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Western blotting: evolution of an old analytical method to a new quantitative tool for biomarker measurements. Western 印迹法：从一种古老的分析方法演变为一种新的生物标志物定量测量工具。
IF 1.8 4区 医学
Bioanalysis Pub Date : 2024-03-01 Epub Date: 2024-02-13 DOI: 10.4155/bio-2023-0212
Carolina Owen, Kelly A Fader, Mohamed Hassanein
{"title":"Western blotting: evolution of an old analytical method to a new quantitative tool for biomarker measurements.","authors":"Carolina Owen, Kelly A Fader, Mohamed Hassanein","doi":"10.4155/bio-2023-0212","DOIUrl":"10.4155/bio-2023-0212","url":null,"abstract":"<p><p>Western blotting (WB) is a widely used laboratory technique for detecting specific proteins in biological matrices. Recent advances in antibody production, automation, gel and membrane manufacturing and highly sensitive detection platforms have transformed WB from a labor-intensive and qualitative method into a highly reproducible and quantitative assay suitable for biomarker detection. Despite these significant improvements in the capabilities and efficiency of WB, there remain challenges that hinder its widespread application as a research, diagnostic (in two-tiered assays like Lyme disease testing) and drug development tool. This article describes recent innovations introduced to WB methodology and the remaining challenges that prevent its wider adoption for biomarker measurements throughout the drug development process.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"319-328"},"PeriodicalIF":1.8,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139721447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Recommendations and feedback from the European Bioanalysis Forum Workshop: 1 year into ICH M10 - keeping our finger on the pulse. 欧洲生物分析论坛研讨会的建议和反馈：ICH M10 实施 1 年--把握脉搏。
IF 1.8 4区 医学
Bioanalysis Pub Date : 2024-03-01 Epub Date: 2024-02-05 DOI: 10.4155/bio-2024-0013
Philip Timmerman, Matthew Barfield, Enric Bertran Portabella, Salvatore Calogero, Kyra Cowan, Jörg Faber, Luca Ferrari, Michaela Golob, Jo Goodman, Lee Goodwin, Mark Jean Gnoth, Richard Hughes, Tsvetelina Ivanova, Gregor Jordan, Anna Laurén, Delphine Maux, Stuart McDougall, Petya Milushewa, Robert Nelson, Gwenda Pynaert, Kamil Sklodowski, Rebecca Sleigh, Petra Struwe, Tom Verhaeghe, Robert Wheller, Steve White, Katja Zeiser
{"title":"Recommendations and feedback from the European Bioanalysis Forum Workshop: 1 year into ICH M10 - keeping our finger on the pulse.","authors":"Philip Timmerman, Matthew Barfield, Enric Bertran Portabella, Salvatore Calogero, Kyra Cowan, Jörg Faber, Luca Ferrari, Michaela Golob, Jo Goodman, Lee Goodwin, Mark Jean Gnoth, Richard Hughes, Tsvetelina Ivanova, Gregor Jordan, Anna Laurén, Delphine Maux, Stuart McDougall, Petya Milushewa, Robert Nelson, Gwenda Pynaert, Kamil Sklodowski, Rebecca Sleigh, Petra Struwe, Tom Verhaeghe, Robert Wheller, Steve White, Katja Zeiser","doi":"10.4155/bio-2024-0013","DOIUrl":"10.4155/bio-2024-0013","url":null,"abstract":"<p><p>The ICH M10 guideline on bioanalytical method validation and sample analysis is being adopted since 2023. However, and inevitably, some paragraphs or requirements remain ambiguous and are open for different interpretations. In support of a harmonized interpretation by the industry and health authorities, the European Bioanalysis Forum organized a workshop on 14 November 2023 in Barcelona, Spain, to discuss unclear and/or ambiguous paragraphs which were identified by the European Bioanalysis Forum community and delegates of the workshop prior to the workshop. This manuscript reports back from the workshop with recommendations and aims at continuing an open scientific discussion within the industry and with regulators in support of a science-driven guideline for the bioanalytical community and in line with the ICH mission - that is, achieve greater harmonization worldwide to ensure that safe, effective and high-quality medicines are developed and registered in the most resource-efficient manner.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"259-270"},"PeriodicalIF":1.8,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139691107","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Highlights from the 25th International Reid Bioanalytical Forum. 第 25 届国际里德生物分析论坛花絮。
IF 1.8 4区 医学
Bioanalysis Pub Date : 2024-03-01 Epub Date: 2024-02-13 DOI: 10.4155/bio-2024-0018
