AIM: To separate and identify differentially expressed nuclear matrix proteins (NMPs) between the immortalized human esophageal epithelial cell line (SHEE) and the malignantly transformed esophageal carcinoma cell lin...AIM: To separate and identify differentially expressed nuclear matrix proteins (NMPs) between the immortalized human esophageal epithelial cell line (SHEE) and the malignantly transformed esophageal carcinoma cell line (SHEEC), and to provide new ways for finding specific markers and the pathogenesis of esophageal carcinoma.METHODS: SHEE and SHEEC cell lines were used to extract NMPs. The quality of NMPs was monitored by Western blot analysis including DNA topoisomerase Ⅱα, proliferation cell nuclear antigen (PCNA) and histone. NMPs of SHEE and SHEEC were analyzed by two-dimensional electrophoresis (2-DE), silver staining and PDQuest6.2 image analysis software. Three spots in which the differentially expressed NMlPs were more obvious, were selected and analyzed with matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI- TOF-MS) and database search.RESULTS: Western blot analysis revealed that DNA topoisomerase Ⅱα and PCNA were detected, and the majority of histones were deleted in NMPs of SHEE and SHEEC. After 2-DE image analysis by PDQuest6.2 software, the 2-DE maps were detected with an average of 106±7.1 spots in SHEE and 132±5.0 spots in SHEEC. Most of them were matched one another (r=0.72), only 16 protein spots were found differing in intensity. Three NMPs including cytoskeletal tropomyosin,FK506bindingprotein6,similartoretinoblastoma binding protein 8 were preliminarily identified by MALDI- TOF-MS.CONCLUSION: These differentially expressed NMPs may play an important role during malignant transformation from SHEE to SHEEC. Their separation and identification will contribute to searching for specific markers and probing into the pathogenesis of esophageal carcinoma.展开更多
Aim: To evaluate the comparative effectiveness of real-time sperm separation technique (Wang's tube method) andother two conventional methods in isolating high-quality sperm preparation, and to compare the spouse ...Aim: To evaluate the comparative effectiveness of real-time sperm separation technique (Wang's tube method) andother two conventional methods in isolating high-quality sperm preparation, and to compare the spouse pregnancy ratein intrauterine insemination (IUI) with sperm preparations isolated by these methods. Methods: The effectivenessof the real-time sperm separation technique, the conventional swim-up and the Percoll discontinuous density gradientmethods in isolating sperm preparations from 60 infertile patients (20 with apparently normal semen and 40, abnormalsemen contaminated with microorganisms and other impurities) was evaluated and compared. The microorganisms to beremoved included bacteria, vires, Chlamydia trachomaticum, Ureaplsama urealyticum, etc. The spouse pregnancyrates in IUI with sperm preparations isolated by these three techniques from 80 oligoasthenoteratospermic patients werealso compared. Results: The quality (including the percentages of normal form, normal-chromatin and motilesperm, and the grade of motility) of sperm obtained by the real-time sperm separation technique was much higher ( P< 0.01) as compared with those by the other two methods. The Wang's tube method was also more effective in remov-ing microorganisms and other impurities. The method provided a higher IUI pregnancy rate than the other two spermseparation techniques ( P < 0.05). Conclusion: The real-time sperm separation technique is the most effectivemethod so far available in isolating high-quality sperm samples to be used in assisted reproduction.展开更多
OBJECTIVE To investigate the compatibility of the prescription Rhodiolarosea,Cordycepsmilitaris,and Rhubarb root.METHODS The DIO mice accompanying with insulin resistance,hypercholesterolemia and obesity were induced ...OBJECTIVE To investigate the compatibility of the prescription Rhodiolarosea,Cordycepsmilitaris,and Rhubarb root.METHODS The DIO mice accompanying with insulin resistance,hypercholesterolemia and obesity were induced by high fat diet(HFD)in C57BL/6mice.For evaluating the insulin sensitivity in vivo,the values of serum insulin,HOMA-IR,and the glucose infuse rate(GIR)in hyperinsulinemiceuglycemic clamp test were determined;the time-dependent blood glucose changes were measured after insulin loading in insulin tolerance test(ITT),or after glucose loading in glucose tolerance test(GTT),respectively.Recombinant human protein tyrosine phosphatase 1B(PTP1B)was overexpressed by hGST-PTP1BBL21 E.coli and purified by GST affinity chromatography.The activities of PTP1 Band glutamine:fructose 6-phosphate amidotransferase(GFAT)were measured by the pNPP and GDH method,respectively.The