Application of FAM127A in pregnancy diseases

A technology of FAM127A and reagents, applied in the field of biomedicine, can solve the problems of slow progress in clinical management and treatment, limited curative effect, etc.

Active Publication Date: 2018-11-13
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The details of its pathogenesis are still to be discovered, resulting in slow progress in its clinical management and treatment in recent years, its diagnosis is still based on the full exposure of the patient's clinical symptoms, and all treatments are symptomatic, with relatively limited efficacy , repeatedly putting obstetricians and neonatologists into a passive situation
Although the current preventive measures against PE have made some progress, their extensive clinical application needs further verification

Method used

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  • Application of FAM127A in pregnancy diseases
  • Application of FAM127A in pregnancy diseases
  • Application of FAM127A in pregnancy diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1 Screening for Gene Markers Related to Preeclampsia

[0069] 1. Sample collection

[0070] 1) Collection of serum samples

[0071] The blood of 45 cases of normal pregnant women and untreated preeclampsia patients was collected, and the EDTA anticoagulant tube was left standing for 10 minutes, and the serum was separated by centrifugation, and stored at -20°C for later use.

[0072] 2) Collection of placenta specimens

[0073] The placenta tissues of 45 cases of preeclampsia and normal pregnant women were collected respectively, rinsed with normal saline twice, after removing water, they were divided into cryopreservation tubes, and stored at -80°C for later use.

[0074] Multiple pregnancies, infectious diseases, chemical drug dependence, maternal smoking, fetal congenital malformations and other pregnancy complications and complications were excluded in both groups. All included subjects signed informed consent before collecting specimens. All the above sp...

Embodiment 2

[0089] Example 2 QPCR sequencing to verify the differential expression of the FAM127A gene

[0090] 1. Large-sample QPCR verification of differential expression of FAM127A gene.

[0091] 2. RNA extraction

[0092] Use QIAGEN tissue RNA extraction kit to extract total RNA in placental tissue, and blood RNA extraction kit to extract RNA in serum. For specific steps, refer to the instruction manual.

[0093] 3. Reverse transcription:

[0094]1) Add dNTP mixture 1μl, Oligo dT primer 1μl, total RNA 2μg, add RNase FreeddH 2 O Make the total volume to 10 μl, carry out denaturation and annealing reaction on the PCR instrument, 65°C, 5min, place at 4°C after the reaction is completed.

[0095] 2) Construct a 20 μl reaction system, continue to add 4 μl of 5×Primer Script Buffer, 0.5 μl of RNase Inhibitor, 0.5 μl of Prime Script RTase, RNase Free ddH 2 O 5.0 μl, carry out the reverse transcription reaction on the PCR instrument according to the following conditions: 42°C for 15-30 mi...

Embodiment 3

[0113] Overexpression of embodiment 3FAM127A gene

[0114] 1. Cell culture

[0115] Human early pregnancy villous trophoblast cell line (HTR-8 / SVneo) was incubated in RPIM-1640 medium containing 10% fetal bovine serum at 37°C, 5% CO 2 cultured in an incubator. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0116] 2. Transfection

[0117] 1) Treatment of cells before transfection

[0118] The trophoblast HTR-8 / SVneo in the logarithmic phase was treated with 1×10 5 The densities were planted in six-well plates at 37°C in 5% CO 2 cultured in an incubator.

[0119] 2) Construction of gene overexpression vector

[0120] Synthesize specific PCR amplification primers according to the sequence of FAM127A in GeneBank, and the primer sequences are as follows:

[0121] Forward primer: 5'-CCGGAAGCTTGCCACCATGGACGGTCGGGTGC-3' (SEQ ID NO.5)

[0122] Reverse primer: 5'-CGGGCGGCCGCGAAGTCCTCGTCTCTCCTCCCA-3' (SEQ ID NO.6...

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Abstract

The invention disclosed application of FAM127A in a pregnancy disease. Specifically, the disease is pre-eclampsia. The expression of FAM127A is down-regulated in a pregnant womon with pre-eclampsia. It is found through large sample verification that the gene has higher positive detection rate, and it is proposed that the gene can be applied to clinical diagnosis of pre-eclampsia. By overexpressingthe FAM127A in trophoblast cells, it is found that the proliferation ability and invasion ability of the trophoblast cells are changed, and it is prompted that the FAM127A can serve as a potential target for the treatment of pre-eclampsia.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to the application of FAM127A in pregnancy diseases, and the specific diseases are preeclampsia. Background technique [0002] Preeclampsia (PE) is a more serious type of hypertensive disorders during pregnancy, mainly manifested as hypertension and proteinuria that occur after 20 weeks of pregnancy, and can be secondary to eclamptic convulsions, cerebral hemorrhage, cerebral edema, multiple organ Insufficiency, HELLP syndrome, placental abruption, fetal growth restriction and other serious maternal and fetal complications. It is the main cause of maternal and child morbidity and mortality, and it also has adverse effects on the long-term prognosis of the mother and her offspring. The incidence of PE varies significantly between different countries and regions, and the incidence of PE in developed countries such as Europe and the United States is between 1% and 5% (Abalos E, Cuesta C, Gros...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/68A61K45/00A61P15/00
CPCA61K45/00A61P15/00C12Q1/6883C12Q2600/136C12Q2600/158G01N33/6893G01N2800/368
Inventor 杨承刚孙耀兰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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