AIM: To investigate the association between loss of heterozygosity (LOH) on chromosome 18 and sporadic gastric cancer.METHODS: Multiplex PCR was used to screen 14 highly polymorphic microsatellite markers on chromosom...AIM: To investigate the association between loss of heterozygosity (LOH) on chromosome 18 and sporadic gastric cancer.METHODS: Multiplex PCR was used to screen 14 highly polymorphic microsatellite markers on chromosome 18 in 45 cases of primary gastric cancer. PCR products were separated on polyacrylamide gels and the electrophoresis maps were analyzed with Genescan and Genotyper.RESULTS: The LOH frequencies in gastric cancer at all 14 markers ranged from 10% to 58%. Eleven markers were found with over 20% LOH frequencies, in which 9 markers located in 18q, and 2 markers in 18p. Two overlapping deleted regions were identified: R1 between D18S61-D18S1161 at 18q22 (9cM) with 24% LOH frequency; R2 between D18S462D18570 at 18q22-23(6cM) with 32% LOH frequency.CONCLUSION: LOH of chromosome 18 (18q and 18p) may be involved in gastric tumorigenesis. Two overlapping deleted fragments suggested that there might be unidentified tumor suppressor genes in those two regions.展开更多
Aim To determine the effects of glucose oi1APD, IK1, IK, ICa-L of ventricular myocytes in guinea pigs. Methods Whole-cell patch-clamp technique was used to record the changed action potential ionic current induced by ...Aim To determine the effects of glucose oi1APD, IK1, IK, ICa-L of ventricular myocytes in guinea pigs. Methods Whole-cell patch-clamp technique was used to record the changed action potential ionic current induced by glucose of single cell in guinea pig ventricular myocytes, to compare the action of 0, 10 and 20 mmol·L^-1 glucoses on transmembrane ionic current. Results (1) Compared with 10 mmol·L^-1 glucose concentrations, 0 and 20 mmol·L^-1 glucose both shortened APD of ventricular myocytes (P<0.05). (2) The inward components of IK1 density were maximal when the glucose concentration was at 10 mmol·L^-1. Normalized 1-V relationships showed that both 0 and 20 mmol·L^-1 glucose produced a left-shift of I-V curve. The reverse potential changed from-72.4 mV to-64.6 mV. (3) Compared with 10 mmol·L^-1, both 0 and 20 mmol·L^-1 glucose markedly increased the ICa-L amplitude and density. The ICa-L current density was (-8.0350.82) pA/pF (n=8) at a test potential of 10 mV when the glucose concentration was 10 mmol·L^-1. But its current density decreased to (-5.45±0.67) pA/pF and (-6.50±0.56) pA/pF when glucose concentrations were 0 and 20 mmol·L^-1, respectively. (4) The current densities of IK were (18.96±2.86) pA/pF, (8.66±1.87) pA/pF, and(15.32:1:3.12) pA/pF, at ±70mV for 0, 10 and 20 mmol·L^-1 glucoses, respectively. Conclusion Glucose in different concentrations has different effects on APD, IK1, IK, and ICa-L of single ventricular myocyte in guinea pigs. There are similar actions of 0 and 20 mmol·L^-1 glucoses on the transmembrane ionic current of ventricular myocytes in guinea pigs.展开更多
AIM To study the membrane trafficking modulation of ROMK1 channel by ubiquitination. METHODS we immunoprecipitated protein with ROMK antibody from rat renal cortex and outer medulla, and apply western blot with ubiqui...AIM To study the membrane trafficking modulation of ROMK1 channel by ubiquitination. METHODS we immunoprecipitated protein with ROMK antibody from rat renal cortex and outer medulla, and apply western blot with ubiquitin antibody to detect that ROMK protein is being ubiquitination under physiological condition in vivo. Furthermore, we used His-antibody and ubiquitin antibodies to recongnize proteins in the cotransfected HEK293 cells with hemagglutinin-ubiquitin and His- tagged ROMK1 constructs, respectively. After all intracellular lysine (Lys) residues of ROMK1 had been individually mutated to arginine (Arg), two -electrode voltage clamp was used to measure the ROMK1 channel activity in Xenopus oocytes. To detect the effect of ubiquitin on ROMK1, we used Western blotting technique to search the difference on total protein expression levelspus oocytes.展开更多
