Objective:To build GPC3 gene short hairpin interference RNA(shRNA)slow virus veclor.observe expression of Huh-7 GPC3 gene in human liver cell line proliferation apoptosis and the effect of GPC3 gene influencing on liv...Objective:To build GPC3 gene short hairpin interference RNA(shRNA)slow virus veclor.observe expression of Huh-7 GPC3 gene in human liver cell line proliferation apoptosis and the effect of GPC3 gene influencing on liver cancer cell growth,and provide theoretical basis for genc therapy of liver cancer.Methods:Hepatocellular carcinoma cell line Huh-7 wsa transfected by a RNA interference technique.GPC3 gene expression in a variety of liver cancer cell lines was detected by fluorescence quantitative PCR.Targeted GPC3 gene seqnences of small interfering RNA(siRNA)PGC-shRNA-GPC3 were restructured.Stable expression cell linse of siRNA were screened and established with the heplp of liposomes(lipofectamine^(TM2000))as carrier transfcetion of human liver cell lines.In order to validate siRNA interference efficiency.GPC3 siRNA mRNA expression was detected after transfection by using RT-PCR and Western blot.The absorbance value of the cells of blank group,untransfection group and transfection group,the cell cycle and cell apoptosis were calculated,and effects of GPC3 gene nn Huh-7 cell proliferation and apoptosis were observed.Results:In the liver cancer cell lines Huh-7 GPC3 gene showed high expression.PGC-shRNA-GPC3 recombinant plasmid was constructde successfully via sequencing validation.Stable recombinant plasmid transfected into liver cancer cell linse Huh-7can obviously inhibit GPC3 mRNA expression level.Conclusions:The targeted GPC3 siRNA can effectively inhibit the expression of GPC3.展开更多
Anion starch nanoparticle (StNP) with a diameter of 50 nm was prepared in wa- ter-in-oil microemulsion, with soluble starch as raw materials and POCl3 as crosslinking agent. PLL-StNP was prepared by linking poly-L-lys...Anion starch nanoparticle (StNP) with a diameter of 50 nm was prepared in wa- ter-in-oil microemulsion, with soluble starch as raw materials and POCl3 as crosslinking agent. PLL-StNP was prepared by linking poly-L-lysine (PLL) on the surface of StNP. At the same time, the size of PLL-StNP and its stability in aqueous solution were checked by AFM. The analysis of plasmid DNA binding, DNase I enzymatic degradation, toxicity and transfection were done. We discovered that PLL-StNP may be used as non-virus nanoparticle gene carrier. And we devel- oped the method of preparing PLL-StNP gene carrier and used it in cell transfection. As non-virus gene carrier, PLL-StNP has some advantages, such as large load of DNA, high transfection effi- ciency, low cell toxicity and biodegradability.展开更多
Hepatitis B virus(HBV) reactivation(HBVr) in patients undergoing immunosuppressive therapy is still a hot topic worldwide. Its prevention and management still represents a challenge for specialists dealing with immuno...Hepatitis B virus(HBV) reactivation(HBVr) in patients undergoing immunosuppressive therapy is still a hot topic worldwide. Its prevention and management still represents a challenge for specialists dealing with immunosuppressed patients. Aim of this paper is to provide a critical review of the relevant information emerged in the recent literature regarding HBV reactivation following immunosuppressive treatments for oncohematological tumors. A computerized literature search