Objective To explore the value of CK13 (Rab) and CK10/13(Mab) as objective and specific biomarkers combined with Alcian-Phloxine-Martius yellow (APM) staining for the diagnosis of amniotic fluid embolism (AFE) and amniotic fluid aspiration (AFA). Methods A retrospective study of forensic autopsy cases of 148 neonatal deaths and 19 maternal deaths in the perinatal stage was conducted at the Institute of Pathology and Forensic Medicine, Zhejiang University. Medical records were reviewed and monoclonal antibody for CK13 (Rab) and CK10/13 (Mab) as immunostaining of amniotic fluid squamous cells, APM staining, and Hematoxylin and Eosin (HE) staining were used to diagnose the AFE and AFA. Descriptive statistics of the patient population were analyzed using SPSS 20.0 software. Results Immunoreactivity of CK13 and CK10/13 specifically identified squamous cells of all the AFE and AFA cases. The amniotic fluid squamous cells were stained positive in a deep brown color with the monoclonal antibody to CK 13 and CK10/13 whereas the endothelial cells and alveolar epithelial cells were negative. There was no CK13 or CK10/13 expression in the non-AFE and non-AFA cases. With APM staining keratinized squamous cells were pink and mucus was blue-green in marked contrast with the surrounding tissue, which improved the detection rates of both keratinized squamous cells and mucus. Conclusions CK13 (Rab) and CK10/13 (Mab) are valuable and reliable biomarkers of amniotic fluid squamous cells. APM reveals enriched mucus and keratinized squamous cells of amniotic fluid. Immunohistochemical detection of CK13 and CK10/13 combined with APM staining can improve the accuracy and reduce the difficulty in the diagnosis of AFE and AFA.
In this paper, Au-Ag alloy nanostar (Au-Ag NSt) substrates were prepared by liquid–liquid three-phase self-assembly method, and the influence of preparation conditions on their Raman activity of the substrate was explored. By adjusting the gold-silver ratios of alloy seeds and the concentration of AgNO 3 in the growth solution, it was found that the LSPR peak can be adjusted. Under the conditions of Au 1 -Ag 3 nanoparticles as seeds and 200 μL AgNO 3 solution (2 mM) as epitaxial growth solution, the prepared Au 1 -Ag 3 NSt substrate exhibits excellent SERS activity. The substrate can detect R6G and CV probe molecules with detection limits as low as 10 −11 M and 10 −10 M, respectively. Experiments have confirmed that the substrate has the characteristics of high sensitivity, good uniformity and strong stability. In addition, the substrate can detect APM molecules far below the safety standard, which further proves that the structure has excellent practical application potential as a SERS substrate.