Usage of IL-1 acceptor antagon in the treatment of SARS
A technology of interleukin and receptor antagonists, applied in antiviral agents, medical preparations containing active ingredients, peptide/protein components, etc., can solve low immunity, high mortality rate of severe SARS patients, severe infection, etc. question
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Embodiment 1
[0036] Example 1: Expression and Purification of Recombinant Human Interleukin 1 Receptor Antagonist
[0037] The full length of IL-1Ra cDNA is 462bp (SEQ ID NO: 1), encoding 153 amino acids (SEQ ID NO: 2), which is synthesized by whole gene synthesis method. The sequence was cloned on the prokaryotic expression vector pET32a (Novagen Company) through the NdeI and HindIII sites at both ends, and transformed into Escherichia coli expression host BL21 (DE3) (Novagen Company) for expression, and the target protein IL-1Ra was present in soluble form in In the cytoplasm, the expression amount reaches more than 30% of the total protein of the bacteria.
[0038] Biostat C-20 automatic fermenter is used for high-density fermentation of IL-1Ra production strains, the collected bacteria are resuspended with lysate, and GYB60-6S high-pressure homogenizer pump is used to destroy the bacteria, and the broken bacteria are centrifuged in two steps With cation exchange chromatography and one...
Embodiment 2
[0040] Example 2: Pharmacodynamic study of endotoxin-induced acute respiratory distress syndrome dog model
[0041] Fifteen dogs weighing 10.0-15.0 kg were anesthetized by intravenous administration of 50 mg / kg sodium pentobarbital. After anesthesia, the trachea was exposed and incised, followed by cannulation to allow 30% O 2 and 70%N 2 The mixed gas was artificially ventilated (tidal volume: 180ml / breath; respiratory rate: 15-20 breaths / min) and the pressure of the airway was measured. Instrumentation included insertion of polyethylene catheters into the bilateral central veins and aorta, and insertion of a 7F catheter in the main pulmonary line. Continuously monitor and record aortic pressure, heart rate, central venous pressure, and pulmonary pressure. Cardiac output was measured in duplicate using the thermodilution technique. Analyze samples obtained from the aortic line for various parameters related to lung function, including arterial blood O 2 Tension, hemoglobin ...
Embodiment 3
[0045] Example 3: Viral ARDS Model
[0046] Female CBA / J mice aged 4-5 weeks were divided into 3 groups with 20 mice in each group. Group 3 animals were first injected intraperitoneally with 0.008cc 20% ketamine (ketamine) and 2% PromAce (acepromazine maleate) to lightly anesthetize the animals, and then intranasally inoculated 30ul containing 1×10 7 0.9% sterile saline for injection of pfu reovirus (Reovirus) was inoculated with 15ul in each nostril.
[0047] From day 5 after inoculation, animals in group 1 were subcutaneously injected with IL-1Ra 24 mg / kg / d, once a day, for 7 consecutive days. Animals in the second group received intravenous injection of corticosteroids 30 mg / kg / d for 7 consecutive days. Group 3 animals were subcutaneously injected with 2.5ml / kgml of physiological saline as a control.
[0048] Bronchoalveolar tank washes were collected at 5, 7, 9, 10, 12, and 14 days after inoculation. Use a cannula to inject in situ with a 21-gauge needle followed by as...
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