A 105-k Da polymer lectin was purified from lamprey（Lampetra japonica） serum by chromatography methods including cation ion-exchange chromatography with a SP-Sepharose TM XL column and size exclusion chromatography ...A 105-k Da polymer lectin was purified from lamprey（Lampetra japonica） serum by chromatography methods including cation ion-exchange chromatography with a SP-Sepharose TM XL column and size exclusion chromatography with a Superdex 200 column. The target fractions were collected according to the direction of hemagglutinating activity. The results revealed that the active fractions could adsorb on SP-Sepharose column and showed a 280 nm UV absorbance peak corresponding to molecular weights of 105 k Da in the following size exclusion chromatography. The target fractions with hemagglutinating activity were further checked by NativePAGE and SDS-PAGE. Two single bands at around 105 k Da and 35 k Da were displayed by two electrophoresis methods respectively, indicating that the protein exists as a trimer in solution. After Native-PAGE and SDS-PAGE,two bands were excised from the gels respectively and further identified by MALDI-TOF/TOF as serum lectin（gi：13094239）. The lectin was able to agglutinate rabbit red blood cells（RRBCs） and sheep red blood cells（SRBCs） in vitro. The lectin isolated from lamprey serum in the current study might be helpful for deeply understanding the innate immune molecules dependent immune defence in jawless vertebrates which have been proved recently that they possess a lymphocyte-based system of anticipatory immunity with variable lymphocyte receptors as mediators.展开更多
目的 采用大气压基质辅助激光解吸电离-离子阱-飞行时间质谱(atmospheric pressure matrix assisted laser desorption combined with ion trap-time of flight mass spectrometry, AP-MALDI-IT-TOF/MS)和解吸电喷雾电离-四级杆-飞行时...目的 采用大气压基质辅助激光解吸电离-离子阱-飞行时间质谱(atmospheric pressure matrix assisted laser desorption combined with ion trap-time of flight mass spectrometry, AP-MALDI-IT-TOF/MS)和解吸电喷雾电离-四级杆-飞行时间质谱(desorption electrospray ionization combined with quadrupole-time of flight mass spectrometry, DESI-Q-TOF/MS)两种技术,实现板蓝根的全质谱成像分析,揭示多种成分的组织原位可视化分布,寻找品质特征相关指标群。方法 不同商品规格的板蓝根,分别喷涂2,5-二羟基苯乙酮(2,5-DHAP)和1,5-二氨基萘(1,5-DAN)基质,在正、负离子模式下进行AP-MALDI-IT-TOF/MS质谱成像、成分鉴定和偏最小二乘回归(partial least squares regression, PLSR)分析。不同品质特征的板蓝根,在正、负离子模式下进行DESI-Q-TOF/MS质谱成像、成分鉴定和正交偏最小二乘判别分析(orthogonal partial least squares discrimination analysis, OPLS-DA)分析。结果 板蓝根经质谱成像分析,初步鉴别得到多个类别约100余个化合物,AP-MALDI-IT-TOF/MS和DESI-Q-TOF/MS均可揭示板蓝根化合物的空间分布,并可区分不同规格、品质样品,3-醛基吲哚、前告依春/表前告依春、isatithioetherin C/isatithioetherin E、松柏苷、紫丁香酚苷、直铁线莲宁B、腺苷、腺嘌呤、尿苷、精氨酸、2-羟基丁二酸、顺丁烯二酸/富马酸、枸橼酸、大黄素-8-O-β-D-葡萄糖苷、异牡荆素在高商品规格/品质样品中特定空间分布信号更强,对分类影响显著,组成品质特征相关指标群。结论 质谱成像可将中药材的品质与化学信息建立连接,为其质量评价和进一步开发利用提供实验基础和新的手段。展开更多
目的 探讨使用加速度肌松监测仪（acceleromyography,AMG）定标和4个成串刺激（train of four ratio,TOF）基线值对麻醉后恢复室（post-anesthesia care unit,PACU）患者肌松残余效应监测准确性的影响,为PACU患者残余肌松的处理提供指...目的 探讨使用加速度肌松监测仪（acceleromyography,AMG）定标和4个成串刺激（train of four ratio,TOF）基线值对麻醉后恢复室（post-anesthesia care unit,PACU）患者肌松残余效应监测准确性的影响,为PACU患者残余肌松的处理提供指导。方法 选择择期全麻手术患者151例,静脉注射依托咪酯、芬太尼诱导后,使用TOF-Watch SX加速度肌松监测仪进行定标和连续测量TOF值5 min确定基线值。然后给予非去极化肌松药插管或置入喉罩。手术完成后,拔除气管导管或喉罩后转运至PACU继续肌松监测,记录TOF基线值（TOFb）和进入PACU时的实测TOF值（TOFa）。结果 共145例患者完成观察,TOFb 0.93-1.29,平均1.07±0.06,其中132例TOFb〉1.0。TOFa〈0.9者41例,肌松残余发生率为28.3%;修正后的TOF值（TOFr）〈0.9者61例,肌松残余发生率为42.1%,两种方法计算的肌松残余发生率的差异有统计学意义（掊2=6.049,P=0.019）。结论 使用AMG时,确定TOF基线值可以提高肌松残余效应监测的准确性,有利于发现潜在的肌松残余患者。展开更多
C-terminal sequencing is important for characterized the primary structure of protein and polypeptides and it must do for quality control of recombination protein drugs.A method of MALDI-TOF-TOF-MS with in source deca...C-terminal sequencing is important for characterized the primary structure of protein and polypeptides and it must do for quality control of recombination protein drugs.A method of MALDI-TOF-TOF-MS with in source decay(ISD) to measure C-terminal sequence of protein was established.This method was applied successfully to measure C-terminal sequence of Neuregulin,TP-Ⅱ,ubiquitin,and myoglobin,respectively and 10,24,24 and 36 amino acid residue of C-terminal were measured.展开更多
