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
目的对本院一位患者产后发热的血培养标本进行基质辅助激光解析电离飞行时间质谱(Matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF MS)分析和16S rRNA基因测序,以明确产后发热的原因,为临床...目的对本院一位患者产后发热的血培养标本进行基质辅助激光解析电离飞行时间质谱(Matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF MS)分析和16S rRNA基因测序,以明确产后发热的原因,为临床诊治提供参考。方法对血培养瓶内阳性培养物转种于哥伦比亚血琼脂平板和巧克力琼脂平板,置于35℃、5%CO_(2)培养箱中孵育,同时直接涂片革兰染色镜检;对培养基上的目标菌株补充瑞氏染色,利用MALDI-TOF MS分析技术对目标菌株进行鉴定;根据质谱鉴定结果对目标菌株进行抗菌药物敏感性试验,采用16S rRNA基因测序技术对目标菌株进行验证确认。结果血培养瓶内阳性培养物直接涂片革兰染色镜检未见菌体结构。哥伦比亚血琼脂平板培养48 h可见针尖样透明小菌落,巧克力琼脂平板无细菌生长。哥伦比亚血琼脂上的目标菌株质谱鉴定结果为人型支原体,置信度为99.9%。抗菌药物敏感性试验结果显示该菌对米诺环素、多西环素、交沙霉素和克林霉素敏感,对其他药物耐药。目标菌株的16S rRNA基因序列结果与人型支原体标准菌株ATCC23114的核苷酸同源性为99.71%。结论患者产后发热的原因为人型支原体菌血症,MALDI-TOF MS分析技术对人型支原体的鉴定结果可靠,MALDI-TOF MS分析联合16S rRNA基因测序技术能提高少见病原体的诊断效率和准确性。展开更多
Lipids serve as fundamental constituents of cell membranes and organelles.Recent studies have highlighted the significance of lipids as biomarkers in the diagnosis of breast cancer.Although liquid chromatography coupl...Lipids serve as fundamental constituents of cell membranes and organelles.Recent studies have highlighted the significance of lipids as biomarkers in the diagnosis of breast cancer.Although liquid chromatography coupled with tandem mass spectrometry(LC-MS/MS)is widely employed for lipid analysis in complex samples,it suffers from limitations such as complexity and time-consuming procedures.In this study,we have developed dopamine-modified TiO_(2)nanoparticles(TiO_(2)-DA)and applied the materials to assist the analysis of lipids by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS).The TiO_(2)-DA can provide large specific surface area and acidic environment,well suited for lipid analysis.The method was initially validated using standard lipid molecules.Good sensitivity,reproducibility and quantification performance was observed.Then,the method was applied to the analysis of 90 serum samples from 30 patients with breast cancer,30 patients with benign breast disease and 30 healthy controls.Five lipid molecules were identified as potential biomarkers for breast cancer.We constructed a classification model based on the MALDI-TOF MS signal of the 5 lipid molecules,and achieved high sensitivity,specificity and accuracy for the differentiation of breast cancer from benign breast disease and healthy control.We further collected another 60 serum samples from 20 healthy controls,20 patients with benign breast disease and 20 patients with breast cancer for MALDITOF MS analysis to verify the accuracy of the classification model.This advancement holds great promise for the development of diagnostic models for other lipid metabolism-related diseases.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.
基金supported by National Natural Science Foundation of China(NSFC,Nos.22074022,22374031)the Ministry of Science and Technology of China,National Key R&D Program of China(No.2022YFC2704300).
文摘Lipids serve as fundamental constituents of cell membranes and organelles.Recent studies have highlighted the significance of lipids as biomarkers in the diagnosis of breast cancer.Although liquid chromatography coupled with tandem mass spectrometry(LC-MS/MS)is widely employed for lipid analysis in complex samples,it suffers from limitations such as complexity and time-consuming procedures.In this study,we have developed dopamine-modified TiO_(2)nanoparticles(TiO_(2)-DA)and applied the materials to assist the analysis of lipids by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS).The TiO_(2)-DA can provide large specific surface area and acidic environment,well suited for lipid analysis.The method was initially validated using standard lipid molecules.Good sensitivity,reproducibility and quantification performance was observed.Then,the method was applied to the analysis of 90 serum samples from 30 patients with breast cancer,30 patients with benign breast disease and 30 healthy controls.Five lipid molecules were identified as potential biomarkers for breast cancer.We constructed a classification model based on the MALDI-TOF MS signal of the 5 lipid molecules,and achieved high sensitivity,specificity and accuracy for the differentiation of breast cancer from benign breast disease and healthy control.We further collected another 60 serum samples from 20 healthy controls,20 patients with benign breast disease and 20 patients with breast cancer for MALDITOF MS analysis to verify the accuracy of the classification model.This advancement holds great promise for the development of diagnostic models for other lipid metabolism-related diseases.