Neil Spooner, Sankeetha Nadarajah, Tim Sangster
{"title":"Highlights from the 25th International Reid Bioanalytical Forum.","authors":"Neil Spooner, Sankeetha Nadarajah, Tim Sangster","doi":"10.4155/bio-2024-0018","DOIUrl":"10.4155/bio-2024-0018","url":null,"abstract":"","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"329-330"},"PeriodicalIF":1.8,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139721445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Interlaboratory evaluation of LC-MS-based biomarker assays. 基于液相色谱-质谱联用技术的生物标志物测定的实验室间评估。
IF 1.8 4区 医学
Bioanalysis Pub Date : 2024-03-01 Epub Date: 2024-02-09 DOI: 10.4155/bio-2023-0173
Kosuke Saito, Ryoya Goda, Koji Arai, Kota Asahina, Mitsuhiko Kawabata, Hitoshi Uchiyama, Tomohiro Andou, Hisao Shimizu, Kentaro Takahara, Masaaki Kakehi, Saki Yamauchi, Shin-Ichiro Nitta, Takahiro Suga, Hisashi Fujita, Rika Ishikawa, Yoshiro Saito
{"title":"Interlaboratory evaluation of LC-MS-based biomarker assays.","authors":"Kosuke Saito, Ryoya Goda, Koji Arai, Kota Asahina, Mitsuhiko Kawabata, Hitoshi Uchiyama, Tomohiro Andou, Hisao Shimizu, Kentaro Takahara, Masaaki Kakehi, Saki Yamauchi, Shin-Ichiro Nitta, Takahiro Suga, Hisashi Fujita, Rika Ishikawa, Yoshiro Saito","doi":"10.4155/bio-2023-0173","DOIUrl":"10.4155/bio-2023-0173","url":null,"abstract":"<p><p>Validation of biomarker assays is crucial for effective drug development and clinical applications. Interlaboratory reproducibility is vital for reliable comparison and combination of data from different centers. This review summarizes interlaboratory studies of quantitative LC-MS-based biomarker assays using reference standards for calibration curves. The following points are discussed: trends in reports, reference and internal standards, evaluation of analytical validation parameters, study sample analysis and normalization of biomarker assay data. Full evaluation of these parameters in interlaboratory studies is limited, necessitating further research. Some reports suggest methods to address variations in biomarker assay data among laboratories, facilitating organized studies and data combination. Method validation across laboratories is crucial for reducing interlaboratory differences and reflecting target biomarker responses.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"389-402"},"PeriodicalIF":1.8,"publicationDate":"2024-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139705980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Validation and comprehensive analysis of Streptococcus pneumoniae IgG WHO enzyme-linked immunosorbent assay in an Indian reference laboratory. 印度参考实验室对肺炎链球菌 IgG 世卫组织酶联免疫吸附试验的验证和综合分析。
IF 1.8 4区 医学
Bioanalysis Pub Date : 2024-02-01 Epub Date: 2024-02-05 DOI: 10.4155/bio-2023-0111
Mettingal Ramakrishnan Shincy, Govindan Vandana, Manheri Mavupadi Akhila, Ravindran Shilpa, Kadahalli Lingegowda Ravikumar
{"title":"Validation and comprehensive analysis of <i>Streptococcus pneumoniae</i> IgG WHO enzyme-linked immunosorbent assay in an Indian reference laboratory.","authors":"Mettingal Ramakrishnan Shincy, Govindan Vandana, Manheri Mavupadi Akhila, Ravindran Shilpa, Kadahalli Lingegowda Ravikumar","doi":"10.4155/bio-2023-0111","DOIUrl":"10.4155/bio-2023-0111","url":null,"abstract":"<p><p>Monitoring serotype-specific IgG levels against pneumococci is crucial for assessing immunity, vaccine efficacy, and evaluating vaccination programs. The WHO ELISA for pneumococci is a standardized assay ensuring consistency in testing and comparability of results across laboratories. It involves a rigorous testing process to confirm accurate, precise and reliable detection of antibodies. We validated the protocol for 13 pneumococcal serotypes by assessing its specificity, reproducibility (coefficient of variation ≤15%), repeatability (coefficient of variation ≤20%), accuracy, lower limit of quantification, stability, and robustness. We found these parameters were within acceptable ranges and showed excellent performance. Our findings imply that the method employed is appropriate for evaluating 13 valent pneumococcal conjugate vaccine which is introduced in the national immunization program by comparing pre-and post-vaccination IgG response.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"191-201"},"PeriodicalIF":1.8,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139691108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