基金The National Program on Key Basic Research Project(973 Program)of China under contract No.2013CB835304the National Marine Public Projects under contract No.201305016+1 种基金the National Natural Science Foundation of China under contract Nos31772884 and 31601865the Key Projects of Scientific Research Platform of Liaoning Provincial Education Department under contract No.L201683651
文摘A 105-k Da polymer lectin was purified from lamprey（Lampetra japonica） serum by chromatography methods including cation ion-exchange chromatography with a SP-Sepharose TM XL column and size exclusion chromatography with a Superdex 200 column. The target fractions were collected according to the direction of hemagglutinating activity. The results revealed that the active fractions could adsorb on SP-Sepharose column and showed a 280 nm UV absorbance peak corresponding to molecular weights of 105 k Da in the following size exclusion chromatography. The target fractions with hemagglutinating activity were further checked by NativePAGE and SDS-PAGE. Two single bands at around 105 k Da and 35 k Da were displayed by two electrophoresis methods respectively, indicating that the protein exists as a trimer in solution. After Native-PAGE and SDS-PAGE,two bands were excised from the gels respectively and further identified by MALDI-TOF/TOF as serum lectin（gi：13094239）. The lectin was able to agglutinate rabbit red blood cells（RRBCs） and sheep red blood cells（SRBCs） in vitro. The lectin isolated from lamprey serum in the current study might be helpful for deeply understanding the innate immune molecules dependent immune defence in jawless vertebrates which have been proved recently that they possess a lymphocyte-based system of anticipatory immunity with variable lymphocyte receptors as mediators.
文摘目的 采用大气压基质辅助激光解吸电离-离子阱-飞行时间质谱(atmospheric pressure matrix assisted laser desorption combined with ion trap-time of flight mass spectrometry, AP-MALDI-IT-TOF/MS)和解吸电喷雾电离-四级杆-飞行时间质谱(desorption electrospray ionization combined with quadrupole-time of flight mass spectrometry, DESI-Q-TOF/MS)两种技术,实现板蓝根的全质谱成像分析,揭示多种成分的组织原位可视化分布,寻找品质特征相关指标群。方法 不同商品规格的板蓝根,分别喷涂2,5-二羟基苯乙酮(2,5-DHAP)和1,5-二氨基萘(1,5-DAN)基质,在正、负离子模式下进行AP-MALDI-IT-TOF/MS质谱成像、成分鉴定和偏最小二乘回归(partial least squares regression, PLSR)分析。不同品质特征的板蓝根,在正、负离子模式下进行DESI-Q-TOF/MS质谱成像、成分鉴定和正交偏最小二乘判别分析(orthogonal partial least squares discrimination analysis, OPLS-DA)分析。结果 板蓝根经质谱成像分析,初步鉴别得到多个类别约100余个化合物,AP-MALDI-IT-TOF/MS和DESI-Q-TOF/MS均可揭示板蓝根化合物的空间分布,并可区分不同规格、品质样品,3-醛基吲哚、前告依春/表前告依春、isatithioetherin C/isatithioetherin E、松柏苷、紫丁香酚苷、直铁线莲宁B、腺苷、腺嘌呤、尿苷、精氨酸、2-羟基丁二酸、顺丁烯二酸/富马酸、枸橼酸、大黄素-8-O-β-D-葡萄糖苷、异牡荆素在高商品规格/品质样品中特定空间分布信号更强,对分类影响显著,组成品质特征相关指标群。结论 质谱成像可将中药材的品质与化学信息建立连接,为其质量评价和进一步开发利用提供实验基础和新的手段。
文摘目的 探讨使用加速度肌松监测仪（acceleromyography,AMG）定标和4个成串刺激（train of four ratio,TOF）基线值对麻醉后恢复室（post-anesthesia care unit,PACU）患者肌松残余效应监测准确性的影响,为PACU患者残余肌松的处理提供指导。方法 选择择期全麻手术患者151例,静脉注射依托咪酯、芬太尼诱导后,使用TOF-Watch SX加速度肌松监测仪进行定标和连续测量TOF值5 min确定基线值。然后给予非去极化肌松药插管或置入喉罩。手术完成后,拔除气管导管或喉罩后转运至PACU继续肌松监测,记录TOF基线值（TOFb）和进入PACU时的实测TOF值（TOFa）。结果 共145例患者完成观察,TOFb 0.93-1.29,平均1.07±0.06,其中132例TOFb〉1.0。TOFa〈0.9者41例,肌松残余发生率为28.3%;修正后的TOF值（TOFr）〈0.9者61例,肌松残余发生率为42.1%,两种方法计算的肌松残余发生率的差异有统计学意义（掊2=6.049,P=0.019）。结论 使用AMG时,确定TOF基线值可以提高肌松残余效应监测的准确性,有利于发现潜在的肌松残余患者。
文摘C-terminal sequencing is important for characterized the primary structure of protein and polypeptides and it must do for quality control of recombination protein drugs.A method of MALDI-TOF-TOF-MS with in source decay(ISD) to measure C-terminal sequence of protein was established.This method was applied successfully to measure C-terminal sequence of Neuregulin,TP-Ⅱ,ubiquitin,and myoglobin,respectively and 10,24,24 and 36 amino acid residue of C-terminal were measured.