in MEDLINE was performed using appropriate terms arrangement, including English-written literature only or additional relevant articles. Articles published only in abstract form and case reports not giving considerable news were excluded. Clinical manifestation of HBVr can be manifold, ranging from asymptomatic self-limiting anicteric hepatitis to life-threatening fulminant liver failure. In clusters of patients adverse outcomes are potentially predictable. Clinicians should be aware of the inherent risk of HBVr among the different virological categories(active carriers, occult HBV carriers and inactive carriers, the most intriguing category), and classes of immunosuppressive drugs. We recommend that patients undergoing immunosuppressive treatments for hematological malignancies should undergo HBV screening. In case of serological sign(s) of current or past infection with the virus, appropriate therapeutic or preventive strategies are suggested, according to both virological categories, risk of HBVr by immunosuppressive drugsand liver status. Either antiviral drug management and surveillance and pre-emptive approach are examined, commenting the current international recommendations about this debated issue.展开更多
【目的】采用 Meta 分析评价他克莫司与环孢素 A 治疗乙型肝炎表面抗原（HBsAg ）阳性患者肾移植术后排斥反应的疗效和安全性。【方法】检索中国期刊全文数据库（CNKI）、维普数据库、万方数据库和中国生物医学文献数据库（检索时间从...【目的】采用 Meta 分析评价他克莫司与环孢素 A 治疗乙型肝炎表面抗原（HBsAg ）阳性患者肾移植术后排斥反应的疗效和安全性。【方法】检索中国期刊全文数据库（CNKI）、维普数据库、万方数据库和中国生物医学文献数据库（检索时间从建库至2014年12月），纳入比较他克莫司与环孢素 A 治疗 HBsAg 阳性患者肾移植术后排斥反应的随机对照试验，其中研究组给予他克莫司，对照组给予环孢素 A ，采用 Rev Man 5．1软件对最终纳入文献的研究结果进行 Meta 分析。【结果】共纳入四项随机对照试验，合计306例患者。 Meta 分析结果显示：观察组存活率高于对照组[RR ＝1．07，95％ CI（1．00，1．14），P ＝0．05]；肝功能异常发生率低于对照组[RR ＝0．35，95％ CI（0．23，0．56），P ＜ 0．01]，但两组急性排斥反应[OR ＝0．51，95％ CI（0．23，1．16），P ＝0．11]和患者肾脏存活率[OR ＝2．74，95％ CI（0．77，9．79），P ＝0．12]差异无统计学意义。【结论】HBsAg 阳性患者肾移植后给予他克莫司抗排斥免疫抑制治疗的患者存活率和肝功能异常发生率均优于给予环孢素 A 治疗的患者。展开更多
Hepatitis B surface antigen prevalence in populations of blood donors in Cameroon is estimated at 6%-16%. As such, the Objective of this study was to determine the prevalence of Hepatitis B surface antigen in a popula...Hepatitis B surface antigen prevalence in populations of blood donors in Cameroon is estimated at 6%-16%. As such, the Objective of this study was to determine the prevalence of Hepatitis B surface antigen in a population of bank employees, who represent a sample closer to the general population. In tests carried out both Yaounde and Douala in December 2011, we detected the hepatitis B surface antigen among 267 workers, including 151 men and 116 women of a median age of 37 years. The Results obtained revealed that the overall prevalence stood at 7.1%, with a 9.9% prevalence ratio in men, three times higher than that in women (3.4%). Subjects aged 20-29 years and senior staffs were more affected (each of them recording 8.1%). Potential nosocomial risks were detected mostly at the level of dental care (52.8%). Among the other risks of exposure, scarification (53.6%) was more frequent. A univariate analysis revealed an insignificant relationship with tattoos (OR 2.6, 95% CI 0.5-10.8, p = 0.2). These results led to the Conclusion that Hepatitis B surface antigen carrier prevalence in a population of workers in Cameroon is not different from that of blood donors. It is mainly due to early vertical and horizontal contamination.展开更多