UHPLC-MS/MS for plasma lamotrigine analysis and comparison with a homogenous enzyme immunoassay. 用于血浆拉莫三嗪分析的超高效液相色谱-质谱/质谱法以及与同源酶免疫分析法的比较。
IF 1.8 4区 医学
Bioanalysis Pub Date : 2024-02-01 Epub Date: 2024-02-09 DOI: 10.4155/bio-2023-0183
Xiaoxu Shi, Dongjie Zhang, Zhigang Zhao, Shenghui Mei
{"title":"UHPLC-MS/MS for plasma lamotrigine analysis and comparison with a homogenous enzyme immunoassay.","authors":"Xiaoxu Shi, Dongjie Zhang, Zhigang Zhao, Shenghui Mei","doi":"10.4155/bio-2023-0183","DOIUrl":"10.4155/bio-2023-0183","url":null,"abstract":"<p><p><b>Aims:</b> To develop and validate a UHPLC-MS/MS method for lamotrigine (LTG) analysis in human plasma and evaluate its agreement with a homogenous enzyme immunoassay (HEIA). <b>Materials & methods:</b> The UHPLC-MS/MS method was developed and validated according to the USFDA/EMA guidelines. A Bland-Altman plot was used to evaluate the agreement between UHPLC-MS/MS and HEIA. <b>Results:</b> Samples were pretreated with one-step protein precipitation and separated in 2.6 min. The intra- and inter-day bias and imprecisions were -15.8 to 15.0% and less than 11.17%, respectively. The recovery and matrix factor were 98.30 to 111.97%. The mean overestimation of UHPLC-MS/MS compared with HEIA was 21.57%. <b>Conclusion:</b> A rapid, sensitive and robust UHPLC-MS/MS method for plasma LTG analysis was developed and validated and was a 21.57% overestimation compared with HEIA.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"233-243"},"PeriodicalIF":1.8,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139705983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Bioanalysis of the Bicycle® toxin conjugate BT5528 and released monomethyl auristatin E via liquid chromatography-tandem mass spectrometry. 通过液相色谱-串联质谱法对自行车®毒素共轭物 BT5528 和释放的单甲基金丝桃素 E 进行生物分析。
IF 1.8 4区 医学
Bioanalysis Pub Date : 2024-02-01 Epub Date: 2023-12-13 DOI: 10.4155/bio-2023-0189
Joseph A Tweed, Fern Adams-Dam, Jane Allanson, Kevin Holmes, Ryan Senior, Hongmei Xu, Mengyao Li, Gavin Bennett, Phil Jeffrey
{"title":"Bioanalysis of the Bicycle<sup>®</sup> toxin conjugate BT5528 and released monomethyl auristatin E via liquid chromatography-tandem mass spectrometry.","authors":"Joseph A Tweed, Fern Adams-Dam, Jane Allanson, Kevin Holmes, Ryan Senior, Hongmei Xu, Mengyao Li, Gavin Bennett, Phil Jeffrey","doi":"10.4155/bio-2023-0189","DOIUrl":"10.4155/bio-2023-0189","url":null,"abstract":"<p><p><b>Background:</b> The Bicycle<sup>®</sup> toxin conjugate BT5528 is a novel peptide therapeutic conjugated to the cytotoxic agent monomethyl auristatin E (MMAE). A bioanalytical assay was developed to quantify BT5528 and unconjugated MMAE in human plasma. <b>Methodology:</b> BT5528 quantitation used a protein precipitation procedure followed by LC-MS/MS detection. Quantitation of MMAE required a selective offline and online solid-phase extraction with detection via LC-MS/MS. <b>Results:</b> BT5528 was quantified over the assay range of 5-2500 ng/ml and free MMAE was quantified over the assay range of 0.05-50 ng/ml. <b>Conclusion:</b> Bioanalytical methods were used in the bioanalysis of intact BT5528 and released MMAE, in a phase I/IIa clinical trial; to date, over 2000 human patient samples have been analyzed.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"155-169"},"PeriodicalIF":1.8,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138796653","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
LC-MS/MS method for simultaneous estimation of raloxifene, cladrin in rat plasma: application in pharmacokinetic studies. 用 LC-MS/MS 方法同时估算大鼠血浆中的雷洛昔芬和克拉霉素：在药代动力学研究中的应用
IF 1.8 4区 医学
Bioanalysis Pub Date : 2024-02-01 Epub Date: 2024-01-10 DOI: 10.4155/bio-2023-0206
Divya Chauhan, Shailesh Dadge, Pavan K Yadav, Nazneen Sultana, Arun Agarwal, Sachin Vishwakarma, Shivam Rathaur, Shubhi Yadav, Manish K Chourasia, Jiaur R Gayen
