AIM: To determine whether or not a low dose of HB vaccine can be effectively used in the rapid vaccination.METHODS: Rapid vaccination (0, 1, 2 months) with low dose (5 μg) or routine dose (10 μg) HB vaccine was stud...AIM: To determine whether or not a low dose of HB vaccine can be effectively used in the rapid vaccination.METHODS: Rapid vaccination (0, 1, 2 months) with low dose (5 μg) or routine dose (10 μg) HB vaccine was studied in 250 subjects (130 school children and 120 university students). Serum from all the participants was tested for HBsAg, anti-HBs and anti-HBc at 1, 3 and 7 months after the first dose of vaccination and all subjects were serum HBV marks negative before the vaccination. Non-responders to a complete initial vaccination from university students were given an additional vaccination with 10 μg of HB vaccine and their serum anti-HBs was tested again one month later.RESULTS: One month after the third dose of vaccination (third month) sero-conversion rates and geometric mean titer (GMTs) were significantly (P＜0.01) higher in the routine dose (resp. 89 % and 106.8) than in the low dose group (resp. 72 % and 59.5). Sero-conversion rates and GMTs were maintained stable for another 4 months in both groups.After an additional vaccination to non-responders with 10 μg HB vaccine, 17/23 subjects (13/15 from those vaccinated with 5 μg vaccine and 4/8 from those vaccinated with 10 μg vaccine) became anti-HBs positive, yielding similar seroconversion rates for both dose groups.CONCLUSION: Higher sero-conversion rates and GMTs were reached in those vaccinated with 10 μg HB vaccine than in those vaccinated with 5 μg HB vaccine after a comp^te vaccination with a 0, 1, 2 month scheme. But the subjects vaccinated with 5 μg vaccine can also reach the similar sero-conversion rate after an additional vaccination.展开更多
AIM: To study the effect of siRNA expressed from DNA vector on HBV replication.METHODS: Human U6 promoter was amplified from genomic DNA and cloned into plasmid pUC18 to construct a mammalian siRNA expression vector p...AIM: To study the effect of siRNA expressed from DNA vector on HBV replication.METHODS: Human U6 promoter was amplified from genomic DNA and cloned into plasmid pUC18 to construct a mammalian siRNA expression vector pUC18U6. Then oligonucleotides coding for a short hairpin RNA against HBV were cloned into pUC18U6 to form pUC18U6HBVsir which was introduced into 2.2.15 cells by using liposome-mediated transfection.2.2.15 cells transfected by pUC18U6 and pUC18U6GFPsir which expressed siRNA against green fluorescent protein and mock-transfected 2.2.15 cells were used as controls.Concentration of HBsAg in the supernatant of the transfected cells was measured by using solid-phase radioimmunoassay.RESULTS: A mammalian siRNA expression vector pUC18U6 was constructed successfully. Compared with controls,pUC18U6HBVsir which expressed siRNA against HBV decreased concentration of HBsAg significantly by 44%(P<0.05).CONCLUSION: HBV replication in 2.2.15 cells is inhibited by siRNA expressed from the DNA vector.展开更多
AIM: To study the distribution pattern of transcription factors NF-kB and AP-1 and their relations with the expression of apoptosis associated-proteins Fas/FasL and ICH-1L/S in human hepatocellular carcinoma (HCC). ME...AIM: To study the distribution pattern of transcription factors NF-kB and AP-1 and their relations with the expression of apoptosis associated-proteins Fas/FasL and ICH-1L/S in human hepatocellular carcinoma (HCC). METHODS: We performed in situ hybridization and immunohistochemical techniques for NF-kB, AP-1, Fas/FasL and ICH-1 in 40 cases of human HCC along with corresponding nontumoral tissues and 7 cases of normal liver tissues. RESULTS: Twenty-two (55%) and 25 (62.5%) of 40 cases for NF-κB and AP-1 were presented for nuclear or both nuclear and cytoplastic staining respectively, while less cases were presented for only cytoplastic staining for NF-κB (18%) and AP-1 (10%) in adjacent nontumoral tissues and negative staining in normal liver tissues. There was no statistically significant difference of NF-κB or AP-1 activation between well differentiated tumors and poorly differentiated tumors (P>0.05). NF-κkB activity is positively corresponded to AP-1 activation. The expression of ICH-1L/S was associated with the activation of NF-κB and AP-1 (P<0.05), but no significant relationship was found between Fas/FasL and NF-κB or AP-l(P>0.05). CONCLUSION: Activation of both NF-κB and AP-1 may be required for ICH-1L/S-induced apoptosis in HCC, but not for Fas/FasL-mediated apoptosis. NF-κB and AP-1 may play important roles in the pathogenesis of human HCC.展开更多