Objective： To explore Chinese medicine （CM） syndrome distribution of chronic hepatitis B virus （HBV） carriers in immunotolerant phase （ITP）. Methods： One hundred and eighty-five chronic HBV carriers in ITP, se...Objective： To explore Chinese medicine （CM） syndrome distribution of chronic hepatitis B virus （HBV） carriers in immunotolerant phase （ITP）. Methods： One hundred and eighty-five chronic HBV carriers in ITP, seen in the Third Affiliated Hospital of Sun Yat-sen University from May 2009 to December 2010, were admitted in an observational study under the guidance of CM. Patients＇ CM symptoms and signs, demographics, liver biochemistries, and qualitative HBV DNA were recorded in the questionnaires. CM syndromes were then differentiated to 15 detailed types and analyzed by generalization. Lastly, the location, pathogenic factors and nature of the disease were also assessed. Results： When CM syndrome patterns were differentiated to 15 types, there were 27 （15%） no syndrome cases, 94 （50%） single syndrome cases and 64 （35%） compound syndromes cases. The main detailed syndromes included Liver （Gan）-qi depression （LQD）, Kidney （Shen）-qi deficiency （KQD）, Spleen （Pi）-qi deficiency （SQD） and Kidney-yang deficiency （KYAD）. After CM syndromes generalized to five types, their frequency was Spleen-Kidney deficiency （SKD）〉LQD〉inner dampness-heat retention （IDHR）〉Liver-Kidney deficiency （LKD）〉blood stasis blocking collateral （BSBC）. SKD and LQD occupied 64%. The disease location included Liver, Gallbladder （Dan）, Spleen, Stomach （Wei） and Kidney. The pathogenic factors were mainly qi stagnation, qi deficiency, yang deficiency, concurrently dampness-heat and blood stasis. The deficiency syndrome was more than excess syndrome in its nature. Conclusions： Most of chronic HBV carriers in ITP have their CM syndrome, and the most common types are SKAD, LQD. This study suggests that the natural history may be improved through breaking the state of immune tolerance or shorten the time of ITP by strengthening Spleen-Kidney and reliving Liver qi.展开更多
目的探讨以多聚赖氨酸为载体的聚合物标记物在丙型肝炎病毒(Hepatitis C virus,HCV)核心抗原检测中的应用。方法以多聚赖氨酸作为载体,将HCV核心抗原单克隆抗体与辣根过氧化物酶(HRP)偶联形成聚合物标记物,同时制备过碘酸钠标记物,比较...目的探讨以多聚赖氨酸为载体的聚合物标记物在丙型肝炎病毒(Hepatitis C virus,HCV)核心抗原检测中的应用。方法以多聚赖氨酸作为载体,将HCV核心抗原单克隆抗体与辣根过氧化物酶(HRP)偶联形成聚合物标记物,同时制备过碘酸钠标记物,比较两种标记物中的抗体相对含量及抗体反应性;将两种标记物分别应用于ELISA和Western blot法检测HCV核心抗原中,比较二者的检测结果,并与市售试剂盒进行比较。结果通过竞争抑制和直接结合试验结果显示,当达到50%抑制率时,聚合物标记物和过碘酸钠标记物的游离抗体浓度分别为0.0019和0.0011mg/ml;当A403值为1.0时,聚合物标记物的稀释度为1:200000左右,而传统标记物为1:20000左右。聚合物标记物和过碘酸钠标记物应用于ELISA法,检测HCV核心抗原的最低检测限分别为2.0和10pg/ml,试验内、试验间变异系数均<10%,特异性良好,临床血清标本检测结果与市售试剂盒的符合率分别为97.0%和98.6%;在Western blot检测中,与过碘酸钠标记物相比,聚合物标记物至少可以提高检测灵敏度10倍。结论以多聚赖氨酸为载体的聚合物标记物可应用于HCV核心抗原的ELISA和Western blot检测,且能提高酶、抗体的比例,进而提高检测的灵敏度,防止漏检。展开更多
基金supported by Wuhan Municipal Science and Technology Bureau of applied basic research project(No.2013062301010823)Wuhan City health planning medieal research project(No.WX14A11)
文摘Objective:To build GPC3 gene short hairpin interference RNA(shRNA)slow virus veclor.observe expression of Huh-7 GPC3 gene in human liver cell line proliferation apoptosis and the effect of GPC3 gene influencing on liver cancer cell growth,and provide theoretical basis for genc therapy of liver cancer.Methods:Hepatocellular carcinoma cell line Huh-7 wsa transfected by a RNA interference technique.GPC3 gene expression in a variety of liver cancer cell lines was detected by fluorescence quantitative PCR.Targeted GPC3 gene seqnences of small interfering