目的探究黄芪注射液对含糖乳酸盐腹膜透析液所致水孔蛋白1表达异常的影响。方法选取70只SD雄性大鼠,分成7组,分别注射不同浓度的黄芪注射液及乳酸盐腹膜透析液,留取引流液及提取腹膜样本进行有关AQP1表达及AQP1 m RNA检测。结果高糖高...目的探究黄芪注射液对含糖乳酸盐腹膜透析液所致水孔蛋白1表达异常的影响。方法选取70只SD雄性大鼠,分成7组,分别注射不同浓度的黄芪注射液及乳酸盐腹膜透析液,留取引流液及提取腹膜样本进行有关AQP1表达及AQP1 m RNA检测。结果高糖高黄芪组、高糖组BUN清除率低于对照组,差异有统计学意义(P<0.05)。高糖高黄芪组AQP1表达(11.01±1.76)低于对照组(13.12±2.02),差异有统计学意义(P<0.05);低糖高黄芪组AQP1 m RNA表达高于低糖组与高糖低黄芪组,差异有统计学意义(P<0.05)。结论适量浓度的黄芪注射液对AQP1表达异常影响不明确。展开更多
Polygalacturonase (PG,EC3.2.1.15) is the key cell wall hydrolase in fruit ripening. The identification and characterization of a full length cDNA (pMT18) encoding for PG from Feicheng peach (Prunus persica (L.) Bat...Polygalacturonase (PG,EC3.2.1.15) is the key cell wall hydrolase in fruit ripening. The identification and characterization of a full length cDNA (pMT18) encoding for PG from Feicheng peach (Prunus persica (L.) Batsch cv. Feicheng) is described. The pMT18 clone is 1188 bp in length, with an open reading frame of 393 amino acids. The homology and phylogenetic analyses indicate a remarkable similarity between peach PG and other ripening related PG. And seven consensus sequences have revealed in peach PG compared to the PG from other plants. However, the profound divergence with other PG and the unique structure features suggest that peach PG probably belongs to a new evolutionary class. In RT PCR analysis, pMT18 related RNA was undetectable in leaves, and was much abundant in ripe fruits. The ripening specific expression pattern of this cDNA will be useful in investigating the roles of PG in fruit ripening and developing a transgenic peach with the improved post harvesting quality in the future.展开更多
Extracellular polymeric substances （EPS） produced by acidophilic bioleaching microorganisms play an important role in the production of acid mine drainage and metal sulfide bioleaching. EPS mediate the contact betwe...Extracellular polymeric substances （EPS） produced by acidophilic bioleaching microorganisms play an important role in the production of acid mine drainage and metal sulfide bioleaching. EPS mediate the contact between microbial cells and growth substrates, having a pivotal role in organic film formation and bacterium-substratum interactions. The production and chemical composition of EPS produced by seven bioleaching strains grown with different substrates were studied. Analysis of the EPS extracted from these strains indicated that the EPS consisted of carbohydrates, proteins and galacturonic acid. The contents of EPS, carbohydrates, proteins and galacturonic acid of EPS were largely related to the kind of strain used and culture condition. The results show that EPS productions of microbes grown with pyrite were significantly higher than those of microbes grown with sulfur or FeSO4·7H2O. The highest EPS production of the seven acidiphilic strains was （159.43±3.93） mg/g, which was produced by Leptospirillum ferriphilum CBCBSUCSU208015 when cultivated with pyrite.展开更多
[Objective] The research aimed to construct the fusion protein expression vector of α-galactosidase-EGFP （enhanced green fluorescent protein） in cucumber controlled by CaMV35S promoter.[Method] CaMV35S promoter seq...[Objective] The research aimed to construct the fusion protein expression vector of α-galactosidase-EGFP （enhanced green fluorescent protein） in cucumber controlled by CaMV35S promoter.[Method] CaMV35S promoter sequence and the coding region of EGFP were amplified by polymerase chain reactions （PCR） with vector pCambia 1303 as the template.Using reverse transcript PCR technology,with total RNAs of cucumber as template,the coding region of acid α-galactosidase Ⅰ in cucumber was amplified.The above three fragments were inserted into the multiple cloning sites of expression vector pCambia 1381c.The fusion expression vector of α-galactosidase-EGFP located at the C-terminal of the target genes was constructed.[Result] After enzyme digestion and sequencing,the fusion expression of α-galactosidase-EGFP in cucumber was constructed successfully.[Conclusion] The research laid the experimental basis for further study on the subcellular localization of α-galactosidase in cucumber.展开更多
