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Screening method for differentially expressed proteins of heterotopic-pregnancy and non-pregnancy decidua tissue

A technology for differentially expressing proteins and decidual tissue, applied in chemical instruments and methods, animal/human peptides, material testing products, etc., can solve problems such as difficulty, difficulty in repeating experiments, and indistinguishable morphological differences, so as to improve the detection ability. , improve the technical level, realize the effect of economic and social benefits

Inactive Publication Date: 2017-05-10
WUHAN LABWAY MEDICAL LAB CO LTD
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Problems solved by technology

[0002] The judgment of pregnant and non-pregnant endometrial tissue is very important clinically. There is no absolute limit for the transition state of endometrium from late secretory stage to early pregnancy stage. Differential diagnosis of decidua-like tissue and non-pregnancy decidua tissue, clinical diagnosis of ectopic pregnancy based on curettage material requirements, etc., there is no satisfactory solution at present, because the endometrial hormone secretion level and endometrial tissue in the process of transition from late secretion to early pregnancy The difference in response to hormones, decidualization of the endometrium caused by some non-pregnancy factors, etc., in the absence of villi, the subtle differences in morphology are difficult to distinguish, and it is necessary to determine whether pregnancy occurs, whether it is intrauterine or Ectopic pregnancy is very difficult
There are few reports on the differences in ultrastructure and molecular biology between the two, and even if there are reports, the specificity is not strong, or the experiment is difficult to repeat

Method used

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  • Screening method for differentially expressed proteins of heterotopic-pregnancy and non-pregnancy decidua tissue
  • Screening method for differentially expressed proteins of heterotopic-pregnancy and non-pregnancy decidua tissue
  • Screening method for differentially expressed proteins of heterotopic-pregnancy and non-pregnancy decidua tissue

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Embodiment 1

[0034] The present invention will respectively carry out tissue culture, light microscope, electron microscope and immune examination to the decidua tissue during intrauterine and ectopic pregnancy, and main contents include:

[0035] 1. For the separation of two kinds of decidua, cell culture, and culture medium;

[0036] 2. Use flow cytometry to examine the distribution of dendritic cells in pregnant, non-pregnant and cycle endometrial tissues;

[0037] 3. Using electron microscopy to study the ultrastructural characteristics of different endometrial tissues, straighten out the relationship between decidual granulosa cells and lymphocytes and dendritic cells containing granules in the cells reported in the literature, and clarify the discovered cell surface What kind of cell is the podocyte-like cell foot process and its function.

[0038] Complete the preparation of protein samples: lyse, freeze-thaw, grind, and centrifuge the decidua separated from ectopic pregnancy, meas...

Embodiment 2

[0041] It was found that the S-100 protein was positive in decidual glands, but also negative in endometrial glands, and positive in some decidual cells (see attached Figure 1-4 shown);

[0042] Masson's trichrome staining can prove the "feather-like collagen inclusions" in the degenerative decidua cells, and suggest that pregnancy has occurred. Because of the false positive results, the accuracy rate is about 85%.

[0043]Electron microscopy revealed that decidual cells were large, round and oval, with rich cytoplasm and organelles, and well-developed rough endoplasmic reticulum, mitochondria, phagosomes, and inclusion bodies. The cell surface has a cellulose-like extracellular plasma membrane similar to the basement membrane. In addition to gap junctions and tight junctions, typical desmosomes were formed between adjacent cells, but this structure was not seen between decidua-like cells. Many scattered and bundled tension filaments were found in some decidual cells, which...

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Abstract

The invention discloses a screening method for differentially expressed proteins of heterotopic-pregnancy and non-pregnancy decidua tissue. The method is characterized by comprising the following steps: carrying out proteomics analysis to discover specific proteins and differential proteins of heterotopic-pregnancy and non-pregnancy decidua tissue; subjecting the specific proteins and differential proteins to separation and purification; and using the purified specific proteins and differential proteins as special biological markers for discrimination of intrauterine and extrauterine pregnancy. After pregnancy, specific proteins can be synthesized in the bodies of both a mother and a fetus. Detection of the specific proteins with different methods is beneficial for diagnosis of pregnancy. Searching for a special marker for pregnancy, especially for distinguishing intrauterine and extrauterine pregnancy, is of critical significance to determination of clinical medical liabilities and medical jurisprudence and is a great breakthrough to the basic theory of medical science.

Description

technical field [0001] The invention relates to a method for screening proteins differentially expressed in ectopic pregnancy and non-pregnant decidua tissues. Background technique [0002] The judgment of pregnant and non-pregnant endometrial tissue is very important clinically. There is no absolute limit for the transition state of endometrium from late secretory stage to early pregnancy stage. Differential diagnosis of decidua-like tissue and non-pregnancy decidua tissue, clinical diagnosis of ectopic pregnancy based on curettage material requirements, etc., there is no satisfactory solution at present, because the endometrial hormone secretion level and endometrial tissue in the process of transition from late secretion to early pregnancy The difference in response to hormones, decidualization of the endometrium caused by some non-pregnancy factors, etc., in the absence of villi, the subtle differences in morphology are difficult to distinguish, and it is necessary to de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N15/14C07K14/47
CPCG01N33/689C07K14/47G01N15/14
Inventor 王成鑫吴玲敏邵俊宗阳如吴笛魏文宁
Owner WUHAN LABWAY MEDICAL LAB CO LTD
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