Plasma metabolism micromolecular marker related to human intestinal canal aganglionosis and application of plasma metabolism micromolecular marker

A technology for ganglion cells and metabolic small molecules, which is applied in the direction of material separation, material analysis, and measurement devices, which can solve the problems of complex experimental steps and low sensitivity, so as to improve sensitivity and specificity, easy detection, and avoid invasiveness The effect of diagnosis

Active Publication Date: 2013-09-04
夏彦恺
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

NMR technology is characterized by no damage to the components to be measured, simple sample pretreatment, but low sensitivity; gas chromatography-mass spectrometr

Method used

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  • Plasma metabolism micromolecular marker related to human intestinal canal aganglionosis and application of plasma metabolism micromolecular marker
  • Plasma metabolism micromolecular marker related to human intestinal canal aganglionosis and application of plasma metabolism micromolecular marker
  • Plasma metabolism micromolecular marker related to human intestinal canal aganglionosis and application of plasma metabolism micromolecular marker

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0088] Example 1 Research object selection and grouping basis

[0089] From July 2009 to September 2011, the inventor collected blood and tissue samples of CA children and normal non-CA children (anorectal pressure measurement, typical imaging examinations) from hospitals such as the Children’s Hospital of Nanjing Medical University that meet the requirements. Behave like figure 1 , figure 2 ). After sorting out the sample data, 100 healthy controls (average age: 89.45±9.1 days) and 100 patients with intestinal agangliosis (average age: 92.3±7.01 days) that meet the requirements were selected as CA metabolism Screening experiment objects for small molecule biomarkers. The specific sample classification standards are as follows:

[0090] Group A: healthy control group (n=100, 85 people for screening, 15 people for independent population verification):

[0091] 1. Age between 0 and 6 months;

[0092] 2. No digestive system diseases;

[0093] 3. No other congenital malformations;

[009...

Example Embodiment

[0101] Example 2 The main diagnosis basis of the research object

[0102] The study aimed at children with repeated bowel movements, abdominal distension, and low intestinal obstruction, using anorectal manometry, imaging examination, and rectal wall histological examination to diagnose congenital intestinal aganglionosis. The main diagnosis is based on: (1) Anorectal manometry shows that the internal anal sphincter relaxation reflex is absent; the rhythmic contraction of the anal canal is significantly reduced; the resting pressure in the rectum and internal sphincter is higher than normal. (2) Enema imaging manifestations: 1. Spasm and stenosis of the distal colon; 2. Dilation of the proximal colon; 3. There is a shifted section between the spasm intestine and the dilated intestine, which is funnel-shaped; (3) Histological examination of the rectal wall The results showed that the submucosal and myenteric plexus ganglion cells were absent or cells developed abnormally. Normal ...

Example Embodiment

[0103] Example 3 UPLC-MS Metabolomics CA Biomarker Screening

[0104] 1. Sample preparation

[0105] 1. Centrifuge the fresh blood in a centrifuge at 3000 rpm for 5 minutes, and take 100 μl of the supernatant into a clean 1.5 ml EP tube.

[0106] 2. 100 μl plasma was used to precipitate protein with 300 μl methanol.

[0107] 3. Aspirate the supernatant, divide equally into 2 parts, blow dry with nitrogen and then vacuum dry.

[0108] 4. Use 50μL of water (containing 0.1% formic acid) to dissolve one part of the dried product (acidic extract), and the other part of the dried product with 50μL of water containing 6.5mM ammonium bicarbonate pH=8 (alkaline extract).

[0109] 2. Instrument testing

[0110] 2.1 Analytical instrument: UPLC series ThermoFisher LTQ-FT linear ion trap cyclotron resonance mass spectrometer from Waters Acquity is used.

[0111] 2.2 Liquid phase conditions:

[0112] 2.2.1 The liquid chromatography column is a Waters BEH C181.7μm100mm chromatographic column, and the colu...

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Abstract

The invention belongs to the fields of analytical chemistry and clinical medicine and discloses a plasma metabolism micromolecular marker related to human intestinal canal aganglionosis and an application of the plasma metabolism micromolecular marker. The plasma metabolism micromolecular marker is one or more than one of phenylpropyl tryptophan, succinyl carnitine and gamma-glutamoyl glutamic acid. The plasma metabolism micromolecular marker has specificity and sensibility on intestinal canal aganglionosis. A kit provided by the invention adopts UPLC-MS (ultra performance liquid chromatography-mass spectrometry) and can accurately detect phenylpropyl tryptophan, succinyl carnitine and gamma-glutamoyl glutamic acid in plasma, better auxiliary information can be provided for diagnosing intestinal canal aganglionosis, invasive diagnosis can be avoided, screening and diagnosis can be carried out in early stage, repeated detection can be realized, and dynamic monitoring is easy to realize.

Description

field of invention [0001] The invention belongs to the field of analytical chemistry and clinical medicine, and relates to plasma metabolic small molecule markers related to human intestinal aganglionosis and applications thereof. Background technique [0002] Congenital intestinal aganglionosis (Colonic Aganglionosis, CA) is a developmental malformation of the digestive tract characterized by the absence of ganglion cells in the distal intestine during the development of the digestive tract. The main pathogenic mechanism is that the enteric nervous system migrates to the caudal dysfunction during the development of the enteric nervous system, which leads to the absence of distal intestinal mucosal muscle and submucosal ganglion cells, the functional limitation of the distal intestinal tract, and the compensation of the proximal intestinal tract. sexual expansion. Its clinical manifestations are delayed meconium excretion caused by difficult defecation, and abdominal disten...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 夏彦恺唐维兵陈敏健唐俊伟乔善磊吴炜陆春城王心如
Owner 夏彦恺
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