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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 used in material separation, analysis materials, measurement devices, etc., can solve the problems of complex experimental steps and low sensitivity, and achieves improved sensitivity and specificity, easy detection, and avoidance of invasiveness. effect of diagnosis

Active Publication Date: 2015-02-11
夏彦恺
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  • Summary
  • 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 spectrometry has good sensitivity and reproducibility, but derivatization methods are generally used for sample pretreatment , making the experimental steps more complicated

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

Embodiment 1

[0088] Embodiment 1 Research Object Selection and Grouping Basis

[0089] From July 2009 to September 2011, the inventor collected blood and tissue samples from children with CA and normal non-CA children who met the requirements from the Children's Hospital Affiliated to Nanjing Medical University and other hospitals (typical anorectal manometry and imaging examinations). behave as figure 1 , figure 2 ), and selected 100 healthy controls (average age: 89.45±9.1 days) and 100 patients with intestinal aganglionosis (average age: 92.3±7.01 days) who met the requirements as CA metabolism Screening subjects for small molecule biomarkers. Specific sample classification criteria are as follows:

[0090] Group A: healthy control group (n=100, 85 people screened, 15 people verified by independent population):

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

[0092] 2. No digestive system disease;

[0093] 3. No other congenital malformations;

[0094] 4. No other major systemic diseas...

Embodiment 2

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

[0102] The study used anorectal manometry, imaging examination, and rectal wall histological examination to diagnose congenital intestinal aganglionosis in children with repeated defecation difficulties, abdominal distension, and low intestinal obstruction. The main diagnostic basis is: (1) Anorectal manometry shows absence of internal anal sphincter relaxation reflex; rhythmic contraction of anal canal is significantly reduced; resting pressure of rectum and internal sphincter is higher than normal. (2) Enema contrast showed: 1. Spasm and stenosis of the distal colon; 2. Proximal expansion of the colon; 3. There was a funnel-shaped transition section between the spastic bowel and dilated bowel; (3) Histological examination of the rectal wall The results showed absence or abnormal cell development of submucosal and myenteric plexus ganglion cells. The normal ganglion cells have large nuclei, deep staining, ce...

Embodiment 3

[0103] Example 3 UPLC-MS metabolomics CA biomarker screening

[0104] 1. Sample pretreatment

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

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

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

[0108] 4. Dissolve one part of the dry product (acidic extract) in 50 μL of water (containing 0.1% formic acid), and dissolve the other part of the dry product in 50 μL of water containing 6.5 mM ammonium bicarbonate pH=8 (basic 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 Company was used.

[0111] 2.2 Liquid phase conditions:

[0112] 2.2.1 The liquid chromatographic column is a Waters BEH C18 1.7μm ...

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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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Patent Type & Authority Patents(China)
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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