Application of SCML2 in diagnosis and treatment of gastroenteropancreatic neuroendocrine tumors
A neuroendocrine and gastrointestinal technology, applied in the field of biomedicine, can solve problems such as no relevant literature reports, and achieve highly specific effects
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Embodiment 1
[0092] Example 1: Research materials and specimens
[0093] 1. Tissue specimen collection
[0094] Sixty-four paraffin sections of GEP-NETs and corresponding paracancerous tissues were collected from the Affiliated Hospital of Nantong University between January 2009 and January 2014, which were confirmed by pathology. All patients did not receive chemotherapy, radiotherapy and hormone therapy before operation. Tumor classification was in accordance with WHO (2010) classification criteria for neuroendocrine tumors, and pathological grading was based on cell proliferation activity and mitotic index. The clinical data of the patients were collected in the medical record room archives, including age, tumor size, clinical stage, pathological features, and operation time. Among them, there were 36 males and 28 females; 38 patients were ≤60 years old, 26 patients were >60 years old; 10 patients had tumors in the esophagus / stomach, 40 intestinal tracts, and 14 pancreatic tumors; 50 ...
Embodiment 2
[0098] Example 2: Immunohistochemical detection of SCML2 in tissues
[0099] 1. Immunohistochemical experiment process (two-step method)
[0100] (1) Treatment of new slides and coverslips: Soak the new slides and coverslips in the acid tank for at least 24 hours, fully rinse with running water and ddH 2 After rinsing three times, soak it in 95% alcohol for at least 24 hours, and dry the cleaned glass slides in an electric constant temperature drying oven. In addition, the glass slides should be treated as follows to prevent detachment: immerse them in the APES working solution (that is, the APES concentrated solution is diluted with acetone at 1:50) and stay for 20-30 seconds, and use ddH 2 O wash away the residual APES, and then put it in a fume hood to dry. The processed slides are stored in slide boxes for future use.
[0101] (2) Slicing and baking: the required tumor tissue is dipped in wax, embedded into a wax block, and cut into thin slices with a thickness of 4-5 μ...
Embodiment 3
[0131] Embodiment 3: ELISA method detects the SCML2 in serum
[0132] 1. The steps of detecting CSML2 in serum by direct ELISA method
[0133] (I) Coating: Take the serum to be tested and add it to a 96-well microtiter plate, 50 μl / well, overnight at 4°C;
[0134] (II) Blocking: Discard the coated serum, wash 3 times with 300 μl / well of washing solution (PBS-0.05% Tween20), add 200 μl / well of blocking solution (PBS-1% BSA), and incubate at 37°C for 1 hour;
[0135] (Ⅲ) Discard the blocking solution, wash once with 300 μl / well of washing solution (PBS-0.05% Tween20), add 100 μl / well of mouse anti-human SCML2 monoclonal antibody (diluted 1:30), and incubate at 37°C for 1 hour;
[0136] (IV) Discard the antibody, wash 3 times with 300 μl / well of washing solution (PBS-0.05% Tween20), add diluted goat anti-mouse IgG-HRP (1:1000 dilution) 100 μl / well, and incubate at 37°C for 1 hour ;
[0137] (V) Discard the antibody, wash 3 times with 300 μl / well of washing solution (PBS-0.05% ...
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