Application of human derived HSF2 as specific diagnosis molecular marker of ulcerative colitis

A technology of ulcerative colitis and molecular markers, applied in the field of biomedicine, can solve problems such as no reports and no specific diagnostic markers for UC.

Inactive Publication Date: 2014-08-13
缪应雷
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that heat shock factor 1 (Heat shock factor 1, HSF1) and heat shock proteins (heat shock proteins, HSPs) mediate the protective effect of inflammatory bowel disease, they pass the heat shock elements in inflammatory factor genes and affect inflammation-related The pathway of transcription facto

Method used

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  • Application of human derived HSF2 as specific diagnosis molecular marker of ulcerative colitis
  • Application of human derived HSF2 as specific diagnosis molecular marker of ulcerative colitis
  • Application of human derived HSF2 as specific diagnosis molecular marker of ulcerative colitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: Difference analysis of protein expression in serum of patients with ulcerative colitis (UC) and healthy volunteers.

[0014] 1. Collection and processing of serum samples

[0015] Take 5ml of fasting venous blood from the research subjects, place it at room temperature for 30min, centrifuge at 2000rpm for 20min, and collect serum (cannot be hemolyzed). Serum samples from 40 cases in the same group were mixed uniformly with 30 ul from each case. Albumin and IgG were then removed from serum samples using an Albumin / IgG Removal Kit. The sample protein was then concentrated using a protein precipitation kit.

[0016] 2. The first solid-phase gradient isoelectric focusing electrophoresis and the balance of the gel strip:

[0017] (1) Take a small tube (1ml / tube) of Hydration Loading Buffer (I) (no DTT, no Bio-Lyte) stored at -20°C from the refrigerator, and dissolve at room temperature. Add 0.01g DTT and 25μl Bio-Lyte to the small tube, and mix thoroughly. Ta...

Embodiment 2

[0051] Example 2: Immunohistochemical detection and analysis of differential expression of HSF2 in mucosal tissue of patients with different degrees of UC. 1. Slicing

[0052] All biopsy tissues were thawed after being taken out from the -80°C refrigerator, embedded with frozen section embedding medium, frozen in liquid nitrogen, and quickly sectioned on a constant temperature cryostat at -20°C. A tissue slice, pasted on a detachment-proof glass slide, fixed in acetone at 4°C for 15 minutes, then taken out, and quickly placed in a -20°C refrigerator for storage after the acetone was completely evaporated.

[0053] 2. Immunohistochemical experimental steps

[0054] (1) Place the slides in a 60°C oven overnight. The next day, the slices were taken out and placed in xylene liquid I and II for deparaffinization while the temperature was still high, each for 15 minutes.

[0055] (2) Then soak in absolute alcohol I and II, 95%, 80%, and 70% alcohol respectively, for two minutes e...

Embodiment 3

[0072] Example 3: Analysis of differential expression of HSF2 in mucosal tissues of UC patients with different degrees detected by real-time quantitative PCR and Western blot.

[0073] (1) Detection of differential expression of HSF2 by real-time fluorescent quantitative PCR

[0074] 1. Extraction of sample RNA

[0075] ① Take frozen and stored lysed intestinal mucosal cells, and let them dissolve completely at room temperature for 5 minutes.

[0076] ②Two-phase separation Add 0.2ml of chloroform to every 1ml of TRIZOL reagent lysed sample, and cap the tube tightly. Shake the tube vigorously by hand for 15 seconds, and incubate at 15 to 30°C for 2 to 3 minutes. Centrifuge at 12000 rpm for 15 minutes at 4°C. After centrifugation, the mixed liquid will be divided into the lower red phenol chloroform phase, the middle layer and the upper layer of the colorless aqueous phase. The RNA was all partitioned into the aqueous phase. The volume of the upper layer of the aqueous phas...

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Abstract

The invention relates to an application of human derived HSF2 as the specific diagnosis molecular marker of ulcerative colitis, and belongs to the technical field of biomedicine. The technical scheme comprises three parts: (1) detecting differential proteins in serums of UC patients and healthy volunteers through a proteomics two-dimensional gel electrophoresis technology and a mass spectrum technology; (2) detecting the HSF2 expression level in colonic mucosa of UC patients in different degrees through an immunohistochemical method; (3) detecting the HSF2 expression level in colonic mucosa of UC patients in different degrees through a real-time quantitative PCR technology and a western blotting technology. 40 UC patient serums and 40 healthy volunteer serums have been compared by the method mentioned above, and the results show that the expression of HSF2 in the UC patient serums prominently increases. Moreover, an immunohistochemical method, a real-time quantitative PCR technology and a western blotting technology are adopted to detect the HSF2 expression in the colonic mucosa of UC patients, and the results show that the HSF2 expression increases as the UC disease goes worse. Thus, the HSF2 can be used as a molecular marker to specifically diagnosing ulcerative colitis.

Description

Technical field: [0001] The invention relates to the application of human HSF2 as a specific diagnostic molecular marker for ulcerative colitis, belonging to the technical field of biomedicine. Background technique: [0002] Ulcerative colitis (UC) is a chronic non-specific colonic inflammation characterized by recurrent abdominal pain, mucus, pus and blood in the stool. In the past 15 years, the number of UC patients in my country has increased year by year, at least over 200,000. The disease is now becoming a common disease of the digestive system and the main cause of chronic diarrhea in my country. Most of the patients are young and middle-aged, and the course of the disease is protracted, and there is a certain relationship with the incidence of colon cancer, which seriously affects the quality of life of patients and social productivity. Studies have shown that patients with a medical history of more than 30 years have an annual risk of colorectal cancer as high as 2...

Claims

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Application Information

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IPC IPC(8): G01N27/64G01N33/68C12Q1/68
Inventor 缪应雷王昆华牛俊坤杜艳周丽峰杨刚
Owner 缪应雷
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