Application of HIPK2 in prediction and diagnosis of septicemia
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A sepsis and risk prediction technology, applied in the fields of biomedicine and disease diagnosis, can solve problems such as use limitations
Pending Publication Date: 2021-07-30
SHANGHAI UNIV
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Embodiment 1
[0030] 1. Experimental method
[0031] 1.1 Isolation of human peripheral blood mononuclear cells
[0032] (1) Prepare a 15ml (or 50ml) centrifuge tube according to the volume of the blood sample, and add blood lymphocyte separation solution LSM equal to the volume of the blood to the bottom of the tube;
[0033] (2) Blow the peripheral blood sample in the anticoagulant tube evenly with a gun, slowly suck the blood and add it to the surface of the separation liquid;
[0034] (3) 2200rpm, horizontal centrifugation at room temperature for 25-30 minutes, pay attention to the slow rise and fall of the centrifuge;
[0035] (4) After centrifugation, red blood cells can be seen sinking to the bottom of the tube, the white layer in the middle is mononuclear cells, and the uppermost layer is plasma. Absorb the cells in the middle layer into a new 15ml tube;
[0036] (5) Fill up the centrifuge tube with PBS and centrifuge at 1300rpm for 5 minutes;
[0037] (6) Discard the supernatant....
Embodiment 2
[0057] 1. Experimental method
[0058] 1.1 LPS endotoxic shock model
[0059] The endotoxic shock model uses 8-12 week old littermate pairs (same age and sex) of wild-type and Hipk2-deficient mice. By intraperitoneal injection of LPS at a dose of 36mg / kg, the general mouse injection volume is 150-200μl. The changes in body weight and death of mice were observed 4 hours after the injection. Most mice died within 12-48 hours of LPS injection. Blood collection and organ experiments were carried out within 3-6 hours of LPS injection.
[0060] 1.2 Cecal ligation and puncture (CLP) model
[0061] (1) The mice were weighed, and the mice were fully anesthetized by intraperitoneally injecting 20 μl / g of tribromoethanol according to the body weight;
[0062] (2) Fix the mice supine on the experimental table, fix the limbs with medical tape, remove the hair from the abdomen, and sterilize with 75% ethanol;
[0063] (3) Open the abdominal skin along the white line of the mouse abdomen...
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Abstract
The invention provides application of HIPK2 in sepsis prediction and diagnosis, belongs to the technical field of biomedicine and disease diagnosis, and provides application of HIPK2 in preparation of a sepsis risk prediction, diagnosis and disease course monitoring reagent. The invention discloses an application of HIPK2 in preparation of a septicemia prediction and diagnosis reagent, and verifies that the transcriptional level of HIPK2 in peripheral blood mononuclear cells of a septicemia patient is significantly reduced, and HIPK2 is negatively correlated with a pro-inflammatory gene IL1B and positively correlated with an anti-inflammatory gene IL10; the cellular level experiments find that HIPK2 can inhibit generation of pro-inflammatory cytokines; therefore, the probability that healthy people with low expression of the HIPK2 develop into septicemia during microbial infection can be increased, so that the HIPK2 can be used as a potential septicemia prediction and diagnosis biomarker. According to the invention, a theoretical basis and a new detection method are provided for septicemia prediction and diagnosis.
Description
technical field [0001] The invention relates to the technical fields of biomedicine and disease diagnosis, and in particular to the application of HIPK2 in the prediction and diagnosis of sepsis. Background technique [0002] Sepsis is a systemic inflammatory response caused by pathogenic microorganisms invading the blood circulation, proliferating and producing toxins in the blood. Sepsis includes the entire infection process from systemic inflammatory response syndrome (SIRS) to septic shock, and the course of the disease ranges from fever, leukocytosis, etc. to multiple organ dysfunction and even death. It is a disease with a high mortality rate ranging from 25%-30% in patients with general and severe sepsis and increasing to 40%-50% in patients with septic shock. Although great progress has been made in the continuous research on sepsis worldwide, there is still no specific treatment for sepsis, so early diagnosis of sepsis is the key to prevent its malignant developmen...
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