Soft tissue separation preparation prepared from elastin-like polypeptide and application of soft tissue separation preparation
An elastin-like and elastin-like technology, which is applied in the fields of peptide/protein components, drug combinations, and medical preparations containing active ingredients, etc., can solve the problem of short maintenance time, high repeated injection rate, and inability to effectively reduce the complications of endoscopic treatment. Incidence and other problems, to achieve the effect of increasing separation distance, safe and effective operation, and reducing operation time
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[0029] 1. Preparation of elastin-like polypeptides
[0030] 1. The preparation method of elastin-like polypeptide:
[0031] ① Transform the vector expressing the elastin-like polypeptide into the expression strain of Escherichia coli;
[0032] ② The BL21 engineered bacteria containing the pET-22b-ELPs recombinant plasmid were inoculated into TB medium containing 100 μg / mL ampicillin at an inoculum size of 1:100 (12 g peptone per liter of TB medium, 24 g yeast extract extract, 2.31 g dipotassium hydrogen phosphate, 12.54 g potassium dihydrogen phosphate, 4 mL glycerol), cultured at 250 r / min at 37°C for 24 h;
[0033] ③ Centrifuge at 4 ℃, 4 000 r / min for 15 min, harvest the cells, and use pre-cooled PBS buffer solution (137 mmol / L NaCl, 2.7 mmol / L KCl, 4.2 mmol / L NaH2PO4, 1.4 mmol / L KH2PO4, pH 7.3) Resuspend the bacterial cells, and then put them in an ice bath to sonicate (sonicate for 2 seconds, with an interval of 3 seconds, for a total of 5 minutes);
[0034] ④ The elast...
Embodiment 1
[0048] Example 1: Study on the effectiveness and safety of soft tissue separation preparation prepared by elastin-like polypeptide in rat gastric submucosal dissection
[0049] 1.1 Experimental method:
[0050] 54 male SD rats were randomly divided into three parts (n=18). For each experiment, one of the three groups was randomly selected to carry out the following experiments at the same time, and the experiment was repeated. After fasting for 24 hours, they were weighed and anesthetized by intraperitoneal injection of 5% chloral hydrate (0.7ml / 100g). Rats were placed in a supine position, and their limbs were fixed on a sterile test bench. The abdomen was cut open, and the gauze pad was used to expose the rat stomach. The iris scissors were used to cut the appetizing tissue along the greater curvature of the stomach, and the stomach wall was opened to expose the gastric mucosa and remove residues. During the whole experimental operation, the breathing and heart rate of the...
Embodiment 2
[0061] Example 2: Study on the effectiveness and safety of soft tissue separation preparation prepared by elastin-like polypeptide in endoscopic mucosal dissection
[0062] 2.1 Experimental method:
[0063] Piglets (35.1±3.5kg) were fasted for 24 hours, prepared the skin on the legs of the piglets, took the left side lying position, intramuscularly injected ketamine (20mg / kg), intubated and connected to a ventilator, and intravenously induced with propofol. Maintain anesthesia. At the same time, turn on the endoscope host, adjust the workstation and recorder, install the gastroscope, connect the suction device, connect the electric cutting device, and prepare disposable injection needles and syringes. Use the endoscope to enter the endoscope through the cavity, slowly supply a small amount of air, and reach the stomach; use the argon knife to mark the APC, with a range of about 3.0cm; insert a disposable injection needle from the biopsy hole, and inject 0.5% methylene blue on...
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