AIM: To study the effect of short hairpin RNAs (shRNAs)expressed from DNA vector on hTERT expression.METHODS: Oligonucleotides coding for four shRNAs against hTERT were cloned into a mammalian shRNA expressionvector p...AIM: To study the effect of short hairpin RNAs (shRNAs)expressed from DNA vector on hTERT expression.METHODS: Oligonucleotides coding for four shRNAs against hTERT were cloned into a mammalian shRNA expressionvector pUC18U6 to form pUC18U6ht1-4, which were thenintroduced into HepG2 cells by using liposome-mediated transfection. HepG2 cells transfected by pUC18U6 and pUC18U6GFPsir, which expressed shRNA against green fluorescent protein (GFP), were used as controls. hTERT mRNA in the transfected cells were quantified by using real-time fluorescent RT-PCR.RESULTS: Among the four shRNAs against hTERT, two decreased the hTERT mRNA level. Compared with the controls, pUC18U6ht which expressed the two shRNAs reduced hTERT mRNA by 39% and 49% (P＜0.05).CONCLUSION: hTERT expression is inhibited by the shRNAs expressed from the DNA vector.展开更多
AIM:To evaluate if HB vaccination can yield a booster effect on the anti-HBs level of those naturally acquired HBV positive markers.METHODS:Sera were collected from 1399 newly enrolled university students aged between...AIM:To evaluate if HB vaccination can yield a booster effect on the anti-HBs level of those naturally acquired HBV positive markers.METHODS:Sera were collected from 1399 newly enrolled university students aged between 18-20 years at the entrance medical examination in 2001.Forty-four students (28 males and 16 females) with positive serum anti-HBs and anti-HBc markers served as an observation group and another 44 students (24 males and 20 females) without any HBV markers as the control. HB vaccination was given to all the students without positive serum HBsAg according to 0,1, 6 month regimen and the peripheral venous blood was sampled from those of both observation and control groups for anti-HBs detection one month after the second and third doses.Anti-HBs levels were measured by ELISA.RESULTS:The seroconversion rate of anti-HBs in the control group was 100% after the second dose,but the geometric mean titers (GMTs) were low.The tendency of serum anti-HBs changes after the 3^rd dose was completely different between the two groups.Although more than half of those with positive anti-HBs and anti-HBc showed a mild increase of anti-HBs levels after the 2na boosting dose (mean antiHBs level was 320:198 mIU),but the increase of serum anti-HBs titer was much smaller than that in the control group.The averages of their initial serum anti-HBs levels and the levels after the 2^nd and 3^rd doses were 198, 320 and 275 mIU respectively. All the subjects from the control group had an obvious increase in their serum anti-HBs levels which was nearly 4 times the baseline level (302:78 mIU).CONCLUSION: HB vaccination can not enhance anti-HBs levels in those with positive serum anti-HBs and anti-HBc markers.展开更多
文摘AIM: To determine whether or not a low dose of HB vaccine can be effectively used in the rapid vaccination.METHODS: Rapid vaccination (0, 1, 2 months) with low dose (5 μg) or routine dose (10 μg) HB vaccine was studied in 250 subjects (130 school children and 120 university students). Serum from all the participants was tested for HBsAg, anti-HBs and anti-HBc at 1, 3 and 7 months after the first dose of vaccination and all subjects were serum HBV marks negative before the vaccination. Non-responders to a complete initial vaccination from university students were given an additional vaccination with 10 μg of HB vaccine and their serum anti-HBs was tested again one month later.RESULTS: One month after the third dose of vaccination (third month) sero-conversion rates and geometric mean titer (GMTs) were significantly (P＜0.01) higher in the routine dose (resp. 89 % and 106.8) than in the low dose group (resp. 72 % and 59.5). Sero-conversion rates and GMTs were maintained stable for another 4 months in both groups.After an additional vaccination to non-responders with 10 μg HB vaccine, 17/23 subjects (13/15 from those vaccinated with 5 μg vaccine and 4/8 from those vaccinated with 10 μg vaccine) became anti-HBs positive, yielding similar seroconversion rates for both dose groups.CONCLUSION: Higher sero-conversion rates and GMTs were reached in those vaccinated with 10 μg HB vaccine than in those vaccinated with 5 μg HB vaccine after a comp^te vaccination with a 0, 1, 2 month scheme. But the subjects vaccinated with 5 μg vaccine can also reach the similar sero-conversion rate after an additional vaccination.