activity ofα-glucosidase was determined with the substrates of p-nitrophenyl-alpha-D-glucopyranoside(pNPG).The IC50 value was calculated using the software GraphPad Prism 5.RESULTS 1st,the standard for the raw materials and for the extraction process of Rhodiolarosea,Cordycepsmilitaris,and Rhubarbroot are formulated according to the inhibition on PTP1 B,α-glucosidase,and GFAT in vitro,and the improvement on impaired glucose tolerance(IGT),hypercholesterolemia,and obesity in vivo,respectively.2nd,the composition was determined by the synergistic effects of Rhodiolarosea,Cordycepsmilitaris,and Rhubarb root on insulin resistance in DIO mice.3rd,in accordance with the IC50 values from the L9(34)and L8(27)orthogonal experiments targeting on PTP1 B and α-glucosidase,respectively,the best rate of Rhodiolarosea∶Cordycepsmilitaris∶Rhubarbroot was selected.4th,the effective dose for the treatment of metabolic syndrome was assessed based on the effects on IGT,hypercholesterolemia,obesity,insulin resistance,respectively,in DIO mice.CONCLUSION The prescription compatibility of Rhodiolarosea,Cordycepsmilitaris,and Rhubarbroot with the rate of 20∶1∶1was formed to treat the metabolic syndrome,such as IGT,hypercholesterolemia,and obesity,with the effective dose of 200 mg·kg-1 by the major mechanism of the improvement on insulin sensitivity.Monarch drug,Rhodiola,can clear away the lung-heat,tonify Qi,resolve stasis and nourish the heart.Adjuvant drug,Cordycepsmilitaris,can tonify the lung Qi and the kidney essence,strong waist and knee,compatibility with Rhodiola to enhance the function of invigorating lung and kidney.Assistant drug,Rhubarb,can clear heat,detoxify,remove blood stasis.These three herbal are compatible to show the effects of tonifying Qi,nourishing essence,cleaning heat,reducing phlegm and resolving masses for the treatment of metabolic syndrome.展开更多
A modified ant colony algorithm for solving optimization problem with continuous parameters is presented. In the method, groups of candidate values of the components are constructed, and each value in the group has it...A modified ant colony algorithm for solving optimization problem with continuous parameters is presented. In the method, groups of candidate values of the components are constructed, and each value in the group has its trail information. In each iteration of the ant colony algorithm, the method first chooses initial values of the components using the trail information. Then GA operations of crossover and mutation can determine the values of the components in the solution. Our experimental results on the problem of nonlinear programming show that our method has a much higher convergence speed and stability than those of simulated annealing (SA) and GA.展开更多
基金the National Natural Science Foundation of China,No.39900069,No.30170428Natural Science Foundation of Guangdong Province,No.990799,No.010431+2 种基金College Natural Science Foundation of Guangdong province,No.200033Medical Scientific Foundation of Guangdong Province,No.A2001419Research and Development Foundation of Shantou University,No.L0004,No.L00012
文摘AIM: To separate and identify differentially expressed nuclear matrix proteins (NMPs) between the immortalized human esophageal epithelial cell line (SHEE) and the malignantly transformed esophageal carcinoma cell line (SHEEC), and to provide new ways for finding specific markers and the pathogenesis of esophageal carcinoma.METHODS: SHEE and SHEEC cell lines were used to extract NMPs. The quality of NMPs was monitored by Western blot analysis including DNA topoisomerase Ⅱα, proliferation cell nuclear antigen (PCNA) and histone. NMPs of SHEE and SHEEC were analyzed by two-dimensional electrophoresis (2-DE), silver staining and PDQuest6.2 image analysis software. Three spots in which the differentially expressed NMlPs were more obvious, were selected and analyzed with matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI- TOF-MS) and database search.RESULTS: Western blot analysis revealed that DNA topoisomerase Ⅱα and PCNA were detected, and the majority of histones were deleted in NMPs of SHEE and SHEEC. After 2-DE image analysis by PDQuest6.2 software, the 2-DE maps were detected with an average of 106±7.1 spots in SHEE and 132±5.0 spots in SHEEC. Most of them were matched one another (r=0.72), only 16 protein spots were found differing in intensity. Three NMPs including cytoskeletal tropomyosin,FK506bindingprotein6,similartoretinoblastoma binding protein 8 were preliminarily identified by MALDI- TOF-MS.CONCLUSION: These differentially expressed NMPs may play an important role during malignant transformation from SHEE to SHEEC. Their separation and identification will contribute to searching for specific markers and probing into the pathogenesis of esophageal carcinoma.
