Long term antiviral therapy with nucleos(t)ide analogs(NAs) may lead to the emergence of drug-resistance viral mutants in chronic hepatitis B virus(HBV) patient.The purpose of this study was to identify adefovir dipiv...Long term antiviral therapy with nucleos(t)ide analogs(NAs) may lead to the emergence of drug-resistance viral mutants in chronic hepatitis B virus(HBV) patient.The purpose of this study was to identify adefovir dipivoxil(ADV) resistance mutations of HBV polymerase and determine effective drugs to replace ADV.The reverse transcriptase(RT) coding region was PCR-amplified using HBV DNA extracted from patient blood samples and sequenced.Nineteen substitution mutations were detected.Among them,rtN238R,rtT240Y and rtN248H were often observed in patients receiving ADV administration.These three potential drug resistant sites were introduced into HBV replication-competent plasmids.The in vitro susceptibility of both wild-type(WT) and mutant-type(MT) HBV to NAs was analyzed by Southern blotting and quantitative real-time PCR.The rtN238R,rtT240Y and rtN248H substitutions had no obvious effect on HBV DNA replication or gene expression.The in vitro susceptibility analysis showed that rtN238R,rtT240Y and rtN248H substitutions were responsible for the reduced susceptibility to ADV,and demonstrated a 5.42-,2.89-and 5.72-fold increase in resistance towards ADV,respectively.However,HBV harbored these mutations retained normal susceptibility to LMV,LdT,ETV and TDF.展开更多
Herein,solid lipid nanoparticles(SLN)were proposed as a new drug delivery system for adefovir dipivoxil(ADV). The octadecylamine-fluorescein isothiocynate(ODA-FITC)was synthesized and used as a fluorescence maker to b...Herein,solid lipid nanoparticles(SLN)were proposed as a new drug delivery system for adefovir dipivoxil(ADV). The octadecylamine-fluorescein isothiocynate(ODA-FITC)was synthesized and used as a fluorescence maker to be incorporated into SLN to investigate the time-dependent cellular uptake of SLN by HepG2.2.15.The SLN of monostearin with ODA-FITC or ADV were prepared by solvent diffusion method in an aqueous system.About 15 wt%drug entrapment efficiency(EE)and 3 wt% drug loading(DL)could be reached in SLN loading ADV.Comparing with free ADV,the inhibitory effects of ADV loaded in SLN on hepatitis B surface antigen(HBsAg),hepatitis B e antigen(HBeAg)and hepatitis B virus(HBV)DNA levels in vitro were significantly enhanced.展开更多
目的了解上海市徐汇区急性呼吸道感染病毒病原学现况。方法 2012-2016年收集监测点医院流感样病例咽拭子标本,采用Real-time RT-PCR方法检测。结果对4 660份标本进行流感检测:flu A 18.80%,flu B 6.78%。对364份标本中进行7种呼吸道病毒...目的了解上海市徐汇区急性呼吸道感染病毒病原学现况。方法 2012-2016年收集监测点医院流感样病例咽拭子标本,采用Real-time RT-PCR方法检测。结果对4 660份标本进行流感检测:flu A 18.80%,flu B 6.78%。对364份标本中进行7种呼吸道病毒PCR阳性率18.68%,其中flu A+flu B 10.44%,HRV 2.75%,ADV、HBo V和PV1型均为0.82%;RSVB 0.55%,PV2、PV4均为0.27%,hm PV未检出。比较一个流行年不同监测点流感检出率无差异。2012-2016年聚集性流感样病例疫情11起,集中在冬春季,流感占90.91%,无暴发疫情。结论徐汇地区呼吸道病毒多样化,流感病毒最常见,且呈冬春季和夏季双峰流行,B型、季H3型、新H1H1型交替,夏季高峰多以季H3型为优势毒株。流感样病例就诊百分比与流感病毒检出率呈正相关。学校、托幼机构为呼吸道病毒感染聚集疫情重点防控单位。应长期监测主要呼吸道病毒活动水平。展开更多
基金supported in part by the National Basic Research Program of China(2007CB512901)
文摘Long term antiviral therapy with nucleos(t)ide analogs(NAs) may lead to the emergence of drug-resistance viral mutants in chronic hepatitis B virus(HBV) patient.The purpose of this study was to identify adefovir dipivoxil(ADV) resistance mutations of HBV polymerase and determine effective drugs to replace ADV.The reverse transcriptase(RT) coding region was PCR-amplified using HBV DNA extracted from patient blood samples and sequenced.Nineteen substitution mutations were detected.Among them,rtN238R,rtT240Y and rtN248H were often observed in patients receiving ADV administration.These three potential drug resistant sites were introduced into HBV replication-competent plasmids.The in vitro susceptibility of both wild-type(WT) and mutant-type(MT) HBV to NAs was analyzed by Southern blotting and quantitative real-time PCR.The rtN238R,rtT240Y and rtN248H substitutions had no obvious effect on HBV DNA replication or gene expression.The in vitro susceptibility analysis showed that rtN238R,rtT240Y and rtN248H substitutions were responsible for the reduced susceptibility to ADV,and demonstrated a 5.42-,2.89-and 5.72-fold increase in resistance towards ADV,respectively.However,HBV harbored these mutations retained normal susceptibility to LMV,LdT,ETV and TDF.
文摘Herein,solid lipid nanoparticles(SLN)were proposed as a new drug delivery system for adefovir dipivoxil(ADV). The octadecylamine-fluorescein isothiocynate(ODA-FITC)was synthesized and used as a fluorescence maker to be incorporated into SLN to investigate the time-dependent cellular uptake of SLN by HepG2.2.15.The SLN of monostearin with ODA-FITC or ADV were prepared by solvent diffusion method in an aqueous system.About 15 wt%drug entrapment efficiency(EE)and 3 wt% drug loading(DL)could be reached in SLN loading ADV.Comparing with free ADV,the inhibitory effects of ADV loaded in SLN on hepatitis B surface antigen(HBsAg),hepatitis B e antigen(HBeAg)and hepatitis B virus(HBV)DNA levels in vitro were significantly enhanced.
文摘目的了解上海市徐汇区急性呼吸道感染病毒病原学现况。方法 2012-2016年收集监测点医院流感样病例咽拭子标本,采用Real-time RT-PCR方法检测。结果对4 660份标本进行流感检测:flu A 18.80%,flu B 6.78%。对364份标本中进行7种呼吸道病毒PCR阳性率18.68%,其中flu A+flu B 10.44%,HRV 2.75%,ADV、HBo V和PV1型均为0.82%;RSVB 0.55%,PV2、PV4均为0.27%,hm PV未检出。比较一个流行年不同监测点流感检出率无差异。2012-2016年聚集性流感样病例疫情11起,集中在冬春季,流感占90.91%,无暴发疫情。结论徐汇地区呼吸道病毒多样化,流感病毒最常见,且呈冬春季和夏季双峰流行,B型、季H3型、新H1H1型交替,夏季高峰多以季H3型为优势毒株。流感样病例就诊百分比与流感病毒检出率呈正相关。学校、托幼机构为呼吸道病毒感染聚集疫情重点防控单位。应长期监测主要呼吸道病毒活动水平。
