PEGylation,the controlled covalent conjugation of polyethylene glycol to therapeutics,enhances therapeutic efficacy through optimized pharmacokinetics.However,to date no high-molecular-weight PEGylated small-molecule ...PEGylation,the controlled covalent conjugation of polyethylene glycol to therapeutics,enhances therapeutic efficacy through optimized pharmacokinetics.However,to date no high-molecular-weight PEGylated small-molecule prodrugs have received regulatory approval.This technological gap can be partially attributed to the exponential proliferation of metabolic intermediates resulting from multi-payload conjugation strategies,which introduces unprecedented analytical complexities in metabolite profiling and pharmacokinetic characterization.To address this challenge,we developed a liquid chromatography-triple-quadrupole/time-of-flight mass spectrometry platform for PEG20k-(irinotecan)3,a Phase III clinical candidate.Our methodology employs payload stoichiometry-based chromatographic resolution for clustering isomeric PEG species.Complementarily,diagnostic product ions at m/z 699.83,569.27,and 587.28 enable systematic differentiation between double-loaded,single-loaded,and released irinotecan payload.This approach successfully identifies eight metabolic clusters spanning from PEG-conjugates,cleaved PEG segments,and released small-molecule species.Its demonstrated capacity to deconvolute complex metabolic profiles—through payload-stoichiometry based chromatographic resolution coupled with diagnostic ion analysis—positions this workflow as an attractive tool for accelerating the development of PEGylated small-molecule therapeutics.展开更多
目的比较硫酸镁钠钾口服液(OSS)与复方聚乙二醇电解质散(PEG)在结肠镜检查前肠道准备中的安全性和有效性。方法回顾性分析2023月9月-2024年9月该院采用OSS与PEG在肠镜检查行前肠道准备的患者2000例。根据不同肠道准备药物,将患者分为OSS...目的比较硫酸镁钠钾口服液(OSS)与复方聚乙二醇电解质散(PEG)在结肠镜检查前肠道准备中的安全性和有效性。方法回顾性分析2023月9月-2024年9月该院采用OSS与PEG在肠镜检查行前肠道准备的患者2000例。根据不同肠道准备药物,将患者分为OSS组(1000例,口服OSS)和PEG组(1000例,口服4 L PEG)。比较两组患者肠道清洁度、清洁效果、肠腔气泡、结肠镜检查阳性率和不良反应发生率。结果OSS组肠道准备成功率为92.5%(925/1000),PEG组肠道准备成功率为91.7%(917/1000),两组患者比较,差异无统计学意义(P>0.05);两组患者左侧结肠波士顿肠道准备量表(BBPS)评分比较,差异无统计学意义(P>0.05),OSS组BBPS总分,以及中段和右侧结肠BBPS评分明显高于PEG组,差异均有统计学意义(P<0.05);OSS组气泡评估满意率为96.2%,高于PEG组的90.3%,差异有统计学意义(P<0.05);两组患者结肠镜检查阳性率和不良反应发生率比较,差异均无统计学意义(P>0.05)。结论OSS在肠道准备方面具有较好的去泡效果,清洁效果与PEG相当,且有较高的安全性。展开更多
DNA结合单锌指蛋白(DNA binding with one finger,Dof)是植物中特有的一类转录因子,参与调控植物组织分化、种子萌发、植物抗逆及代谢等生理生化过程。为鉴定并分析高粱Dof转录因子家族成员,利用生物信息学方法从全基因组水平对高粱Dof...DNA结合单锌指蛋白(DNA binding with one finger,Dof)是植物中特有的一类转录因子,参与调控植物组织分化、种子萌发、植物抗逆及代谢等生理生化过程。为鉴定并分析高粱Dof转录因子家族成员,利用生物信息学方法从全基因组水平对高粱Dof转录因子家族成员的基本基因结构、理化性质、保守基序、启动子顺式作用元件、组织特异性表达及干旱胁迫表达等进行分析。20%PEG6000模拟干旱胁迫处理豫粱10号的幼苗后,利用qPCR技术分析Dof基因家族成员响应不同时间段干旱胁迫的表达情况。结果共鉴定了30个高粱Dof基因,分布在9条染色体上,基因保守性分析结果显示,Motif1(即锌指结构)是高粱Dof基因家族中保守的蛋白质序列;其中29个基因中都含有CX2CX21CX2C保守基序,而SbDof15基因保守序列为CX20CX2C;启动子结构分析表明,Dof基因的启动子区域中含有ABA、MYB、MeJA、低温等逆境胁迫响应元件;进化树分析结果表明,高粱Dof蛋白与玉米的同源性更高,热图分析表明Dof家族成员在高粱的不同部位都有表达。qRT-PCR结果表明,高粱Dof基因家族成员可能参与高粱应对干旱胁迫的响应,不同的Dof基因成员在应对干旱胁迫时的调控机制可能不一样。展开更多
Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene g...Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene glycol(PEG),to immediately fuse closely apposed open ends of severed proximal and distal axons in rat sciatic nerves.We have previously reported that sciatic nerve axons repaired by PEG-fusion do not undergo Wallerian degeneration,and PEG-fused animals exhibit rapid(within 2–6 weeks)and extensive locomotor recovery.Furthermore,our previous report showed that PEG-fusion of severed sciatic motor axons was non-specific,i.e.,spinal motoneurons in PEG-fused animals were found to project to appropriate as well as inappropriate target muscles.In this study,we examined the consequences of PEG-fusion for sensory axons of the sciatic nerve.Young adult male and female rats(Sprague–Dawley)received either a unilateral single cut or ablation injury to the sciatic nerve and subsequent repair with or without(Negative Control)the application of PEG.Compound action potentials recorded immediately after PEG-fusion repair confirmed conduction across the injury site.The success of PEG-fusion was confirmed through Sciatic Functional Index testing with PEG-fused animals showing improvement in locomotor function beginning at 35 days postoperatively.At 2–42 days postoperatively,we anterogradely labeled sensory afferents from the dorsal aspect of the hindpaw following bilateral intradermal injection of wheat germ agglutinin conjugated horseradish peroxidase.PEG-fusion repair reestablished axonal continuity.Compared to unoperated animals,labeled sensory afferents ipsilateral to the injury in PEG-fused animals were found in the appropriate area of the dorsal horn,as well as inappropriate mediolateral and rostrocaudal areas.Unexpectedly,despite having intact peripheral nerves,similar reorganizations of labeled sensory afferents were also observed contralateral to the injury and repair.This central reorganization may contribute to the improved behavioral recovery seen after PEG-fusion repair,supporting the use of this novel repair methodology over currently available treatments.展开更多
基金support from the National Natural Science Foundation of China(Nos.82030107,82304443,82373944)the Hainan Provincial Natural Science Foundation of China(No.825QN288).
文摘PEGylation,the controlled covalent conjugation of polyethylene glycol to therapeutics,enhances therapeutic efficacy through optimized pharmacokinetics.However,to date no high-molecular-weight PEGylated small-molecule prodrugs have received regulatory approval.This technological gap can be partially attributed to the exponential proliferation of metabolic intermediates resulting from multi-payload conjugation strategies,which introduces unprecedented analytical complexities in metabolite profiling and pharmacokinetic characterization.To address this challenge,we developed a liquid chromatography-triple-quadrupole/time-of-flight mass spectrometry platform for PEG20k-(irinotecan)3,a Phase III clinical candidate.Our methodology employs payload stoichiometry-based chromatographic resolution for clustering isomeric PEG species.Complementarily,diagnostic product ions at m/z 699.83,569.27,and 587.28 enable systematic differentiation between double-loaded,single-loaded,and released irinotecan payload.This approach successfully identifies eight metabolic clusters spanning from PEG-conjugates,cleaved PEG segments,and released small-molecule species.Its demonstrated capacity to deconvolute complex metabolic profiles—through payload-stoichiometry based chromatographic resolution coupled with diagnostic ion analysis—positions this workflow as an attractive tool for accelerating the development of PEGylated small-molecule therapeutics.
