BACKGROUND: 10-23 DNA enzyme is one kind of de-oxyribozymes for RNA cleavage. The inhibition effects of 10-23 DNA enzyme on the expression of the HBV C gene in HepG2. 2. 15 cells were demonstrated previously. The aim ...BACKGROUND: 10-23 DNA enzyme is one kind of de-oxyribozymes for RNA cleavage. The inhibition effects of 10-23 DNA enzyme on the expression of the HBV C gene in HepG2. 2. 15 cells were demonstrated previously. The aim of this study was to further explore the cleavage activities of 10-23 DNA enzyme targeting at HBV C gene mRNA in vitro. METHODS: 10-23 DNA enzyme named Drz-HBV-C-9 specific to HBV C gene ORF A1816UG was designed and synthesized. HBV C gene mRNA was obtained by the in vitro transcription method. Cleavage activities of Drz-HBV-C-9 were observed in vitro. Values of kinetic parameters including Km,Kcat and Kcat/Km were calculated accordingly. RESULTS: Under the certain cleavage conditions, Drz-HBV-C-9 could efficiently cleave target mRNA at specific sites in vitro. Cleavage products of 109nt plus 191nt were obtained. The kinetic parameters, Km,Kcat and Kcat/ Km for Drz-HBV-C-9, were 1.4 ×10-9 mol, 1.6 min-1 and 1.1 × 109 mol-1 · min-1, respectively. CONCLUSIONS: 10-23 DNA enzyme targeting at HBV C gene mRNA possesses specific cleavage activities in vitro. This would be a potent antiviral strategy with respect to HBV gene therapy.展开更多
AIM: To analyze loss of heterozygosity (LOH) and homozygous deletion on p53 gene (exon2-3, 4 and 11), chromosome 10q22-10q23 and 22q11.2 -22q12.1 in human hepatocellular carcinoma (HCC).METHODS: PCR and PCR-based micr...AIM: To analyze loss of heterozygosity (LOH) and homozygous deletion on p53 gene (exon2-3, 4 and 11), chromosome 10q22-10q23 and 22q11.2 -22q12.1 in human hepatocellular carcinoma (HCC).METHODS: PCR and PCR-based microsatellite polymorphism analysis techniques were used.RESULTS: LOH was observed at D10S579 (10q22-10q23) in 4 of 20 tumors (20%), at D22S421 (22q11.2-22q12.1) in 3 of 20(15%), at TP53.A (p53gene exon 2-3) in 4 of 20 (20%), at TP53.B (p53gene exon 4) in 6 of 20(30%), and at TP53.G (p53gene exon 11)in 0 of 20(0%). Homozygous deletion was detected at 10q22-10q23(8/20; 40%), 22q11.2-22q12.1(8/20; 40%), p53 gene exon 2-3(0/20;0%), p53gene exon 4(6/20; 30%), and p53gene exon 11(2/20; 10%).CONCLUSION: There might be unidentified tumor suppressor genes on chromosome 10q22-10q23 and 22q11.2-22q12.1 that contribute to the pathogenesis and development of HCC.展开更多
目的:连续监测"冰火两重灸"疗法对不同淋巴细胞计数值的AIDS免疫重建不良患者的白介素-23(Interleukin-23,IL-23)及白介素-10(Interleukin-10,IL-10)的影响。方法:选取云南中医药大学第三附属医院的AIDS免疫重建不良患者30例...目的:连续监测"冰火两重灸"疗法对不同淋巴细胞计数值的AIDS免疫重建不良患者的白介素-23(Interleukin-23,IL-23)及白介素-10(Interleukin-10,IL-10)的影响。方法:选取云南中医药大学第三附属医院的AIDS免疫重建不良患者30例,按淋巴细胞计数值进行分组,分为两组,运用冰火两重灸联合高效抗逆转录病毒治疗(Highly Active Antiretroviral Therapy,HAART)治疗,观察0、3、6、9、12个月的IL-23及IL-10的数值变化。结果:两组在不同测定时间的IL-10及IL-23(对数值)均数的差异均有统计学意义(P<0.05),且都有随时间推移而下降的趋势(P<0.05)。结论:在不同免疫重建不良的AIDS患者中,因淋巴细胞数值的不同,在同一联合治疗方案下,均呈现有效治疗趋势,中医治疗联合国家推荐的一线方案治疗值得长期观察。展开更多
Objective To prepare fluorescent silica nanoparticles(FSNPs) carrying 10-23 deoxyribozymes(10-23DRzs) and to evaluate their inhibitory effect on human hepatitis B virus(HBV).Methods The FSNPs were prepared by microemu...Objective To prepare fluorescent silica nanoparticles(FSNPs) carrying 10-23 deoxyribozymes(10-23DRzs) and to evaluate their inhibitory effect on human hepatitis B virus(HBV).Methods The FSNPs were prepared by microemulsion method and further modified by NaCl.Their diameter and the Zeta potential were tested.The HBV-specific 10-23DRzs were designed according to the sequences of S and C genes of HBV.The 10-23DRzs were connected to FSNPs,which constituted the FSNPs-DNA.The connecting efficiency and the protective effect of nanoparticles on 10-23DRzs were tested by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE).The HepG2.2.15 cells were transfected by FSNPs-DNA,of which the inhibitory effects on HBsAg and HBeAg were analyzed by enzyme-linked immunosorbent assay(ELISA).Results The nanoparticles were spherical and uniform in size,with a diameter of 220 nm and the surface Zeta potential of +15.2 mV.The combination of DNA and FSNPs effectively protected DNA from nuclease degradation.Transfection of FSNPs-DNA significantly inhibited the expression of HBV S and C genes compared to the liposome control group.Conclusion The FSNPs have been successfully prepared and efficiently connected to HBV-specific 10-23DRzs,which significantly inhibit the expression of HBV S and C genes in cell culture.展开更多
基金This study was supported by a grant from the National Natural Science Foundation of China (No. 30271183).