Objective To investigate the role of poly-lactic acid and agarose gelatin in promoting the functional recovery of the injured spinal cord. Methods Poly-lactic acid （PLA） or agarose was embedded in the space between ...Objective To investigate the role of poly-lactic acid and agarose gelatin in promoting the functional recovery of the injured spinal cord. Methods Poly-lactic acid （PLA） or agarose was embedded in the space between two stumps of the hemisectioned spinal cord. Immunohistochemistry was used to show astroglia proliferation and the infiltration of RhoA-positive cells. Locomotor activity recovery was evaluated by testing the function of hindlimbs. Results Astroglias and RhoA labeled non-neuronal cells accumulated in the area adjacent to the implant, while the number of RhoA-posirive cells was decreased dramatically in the absence of implant. Animals implanted with agarose gelatin recovered more quickly than those with PLA, concomitant with a higher survival rate of the neurons. Conclusion Both PLA and agarose gelatin benefited the recovery of spinal cord after injury by providing a scaffold for astroglia processes. Modulation of the rigidity, pore size and inner structure of PLA and agarose gelatin might make these biodegradable materials more effective in the regeneration of the central nervous system （CNS）.展开更多
AIM： To investigate the effect of Lactobacillus bulgaricus （LBG） on the Toll-like receptor 4 （TLR4） pathway and interleukin-8 （IL-8） production in SGC-7901 cells treated with Helicobacter pyloriSydney strain 1 ...AIM： To investigate the effect of Lactobacillus bulgaricus （LBG） on the Toll-like receptor 4 （TLR4） pathway and interleukin-8 （IL-8） production in SGC-7901 cells treated with Helicobacter pyloriSydney strain 1 lipopolysaccharide （HpyloriSS1-LPS）. METHODS： SGC-7901 cells were treated with HpyloriSS1-LPS in the presence or absence of pretreatment for 1 h with viable LBG or supernatant recovered from LBG culture MRS broth （LBG-s）. Cellular lysates were prepared for Western blot with anti-TLR4, anti-transforming growth factor β-activated kinase 1 （TAK1）, anti-phospho-TAK1, anti-nuclear factor κB （NF-κB）, anti-p38 mitogen-activated protein kinase （p38MAPK）, and anti-phospho-p38MAPK antibodies. The amount of IL-8 in cell culture medium was measured by ELISA. RESULTS： H pyloriSS1-LPS up-regulated the expression of TLR4, stimulated the phosphorylation of TAKI, subsequently enhanced the activation of NF- κB and the phosphorylation of p38MAPK in a time- dependent manner, leading to augmentation of IL-8 production in SGC-7901 cells. Viable LBG or LBG-s pretreatment attenuated the expression of TLR4, inhibited the phosphorylation of TAK1 and p38MAPK, prevented the activation of NF-κB, and consequently blocked IL-8 production.CONCLUSION： H py/oriSS1-LPS induces IL-8 production through activating TLR4 signaling in SGC-7901 cells and viable LBG or LBG-s prevents H pyloriSS1-LPS-mediated IL-8 production via inhibition of the TLR4 pathway.展开更多
AIM： To analyze the distinct immune responses induced by Lactobacillus peptidoglycan （PG）. METHODS： BALB/c mice were intraperitoneally injected with PG once a day for three consecutive days, Peritoneal macrophage ...AIM： To analyze the distinct immune responses induced by Lactobacillus peptidoglycan （PG）. METHODS： BALB/c mice were intraperitoneally injected with PG once a day for three consecutive days, Peritoneal macrophage and splenocyte mRNA was extracted and the gene expression profile was studied using high-density oligonudeotide microarrays. Inhibitory effects of Lactobacillus PG on colon tumor tissue were studied in vitro and in vivo, RESULTS： The gene expression profiles revealed that the TLR-NF-kB and Jak-STAT signaling pathways were highly activated. An inflammatory phenotype was induced when peritoneal macrophages were initially exposed to