文摘目的比较硫酸镁钠钾口服液(OSS)与复方聚乙二醇电解质散(PEG)在结肠镜检查前肠道准备中的安全性和有效性。方法回顾性分析2023月9月-2024年9月该院采用OSS与PEG在肠镜检查行前肠道准备的患者2000例。根据不同肠道准备药物,将患者分为OSS组(1000例,口服OSS)和PEG组(1000例,口服4 L PEG)。比较两组患者肠道清洁度、清洁效果、肠腔气泡、结肠镜检查阳性率和不良反应发生率。结果OSS组肠道准备成功率为92.5%(925/1000),PEG组肠道准备成功率为91.7%(917/1000),两组患者比较,差异无统计学意义(P>0.05);两组患者左侧结肠波士顿肠道准备量表(BBPS)评分比较,差异无统计学意义(P>0.05),OSS组BBPS总分,以及中段和右侧结肠BBPS评分明显高于PEG组,差异均有统计学意义(P<0.05);OSS组气泡评估满意率为96.2%,高于PEG组的90.3%,差异有统计学意义(P<0.05);两组患者结肠镜检查阳性率和不良反应发生率比较,差异均无统计学意义(P>0.05)。结论OSS在肠道准备方面具有较好的去泡效果,清洁效果与PEG相当,且有较高的安全性。
文摘DNA结合单锌指蛋白(DNA binding with one finger,Dof)是植物中特有的一类转录因子,参与调控植物组织分化、种子萌发、植物抗逆及代谢等生理生化过程。为鉴定并分析高粱Dof转录因子家族成员,利用生物信息学方法从全基因组水平对高粱Dof转录因子家族成员的基本基因结构、理化性质、保守基序、启动子顺式作用元件、组织特异性表达及干旱胁迫表达等进行分析。20%PEG6000模拟干旱胁迫处理豫粱10号的幼苗后,利用qPCR技术分析Dof基因家族成员响应不同时间段干旱胁迫的表达情况。结果共鉴定了30个高粱Dof基因,分布在9条染色体上,基因保守性分析结果显示,Motif1(即锌指结构)是高粱Dof基因家族中保守的蛋白质序列;其中29个基因中都含有CX2CX21CX2C保守基序,而SbDof15基因保守序列为CX20CX2C;启动子结构分析表明,Dof基因的启动子区域中含有ABA、MYB、MeJA、低温等逆境胁迫响应元件;进化树分析结果表明,高粱Dof蛋白与玉米的同源性更高,热图分析表明Dof家族成员在高粱的不同部位都有表达。qRT-PCR结果表明,高粱Dof基因家族成员可能参与高粱应对干旱胁迫的响应,不同的Dof基因成员在应对干旱胁迫时的调控机制可能不一样。
基金supported by the Department of Defense AFIRMⅢW81XWH-20-2-0029 grant subcontractLone Star Paralysis gift,UT POC19-1774-13 grant+1 种基金Neuraptive Therapeutics Inc.26-7724-56 grantNational Institutes of Health R01-NS128086(all to GDB)。
文摘Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene glycol(PEG),to immediately fuse closely apposed open ends of severed proximal and distal axons in rat sciatic nerves.We have previously reported that sciatic nerve axons repaired by PEG-fusion do not undergo Wallerian degeneration,and PEG-fused animals exhibit rapid(within 2–6 weeks)and extensive locomotor recovery.Furthermore,our previous report showed that PEG-fusion of severed sciatic motor axons was non-specific,i.e.,spinal motoneurons in PEG-fused animals were found to project to appropriate as well as inappropriate target muscles.In this study,we examined the consequences of PEG-fusion for sensory axons of the sciatic nerve.Young adult male and female rats(Sprague–Dawley)received either a unilateral single cut or ablation injury to the sciatic nerve and subsequent repair with or without(Negative Control)the application of PEG.Compound action potentials recorded immediately after PEG-fusion repair confirmed conduction across the injury site.The success of PEG-fusion was confirmed through Sciatic Functional Index testing with PEG-fused animals showing improvement in locomotor function beginning at 35 days postoperatively.At 2–42 days postoperatively,we anterogradely labeled sensory afferents from the dorsal aspect of the hindpaw following bilateral intradermal injection of wheat germ agglutinin conjugated horseradish peroxidase.PEG-fusion repair reestablished axonal continuity.Compared to unoperated animals,labeled sensory afferents ipsilateral to the injury in PEG-fused animals were found in the appropriate area of the dorsal horn,as well as inappropriate mediolateral and rostrocaudal areas.Unexpectedly,despite having intact peripheral nerves,similar reorganizations of labeled sensory afferents were also observed contralateral to the injury and repair.This central reorganization may contribute to the improved behavioral recovery seen after PEG-fusion repair,supporting the use of this novel repair methodology over currently available treatments.