文摘BACKGROUND: 10-23 DNA enzyme is one kind of de-oxyribozymes for RNA cleavage. The inhibition effects of 10-23 DNA enzyme on the expression of the HBV C gene in HepG2. 2. 15 cells were demonstrated previously. The aim of this study was to further explore the cleavage activities of 10-23 DNA enzyme targeting at HBV C gene mRNA in vitro. METHODS: 10-23 DNA enzyme named Drz-HBV-C-9 specific to HBV C gene ORF A1816UG was designed and synthesized. HBV C gene mRNA was obtained by the in vitro transcription method. Cleavage activities of Drz-HBV-C-9 were observed in vitro. Values of kinetic parameters including Km,Kcat and Kcat/Km were calculated accordingly. RESULTS: Under the certain cleavage conditions, Drz-HBV-C-9 could efficiently cleave target mRNA at specific sites in vitro. Cleavage products of 109nt plus 191nt were obtained. The kinetic parameters, Km,Kcat and Kcat/ Km for Drz-HBV-C-9, were 1.4 ×10-9 mol, 1.6 min-1 and 1.1 × 109 mol-1 · min-1, respectively. CONCLUSIONS: 10-23 DNA enzyme targeting at HBV C gene mRNA possesses specific cleavage activities in vitro. This would be a potent antiviral strategy with respect to HBV gene therapy.
基金Supported by the Natural Science Foundation of Anhui Province,No.99044312(YW),No.01043716(SYG)and Natural Science Foundation of Anhui Educational Commission,No.JL-97-077(YW)
文摘AIM: To analyze loss of heterozygosity (LOH) and homozygous deletion on p53 gene (exon2-3, 4 and 11), chromosome 10q22-10q23 and 22q11.2 -22q12.1 in human hepatocellular carcinoma (HCC).METHODS: PCR and PCR-based microsatellite polymorphism analysis techniques were used.RESULTS: LOH was observed at D10S579 (10q22-10q23) in 4 of 20 tumors (20%), at D22S421 (22q11.2-22q12.1) in 3 of 20(15%), at TP53.A (p53gene exon 2-3) in 4 of 20 (20%), at TP53.B (p53gene exon 4) in 6 of 20(30%), and at TP53.G (p53gene exon 11)in 0 of 20(0%). Homozygous deletion was detected at 10q22-10q23(8/20; 40%), 22q11.2-22q12.1(8/20; 40%), p53 gene exon 2-3(0/20;0%), p53gene exon 4(6/20; 30%), and p53gene exon 11(2/20; 10%).CONCLUSION: There might be unidentified tumor suppressor genes on chromosome 10q22-10q23 and 22q11.2-22q12.1 that contribute to the pathogenesis and development of HCC.
文摘目的:连续监测"冰火两重灸"疗法对不同淋巴细胞计数值的AIDS免疫重建不良患者的白介素-23(Interleukin-23,IL-23)及白介素-10(Interleukin-10,IL-10)的影响。方法:选取云南中医药大学第三附属医院的AIDS免疫重建不良患者30例,按淋巴细胞计数值进行分组,分为两组,运用冰火两重灸联合高效抗逆转录病毒治疗(Highly Active Antiretroviral Therapy,HAART)治疗,观察0、3、6、9、12个月的IL-23及IL-10的数值变化。结果:两组在不同测定时间的IL-10及IL-23(对数值)均数的差异均有统计学意义(P<0.05),且都有随时间推移而下降的趋势(P<0.05)。结论:在不同免疫重建不良的AIDS患者中,因淋巴细胞数值的不同,在同一联合治疗方案下,均呈现有效治疗趋势,中医治疗联合国家推荐的一线方案治疗值得长期观察。
基金Supported by National Hi-tech Project of China(No.2007AA02-1803 and 2007AA021901)
文摘Objective To prepare fluorescent silica nanoparticles(FSNPs) carrying 10-23 deoxyribozymes(10-23DRzs) and to evaluate their inhibitory effect on human hepatitis B virus(HBV).Methods The FSNPs were prepared by microemulsion method and further modified by NaCl.Their diameter and the Zeta potential were tested.The HBV-specific 10-23DRzs were designed according to the sequences of S and C genes of HBV.The 10-23DRzs were connected to FSNPs,which constituted the FSNPs-DNA.The connecting efficiency and the protective effect of nanoparticles on 10-23DRzs were tested by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE).The HepG2.2.15 cells were transfected by FSNPs-DNA,of which the inhibitory effects on HBsAg and HBeAg were analyzed by enzyme-linked immunosorbent assay(ELISA).Results The nanoparticles were spherical and uniform in size,with a diameter of 220 nm and the surface Zeta potential of +15.2 mV.The combination of DNA and FSNPs effectively protected DNA from nuclease degradation.Transfection of FSNPs-DNA significantly inhibited the expression of HBV S and C genes compared to the liposome control group.Conclusion The FSNPs have been successfully prepared and efficiently connected to HBV-specific 10-23DRzs,which significantly inhibit the expression of HBV S and C genes in cell culture.