基金Project suppocted by the Youth Science Research Foundation of the Department of Public Health,Guangdong Province(No.B199121)
文摘Aim: To evaluate the comparative effectiveness of real-time sperm separation technique (Wang's tube method) andother two conventional methods in isolating high-quality sperm preparation, and to compare the spouse pregnancy ratein intrauterine insemination (IUI) with sperm preparations isolated by these methods. Methods: The effectivenessof the real-time sperm separation technique, the conventional swim-up and the Percoll discontinuous density gradientmethods in isolating sperm preparations from 60 infertile patients (20 with apparently normal semen and 40, abnormalsemen contaminated with microorganisms and other impurities) was evaluated and compared. The microorganisms to beremoved included bacteria, vires, Chlamydia trachomaticum, Ureaplsama urealyticum, etc. The spouse pregnancyrates in IUI with sperm preparations isolated by these three techniques from 80 oligoasthenoteratospermic patients werealso compared. Results: The quality (including the percentages of normal form, normal-chromatin and motilesperm, and the grade of motility) of sperm obtained by the real-time sperm separation technique was much higher ( P< 0.01) as compared with those by the other two methods. The Wang's tube method was also more effective in remov-ing microorganisms and other impurities. The method provided a higher IUI pregnancy rate than the other two spermseparation techniques ( P < 0.05). Conclusion: The real-time sperm separation technique is the most effectivemethod so far available in isolating high-quality sperm samples to be used in assisted reproduction.
基金The project supported by National Major Special Project on New Drug Innovation of China(2009ZX09103-4322012ZX09103-101-063+1 种基金2012ZX09301002-004)We also appreciate the support of Novo Nordisk Union Diabetes Research Talent Fund
文摘OBJECTIVE To investigate the compatibility of the prescription Rhodiolarosea,Cordycepsmilitaris,and Rhubarb root.METHODS The DIO mice accompanying with insulin resistance,hypercholesterolemia and obesity were induced by high fat diet(HFD)in C57BL/6mice.For evaluating the insulin sensitivity in vivo,the values of serum insulin,HOMA-IR,and the glucose infuse rate(GIR)in hyperinsulinemiceuglycemic clamp test were determined;the time-dependent blood glucose changes were measured after insulin loading in insulin tolerance test(ITT),or after glucose loading in glucose tolerance test(GTT),respectively.Recombinant human protein tyrosine phosphatase 1B(PTP1B)was overexpressed by hGST-PTP1BBL21 E.coli and purified by GST affinity chromatography.The activities of PTP1 Band glutamine:fructose 6-phosphate amidotransferase(GFAT)were measured by the pNPP and GDH method,respectively.The activity ofα-glucosidase was determined with the substrates of p-nitrophenyl-alpha-D-glucopyranoside(pNPG).The IC50 value was calculated using the software GraphPad Prism 5.RESULTS 1st,the standard for the raw materials and for the extraction process of Rhodiolarosea,Cordycepsmilitaris,and Rhubarbroot are formulated according to the inhibition on PTP1 B,α-glucosidase,and GFAT in vitro,and the improvement on impaired glucose tolerance(IGT),hypercholesterolemia,and obesity in vivo,respectively.2nd,the composition was determined by the synergistic effects of Rhodiolarosea,Cordycepsmilitaris,and Rhubarb root on insulin resistance in DIO mice.3rd,in accordance with the IC50 values from the L9(34)and L8(27)orthogonal experiments targeting on PTP1 B and α-glucosidase,respectively,the best rate of Rhodiolarosea∶Cordycepsmilitaris∶Rhubarbroot was selected.4th,the effective dose for the treatment of metabolic syndrome was assessed based on the effects on IGT,hypercholesterolemia,obesity,insulin resistance,respectively,in DIO mice.CONCLUSION The prescription compatibility of Rhodiolarosea,Cordycepsmilitaris,and Rhubarbroot with the rate of 20∶1∶1was formed to treat the metabolic syndrome,such as IGT,hypercholesterolemia,and obesity,with the effective dose of 200 mg·kg-1 by the major mechanism of the improvement on insulin sensitivity.Monarch drug,Rhodiola,can clear away the lung-heat,tonify Qi,resolve stasis and nourish the heart.Adjuvant drug,Cordycepsmilitaris,can tonify the lung Qi and the kidney essence,strong waist and knee,compatibility with Rhodiola to enhance the function of invigorating lung and kidney.Assistant drug,Rhubarb,can clear heat,detoxify,remove blood stasis.These three herbal are compatible to show the effects of tonifying Qi,nourishing essence,cleaning heat,reducing phlegm and resolving masses for the treatment of metabolic syndrome.
基金This research was supported in part Chinese National Science Foundation under contract 60074013,Chinese National Foundation of High Performance Computing under contract 00219 and Science Foundation of Jiangsu Educational Commission,China
文摘A modified ant colony algorithm for solving optimization problem with continuous parameters is presented. In the method, groups of candidate values of the components are constructed, and each value in the group has its trail information. In each iteration of the ant colony algorithm, the method first chooses initial values of the components using the trail information. Then GA operations of crossover and mutation can determine the values of the components in the solution. Our experimental results on the problem of nonlinear programming show that our method has a much higher convergence speed and stability than those of simulated annealing (SA) and GA.
