Biomarker for auxiliary diagnosis of recurrent abortion, and application thereof

A recurrent miscarriage, biomarker technology, applied in disease diagnosis, biological testing, biomaterial analysis, etc., can solve problems such as unclear pathogenesis, and achieve the effect of promoting migration, invasion and proliferation

Pending Publication Date: 2022-02-15
DALIAN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the pathogenesis of RSA is still unclear
At present, RSA can only be diagnosed by exclusion method, including:

Method used

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  • Biomarker for auxiliary diagnosis of recurrent abortion, and application thereof
  • Biomarker for auxiliary diagnosis of recurrent abortion, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The expressions of FUT4 and α1,3 fucose were detected in the villi of normal women and patients with recurrent miscarriage, and the villi were processed by paraffin embedding.

[0025] 1. Detection of localization and expression of FUT4 and α1,3 fucose in villous tissue by immunohistofluorescence

[0026] (1) Dewaxing: Put the prepared paraffin sections into the following solutions in turn: soak in xylene I for 10 minutes, then soak in xylene II for 10 minutes, soak in 100% ethanol I for 5 minutes, soak in 100% ethanol II for 5 minutes, and soak in 95% ethanol Soak for 5 minutes, soak in 85% ethanol for 5 minutes, soak in 70% ethanol for 5 minutes; then rinse with slow water for 10 minutes, do not directly wash the slices. Dip in PBS 3 times, 5min each time.

[0027] (2) Antigen retrieval: Put the slices into citrate buffer, cover them, and thaw in a microwave oven for 20 minutes. Take it out and let it cool down to room temperature naturally.

[0028] (3) Rinse with...

Embodiment 2

[0054] 1. Immunofluorescence detection of FUT4 and α1,3 fucose expression in trophoblast cells of different experimental groups (Scramble control group, FUT4 siRNA transfection group, FUT4 siRNA and FUT4 cDNA transfection group simultaneously)

[0055] Slide: After the cells are digested with trypsin, centrifuge at 800 rpm for 4 minutes, resuspend in fresh medium, put the cell suspension into a petri dish with slide, and let the cells grow on the slide.

[0056] Collection: remove the culture medium in the culture dish, wash with PBS 3 times, 3 minutes each time.

[0057] Fixation: add 4% paraformaldehyde to the petri dish obliquely, and fix it for 20 minutes.

[0058] Paraformaldehyde was discarded, washed 3 times with PBS, 3 min each time.

[0059] Sealing: Select slides and place them in a wet box, add immunostaining blocking solution to seal for 1 hour.

[0060] Primary antibody incubation: clip the slices from the blocking solution, blot dry on filter paper, incubate wi...

Embodiment 3

[0067] In Vitro Migration Experiment of Early Pregnancy Villi Explants

[0068] (1) Laying gel: melt matrigel in a refrigerator at 4°C, dilute matrigel with serum-free medium DMEM / F-12 at a ratio of 1:2, gently blow and mix, add 50 μL to each well of a 96-well plate matrigel, try not to generate air bubbles, and place in a 37°C incubator.

[0069] (2) The villous tissue in the first trimester of pregnancy was obtained from healthy women undergoing artificial abortion in the obstetrics and gynecology operating room of the affiliated hospital for non-medical reasons, and the blood clots were rinsed with pre-cooled sterile PBS immediately.

[0070] (3) Put the villous tissue into a large dish containing PBS in a sterile operating table, and cut into 2-5mm fragments.

[0071] (4) Carefully transfer to the matrigel of the 96-well plate so that it is anchored in the center of the matrigel.

[0072] (5) After anchoring for 4-6 hours, add complete medium, observe the morphology unde...

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Abstract

The invention discloses a biomarker for auxiliary diagnosis of recurrent abortion, and application thereof, and belongs to the technical field of biology. According to the invention, the fact that alpha1, 3 catalyzed and synthesized by fucosyltransferase 4 (FUT4) is connected with fucosylation is found for the first time, the migration, invasion and multiplication capacity of trophoblast cells is promoted, and the alpha1, 3 can be used as a biological marker for auxiliary diagnosis of recurrent abortion. It is proved that the influence of FUT4 and alpha 1, 3 connected fucosylation catalytically synthesized by FUT4 on migration invasion and multiplication capacity of trophoblast cells, which is beneficial to prediction and diagnosis of recurrent abortion and deep understanding of an embryo implantation mechanism, and also can provide a new theoretical basis for clinical treatment of recurrent abortion taking sugar as a target spot.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a biomarker for auxiliary diagnosis of recurrent miscarriage and its application. Background technique [0002] In women's early pregnancy (≤20 weeks), the natural loss of less than 500g of pregnancy is called spontaneous abortion (Spontaneous Abortion), which is a common adverse outcome in obstetrics and gynecology. However, two or more consecutive spontaneous abortions in early pregnancy are recurrent spontaneous abortions (RSA). The etiology of recurrent miscarriage is very complicated, and the common ones include immune factors, endocrine factors, genetic factors, thrombotic diseases and infection, etc. Excluding the above-mentioned causes, more than 50% of RSA patients have unknown etiology, which is called unexplained recurrent spontaneous miscarriage (unexplained recurrent spontaneous abortion) abortion, URSA). At present, there is no clinical method to diagnose ...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/68
CPCG01N33/573G01N33/6893G01N2333/91102G01N2800/368G01N2800/54G01N2333/42
Inventor 燕秋刘帅王浩崔馨元王录尧
Owner DALIAN MEDICAL UNIVERSITY
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