TPA gene thread and its preparation method

A suture and gene technology, applied in the medical field, can solve problems such as 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: 2007-08-01
姬尚义
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chinese patent CN94100044.3 discloses gene and drug sutures, bu...

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 a kind of tPA gene suture as well as preparing method, the characteristic of the invention is that the eukaryotic expression vector pSecTag2B-t-PA of modified gene tPA is attached to the suture. The preparing procedure includes the following steps: labeling the pSecTag2B-t-PA recombinant plasmid with basic ion bangosome; soaking the suture into bangosome/plasmid DNA compound solution repeatedly to get the tPA gene suture. The invention can be used in the saturation of cardiac muscle in cardiac valve replacement; it can remove or prevent the sludged blood so that it can prevent the form of thrombus around the valvula which can impact the valvula function after cardiac valve replacement.

Description

technical field [0001] The invention relates to the medical field, in particular to a tP4 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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IPC IPC(8): A61L17/14A61L17/00C12N15/79C12N15/58
Inventor 姬尚义
Owner 姬尚义
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