Low-ion-density discontinuous polyacrylamide electrophoresis (pH 8.0) of LDH from different tissuse of four species (Tetrastes bonasia,Chrysolophus pictus, Phasians colchicus, Gallus gallus domesticus) in Galliformes...Low-ion-density discontinuous polyacrylamide electrophoresis (pH 8.0) of LDH from different tissuse of four species (Tetrastes bonasia,Chrysolophus pictus, Phasians colchicus, Gallus gallus domesticus) in Galliformes showed that this electrophoresis system was suitable for analysing avian LDH. Thermostabilty to heat and nonsusceptibility to urea inhibition of low-density urea of the four liver LDHs were in such order as LDH1> LDH2> LDH3> LDH4> LDH5. And it appeared. by comparing relative movable ratio of A and B subunits of LDHs, that difference exists between Phasianidae and Tetraonidae and in Phasianidae Phasianus was closer to Chrysolophus than to Gallus.展开更多
Objective. To study the characteristics of changes of LDH enzyme patterns of mice under slight hypoxia.Methods. Mice treated with artificial hypoxia, various tissues were made for the test of LDH enzymatic activity by...Objective. To study the characteristics of changes of LDH enzyme patterns of mice under slight hypoxia.Methods. Mice treated with artificial hypoxia, various tissues were made for the test of LDH enzymatic activity by the specific staining technique. LDH (1 -5) relative percentage enzymatic activity (RPEA) were measured with CS-910 dual-wavelength thin layer chromatography scanner.Results. The RPEA of LDH isozymes of various tissues after slight hypoxia shifted to the isozymes LDH1 and LDH2, whose principal subunits are H subunits, and the RPEA of LDH,(H4), LDH2(H3M) increased, while RPEA of LDH5(M4) in various tissues decreased prominently except the cardiac muscle, and that of LDH4(HM3) decreased as well. After polyacrylamide gel electrophoresis (PAGE) of the hypoxia treated cardiac muscle specimen was made, activity subbands originated regularly in the isozyme patterns of LDH, with the regularity of LDH1 (0 subband), LDH2 (0-1 subbands), LDH3 (0-2 subbands), LDH4 (1-3 subbands), LDH5 (2-4 subbands). After adding appropriate amount of NAD+ to the hypoxia treated cardiac muscle specimen, PAGE showed the subbands of four isozymes (LDH2-LDH5) reduced or even totally disappeared in the isozyme patterns.Conclusions. The negative feedback regulation of coenzymization and decoenzymization of LDH isozymes is one of the mouse stress responses to slight hypoxia.展开更多
目的研究敲低和过表达LDHB(lactate dehydrogenase B)对HeLa细胞生长的影响。方法根据慢病毒质粒pLn(pLL3.7-neo)特点,设计三对分别靶向LDHB c DNA 48,201,443位核苷酸序列的shRNA(short hairpin RNA)引物,退火后与pLn重组。同时,将LDHB...目的研究敲低和过表达LDHB(lactate dehydrogenase B)对HeLa细胞生长的影响。方法根据慢病毒质粒pLn(pLL3.7-neo)特点,设计三对分别靶向LDHB c DNA 48,201,443位核苷酸序列的shRNA(short hairpin RNA)引物,退火后与pLn重组。同时,将LDHB cDNA与慢病毒质粒pIp(pBOBI-IRES-puro)重组,获得的重组质粒与慢病毒包装质粒共转染293T细胞,将获得的病毒上清感染HeLa细胞,Western blot鉴定LDHB的敲低和过表达对LDHB蛋白水平的影响,并观察其对HeLa细胞生长的影响。结果靶向LDHB第48位核苷酸的shRNA显著敲低LDHB(P<0.01),443位次之,201位无敲低作用。shRNA序列对HeLa细胞生长的抑制作用与其敲低效果成正相关。转染pIp-LDHB显著升高LDHB在HeLa细胞中的表达水平(P<0.01),但不影响其生长。结论敲低LDHB抑制HeLa细胞生长,下调LDHB的表达可能有助于宫颈癌的治疗。展开更多
文摘Low-ion-density discontinuous polyacrylamide electrophoresis (pH 8.0) of LDH from different tissuse of four species (Tetrastes bonasia,Chrysolophus pictus, Phasians colchicus, Gallus gallus domesticus) in Galliformes showed that this electrophoresis system was suitable for analysing avian LDH. Thermostabilty to heat and nonsusceptibility to urea inhibition of low-density urea of the four liver LDHs were in such order as LDH1> LDH2> LDH3> LDH4> LDH5. And it appeared. by comparing relative movable ratio of A and B subunits of LDHs, that difference exists between Phasianidae and Tetraonidae and in Phasianidae Phasianus was closer to Chrysolophus than to Gallus.
文摘Objective. To study the characteristics of changes of LDH enzyme patterns of mice under slight hypoxia.Methods. Mice treated with artificial hypoxia, various tissues were made for the test of LDH enzymatic activity by the specific staining technique. LDH (1 -5) relative percentage enzymatic activity (RPEA) were measured with CS-910 dual-wavelength thin layer chromatography scanner.Results. The RPEA of LDH isozymes of various tissues after slight hypoxia shifted to the isozymes LDH1 and LDH2, whose principal subunits are H subunits, and the RPEA of LDH,(H4), LDH2(H3M) increased, while RPEA of LDH5(M4) in various tissues decreased prominently except the cardiac muscle, and that of LDH4(HM3) decreased as well. After polyacrylamide gel electrophoresis (PAGE) of the hypoxia treated cardiac muscle specimen was made, activity subbands originated regularly in the isozyme patterns of LDH, with the regularity of LDH1 (0 subband), LDH2 (0-1 subbands), LDH3 (0-2 subbands), LDH4 (1-3 subbands), LDH5 (2-4 subbands). After adding appropriate amount of NAD+ to the hypoxia treated cardiac muscle specimen, PAGE showed the subbands of four isozymes (LDH2-LDH5) reduced or even totally disappeared in the isozyme patterns.Conclusions. The negative feedback regulation of coenzymization and decoenzymization of LDH isozymes is one of the mouse stress responses to slight hypoxia.
文摘目的研究敲低和过表达LDHB(lactate dehydrogenase B)对HeLa细胞生长的影响。方法根据慢病毒质粒pLn(pLL3.7-neo)特点,设计三对分别靶向LDHB c DNA 48,201,443位核苷酸序列的shRNA(short hairpin RNA)引物,退火后与pLn重组。同时,将LDHB cDNA与慢病毒质粒pIp(pBOBI-IRES-puro)重组,获得的重组质粒与慢病毒包装质粒共转染293T细胞,将获得的病毒上清感染HeLa细胞,Western blot鉴定LDHB的敲低和过表达对LDHB蛋白水平的影响,并观察其对HeLa细胞生长的影响。结果靶向LDHB第48位核苷酸的shRNA显著敲低LDHB(P<0.01),443位次之,201位无敲低作用。shRNA序列对HeLa细胞生长的抑制作用与其敲低效果成正相关。转染pIp-LDHB显著升高LDHB在HeLa细胞中的表达水平(P<0.01),但不影响其生长。结论敲低LDHB抑制HeLa细胞生长,下调LDHB的表达可能有助于宫颈癌的治疗。
