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In assisted reproduction, the seminal plasma\u2010based characteristics of normozoospermic men achieving successful clinical pregnancy through rescue intracytoplasmic sperm injection after <jats:italic>in vitro<\/jats:italic> fertilization failure remain unclear.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>To identify potential seminal plasma proteins to contribute to a new understanding of unexplained male factor infertility.<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and Methods<\/jats:title><jats:p>An approach with isobaric tags for relative and absolute quantification labeling coupled with liquid chromatography matrix\u2010assisted laser desorption ionization mass spectrometry was applied to investigate differentially expressed proteins in the seminal plasma of a rescue intracytoplasmic sperm injection pregnancy group versus an <jats:italic>in vitro<\/jats:italic> fertilization pregnancy group of normozoospermic men.<\/jats:p><\/jats:sec><jats:sec><jats:title>Result(s)<\/jats:title><jats:p>The present work revealed seventy\u2010three differentially expressed seminal plasma proteins between the <jats:italic>in vitro<\/jats:italic> fertilization and rescue intracytoplasmic sperm injection groups. Forty\u2010five proteins were upregulated, and 28 proteins were downregulated in the rescue intracytoplasmic sperm injection group compared with the <jats:italic>in vitro<\/jats:italic> fertilization group. Bioinformatics analyses showed that these altered proteins were involved in various functions, including the kallikrein\u2010related proteolytic cascade, immune response, and heparin binding. Furthermore, the validity of the proteomic results was verified by Western blot analysis of the proteins (lactoferrin [LTF], fibronectin [FN1], creatine kinase B type [CKB], kallikrein\u20102 [KLK2], aminopeptidase N [ANPEP], extracellular matrix protein 1 [ECM1], glycodelin [PAEP], alpha\u20101\u2010antitrypsin [SERPINA1], and semenogelin\u20101 [SEMG1]) and immunofluorescence. Moreover, 16% of the seminal plasma proteins identified in the present work have not been reported in previous studies.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>This panel of altered seminal plasma proteins associated with unexplained male factor infertility might have clinical relevance and may be useful in the diagnosis and prognosis of idiopathic infertility in <jats:italic>in vitro<\/jats:italic> fertilization.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Our work not only provides a new complementary high\u2010confidence dataset of seminal plasma proteins but also shines new light onto the molecular characteristics of seminal plasma from normozoospermic men with different assisted reproductive outcomes.<\/jats:p><\/jats:sec>","DOI":"10.1111\/andr.12687","type":"journal-article","created":{"date-parts":[[2019,7,31]],"date-time":"2019-07-31T07:25:05Z","timestamp":1564557905000},"page":"407-420","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Characterization of seminal plasma proteomic alterations associated with the IVF and rescue\u2010ICSI pregnancy in assisted reproduction"],"prefix":"10.1111","volume":"8","author":[{"given":"X.","family":"Liu","sequence":"first","affiliation":[{"name":"Central Laboratory The Affiliated Yantai Yuhuangding Hospital of Qingdao University Yantai Shandong China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G.","family":"Liu","sequence":"additional","affiliation":[{"name":"Reproductive Center Tianjin Aiwei Hospital Tianjin 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