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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.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| Sepsis with type 2 diabetes | Sepsis patients with type 2 diabetes |
| |
| Sepsis without type 2 diabetes | Sepsis patients without type 2 diabetes |
| |
| Volunteers | Healthy volunteers |
|
| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Metabolic level of BCAAs in monocytes and macrophages | Other | Metabolic level of BCAAs in monocytes and macrophages |
|
| Measure | Description | Time Frame |
|---|---|---|
| Intracellular levels of branched-chain amino acids in monocytes | BCAAs Assay Kit will be used to measure the intracellular levels of BCAAs in monocytes. | up to 14 day after diagnose of sepsis |
| Measure | Description | Time Frame |
|---|---|---|
| Plasma levels of branched-chain amino acids | BCAAs Assay Kit will be used to measure the plasma levels of BCAAs. | 1,3,5,7,10,14 day after diagnose of sepsis |
| The expression of branched-chain amino acid aminotransferase in monocytes |
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Inclusion Criteria:
Exclusion Criteria:
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Patients who are diagnosed as sepsis, with or without type 2 diabetes.
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| ID | Term |
|---|---|
| D018746 | Systemic Inflammatory Response Syndrome |
| ID | Term |
|---|---|
| D007249 | Inflammation |
| D010335 | Pathologic Processes |
| D013568 | Pathological Conditions, Signs and Symptoms |
| D012769 | Shock |
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The expression of branched-chain amino acid aminotransferase in monocytes will be determined by qPCR and western blot.
| 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. | 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. | 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. | 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. | 1,3,5,7,10,14 day after diagnose of sepsis. |
| The plasma level of IL-10 | The level of IL-10 will be determined by ELISA kit. | 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. | 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. | 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. | 1,3,5,7,10,14 day after diagnose of sepsis. |