhotic rats with BT (22.2±7.8) was greater than these without BT (10.5±2.9). Intestinal transit (geometric center ratio) was significantly delayed in cirrhotic rats (0.31±0.06) and further more delayed in cirrhotic rats with BT (0.24±0.06) than these without BT (0.38±0.11). Cirrhotic rats with IBO had significantly higher rates of intestinal bacterial and endotoxin translocation, slower intestinal transit time and higher intestinal permeability than those without IBO. It was also found that BT was closely associated with IBO and the injury of intestinal barrier. Compared with the placebo group,cisapride-treated rats had lower rates of bacterial/endotoxin translocation and IBO, which was closely associated with increased intestinal transit and improved intestinal permeability by cisapride.CONCLUSION: These results indicate that endotoxin and bacterial translocation in cirrhotic rats may be attributed to IBO and increased intestinal permeability. Cisapride that accelerates intestinal transit and improve intestinal permeability might be helpful in preventing intestinal bacterial and endotoxin translocation.
基金Supported by the National Key Basic Research Special Funds (NKBRSF), No. 2001CB510004
文摘AEM: To investigate the expression pattern of epithelial cell adhesion molecule (Ep-CAM) on normal and malignant colon tissues to evaluate its diagnostic and therapeutic significance. METHODS: cDNA encoding Ep-CAM extracellular domain was doned by reverse transcription-polymerase chain reaction (RT-PCR) from excised malignant colon tissues and inserted into a glutathione S-transferase (GST)-tagged vector. Ep-CAM-GST fusion protein was induced by isopropyl-p-D-thiogalactopyranoside (IPTG) and purified with glutathione-sepharose. The Ep-CAM-GST fusion protein was mixed with Freund's adjuvant and Balb/c mice were immunized with it. Sp2/0 myeloma cells were fused with the spleen cells of the immunized mice. After having selected by indirect ELISA, the anti-Ep-CAM monoclonal antibodies (MAbs) were generaled and the corresponding ascites were obtained. Finally, the human colon carcinoma tissue array prepared from seventy individual patients was stained with the anti-Ep-CAM MAbs. RESULTS: The isdated Ep-CAM cDNA sequence was identical to the data in GenBank. The expressed fusion protein was almost soluble and had a molecular weight (MW) of 53 ku. Four MAbs against Ep-CAM were obtained and designated as FMU-Epl, FMU-Ep2, FMU-Ep3 and FMU-Ep4 respectively. Among them, FMU-Ep4 could recognize the natural Ep-CAM on Colo205 and SW480 cells, and all of them could be used for immunohistochemical staining of tissue sections. It was fbund that Ep-CAM was distributed differently in normal and various malignant colon tissues, induding squamous cell carcinoma, signet-ring cell carcinoma and adenocarcinoma. In normal colon gland epithelia, Ep-CAM antigen was mainly distributed on the basolateral membrane and in the region between the basolateral membrane and the cytoplastic part near the nuclei, whereas the expression pattern of colon malignancies was mainly on the whole surface of epithelia and the expression was much higher than the normal colon tissues. The staining pattern of tissue array showed in adenocarcinoma and papillary adenocarcinoma, and the expression of Ep-CAM was increased from grade I to grade Ⅲ. CONCLUSION: MAbs against Ep-CAM might be useful for research on the structure and function of Ep-CAM and may have diagnostic and therapeutic value to various colon carcinomas.
基金supported by the National Key Research and Development Program of China(2016YFC0204300)National Natural Science Foundation of China(21207037,21333003,21571061)+1 种基金the "Shu Guang" Project of the Shanghai Municipal Education Commission(12SG29)the Commission of Science and Technology of Shanghai Municipality(15DZ1205305)~~
文摘By taking advantage of silylanization, Al2O3 support was modified by organosilane and supported Pd-Cu-Clx/Al2O3 catalysts were prepared. The effects of hydrophobicity on catalyst stability during CO oxidation were investigated. The physicochemical properties and redox potential of the catalyst were characterized by N2 adsorption-desorption, XRD, H2-TPR, and XPS. In order to understand the relationship between the oxidation stability of CO and the presence of water, the CO oxidation mechanism was studied by in situ DRIFT. Support pretreatment markedly promoted catalyst stability during CO oxidation; CO conversion was 78% after 150 h at saturated humidity and freezing point. Modification led to an obvious decrease in chloride ion concentration and enhancement in hydrophobicity. The role of water in CO oxidation was complicated. The presence of water favored CO oxidation over active Pd~＋ species and Pd0 reoxidation by Cu^（2＋） species. Meanwhile, water also inhibited the formation of the active Pd~＋ species and helped to produce carbonate species. Compared with the form of the carbonate species, the inhibition of water to produce active Pd~＋ species played the main detrimental role in catalyst stability.
基金supported by the National Natural Science Foundation of China(Grant NO.31471851,31672133 and 31872086)the 111 Project(Grant No.B17043).
文摘Strawberry is increasingly used as a model plant for research on fruit growth and development.The transient gene manipulation(TGM)technique is widely used to determine the function of plant genes,including those in strawberry fruits.However,its reliable application for the precise identification of gene function has been difficult owing to the lack of conditional optimization.In this study,we found that successful transient gene manipulation requires optimization,with the vector type,temperature,and fruit developmental stage being three major factors determining success.Notably,we found that transient gene manipulation was feasible only from the large green fruit stage onwards,making it especially suitable for identifying genes involved in strawberry fruit ripening.Furthermore,we established a method called percentage difference of phenotype(PDP),in which the functional effect of a gene could be precisely and efficiently identified in strawberry fruits.This method can be used to estimate the functional effect of a gene as a value from 0 to 100%,such that different genes can be quantitatively compared for their relative abilities to regulate fruit ripening.This study provides a useful tool for accelerating research on the molecular basis of strawberry fruit ripening.
