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To exam the effects of Phyllanthus niruri extracts(corilagin, phyllanthin and brevifolin) on human neutrophils
Phyllanthus niruri extracts have anti-inflammatory effects in various cellular systems (monocytes, neutrophil). However, the underlying effects and mechanisms of Phyllanthus niruri extracts on neutrophils have not yet been investigated in detail.
In this study, first we would like to systemic exam the effects of corilagin, phyllanthin and brevifolin, three major active components, on human neutrophils.
Further to clarify whether Phyllanthus niruri extracts have any effect on neutrophils that may provide some insights on the ability of these compounds to modulate the innate immune response in acute organ injury.
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| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| venipuncture | Procedure | Plan to enroll 240 health people who are 20-35 years of age. People with received veinpuncture. Blood will be collected at time and drawn into Heparin-coated tube. |
| Measure | Description | Time Frame |
|---|---|---|
| Measurement of superoxide anion release | Superoxide anion released from human neutrophils were determined by measuring ferricytochrome c reduction.The neutrophils were activated with fMLF (30 nM) and cytochalasin B(0.5 μg/mL) (fMLF/CB). Changes in absorbance that occurred at 550 nm were observed continuously using a double beam spectrophotometer. Superoxide anion level was calculated using the methods described in previous report. | After neutrophil isolation, an average of 3 months |
| Measurement of intracellular ROS formation | Neutrophil ROS production was determined from the conversion of non-fluorescent DHR 123 to fluorescent rhodamine 123, detected using flow cytometry. Neutrophils (2 × 106 cells/ml) were incubated with DHR 123 (2 μM) for 15 min at 37°C, and then treated with honokiol (0.1-10 μM) for 5 min before the addition of fMLP/CB (0.5 μg/ml) for a further 5 min. The change in fluorescence was analysed using flow cytometry | After neutrophil isolation, an average of 3 months |
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Inclusion Criteria:
Exclusion Criteria:
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Voluntary healthy donor (20-35 years old) do not have coagulopathy, systemic infective disease and severe liver and renal function impairment.
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| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| Chang Gung Memorial Hospital | Taoyuan | 333 | Taiwan |
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| ID | Term |
|---|---|
| D018962 | Phlebotomy |
| ID | Term |
|---|---|
| D001800 | Blood Specimen Collection |
| D013048 | Specimen Handling |
| D019411 | Clinical Laboratory Techniques |
| D019937 | Diagnostic Techniques and Procedures |
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Seperation of human neutrophils were according to a standardized procedures and following steps: using 3%(w/v) dextran sedimentation for 30 min, then gradient centrifugation in Ficoll-Hypaque, and hypotonic lysis of erythrocytes. The purified neutrophils were determined by the Trypan blue exclusion method and need to contained >98% viable cells. Then they were suspended in without calcium ion HBSS buffer (pH 7.4), and stored at 4 °C for analysis
| D003933 | Diagnosis |
| D011677 | Punctures |
| D013812 | Therapeutics |
| D013514 | Surgical Procedures, Operative |
| D008919 | Investigative Techniques |