Lactobacillus PG and switched to a more complex phenotype when BALB/c mice were treated with three doses of Lactobacillus PG. A protective physiological inflammatory response was induced after three consecutive days of PG treatment. It was tending toward Thl dominant immune response. Lactobacillus PG also appeared to induce a significant in vivo anti-colon tumor effect. CONCLUSION： Lactobacillus PG is responsible for certain immune responses induced by Lactobacilli. Anti-tumor effects of Lactobacilliare likely to attribute to the activation of macrophages by PG expressed on the bacterial cell surface.展开更多
Base excision repair （BER） is an evolutionarily conserved process for maintaining genomic integrity by eliminating several dozen damaged （oxidized or aikylated） or inappropriate bases that are generated endogenous...Base excision repair （BER） is an evolutionarily conserved process for maintaining genomic integrity by eliminating several dozen damaged （oxidized or aikylated） or inappropriate bases that are generated endogenously or induced by genotoxicants, predominantly, reactive oxygen species （ROS）. BER involves 4-5 steps starting with base excision by a DNA glycosylase, followed by a common pathway usually involving an AP-endonuclease （APE） to generate 3＇ OH terminus at the damage site, followed by repair synthesis with a DNA polymerase and nick sealing by a DNA iigase. This pathway is also responsible for repairing DNA single-strand breaks with blocked termini directly generated by ROS. Nearly all glycosylases, far fewer than their substrate lesions particularly for oxidized bases, have broad and overlapping substrate range, and could serve as back-up enzymes in vivo. In contrast, mammalian cells encode only one APE, APEI, unlike two APEs in lower organisms. In spite of overall similarity, BER with distinct subpathways in the mammals is more complex than in E. coli. The glycosylases form complexes with downstream proteins to carry out efficient repair via distinct subpathways one of which, responsible for repair of strand breaks with 3＇ phosphate termini generated by the NEIL family glycosylases or by ROS, requires the phosphatase activity of polynucleotide kinase instead of APE1. Different complexes may utilize distinct DNA polymerases and iigases. Mammalian glycosylases have nonconserved extensions at one of the termini, dispensable for enzymatic activity but needed for interaction with other BER and non-BER proteins for complex formation and organeile targeting. The mammalian enzymes are sometimes covalently modified which may affect activity and complex formation. The focus of this review is on the early steps in mammalian BER for oxidized damage.展开更多
文摘目的探究黄芪注射液对含糖乳酸盐腹膜透析液所致水孔蛋白1表达异常的影响。方法选取70只SD雄性大鼠,分成7组,分别注射不同浓度的黄芪注射液及乳酸盐腹膜透析液,留取引流液及提取腹膜样本进行有关AQP1表达及AQP1 m RNA检测。结果高糖高黄芪组、高糖组BUN清除率低于对照组,差异有统计学意义(P<0.05)。高糖高黄芪组AQP1表达(11.01±1.76)低于对照组(13.12±2.02),差异有统计学意义(P<0.05);低糖高黄芪组AQP1 m RNA表达高于低糖组与高糖低黄芪组,差异有统计学意义(P<0.05)。结论适量浓度的黄芪注射液对AQP1表达异常影响不明确。
文摘Polygalacturonase (PG,EC3.2.1.15) is the key cell wall hydrolase in fruit ripening. The identification and characterization of a full length cDNA (pMT18) encoding for PG from Feicheng peach (Prunus persica (L.) Batsch cv. Feicheng) is described. The pMT18 clone is 1188 bp in length, with an open reading frame of 393 amino acids. The homology and phylogenetic analyses indicate a remarkable similarity between peach PG and other ripening related PG. And seven consensus sequences have revealed in peach PG compared to the PG from other plants. However, the profound divergence with other PG and the unique structure features suggest that peach PG probably belongs to a new evolutionary class. In RT PCR analysis, pMT18 related RNA was undetectable in leaves, and was much abundant in ripe fruits. The ripening specific expression pattern of this cDNA will be useful in investigating the roles of PG in fruit ripening and developing a transgenic peach with the improved post harvesting quality in the future.
