A group of serum proteins as potential diagnostic biomarkers for Yin‐deficiency‐heat syndrome

Abstract

Yin‐deficiency‐heat (YDH) syndrome is a very common subhealth status in Traditional Chinese Medicine. However, currently, there is no unified standard for diagnosing YDH syndrome. We applied the iTRAQ‐2D LC–MS/MS method to explore the potential of serum protein profiles as biomarker for YDH syndrome. A total of 120 differentially expressed proteins (79 downregulated and 41 upregulated) were identified by the proteomic profiling. The results of KEGG pathway analysis showed that the functions of the differentially expressed proteins were mainly involved in complement and coagulation cascades. The clinical data showed that YDH syndrome was closely related to inflammation and coagulation, compared with the healthy controls. The ELISA validation results indicated that the expression levels of ALB, CFI, and KLKB1 were downregulated in the YDH syndrome group (p < .05). Moreover, we established a decision tree model based on the combination of these three proteins and achieved a sensitivity of 87.5%, a specificity of 84.4%, and AUC of 0.891. The results indicated that the combination of ALB, CFI, and KLKB1 may serve as potential biomarkers for diagnosing YDH syndrome. Our study can provide a new method for YDH syndrome diagnosis, and may also provide an experimental basis to understand the molecular mechanism of YDH syndrome.

iTRAQ-based quantitative proteomics analysis of cantaloupe (Cucumis melo var. saccharinus) after cold storage

“…The Shimadzu LC-20AB liquid phase system was used to separate the peptides, with 5 μm of each sample being added to a 4.6 × 250 mm Gemini C18 column [12]. The extracted peptides were reconstituted with 2 mL of mobile phase A (5% ACN pH 9.8) and then injected and eluted at a flow rate gradient of 1 mL/min: 5% mobile phase B (95% CAN, pH 9.8) for 10 min, 5 to 35% mobile phase B for 40 min, 35 to 95% mobile phase B for 1 min, mobile phase B for 3 min, and 5% mobile phase B for 10 min [57]. The elution peak was monitored at a wavelength of 214 nm and one component was collected per minute.…”

“…The elution peak was monitored at a wavelength of 214 nm and one component was collected per minute. The samples were combined with a chromatographic elution peak map to obtain 20 components, which were then freeze-dried [57].…”

“…Separation was carried out by a Shimadzu LC-20 AD model nanoliter liquid chromatograph. The sample was first enriched and desalted in a trap column, and then connected in series with a self-packed C18 column (75 μm inner diameter, 3.6 μm column particle size, 15 cm column length) and separated at a flow rate of 300 nl/min through the following effective gradient: 0-8 min, 5% mobile phase B (98% ACN, 0.1% FA); 8-43 min, the mobile phase B linearly increased from 8 to 35%; 43-48 min, the mobile phase B increased from 35 to 60%; 48-50 min, mobile phase B increased from 60 to 80%; 50-55 min, 80% mobile phase B; 55-65 min, 5% mobile phase B [57].…”

iTRAQ-based quantitative proteomics analysis of cantaloupe (Cucumis melo var. saccharinus) after cold storage

“…The Shimadzu LC-20AB liquid phase system was used to separate the peptides, with 5 μm of each sample being added to a 4.6 × 250 mm Gemini C18 column [12]. The extracted peptides were reconstituted with 2 mL of mobile phase A (5% ACN pH 9.8) and then injected and eluted at a flow rate gradient of 1 mL/min: 5% mobile phase B (95% CAN, pH 9.8) for 10 minutes, 5% to 35% mobile phase B for 40 minutes, 35% to 95% mobile phase B for 1 minute, mobile phase B for 3 minutes, and 5% mobile phase B for 10 minutes [57]. The elution peak was monitored at a wavelength of 214 nm and one component was collected per minute.…”

“…Separation was carried out by a Shimadzu LC-20AD model nanoliter liquid chromatograph. The sample was first enriched and desalted in a trap column, and then connected in series with a self-packed C18 column (75 micron inner diameter, 3.6 micron column particle size, 15 cm column length) and separated at a flow rate of 300 nl/min through the following effective gradient: 0-8 minutes, 5% mobile phase B (98% ACN, 0.1% FA); 8-43 minutes, the mobile phase B linearly increased from 8% to 35%; 43-48 minutes, the mobile phase B increased from 35% to 60%; 48-50 minutes, mobile phase B increased from 60% to 80%; 50-55 minutes, 80% mobile phase B; 55-65 minutes, 5% mobile phase B [57].…”