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Application of urine IGKC protein and polypeptide fragment thereof in normal pregnancy or gestational diabetes

A technology of gestational diabetes and polypeptide fragments, applied in immunoglobulins, anti-animal/human immunoglobulins, peptides, etc., can solve the problems of frequent monitoring and achieve the effect of convenient storage

Active Publication Date: 2021-06-04
张曼
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For patients with diagnosed gestational diabetes, the standards and requirements for blood glucose control will be stricter and the monitoring will be more frequent

Method used

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  • Application of urine IGKC protein and polypeptide fragment thereof in normal pregnancy or gestational diabetes
  • Application of urine IGKC protein and polypeptide fragment thereof in normal pregnancy or gestational diabetes
  • Application of urine IGKC protein and polypeptide fragment thereof in normal pregnancy or gestational diabetes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Urine Specimen Collection and Processing

[0025] A total of 60 patients with gestational diabetes mellitus (GDM) were clinically diagnosed in the Obstetrics Department of Beijing Shijitan Hospital Affiliated to Capital Medical University, including 30 samples from the second trimester (GDM-M, 13-27 weekends) and 30 samples from the third trimester (GDM-M). L, 28 weeks and later) samples; 31 cases of normal pregnant women (N) with routine outpatient check-ups; 31 cases of healthy middle-aged urine samples from normal pregnant women (T), the control group, all aged 15 to 10 years old 49 years old. After the urine was collected, it was centrifuged at 400×g for 5 minutes, the supernatant was taken, aliquoted, and frozen at -80°C. Inclusion criteria: Patients with gestational diabetes mellitus and normal pregnant women have their first pregnancy examination and file in our hospital since the early pregnancy (8-12 weeks), and the data are complete; A 75-g o...

Embodiment 2

[0029] Example 2 Magnetic bead purification and screening of urine peptides

[0030]Weak cation-exchange magnetic beads were used to enrich the peptides in urine, and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to analyze the peptide spectrum in urine. After calibrating the instrument with a standard, mix 1 μl of the eluate with 10 μl of matrix (0.3 % α-cyano-4-hydroxycinnamic acid, HCCA), and take 1 μl to spot on the Anchorchip (Autoflex MALDITOF, Bruker-Dalton) target plate on, dry at room temperature. The sample is ionized by nitrogen laser irradiation and then subjected to mass spectrometry analysis, collecting data in the range of 1000-10000 Da, and obtaining a mass spectrogram composed of protein peaks with different mass-to-charge ratios. For each MALDI crystallization site, a total of 400 laser irradiations (50 irradiations at 8 different positions of each crystallization site), the average value represents a...

Embodiment 4

[0031] Example 4 Identification and Analysis of Differential Peptides

[0032] Take 20ul from the sample tube for LC-MS / MS analysis. EASY-nLC1000 HPLC system was used for separation. Liquid phase A was 0.1% formic acid in acetonitrile (2% acetonitrile), and liquid phase B was 0.1% formic acid in acetonitrile (98% acetonitrile). 100% solution A equilibrates a 100×100mm BEHnanoACquity column with a flow rate of 400nl / min. The chromatographic spray needle is PN: FS360-20-10-N-20-C12. Samples were separated by capillary high-performance liquid chromatography and analyzed by a Q-Exactive mass spectrometer (Thermo). The ion source voltage is 3.5kv, the analysis time is 120min, and the precursor ion scanning range is 300-2000m / z. Mass-to-charge ratios of peptides and peptide fragments were collected as follows: 20 fragments were collected after each full scan (MS2scan, HCD). At M / Z 200, MS1 has a resolution of 70,000 and MS2 has a resolution of 17,500. The data analysis soft...

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Abstract

The invention provides an application of a urine immunoglobulin kappa constant region (IGKC) and a polypeptide fragment of the IGKC, and particularly relates to an application of a urine IGKC protein and a polypeptide fragment of the IGKC protein in preparation of a preparation for monitoring normal pregnancy or gestational diabetes mellitus. Researches prove that compared with a normal childbearing age woman group (normal control), the expression of IGKC and polypeptide fragments thereof in normal pregnancy and gestational diabetes mellitus groups is increased, and the IGKC and polypeptide fragments thereof can be used for monitoring and auxiliary diagnosis of gestational state or gestational diabetes mellitus patients. According to the invention, the advantages of noninvasive acquisition, large-scale repeated sampling and convenient preservation of a urine sample are exerted, and the urine IGKC protein and the polypeptide fragment thereof are detected by using the urine sample.

Description

technical field [0001] The invention relates to the new application of urine immunoglobulin kappa constant region (IGKC) protein and its polypeptide fragment, in particular to the application of urine IGKC protein and its polypeptide fragment in monitoring normal pregnancy or gestational diabetes. Background technique [0002] During pregnancy, in order to meet the needs of fetal growth and development, a series of changes will occur in the reproductive, blood circulation, urinary, respiratory, digestive and endocrine systems of pregnant women, especially in the endocrine system, the secretion of gonadotropins decreases, prolactin, thyroid stimulating hormone, etc. And the secretion of adrenocorticotropic hormone, glucocorticoid, aldosterone, and thyroid hormone increases. In the second and third trimesters of pregnancy, some pregnant women are less sensitive to insulin, which decomposes and antagonizes insulin and reduces insulin activity. Pregnant women cannot normally com...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68C07K16/18
CPCG01N33/6893G01N33/6854G01N33/6857C07K16/18G01N2800/368G01N2800/042C07K2317/52
Inventor 张曼胡智颖
Owner 张曼
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