Recombination plasmid and application in disease prevention and control

A technology of recombining plasmids and diseases, applied in the field of biomedicine, to achieve the effect of preventing excessive scar formation, simple and convenient application, and promoting wound healing

Inactive Publication Date: 2004-05-19
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology can be used to improve blood flow during surgery or reduce scars caused by blockages in arteries. It involves introducing certain genes called hgf (human follicle-stimulating factors) onto affected areas where they are damaged due to injury or disease. These genes help stimulate neovascularization within these regions while also reducing their ability to form connective tissue structures like ligaments on surrounding bone surfaces. By adding this type of material, patients may have better outcomes from medical procedures involving blocked vessels.

Problems solved by technology

This patents discusses different ways to deliver medicine safely and efficiently during heart attacks such as vasculitis caused by plaque rupture syndrome. There currently exist several techniques like injecting stem cells directly onto damaged areas, applying external factors like pressure waves, electricity shock waveings, ultrasonics, laser irradiation, etc., but these approaches face challenges due to their invasiveness and potential harmful effects associated with them. To address this issue, we propose introducing biology technology called gene therapies, specifically fusion vectors containing exonuous hair follicle angioplasty agents, to provide better safety and efficacy over existing strategies.

Method used

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  • Recombination plasmid and application in disease prevention and control
  • Recombination plasmid and application in disease prevention and control
  • Recombination plasmid and application in disease prevention and control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Experimental Study of Using Recombinant Plasmid pUDKH Gene to Treat Limb Ischemia in Rats

[0059] 1. Construction and preparation of recombinant plasmids

[0060] (1) Construction of plasmid pUDKH

[0061] Build process like figure 1 : The pUC18 cloning vector was digested with PvuII, and a 2.3kb DNA fragment was recovered. pcDNA3 was digested with NruI and PvuII, and a 1 kb DNA fragment was recovered, which contained cytomegalovirus promoter, multiple cloning site and growth hormone terminator. The two fragments were connected to obtain plasmid pUD (3.3kb). Digest pUD with AvaII and SspI to recover a 2.7kb DNA fragment, and digest pEGFP-N1 with AvrII to recover a 1.1kb DNA fragment, which contains the complete kanamycin gene; connect the two recovered DNA fragments to obtain Plasmid pUDK (3.8kb). Then insert the cDNA of the human hepatocyte growth factor coding region into the multiple cloning site of the pUDK vector to obtain the recombinant vector pUD...

Embodiment 2

[0083] Example 2 Prediction of scar formation by recombinant plasmid pUDKH carrying human hepatocyte growth factor gene

[0084] Anti-effect

[0085] 1. Construction and large-scale purification of recombinant plasmid pUDKH

[0086] Same as Example 1

[0087] 2. Transfection of recombinant plasmid DNA into NIH3T3 cells

[0088] The plasmid DNA was transfected into NIH3T3 cells using the lipofectin method, and 5 micrograms of DNA + 20 microliters of liposomes were used for each transfection. After transfecting pUDK-lacZ, its transfection efficiency was 7.96%; after transfecting pUDKH (number of cells was 1.0×10 6 ), samples were taken at 24 hours, 48 ​​hours and 72 hours after transfection, and the expression level of HGF was detected by ELISA method. The primary antibody was mouse anti-human HGF monoclonal antibody (R&D). The results showed that the expression of HGF could be detected in the supernatant of NIH3T3 cells transfected with pUDKH (such as Figure 4 , Figure...

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Abstract

The present invention relates to a recombinant plasmid with human hepatocyte growth factor gene and its application for preventing and curing diseases. The human hepatocyte growth factor gene is inserted into pUDK plasmid vector to obtain recombinant plasmid pUDKH. After said recombinant plasmid is transferred into ischemic region of rat or rabbit member by means of local intramuscular injection or gene gun method, said recombinant plasmid can effectively express and obviously promote vascularization. Said recombinant plasmid can be applied to surface of wound of rabbit ear of transferred into wound position by means of gene gun, it possesses the function of preventing and curing scar, so that aid recombinant plasmid possesses good application for preventing and curing ischmic diseases and pathological scar.

Description

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Claims

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Application Information

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Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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