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Ultrasonic microbubbles for treating arthritis and use thereof

A technology of ultrasonic microbubbles and arthritis, applied in the treatment of inflammatory arthritis, in the field of therapeutic ultrasonic microbubbles, to reduce the burden of life, improve the overall treatment effect, and reduce the economic and living burden

Inactive Publication Date: 2013-04-03
叶琳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no relevant literature report on the combination of tumor necrosis factor receptor gene and ultrasonic microbubbles at home and abroad to prepare therapeutic ultrasonic microbubbles for the treatment of inflammatory arthritis

Method used

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  • Ultrasonic microbubbles for treating arthritis and use thereof
  • Ultrasonic microbubbles for treating arthritis and use thereof
  • Ultrasonic microbubbles for treating arthritis and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Evaluation of the properties of the therapeutic microbubbles of the present invention

[0042] figure 1 Shown is the model diagram of therapeutic ultrasonic microbubbles combined with tumor necrosis factor receptor gene. The tumor necrosis factor receptor gene plasmid is added before the formation of ultrasound contrast agent microbubbles. The specific method is: combine the tumor necrosis factor receptor gene plasmid with The film-forming material of lipid ultrasound contrast agent microbubbles is added before the lipid suspension is oscillated into microbubbles, and a suspension containing a large number of microbubbles is formed after oscillation by mechanical oscillation. After oscillation, the tumor necrosis factor receptor gene Plasmids are electrostatically bound directly to the inner or outer surface of the microvesicle shell. This method is mainly aimed at lipid ultrasound contrast agent microbubbles. Before the formation of ultrasound contrast agent...

Embodiment 2

[0048] Example 2 The present invention competitively blocks the binding of TNF and receptors in vitro

[0049] 1. Experimental method (taking sTNFR-Fc / pcDNA3.1 as an example)

[0050] LB liquid medium (tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L) at 37 degrees, shaking culture transformed into sTNFR-Fc / pcDNA3.1, pcDNA3.1, sTNFR-4HA / pcDNA3 respectively .1 and 4HA / pcDNA3.1 plasmids for the four DH5α strains. 10000g high-speed centrifugation to collect 3ml of bacterial liquid, using commercial plasmid extraction kit E.Z.N.A.? Plasmid Mini Kit I ( Omega Bio , D6943) using the classic alkaline lysis method and silica gel column purification, and extract the plasmid according to the instructions. Plasmid concentration and purity were determined by UV spectrophotometer.

[0051] Human embryonic kidney cells (HEK293) at 1×10 5 Cells were seeded in a six-well plate and cultured overnight in DMEM high-glucose medium containing 10% fetal bovine serum. After the cell fu...

Embodiment 3

[0074] Example 3 The present invention competitively blocks the binding of TNF and receptors in vivo to alleviate the effect of rheumatoid arthritis

[0075] 1. Experimental method

[0076] Male DBA / 1j mice aged 6-8 weeks, weighing 20g, were randomly divided into groups, and the CIA model was established: 100 μg of bovine type II collagen was dissolved in 100 μl of acetic acid and fully emulsified with complete Freund’s adjuvant. Choose mice for multi-point injection at the base of the tail, and 21 days later, use the same dose to enhance immunity at the same location again. Mice received an injection of the same volume (100 μl) of PBS as a control. Among them, 24 days after the CIA model was established, the mice were randomly divided into groups and treated.

[0077] 2. Experimental grouping

[0078] ① sTNFR-Fc / pcDNA3.1 microbubble treatment ultrasound group: mix microbubbles and sTNFR-Fc / pcDNA3.1 plasmid solution according to a certain ratio (30μl microbubbles contain 15...

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Abstract

The invention discloses ultrasonic microbubbles for treating arthritis and a use thereof. The microbubbles contain gene plasmids capable of expressing tumor necrosis factor receptor (TNFR) protein, wherein the gene plasmids are adsorbed to the surface of the microbubbles and / or packaged in the microbubbles; the gene plasmids are expression plasmids; the gene plasmids of the tumor necrosis factor receptor protein contain extracellular domain gene of the tumor necrosis factor receptor (TNFR) or contain gene segment of the protein or peptide fragment, which can be combined with a tumor necrosis factor (TNF); and the gene plasmids express soluble protein or peptide fragment which can prevent the tumor necrosis factor from combining with the tumor necrosis factor receptor on a cell membrane after transfecting the body cells. The ultrasonic microbubbles are not only low in manufacturing cost for greatly relieving the economic burden of the patients and families, but also have breakthrough development in the field of treating anthritis, and can be extensively popularized and used in the field.

Description

technical field [0001] The invention relates to the field of inflammatory arthritis treatment, specifically a therapeutic ultrasonic microbubble containing a plasmid capable of producing soluble tumor necrosis factor receptor gene, and its use in the treatment of inflammatory arthritis. Background technique [0002] Tumor necrosis factor (English abbreviation: TNF) is a cytokine secreted by macrophages and lymphocytes, TNF-α is mainly secreted by mononuclear macrophages, and TNF-β is mainly secreted by activated T lymphocytes. TNF got its name from early research finding that it can directly dissolve and inhibit proliferation of tumors. Recent studies have proved that TNF-α is involved in various inflammatory reactions and is a very important pro-inflammatory factor. TNF-α plays a very important role in the pathogenesis of inflammatory arthritis. In clinical diseases, the level of TNF-α in the body is too high, which mediates the inflammatory response and causes the destruc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/00A61K48/00A61K49/00A61K41/00A61P19/02
Inventor 叶琳
Owner 叶琳
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