基金Supported by National Natural Science Foundation of China,No.30100157Medical Research Fund of Chinese PLA,No.01MA184and Innovation Project of FMMU,No.CX99005
文摘AIM: To study the effect of siRNA expressed from DNA vector on HBV replication.METHODS: Human U6 promoter was amplified from genomic DNA and cloned into plasmid pUC18 to construct a mammalian siRNA expression vector pUC18U6. Then oligonucleotides coding for a short hairpin RNA against HBV were cloned into pUC18U6 to form pUC18U6HBVsir which was introduced into 2.2.15 cells by using liposome-mediated transfection.2.2.15 cells transfected by pUC18U6 and pUC18U6GFPsir which expressed siRNA against green fluorescent protein and mock-transfected 2.2.15 cells were used as controls.Concentration of HBsAg in the supernatant of the transfected cells was measured by using solid-phase radioimmunoassay.RESULTS: A mammalian siRNA expression vector pUC18U6 was constructed successfully. Compared with controls,pUC18U6HBVsir which expressed siRNA against HBV decreased concentration of HBsAg significantly by 44%(P<0.05).CONCLUSION: HBV replication in 2.2.15 cells is inhibited by siRNA expressed from the DNA vector.
文摘AIM: To study the distribution pattern of transcription factors NF-kB and AP-1 and their relations with the expression of apoptosis associated-proteins Fas/FasL and ICH-1L/S in human hepatocellular carcinoma (HCC). METHODS: We performed in situ hybridization and immunohistochemical techniques for NF-kB, AP-1, Fas/FasL and ICH-1 in 40 cases of human HCC along with corresponding nontumoral tissues and 7 cases of normal liver tissues. RESULTS: Twenty-two (55%) and 25 (62.5%) of 40 cases for NF-κB and AP-1 were presented for nuclear or both nuclear and cytoplastic staining respectively, while less cases were presented for only cytoplastic staining for NF-κB (18%) and AP-1 (10%) in adjacent nontumoral tissues and negative staining in normal liver tissues. There was no statistically significant difference of NF-κB or AP-1 activation between well differentiated tumors and poorly differentiated tumors (P>0.05). NF-κkB activity is positively corresponded to AP-1 activation. The expression of ICH-1L/S was associated with the activation of NF-κB and AP-1 (P<0.05), but no significant relationship was found between Fas/FasL and NF-κB or AP-l(P>0.05). CONCLUSION: Activation of both NF-κB and AP-1 may be required for ICH-1L/S-induced apoptosis in HCC, but not for Fas/FasL-mediated apoptosis. NF-κB and AP-1 may play important roles in the pathogenesis of human HCC.
基金Supported by the Shaanxi Province Natural Science Foundation,No. 2003C217
文摘AIM: To study the effect of short hairpin RNAs (shRNAs)expressed from DNA vector on hTERT expression.METHODS: Oligonucleotides coding for four shRNAs against hTERT were cloned into a mammalian shRNA expressionvector pUC18U6 to form pUC18U6ht1-4, which were thenintroduced into HepG2 cells by using liposome-mediated transfection. HepG2 cells transfected by pUC18U6 and pUC18U6GFPsir, which expressed shRNA against green fluorescent protein (GFP), were used as controls. hTERT mRNA in the transfected cells were quantified by using real-time fluorescent RT-PCR.RESULTS: Among the four shRNAs against hTERT, two decreased the hTERT mRNA level. Compared with the controls, pUC18U6ht which expressed the two shRNAs reduced hTERT mRNA by 39% and 49% (P＜0.05).CONCLUSION: hTERT expression is inhibited by the shRNAs expressed from the DNA vector.
文摘AIM:To evaluate if HB vaccination can yield a booster effect on the anti-HBs level of those naturally acquired HBV positive markers.METHODS:Sera were collected from 1399 newly enrolled university students aged between 18-20 years at the entrance medical examination in 2001.Forty-four students (28 males and 16 females) with positive serum anti-HBs and anti-HBc markers served as an observation group and another 44 students (24 males and 20 females) without any HBV markers as the control. HB vaccination was given to all the students without positive serum HBsAg according to 0,1, 6 month regimen and the peripheral venous blood was sampled from those of both observation and control groups for anti-HBs detection one month after the second and third doses.Anti-HBs levels were measured by ELISA.RESULTS:The seroconversion rate of anti-HBs in the control group was 100% after the second dose,but the geometric mean titers (GMTs) were low.The tendency of serum anti-HBs changes after the 3^rd dose was completely different between the two groups.Although more than half of those with positive anti-HBs and anti-HBc showed a mild increase of anti-HBs levels after the 2na boosting dose (mean antiHBs level was 320:198 mIU),but the increase of serum anti-HBs titer was much smaller than that in the control group.The averages of their initial serum anti-HBs levels and the levels after the 2^nd and 3^rd doses were 198, 320 and 275 mIU respectively. All the subjects from the control group had an obvious increase in their serum anti-HBs levels which was nearly 4 times the baseline level (302:78 mIU).CONCLUSION: HB vaccination can not enhance anti-HBs levels in those with positive serum anti-HBs and anti-HBc markers.
