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A kind of preparation method of digestive juice

A technology of digestive juice and solution, applied in biochemical equipment and methods, microorganisms, culture process, etc., can solve the problems of insufficient materials, affecting the patient's treatment plan, prognosis and outcome, long operation time, etc. The effect of accurate lymph node metastasis positive rate and convenient material collection

Active Publication Date: 2021-04-13
SHAOXING PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the ex vivo tissue specimens of tumor lymph node dissection, thorough lymph node sampling is very important for tumor diagnosis and staging, formulation of treatment plans, and judgment of prognosis. However, neither conventional fixative solution nor existing specimen fixation methods can meet the requirements of rapid fixation while improving the quality of lymph nodes. The speed and detection rate of material collection make pathologists find lymph nodes in the long-term isolated adipose tissue with many disadvantages such as insufficient material collection and long operation time, which will ultimately affect the patient's treatment plan, prognosis and outcome

Method used

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  • A kind of preparation method of digestive juice
  • A kind of preparation method of digestive juice
  • A kind of preparation method of digestive juice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: Digestive juice of different compositions constitutes

[0031] The core functional components of the digestive juice provided by this application are digestive enzymes, and the main components are proteinase K, trypsin and EDTA. Under different implementation environments, one of PBS or Tris-HCl can also be used as a buffer, and a CaCl 2 To stabilize the enzyme activity, the specific composition selection method can be found in Table 1.

[0032] Table 1 Digestive fluids with different compositions

[0033] serial number Proteinase K Trypsin EDTA PBS Tris-HCl CaCl 2

1 √ √ √ - - - 2 √ √ √ √ - - 3 √ √ √ - √ 4 √ √ √ - - √ 5 √ √ √ √ √ - 6 √ √ √ √ - √ 7 √ √ √ - √ √ 8 √ √ √ √ √ √

[0034] We tested the digestive juice of the core components (ie, No. 1 in Table 1) and the supplementary components respectively, and obtained the above 8 situations. The sa...

Embodiment 2

[0037] At the same time, under different implementation environments, they were dissolved in different buffers, and the role of the digestive solution will be described below by dissolving in PBS solution and Tris-HCl as representatives.

[0038] (1) PBS buffer: pH7.4, the main function is to buffer and maintain the pH value and salt ion concentration of the reaction system. In addition, 1-10mM CaCl can also be added 2 , to stabilize divalent cations required for enzyme activity.

[0039] (2) pH7.5 50mM Tris-HCl buffer solution, during the preparation process, 1-10mM CaCl can be added 2 , the preparation method is: add 6 grams of Tris-Base to 900ml of water, adjust the pH to 7.5 with HCl, and add CaCl 2 to a final concentration of 10 mM. The main function of Tris is to buffer and maintain the pH value and salt ion concentration of the reaction system, CaCl 2 Divalent cation required for stabilization of enzyme activity.

Embodiment 3

[0040] Embodiment 3: digestion effect contrast

[0041]In this example, six copies of fresh adipose tissue and lymph nodes resected by radical gastrectomy were taken and processed as follows: (1) The fixative was treated at room temperature for 3 hours, and the fixative was composed of components with the following concentration ratios: Formaldehyde 100ml / L, lecithin 50g / L, absolute ethanol 250ml / L, acetone 250ml / L, deoxycholic acid 47.5g / L, methanol 250ml / L, PBS solution 1×, methylene blue 0.1g / L;( 2) Treat with 0.1N HCl / PBS solution at room temperature for 1 hour; (3) Treat with 0.1N NaOH / PBS solution at room temperature for 1 hour; (4) Treat with digestive solution, 25mg trypsin-1%EDTA / ml+Protease K, 37℃ water bath 4 hours, and carry out parallel experiment to six samples with proteinase K of 0.5mg / ml, 0.25mg / ml, 0.1mg / ml, 0.5mg / ml, 0.25mg / ml, 0.1mg / ml respectively, the above treatment each Phase effects are observed.

[0042] Each tissue specimen processed in step (1) is...

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Abstract

The application relates to a method for preparing digestive juice, which belongs to the technical field of test sample preparation. Weigh EDTA powder and dissolve it in PBS, adjust the pH of the solution to be suitable during preparation, stir evenly, and prepare EDTA mother liquor; weigh trypsin and proteinase K respectively, dissolve them in PBS solution, and drop them into EDTA mother liquor in proportion, according to PBS needs to be added dropwise, and the formed digestive juice includes the following components: proteinase K 0.1-2mg / ml, trypsin 1-100mg / ml, EDTA 0.1-2mg / ml. Applying this application to the treatment of tumor lymph node tissue samples, especially combined with fixative and acid and alkali treated tumor lymph node radical treatment specimens, increases the difference in softness and hardness of adipose tissue and lymph nodes, facilitates lymph node sampling, and effectively increases the number of lymph nodes detected and accurate positive rate of lymph node metastasis.

Description

technical field [0001] The application relates to a method for preparing digestive juice, which belongs to the technical field of test sample preparation. Background technique [0002] Existing tumor lymph node dissected tissue specimens in vitro are usually soaked in formalin fixative solution, mainly because formaldehyde can preserve histomorphological and structural characteristics for a long time and has good economic utility at the same time. Although this type of specimen processing method can well fix the shape of the tissue, it also hardens the specimen containing adipose tissue, which greatly increases the difficulty in obtaining materials in pathology departments, especially lymph nodes. In the ex vivo tissue specimens of tumor lymph node dissection, thorough lymph node sampling is very important for tumor diagnosis and staging, formulation of treatment plans, and judgment of prognosis. However, neither conventional fixative solution nor existing specimen fixation ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2500/30C12N2501/734
Inventor 王诚陆晓青余美文侯传玲姚育英孙爱静
Owner SHAOXING PEOPLES HOSPITAL
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