The current project was designed to examine the metabolic level of branched-chain amino acids in plasm and monocyte/macrophage, and its role in immune dysfunction during sepsis.
In this study, the investigators will collect the peripheral blood of 1,3,7,14 day after diagnose of sepsis. Then, the concentration of branched-chain amino acids(BCAAs) in monocytes and plasma, and the expression of BCAAs metabolic enzymes in monocytes will be determined. Additionally, the inflammatory cytokines, e.g., Interleukin-1β (IL-1β), Tumor necrosis factor-α (TNF-α), Interleukin-10 (IL-10）, the chemokines macrophage inflammatory proteins 1α and 1β (MIP-1α and 1β) in the peripheral blood and the phagocytic function of monocytes will also be detected. The current project was designed to examine the metabolic level of branched-chain amino acids in plasm and monocyte/macrophage, and its role in immune dysfunction during sepsis.
Study Type
OBSERVATIONAL
Enrollment
90
Metabolic level of BCAAs in monocytes and macrophages
Intracellular levels of branched-chain amino acids in monocytes
BCAAs Assay Kit will be used to measure the intracellular levels of BCAAs in monocytes.
Time frame: up to 14 day after diagnose of sepsis
Plasma levels of branched-chain amino acids
BCAAs Assay Kit will be used to measure the plasma levels of BCAAs.
Time frame: 1,3,5,7,10,14 day after diagnose of sepsis
The expression of branched-chain amino acid aminotransferase in monocytes
The expression of branched-chain amino acid aminotransferase in monocytes will be determined by qPCR and western blot.
Time frame: 1,3,5,7,10,14 day after diagnose of sepsis
The expression of branched-chain α-ketoacid dehydrogenase in monocytes
The expression of branched-chain α-ketoacid dehydrogenase in monocytes will be determined by qPCR and western blot.
Time frame: 1,3,5,7,10,14 day after diagnose of sepsis
The expression of PP2Cm in monocytes
The expression of mitochondrial-targeted phosphatase, PP2Cm in monocytes will be determined by qPCR and western blot.
Time frame: 1,3,5,7,10,14 day after diagnose of sepsis
The plasma level of IL-1β
The plasma level of IL-1β will be determined by ELISA kit.
Time frame: 1,3,5,7,10,14 day after diagnose of sepsis.
The plasma level of TNF-α
The plasma level of TNF-α will be determined by ELISA kit.
Time frame: 1,3,5,7,10,14 day after diagnose of sepsis.
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The plasma level of IL-10
The level of IL-10 will be determined by ELISA kit.
Time frame: 1,3,5,7,10,14 day after diagnose of sepsis.
The plasma level of chemokine MIP-1α.
The plasma level of chemokine MIP-1α will be determined by ELISA kit.
Time frame: 1,3,5,7,10,14 day after diagnose of sepsis.
The plasma level of chemokine MIP-1β.
The plasma level of chemokine MIP-1β will be determined by ELISA kit.
Time frame: 1,3,5,7,10,14 day after diagnose of sepsis.
The phagocytic index of monocytes.
The phagocytic index of monocytes will be determined by a E.coli Phagocytosis Assay Kit.
Time frame: 1,3,5,7,10,14 day after diagnose of sepsis.