基金Supported by the Teaching and Research Award Program for Outstanding Young Teachers in Higher Education Institutions by Ministry of Education of China and the National Natural Science Foundation,No.30370783 and the Key Project of Science and Technology o
文摘AIM: To investigate the association between loss of heterozygosity (LOH) on chromosome 18 and sporadic gastric cancer.METHODS: Multiplex PCR was used to screen 14 highly polymorphic microsatellite markers on chromosome 18 in 45 cases of primary gastric cancer. PCR products were separated on polyacrylamide gels and the electrophoresis maps were analyzed with Genescan and Genotyper.RESULTS: The LOH frequencies in gastric cancer at all 14 markers ranged from 10% to 58%. Eleven markers were found with over 20% LOH frequencies, in which 9 markers located in 18q, and 2 markers in 18p. Two overlapping deleted regions were identified: R1 between D18S61-D18S1161 at 18q22 (9cM) with 24% LOH frequency; R2 between D18S462D18570 at 18q22-23(6cM) with 32% LOH frequency.CONCLUSION: LOH of chromosome 18 (18q and 18p) may be involved in gastric tumorigenesis. Two overlapping deleted fragments suggested that there might be unidentified tumor suppressor genes in those two regions.
文摘Aim To determine the effects of glucose oi1APD, IK1, IK, ICa-L of ventricular myocytes in guinea pigs. Methods Whole-cell patch-clamp technique was used to record the changed action potential ionic current induced by glucose of single cell in guinea pig ventricular myocytes, to compare the action of 0, 10 and 20 mmol·L^-1 glucoses on transmembrane ionic current. Results (1) Compared with 10 mmol·L^-1 glucose concentrations, 0 and 20 mmol·L^-1 glucose both shortened APD of ventricular myocytes (P<0.05). (2) The inward components of IK1 density were maximal when the glucose concentration was at 10 mmol·L^-1. Normalized 1-V relationships showed that both 0 and 20 mmol·L^-1 glucose produced a left-shift of I-V curve. The reverse potential changed from-72.4 mV to-64.6 mV. (3) Compared with 10 mmol·L^-1, both 0 and 20 mmol·L^-1 glucose markedly increased the ICa-L amplitude and density. The ICa-L current density was (-8.0350.82) pA/pF (n=8) at a test potential of 10 mV when the glucose concentration was 10 mmol·L^-1. But its current density decreased to (-5.45±0.67) pA/pF and (-6.50±0.56) pA/pF when glucose concentrations were 0 and 20 mmol·L^-1, respectively. (4) The current densities of IK were (18.96±2.86) pA/pF, (8.66±1.87) pA/pF, and(15.32:1:3.12) pA/pF, at ±70mV for 0, 10 and 20 mmol·L^-1 glucoses, respectively. Conclusion Glucose in different concentrations has different effects on APD, IK1, IK, and ICa-L of single ventricular myocyte in guinea pigs. There are similar actions of 0 and 20 mmol·L^-1 glucoses on the transmembrane ionic current of ventricular myocytes in guinea pigs.
文摘AIM To study the membrane trafficking modulation of ROMK1 channel by ubiquitination. METHODS we immunoprecipitated protein with ROMK antibody from rat renal cortex and outer medulla, and apply western blot with ubiquitin antibody to detect that ROMK protein is being ubiquitination under physiological condition in vivo. Furthermore, we used His-antibody and ubiquitin antibodies to recongnize proteins in the cotransfected HEK293 cells with hemagglutinin-ubiquitin and His- tagged ROMK1 constructs, respectively. After all intracellular lysine (Lys) residues of ROMK1 had been individually mutated to arginine (Arg), two -electrode voltage clamp was used to measure the ROMK1 channel activity in Xenopus oocytes. To detect the effect of ubiquitin on ROMK1, we used Western blotting technique to search the difference on total protein expression levelspus oocytes.