RNA(siRNA)PGC-shRNA-GPC3 were restructured.Stable expression cell linse of siRNA were screened and established with the heplp of liposomes(lipofectamine^(TM2000))as carrier transfcetion of human liver cell lines.In order to validate siRNA interference efficiency.GPC3 siRNA mRNA expression was detected after transfection by using RT-PCR and Western blot.The absorbance value of the cells of blank group,untransfection group and transfection group,the cell cycle and cell apoptosis were calculated,and effects of GPC3 gene nn Huh-7 cell proliferation and apoptosis were observed.Results:In the liver cancer cell lines Huh-7 GPC3 gene showed high expression.PGC-shRNA-GPC3 recombinant plasmid was constructde successfully via sequencing validation.Stable recombinant plasmid transfected into liver cancer cell linse Huh-7can obviously inhibit GPC3 mRNA expression level.Conclusions:The targeted GPC3 siRNA can effectively inhibit the expression of GPC3.
文摘Anion starch nanoparticle (StNP) with a diameter of 50 nm was prepared in wa- ter-in-oil microemulsion, with soluble starch as raw materials and POCl3 as crosslinking agent. PLL-StNP was prepared by linking poly-L-lysine (PLL) on the surface of StNP. At the same time, the size of PLL-StNP and its stability in aqueous solution were checked by AFM. The analysis of plasmid DNA binding, DNase I enzymatic degradation, toxicity and transfection were done. We discovered that PLL-StNP may be used as non-virus nanoparticle gene carrier. And we devel- oped the method of preparing PLL-StNP gene carrier and used it in cell transfection. As non-virus gene carrier, PLL-StNP has some advantages, such as large load of DNA, high transfection effi- ciency, low cell toxicity and biodegradability.
文摘Hepatitis B virus(HBV) reactivation(HBVr) in patients undergoing immunosuppressive therapy is still a hot topic worldwide. Its prevention and management still represents a challenge for specialists dealing with immunosuppressed patients. Aim of this paper is to provide a critical review of the relevant information emerged in the recent literature regarding HBV reactivation following immunosuppressive treatments for oncohematological tumors. A computerized literature search in MEDLINE was performed using appropriate terms arrangement, including English-written literature only or additional relevant articles. Articles published only in abstract form and case reports not giving considerable news were excluded. Clinical manifestation of HBVr can be manifold, ranging from asymptomatic self-limiting anicteric hepatitis to life-threatening fulminant liver failure. In clusters of patients adverse outcomes are potentially predictable. Clinicians should be aware of the inherent risk of HBVr among the different virological categories(active carriers, occult HBV carriers and inactive carriers, the most intriguing category), and classes of immunosuppressive drugs. We recommend that patients undergoing immunosuppressive treatments for hematological malignancies should undergo HBV screening. In case of serological sign(s) of current or past infection with the virus, appropriate therapeutic or preventive strategies are suggested, according to both virological categories, risk of HBVr by immunosuppressive drugsand liver status. Either antiviral drug management and surveillance and pre-emptive approach are examined, commenting the current international recommendations about this debated issue.