{"title":"LC-MS/MS method for simultaneous estimation of raloxifene, cladrin in rat plasma: application in pharmacokinetic studies.","authors":"Divya Chauhan, Shailesh Dadge, Pavan K Yadav, Nazneen Sultana, Arun Agarwal, Sachin Vishwakarma, Shivam Rathaur, Shubhi Yadav, Manish K Chourasia, Jiaur R Gayen","doi":"10.4155/bio-2023-0206","DOIUrl":"10.4155/bio-2023-0206","url":null,"abstract":"<p><p><b>Aim:</b> A newer LC-MS/MS method was developed and validated for the simultaneous quantification of raloxifene (RL) and cladrin (CL). <b>Methodology:</b> Both drugs were resolved in RP-18 (4.6 × 50 mm, 5 μ) Xbridge Shield column using acetonitrile and 0.1% aqueous solution of formic acid (FA) (70:30% v/v) as mobile phase by using biological matrices in female Sprague-Dawley rats using-MS/MS. <b>Results:</b> The developed method was found to be linear over the concentration ranges of 1-600 ng/ml, and lower limit of quantification was 1 ng/ml for RL and CL, respectively. Pharmacokinetic results of RL+CL showed C<sub>max</sub> = 4.23 ± 0.61, 26.97 ± 1.14 ng/ml, at T<sub>max</sub>(h) 5.5 ± 1.00 and 3.5 ± 1.00, respectively. <b>Conclusion:</b> Pharmacokinetic study results will be useful in the future for the combined delivery of RL and CL for osteoporosis treatment.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"141-153"},"PeriodicalIF":1.8,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139401632","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Lessons learned from regulatory submissions involving endogenous therapeutic analyte bioanalysis. 从涉及内源性治疗分析生物分析的监管申请中吸取的经验教训。
IF 1.8 4区 医学
Bioanalysis Pub Date : 2024-02-01 Epub Date: 2023-12-13 DOI: 10.4155/bio-2023-0209
Chongwoo Yu, Wenlei Jiang, Murali Matta, Rong Wang, Sam Haidar, Hyeonglim Seo
{"title":"Lessons learned from regulatory submissions involving endogenous therapeutic analyte bioanalysis.","authors":"Chongwoo Yu, Wenlei Jiang, Murali Matta, Rong Wang, Sam Haidar, Hyeonglim Seo","doi":"10.4155/bio-2023-0209","DOIUrl":"10.4155/bio-2023-0209","url":null,"abstract":"<p><p>Endogenous therapeutic analytes include hormones, neurotransmitters, vitamins, fatty acids and inorganic elements that are naturally present in the body because either the body produces them or they are present in the normal diet. The accurate measurement of endogenous therapeutic analytes poses a challenge when the administered exogenous therapeutic analyte and its endogenous counterpart cannot be distinguished. In this article, real case examples with endogenous therapeutic analyte bioanalysis during drug development in support of regulatory submissions are collected and presented. The article highlights common challenges encountered and lessons learned related to bioanalysis of endogenous therapeutic analytes and provides practical tips and strategies to consider from a regulatory perspective.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"171-184"},"PeriodicalIF":1.8,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138796676","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Validation of Mitra® VAMS® as a blood collection technique for trace elements analysis using ICP-MS/MS. 使用 ICP-MS/MS 对 Mitra® VAMS® 作为痕量元素分析的血液采集技术进行验证。
IF 1.8 4区 医学
Bioanalysis Pub Date : 2024-02-01 Epub Date: 2024-01-16 DOI: 10.4155/bio-2023-0180
Anthony Breton, Ciprian Mihai Cirtiu, Cyril Muehlethaler, James Rudge, Normand Fleury
{"title":"Validation of Mitra<sup>®</sup> VAMS<sup>®</sup> as a blood collection technique for trace elements analysis using ICP-MS/MS.","authors":"Anthony Breton, Ciprian Mihai Cirtiu, Cyril Muehlethaler, James Rudge, Normand Fleury","doi":"10.4155/bio-2023-0180","DOIUrl":"10.4155/bio-2023-0180","url":null,"abstract":"<p><p><b>Background:</b> Clinical dosage of toxic and essential elements in blood is well established and the collection method is still by venipuncture. This method has drawbacks and is not suited for everyone. Volumetric absorptive microsampling (VAMS) has been shown to have advantages over venipuncture. <b>Materials & methods:</b> Using inductively coupled plasma tandem mass spectrometry, a method for quantifying elements in whole blood sampled on VAMS was developed/validated. Method's performance was assessed by comparison with whole blood results. <b>Results:</b> Validation and performance assessment tests tend to show that most of the targeted elements provides accurate and reproducible results comparing to a method of reference. <b>Conclusion:</b> Overall, VAMS presents good preliminary results to eventually become an alternative to venipuncture for blood sampling for some trace elements analysis purposes.</p>","PeriodicalId":8797,"journal":{"name":"Bioanalysis","volume":" ","pages":"203-217"},"PeriodicalIF":1.8,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139471810","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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