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Method based on gene/protein as diagnostic marker and therapeutic target for biliary atresia and application thereof

A technology of biliary atresia and treatment target, applied in the biological field, can solve problems such as inability to treat biliary atresia, and achieve the effects of improving hypoxia and bile duct injury, improving liver survival rate, and slowing down the progression of the disease.

Pending Publication Date: 2020-10-16
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The purpose of the present invention is to provide a method based on gene / protein as a diagnostic marker and therapeutic target for biliary atresia and its application to solve the problem that the prior art does not provide a method for diagnosing and treating biliary atresia from the perspective of vascular morphology, and Existing diagnostic methods for biliary atresia cannot be used for the treatment of biliary atresia based on their technical basis

Method used

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  • Method based on gene/protein as diagnostic marker and therapeutic target for biliary atresia and application thereof
  • Method based on gene/protein as diagnostic marker and therapeutic target for biliary atresia and application thereof
  • Method based on gene/protein as diagnostic marker and therapeutic target for biliary atresia and application thereof

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Embodiment 1

[0052] The present invention provides a method based on gene / protein as a diagnostic marker and therapeutic target for biliary atresia, comprising the following steps:

[0053] ① Summarize the structural characteristics of the human BA hepatic artery system. In order to understand the structural characteristics of the human hepatic artery system, the main hepatic artery and the level of resistance arterioles in the intrahepatic portal area were selected for observation. The former was measured by Doppler ultrasound and indocyanine green vessels contrast methods, the latter with the aid of histological measurements;

[0054] ② To study the expression of Notch in the liver of human BA and animal models. In order to study the expression of Notch from multiple levels, the detection methods used RT-PCR, Western-blots, OPAL multispectral staining, and primary culture and staining of smooth muscle cells;

[0055] ③ Diverse interventions targeting Notch3 in animal models. Specificall...

Embodiment 2

[0078]Observation method of spider-like vascular sign (HSST sign) on the liver capsule surface: HSST sign observation and photo collection can be completed under direct vision of the laparoscopic system. In this example, after connecting the laparoscope and before the free gallbladder operation, photos of the liver and diaphragm were collected. Image 6 It indicated that the liver histopathology of BA children less than 1 month old did not show obvious fibrosis, but showed the same typical spider-like vascular signs on the liver capsule surface as 2-month-old BA children.

[0079] Rotavirus (RRV)-induced mouse model method and mouse liver capsule surface vascular sign observation method: adult wild-type SPF grade BALB / c mice (7w) were bred according to the ratio of male to female 2:1 or 3:1, each The body weight changes of pregnant mice were recorded daily, and the production situation was expected to be checked every day during the expected period of delivery for timely proce...

Embodiment 3

[0082] In step ② of Example 1, "Study on the expression of Notch in the liver of human BA and animal models", the expression detection method of Notch3-Hey1 pathway exists in the liver of human BA: RT-PCR and Western-blot are used in the quantitative detection method. Among them, RT-PCR detects Notch1-4 in human liver tissue, and its main downstream target genes Hes1, Hes5, Hey1, Hey2, HeyL. Figure 8 Map showing abnormally high expression of the Notch3-Hey1 pathway in human BA liver. The primer sequences are as follows:

[0083]

[0084] First, total RNA is extracted from the tissue, and cDNA is produced by conventional reverse transcription. The specific steps are:

[0085] 1) Take liver tissue treated with RNAlater and frozen in -80 refrigerator, take 0.05g of it and grind it in a homogenizer, add 1ml Trizol to fully homogenize it, and transfer the homogenate to a 1.5ml EP tube;

[0086] 2) Add 0.2ml of chloroform to each tube, shake rapidly for 15s and let it stand fo...

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Abstract

The invention discloses a method based on a gene / protein as a diagnostic marker and a therapeutic target for biliary atresia and application thereof. Notch3 signaling pathway and remodeling of hepaticartery system mediated thereby are first proposed in the context of biliary atresia (BA), and constitute a perfection to the existing theoretical mechanism. The Notch3-Hey1 gene / protein, which is highly expressed in liver tissue, can be converted into a BA diagnostic marker which is helpful to prediction of prognosis in children and selection of surgical surgical methods as an auxiliary means. Incurrent clinical practice, anti-inflammatory and anti-fibrosis treatment cannot achieve ideal effect, and Kasai operation can not completely stop the progression of BA; however, a new adjuvant therapy strategy is provided by Notch3 signaling pathway inhibitors, thereby beneficially improving hypoxia and bile duct injury in portal area, slowing down disease progression and raising survival rates of patients with BA liver

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a method based on gene / protein as a diagnostic marker and therapeutic target for biliary atresia and its application. Background technique [0002] Biliary atresia (BA) is a severe obstructive jaundice disease unique to newborns. The main histological features are progressive atresia of extrahepatic bile duct and extensive sclerosing cholangitis of intrahepatic bile duct system. The disease progresses rapidly. Its natural survival period is only two years, and the incidence rate in mainland China is 1 / 5000-1 / 12000. Although hepatic portoenterostomy (Kasai operation) can surgically reconstruct and restore the bile flow and prolong the survival time of the autologous liver, it cannot cure the disease. At least 75% of the children's autologous livers will continue to progress in fibrosis, and liver transplantation is the only option. At present, the etiology of biliary atresia ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883G01N33/68C12Q1/02A61K45/00A61P1/16
CPCC12Q1/6883G01N33/6893G01N33/5005A61K45/00A61P1/16G01N2800/08C12Q2600/158
Inventor 汤绍涛常晓盼阳历
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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