文摘Extracellular polymeric substances （EPS） produced by acidophilic bioleaching microorganisms play an important role in the production of acid mine drainage and metal sulfide bioleaching. EPS mediate the contact between microbial cells and growth substrates, having a pivotal role in organic film formation and bacterium-substratum interactions. The production and chemical composition of EPS produced by seven bioleaching strains grown with different substrates were studied. Analysis of the EPS extracted from these strains indicated that the EPS consisted of carbohydrates, proteins and galacturonic acid. The contents of EPS, carbohydrates, proteins and galacturonic acid of EPS were largely related to the kind of strain used and culture condition. The results show that EPS productions of microbes grown with pyrite were significantly higher than those of microbes grown with sulfur or FeSO4·7H2O. The highest EPS production of the seven acidiphilic strains was （159.43±3.93） mg/g, which was produced by Leptospirillum ferriphilum CBCBSUCSU208015 when cultivated with pyrite.
基金Supported by National Basic Research Program of China( 2009CB119000)National Natural Science Foundation(30871721)~~
文摘[Objective] The research aimed to construct the fusion protein expression vector of α-galactosidase-EGFP （enhanced green fluorescent protein） in cucumber controlled by CaMV35S promoter.[Method] CaMV35S promoter sequence and the coding region of EGFP were amplified by polymerase chain reactions （PCR） with vector pCambia 1303 as the template.Using reverse transcript PCR technology,with total RNAs of cucumber as template,the coding region of acid α-galactosidase Ⅰ in cucumber was amplified.The above three fragments were inserted into the multiple cloning sites of expression vector pCambia 1381c.The fusion expression vector of α-galactosidase-EGFP located at the C-terminal of the target genes was constructed.[Result] After enzyme digestion and sequencing,the fusion expression of α-galactosidase-EGFP in cucumber was constructed successfully.[Conclusion] The research laid the experimental basis for further study on the subcellular localization of α-galactosidase in cucumber.
文摘Objective To investigate the role of poly-lactic acid and agarose gelatin in promoting the functional recovery of the injured spinal cord. Methods Poly-lactic acid （PLA） or agarose was embedded in the space between two stumps of the hemisectioned spinal cord. Immunohistochemistry was used to show astroglia proliferation and the infiltration of RhoA-positive cells. Locomotor activity recovery was evaluated by testing the function of hindlimbs. Results Astroglias and RhoA labeled non-neuronal cells accumulated in the area adjacent to the implant, while the number of RhoA-posirive cells was decreased dramatically in the absence of implant. Animals implanted with agarose gelatin recovered more quickly than those with PLA, concomitant with a higher survival rate of the neurons. Conclusion Both PLA and agarose gelatin benefited the recovery of spinal cord after injury by providing a scaffold for astroglia processes. Modulation of the rigidity, pore size and inner structure of PLA and agarose gelatin might make these biodegradable materials more effective in the regeneration of the central nervous system （CNS）.
