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Tissue repair protein CHRD, and coding gene and applications thereof

A technology that encodes genes and proteins, which is applied in the fields of biomedicine and daily necessities, can solve problems such as unsatisfactory application, secondary trauma of wounds, and insufficient fluid absorption, and achieve significant repair effects, effective repair, and improved healing speed Effect

Active Publication Date: 2020-06-19
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional dressing mainly refers to medical absorbent cotton gauze, although it has many advantages, its application is not satisfactory
For example, the amount of liquid absorption is not large enough, so it needs to be packed thickly and replaced frequently; the dressing is easy to dry after absorbing liquid, and it will stick to the wound, causing secondary trauma, and it is inconvenient to use.

Method used

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  • Tissue repair protein CHRD, and coding gene and applications thereof
  • Tissue repair protein CHRD, and coding gene and applications thereof
  • Tissue repair protein CHRD, and coding gene and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1. CHRD gene design and protein expression

[0039] 1. Genetic design

[0040] The inventor designed a gene whose nucleotide sequence is shown in Sequence 2 in the sequence listing, with a full length of 1719bp. The gene encodes a protein whose amino acid sequence is shown in Sequence 1 in the sequence table, with a total length of 572 amino acids, and is named CHRD protein. Sangon Bioengineering (Shanghai) Co., Ltd. was entrusted to carry out the whole gene synthesis of the gene and added Ndel restriction site (CATATG) and Xhol restriction site (CTCGAG) at the 5' end and 3' end of the gene respectively. The gene sequence with restriction sites is shown as sequence 3 in the sequence listing. The synthesized gene was sequenced and verified, and the gene with the correct sequence was used for subsequent vector construction.

[0041] Amino acid sequence of CHRD (572aa)

[0042]MSSGVDLGTENLYFQSNAMSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRRLMEAFAKRKKMKGVQLKL...

Embodiment 2

[0065] Example 2. Wound healing performance evaluation of CHRD protein

[0066] 1. Evaluation of wound healing effect of pure skin

[0067] Male SD rats with a body weight of 250-300 g were selected to create a back full dermal trauma model. Nine SD rats were randomly divided into 3 groups, 3 in each group, wherein 2 groups were used as the experimental group, and 1 group was used as the control group, anesthetized by intraperitoneal injection of chloral hydrate (10wt%) (0.5mL / 100g). Under aseptic conditions, after the back is shaved and routinely disinfected, create a 1.0×1.0cm spine with the spine as the midline 2 The size of the skin excision wound. The two experimental groups were repaired with gauze and CHRD powder respectively, in which the gauze group used sterile gauze to bind the wound (gauze was changed every 3 days), and the CHRD group was evenly applied on the wound with CHRD powder (the dressing was changed every 3 days). ). The control group exposed the wound...

Embodiment 3

[0076] Example 3. Biosafety evaluation of CHRD protein

[0077] MTT method was used to study the toxicity of CHRD protein to mouse fibroblast L929 (Cell Bank of Type Culture Collection Committee, Chinese Academy of Sciences) to evaluate the biological safety of CHRD protein. Mouse fibroblast L929 cells were used at 1×10 4 The concentration / well was inoculated into DMEM medium (Beijing Soleibao) in a 96-well plate. Five experimental groups and one control group were set up, and three replicate holes were set up for each group. After culturing the cells at 37°C for 24 hours, CHRD protein was added to the wells of the experimental group, so that the final concentrations of CHRD protein in the five experimental groups were 50 mg / L, 100 mg / L, 200 mg / L, 400 mg / L, and 800 mg / L. The control group did not add CHRD protein. Continue culturing at 37°C for 24 hours, and then use the MTT method to detect the number of viable cells, as follows: add 10 μl of MTT solution (5 mg / ml) to each...

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Abstract

The invention relates to the field of biological medicines and daily necessities, and specifically relates to tissue repair protein CHRD, and a coding gene and applications thereof. The protein is shown as the following a1 or a2: a1, the protein whose amino acid sequence is shown as a sequence 1 in a sequence table; and a2, the protein formed through the substitution and / or deletion and / or addition of one or several amino acid residues to the amino acid sequence shown as the sequence 1 in the sequence table and related to the repairing of skin wounds. Through the proving of experiments, the CHRD protein has significant repair effects on simple skin wound repair and wounds with radiation and skin damage; and compared with sterile medical gauze, the CHRD protein can make the healing speed ofthe wounds with radiation and skin damage accelerated about 30%.

Description

technical field [0001] The invention relates to the fields of biomedicine and daily necessities, in particular to a recombinant protein CHRD for skin wound repair, its coding gene and its application. Background technique [0002] The skin is an important barrier of the human body. Various operations, trauma, burns, dermatitis, etc. will lead to skin damage and defects, causing various traumas. Wound repair refers to a series of pathophysiological processes in which local tissues are repaired through regeneration, repair, and reconstruction after tissue defects (such as wounds, wounds, etc.) are caused by trauma or other injuries. It is an important field of medical development. It is of great significance to alleviate the pain of patients and avoid scars after repair. [0003] The process of wound repair requires the help of new medical materials, especially biomedical materials. Biomaterials are used to contact and interact with living systems, and to diagnose and treat c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/00C12N15/11C12N15/70C12N1/21A61K38/16A61K9/00A61P17/02
CPCA61K9/0014A61K38/00A61P17/02C07K14/00C12N15/70
Inventor 彭志平陈晓良
Owner CHONGQING MEDICAL UNIVERSITY
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