文摘【目的】采用 Meta 分析评价他克莫司与环孢素 A 治疗乙型肝炎表面抗原（HBsAg ）阳性患者肾移植术后排斥反应的疗效和安全性。【方法】检索中国期刊全文数据库（CNKI）、维普数据库、万方数据库和中国生物医学文献数据库（检索时间从建库至2014年12月），纳入比较他克莫司与环孢素 A 治疗 HBsAg 阳性患者肾移植术后排斥反应的随机对照试验，其中研究组给予他克莫司，对照组给予环孢素 A ，采用 Rev Man 5．1软件对最终纳入文献的研究结果进行 Meta 分析。【结果】共纳入四项随机对照试验，合计306例患者。 Meta 分析结果显示：观察组存活率高于对照组[RR ＝1．07，95％ CI（1．00，1．14），P ＝0．05]；肝功能异常发生率低于对照组[RR ＝0．35，95％ CI（0．23，0．56），P ＜ 0．01]，但两组急性排斥反应[OR ＝0．51，95％ CI（0．23，1．16），P ＝0．11]和患者肾脏存活率[OR ＝2．74，95％ CI（0．77，9．79），P ＝0．12]差异无统计学意义。【结论】HBsAg 阳性患者肾移植后给予他克莫司抗排斥免疫抑制治疗的患者存活率和肝功能异常发生率均优于给予环孢素 A 治疗的患者。
文摘Hepatitis B surface antigen prevalence in populations of blood donors in Cameroon is estimated at 6%-16%. As such, the Objective of this study was to determine the prevalence of Hepatitis B surface antigen in a population of bank employees, who represent a sample closer to the general population. In tests carried out both Yaounde and Douala in December 2011, we detected the hepatitis B surface antigen among 267 workers, including 151 men and 116 women of a median age of 37 years. The Results obtained revealed that the overall prevalence stood at 7.1%, with a 9.9% prevalence ratio in men, three times higher than that in women (3.4%). Subjects aged 20-29 years and senior staffs were more affected (each of them recording 8.1%). Potential nosocomial risks were detected mostly at the level of dental care (52.8%). Among the other risks of exposure, scarification (53.6%) was more frequent. A univariate analysis revealed an insignificant relationship with tattoos (OR 2.6, 95% CI 0.5-10.8, p = 0.2). These results led to the Conclusion that Hepatitis B surface antigen carrier prevalence in a population of workers in Cameroon is not different from that of blood donors. It is mainly due to early vertical and horizontal contamination.
基金Supported by Grants from the National Major Projects for AIDS and Viral Hepatitis of China(No.2008ZX10005-008)
文摘Objective： To explore Chinese medicine （CM） syndrome distribution of chronic hepatitis B virus （HBV） carriers in immunotolerant phase （ITP）. Methods： One hundred and eighty-five chronic HBV carriers in ITP, seen in the Third Affiliated Hospital of Sun Yat-sen University from May 2009 to December 2010, were admitted in an observational study under the guidance of CM. Patients＇ CM symptoms and signs, demographics, liver biochemistries, and qualitative HBV DNA were recorded in the questionnaires. CM syndromes were then differentiated to 15 detailed types and analyzed by generalization. Lastly, the location, pathogenic factors and nature of the disease were also assessed. Results： When CM syndrome patterns were differentiated to 15 types, there were 27 （15%） no syndrome cases, 94 （50%） single syndrome cases and 64 （35%） compound syndromes cases. The main detailed syndromes included Liver （Gan）-qi depression （LQD）, Kidney （Shen）-qi deficiency （KQD）, Spleen （Pi）-qi deficiency （SQD） and Kidney-yang deficiency （KYAD）. After CM syndromes generalized to five types, their frequency was Spleen-Kidney deficiency （SKD）〉LQD〉inner dampness-heat retention （IDHR）〉Liver-Kidney deficiency （LKD）〉blood stasis blocking collateral （BSBC）. SKD and LQD occupied 64%. The disease location included Liver, Gallbladder （Dan）, Spleen, Stomach （Wei） and Kidney. The pathogenic factors were mainly qi stagnation, qi deficiency, yang deficiency, concurrently dampness-heat and blood stasis. The deficiency syndrome was more than excess syndrome in its nature. Conclusions： Most of chronic HBV carriers in ITP have their CM syndrome, and the most common types are SKAD, LQD. This study suggests that the natural history may be improved through breaking the state of immune tolerance or shorten the time of ITP by strengthening Spleen-Kidney and reliving Liver qi.