基金Basic Research Project of Science and Technology Offi ce of Sichuan Province, No. 04JY029-090-1
文摘AIM： To investigate the effect of Lactobacillus bulgaricus （LBG） on the Toll-like receptor 4 （TLR4） pathway and interleukin-8 （IL-8） production in SGC-7901 cells treated with Helicobacter pyloriSydney strain 1 lipopolysaccharide （HpyloriSS1-LPS）. METHODS： SGC-7901 cells were treated with HpyloriSS1-LPS in the presence or absence of pretreatment for 1 h with viable LBG or supernatant recovered from LBG culture MRS broth （LBG-s）. Cellular lysates were prepared for Western blot with anti-TLR4, anti-transforming growth factor β-activated kinase 1 （TAK1）, anti-phospho-TAK1, anti-nuclear factor κB （NF-κB）, anti-p38 mitogen-activated protein kinase （p38MAPK）, and anti-phospho-p38MAPK antibodies. The amount of IL-8 in cell culture medium was measured by ELISA. RESULTS： H pyloriSS1-LPS up-regulated the expression of TLR4, stimulated the phosphorylation of TAKI, subsequently enhanced the activation of NF- κB and the phosphorylation of p38MAPK in a time- dependent manner, leading to augmentation of IL-8 production in SGC-7901 cells. Viable LBG or LBG-s pretreatment attenuated the expression of TLR4, inhibited the phosphorylation of TAK1 and p38MAPK, prevented the activation of NF-κB, and consequently blocked IL-8 production.CONCLUSION： H py/oriSS1-LPS induces IL-8 production through activating TLR4 signaling in SGC-7901 cells and viable LBG or LBG-s prevents H pyloriSS1-LPS-mediated IL-8 production via inhibition of the TLR4 pathway.
基金Supported by the PhD Programs Foundation of Ministry of Education of China, No. 20040295005
文摘AIM： To analyze the distinct immune responses induced by Lactobacillus peptidoglycan （PG）. METHODS： BALB/c mice were intraperitoneally injected with PG once a day for three consecutive days, Peritoneal macrophage and splenocyte mRNA was extracted and the gene expression profile was studied using high-density oligonudeotide microarrays. Inhibitory effects of Lactobacillus PG on colon tumor tissue were studied in vitro and in vivo, RESULTS： The gene expression profiles revealed that the TLR-NF-kB and Jak-STAT signaling pathways were highly activated. An inflammatory phenotype was induced when peritoneal macrophages were initially exposed to Lactobacillus PG and switched to a more complex phenotype when BALB/c mice were treated with three doses of Lactobacillus PG. A protective physiological inflammatory response was induced after three consecutive days of PG treatment. It was tending toward Thl dominant immune response. Lactobacillus PG also appeared to induce a significant in vivo anti-colon tumor effect. CONCLUSION： Lactobacillus PG is responsible for certain immune responses induced by Lactobacilli. Anti-tumor effects of Lactobacilliare likely to attribute to the activation of macrophages by PG expressed on the bacterial cell surface.
文摘Base excision repair （BER） is an evolutionarily conserved process for maintaining genomic integrity by eliminating several dozen damaged （oxidized or aikylated） or inappropriate bases that are generated endogenously or induced by genotoxicants, predominantly, reactive oxygen species （ROS）. BER involves 4-5 steps starting with base excision by a DNA glycosylase, followed by a common pathway usually involving an AP-endonuclease （APE） to generate 3＇ OH terminus at the damage site, followed by repair synthesis with a DNA polymerase and nick sealing by a DNA iigase. This pathway is also responsible for repairing DNA single-strand breaks with blocked termini directly generated by ROS. Nearly all glycosylases, far fewer than their substrate lesions particularly for oxidized bases, have broad and overlapping substrate range, and could serve as back-up enzymes in vivo. In contrast, mammalian cells encode only one APE, APEI, unlike two APEs in lower organisms. In spite of overall similarity, BER with distinct subpathways in the mammals is more complex than in E. coli. The glycosylases form complexes with downstream proteins to carry out efficient repair via distinct subpathways one of which, responsible for repair of strand breaks with 3＇ phosphate termini generated by the NEIL family glycosylases or by ROS, requires the phosphatase activity of polynucleotide kinase instead of APE1. Different complexes may utilize distinct DNA polymerases and iigases. Mammalian glycosylases have nonconserved extensions at one of the termini, dispensable for enzymatic activity but needed for interaction with other BER and non-BER proteins for complex formation and organeile targeting. The mammalian enzymes are sometimes covalently modified which may affect activity and complex formation. The focus of this review is on the early steps in mammalian BER for oxidized damage.
