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Application of neutrophil elastase in the preparation of products for diagnosing biliary atresia

A technology of elastase and neutrophils, applied in the application field of the product, can solve the problems of poor prognosis of children after surgery, unable to meet the needs of children with BA, bile duct damage and other problems

Active Publication Date: 2022-03-18
GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, sometimes the display of gallbladder and bile duct under B-ultrasound does not match the intraoperative display, which leads to a decrease in the accuracy of B-ultrasound diagnosis of BA;
[0006] (3) Intraoperative cholangiography and postoperative pathological examination: Although intraoperative cholangiography and postoperative pathological examination are the "gold standard" for diagnosing BA, they are invasive examinations, and the children diagnosed at this time are all younger Large, may be detrimental to the prognosis of children after surgery
[0008] (1) Kasai operation: Kasai operation is a very traumatic operation in pediatric surgery. Although it can improve the symptoms of cholestasis in some children with BA, most of them are older at the time of operation, resulting in poor postoperative prognosis; And the progressive destruction of bile duct and liver fibrosis will occur in children with BA, which makes Kasai surgery mostly unable to solve the fundamental problem of BA;
[0009] (2) Liver transplantation: Liver transplantation is the only long-term survival treatment for children with BA. Due to the lack of liver sources in my country, it cannot satisfy all children with BA, and the cost of liver transplantation is relatively high

Method used

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  • Application of neutrophil elastase in the preparation of products for diagnosing biliary atresia
  • Application of neutrophil elastase in the preparation of products for diagnosing biliary atresia
  • Application of neutrophil elastase in the preparation of products for diagnosing biliary atresia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1 Detection of the expression of neutrophil elastase in biliary atresia

[0071] (1) Clinical sample collection

[0072] All healthy control children (infants without evidence of liver disease, NC, n=16), children in the cholestatic group (defined as serum direct bilirubin above 17 μmol / L, IHC, n=32) and children in the biliary atresia group (Bile duct atresia was confirmed by intraoperative cholangiography and postoperative histopathological examination of extrahepatic bile ducts, BA, n=39) from the Department of Surgery of Guangzhou Women and Children's Medical Center (Guangzhou, China) between March 2019 and March 2020 )recruit. The diagnosis of children with IHC and BA is based on the clinical characteristics, biochemical indicators of liver and gallbladder, intraoperative cholangiography, and histopathological examination of extrahepatic bile ducts after operation. EDTA anticoagulated blood samples (2ml) were collected from children in each group before o...

Embodiment 2

[0080] Example 2 Demonstrates the importance of neutrophil elastase in BA in biliary atresia mice

[0081](1) Pregnant BALB / c mice (10-12 weeks old, body weight 35-40 g) were purchased from Guangdong Provincial Animal Experiment Center, and CD177- / -Balb / c mice were purchased from Cyagen (USA) Biological Co., Ltd. Mice were reared in air-filtered, specific pathogen-free individual micro-isolation cages (temperature 25°C, dark and light intermittently produced for 12 h), and had free access to sterile water and autoclaved food. Newborn mice within 24 hours of birth (average body weight 1.5-1.6 g) were used to establish the BA mouse model and conduct downstream experiments. The experimental protocol was approved by the Animal Care and Use Committee of the Experimental Animal Center of Guangzhou Medical University (IACUC-DB-16-0602), and all animal experiments were carried out in the center;

[0082] (2) Rhesus monkey rotavirus (RRV) strain MMU 18006 was purchased from ATCC (ATCC...

Embodiment 3

[0087] Example 3 In vitro experiments further confirm that neutrophil elastase plays an important role in BA

[0088] Human neutrophil culture medium and normal fetal bile duct epithelial cells (BEC) were co-cultured in vitro:

[0089] (1) Separation of neutrophils in children with BA and hemangioma: According to the manufacturer's instructions, the blood samples of the BA group were lysed into a single-cell suspension with red blood cell lysate, added with Fc Block and incubated at room temperature for 10 minutes, and then added with anti -human PEconjugated CD177 antibody was incubated at 4°C for 30min, after washing, anti-PE Microbeads were added and incubated at 4°C for 15min, after washing, neutrophils were sorted on a magnetic sorter (Miltenyi Biotec, Germany) using an LD separation column , the sorted neutrophils were tested for cell purity on BD FACS Aria II (BD Biosciences, USA), and cells with cell purity>95% were used for culturing;

[0090] (2) Human biliary epith...

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Abstract

The invention provides an application of neutrophil elastase in the preparation of a product for diagnosing biliary atresia, and relates to the technical field of molecular diagnosis. The invention firstly confirms that by detecting a large number of BA patient samples and combining in vivo and in vitro experiments The expression of NE in liver tissue and peripheral blood of BA can be used as a powerful indicator to improve the diagnosis of BA and evaluate the degree of liver injury. Therefore, detecting the expression of NE can provide a more convenient, direct and accurate method for clinical diagnosis and evaluation of the condition of children with BA. method. In addition, the present invention also confirms that the NE inhibitor has a therapeutic effect in animal experiments and can be used in the development of drugs for the treatment of BA.

Description

technical field [0001] The invention relates to the technical field of molecular diagnosis, in particular to an application of neutrophil elastase in preparing products for diagnosing biliary atresia. Background technique [0002] Biliary atresia (BA) is the most common cause of chronic liver disease in infants and young children, and it is also the main indication for liver transplantation in infants and young children. The etiology and pathogenesis are still unclear. The clinical manifestations of BA are mainly progressive jaundice of the skin and mucous membranes, clay-like stool, tea-like urine, and hepatosplenomegaly. The prognosis is poor and the mortality rate is high. my country is a high-incidence area of ​​BA, with an incidence rate as high as 1 / 5000-8000. At present, the diagnosis of BA is mainly based on intraoperative angiography and postoperative pathology, and the treatment is mainly based on Kasai operation and liver transplantation. Although liver transplan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573A61K45/00A61P1/16
CPCG01N33/573A61K45/00A61P1/16G01N2333/966G01N2800/085G01N2800/52
Inventor 张锐忠付铭童燕陆谭乐东王贺珍陈严夏慧敏
Owner GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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