TPA gene thread and its preparation method

A suture and gene technology, applied in the medical field, can solve the problems of structure and preparation method description, and achieve the effects of eliminating or avoiding the formation of blood clots, enhancing fibrinolytic activity, and prolonging survival time

Inactive Publication Date: 2009-07-15
姬尚义
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent CN94100044.3 discloses gene and drug sutures, but does not describe its own specific structure and preparation method

Method used

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  • TPA gene thread and its preparation method
  • TPA gene thread and its preparation method
  • TPA gene thread and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Preparation of tPA gene suture

[0023] 1. Main reagents

[0024] Plasmid pSecTag2B was purchased from Invitrogen Biological Company of the United States, and competent cells E.ColiJM109 were purchased from Promega Company. Restriction enzymes HindIII, KpnI, BamHI and XhoI were purchased from Bao Biological Engineering Co., Ltd., T 4 - DNA ligase was purchased from New England Biolabs, and QIAquick Gel Extraction Kit and QIAGEN PCR Product Purification Kit were purchased from QIAGEN.

[0025] 2. Method

[0026] Three ESTs were artificially synthesized: tPA-1 as shown in SEQ ID NO.2, tPA-2 as shown in SEQ ID NO.3 and tPA-3 as shown in SEQ ID NO.4, respectively connected to cloning vectors to prepare plasmid pOTB6 , pOTB7 and pCMV-SPORT6.

[0027] Three pairs of primers t-PA-1F (SEQ ID NO.5) and t-PA-1R (SEQ ID NO.6), t-PA-2F (SEQ ID NO.7) and t-PA-2R (SEQ ID NO. NO.8), t-PA-3F (SEQ ID NO.9) and t-PA-3R (SEQ ID NO.10) amplify plasmids pOTB6, pOTB7 and pCMV-...

Embodiment 2

[0036] Example 2 Effect of tPA gene suture in preventing thrombosis after cardiac surgery

[0037] 1. Materials

[0038] Animals: 36 healthy pigs, weighing 40-50KG, female, 8 months old. Among them, 18 were used for blood supply, and the other 18 were randomly divided into 9 in the control group and 9 in the experimental group.

[0039] 2. Method

[0040] 2.1 Pig tricuspid valve replacement and transgene therapy

[0041] The control group underwent simple tricuspid valve replacement under conventional extracorporeal circulation; the experimental group sutured the prepared tPA gene suture serpentinely or circularly between the myocardial wall of the right ventricular outflow tract after tricuspid valve replacement, and the length of the gene suture About 10cm, at the same time, perform ultrasonic treatment on the part of the gene suture, frequency: 1kHz, intensity: 0.5w / cm 2 , time: 30min. At 1 week, 2 weeks, 4 weeks, 8 weeks and 12 weeks after operation, venous blood was ...

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Abstract

The invention relates to the medical field, and provides a tPA gene suture and a preparation method thereof, which is characterized in that the recombinant tPA gene eukaryotic expression vector pSecTag2B-t-PA is attached to the surgical suture. The preparation method comprises: marking pSecTag2B-t-PA recombinant plasmid with cationic liposome; repeatedly soaking surgical suture in liposome / plasmid DNA complex solution to obtain tPA gene suture. The tPA gene suture of the invention is used for myocardial suturing in heart valve replacement, which can eliminate or avoid the formation of blood clots in time, so as to avoid the formation of thrombus around the valve after valve replacement and affect the function of the valve.

Description

technical field [0001] The invention relates to the medical field, in particular to a tPA gene suture and a preparation method thereof. Background technique [0002] Under normal physiological conditions, the coagulation-anticoagulation and fibrinolysis-antifibrinolysis mechanisms balance each other to ensure the normal flow of blood in the body without the formation of thrombus or bleeding. The imbalance between thrombus formation and thrombolysis mechanism makes thrombus not form properly or cannot be effectively cleared after formation, leading to tissue / organ ischemia / necrosis; on the other hand, excessive anticoagulation / fibrinolysis can also lead to bleeding. [0003] Thrombosis is easy to form after artificial heart valve replacement, and patients usually need anticoagulant therapy for life, which brings great inconvenience and danger to patients. Tissue-type plasminogen activator (tissue-type plasminogen activator, tPA) is an important factor in the body to induce f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L17/14A61L17/00C12N15/79C12N15/58
Inventor 姬尚义
Owner 姬尚义
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