Application of CD177 in preparation of product for diagnosing biliary atresia

A technology of biliary atresia and products, which is applied in the application field of products, can solve problems such as irregular arrangement of lobules, necrosis of liver cells, and poor specificity, so as to ensure the timing of diagnosis and treatment, ensure accuracy, and have strong detection specificity Effect

Pending Publication Date: 2020-12-15
GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0009] (6) Histopathological examination of liver biopsy: liver biopsy, or percutaneous liver biopsy and biopsy are generally recommended. Neonatal hepatitis is characterized by irregular arrangement of lobules, hepatocyte necrosis, giant cell degeneration, and portal inflammation The main manifestations of biliary atresia are obvious hyperplasia of bile canaliculi, bile embolism, and fibrosis around the portal area, but multinucleated giant cells can also be seen in some specimens. Therefore, liver biopsy can sometimes cause diagnostic difficulties and even errors, with 10-10 15% of cases can not make a correct diagnosis based on this; and liver biopsy is an invasive examination;
The specificity of other inspection methods is poor
Clinically, there is a lack of diagnostic markers related to biliary atresia, so finding diagnostic markers for biliary atresia is more helpful for the diagnosis of children with biliary atresia

Method used

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  • Application of CD177 in preparation of product for diagnosing biliary atresia
  • Application of CD177 in preparation of product for diagnosing biliary atresia
  • Application of CD177 in preparation of product for diagnosing biliary atresia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 detects the number of CD177+ cells in BA

[0073] (1) Detection of CD177+ neutrophils by flow cytometry

[0074] Clinical sample collection: 1) Liver tissue samples from children with BA and infantile hepatitis syndrome: after ethical approval and the informed consent of the patient’s family members, a fresh tissue sample from the edge of the right lobe of the liver with a diameter of about 1.5 cm was cut during biliary tract exploration or during Kasai’s operation; 2) Peripheral blood samples from children with BA, infantile hepatitis syndrome and hemangioma of the same age: after ethical approval and the informed consent of the patient's family members, fresh peripheral blood samples from children with BA, infant hepatitis syndrome and hemangioma of the same age were collected before operation and without antibiotic intervention. blood sample;

[0075] Collection of mouse liver tissue samples: Using the aforementioned method, a BA mouse model was establishe...

Embodiment 2

[0084] Example 2 further confirms the importance of CD177+ cells in BA on gene knockout mice

[0085] CD177 knockout mice were infected with RRV experiment:

[0086] (1) CD177 knockout mice (CyagenQuote: KOCMS180821DA4+KOCMS190314DA2-B) bred in the previous period were intraperitoneally injected with RRV 20 μL (titer 1.5×10 6 PFU).

[0087] (2) The mice were randomly divided into: ① only RRV injection group; ② normal mice control group. The control group was injected with the same amount of normal saline at the same time period. The liver tissue samples of mice in the above groups were collected on days 3, 6, 9, and 12.

[0088] (3) Observe the changes in the body weight, skin jaundice, survival rate and liver function of the mice; conduct extrahepatic cholangiography by routinely injecting methylene blue into the gallbladder to confirm whether there is extrahepatic biliary atresia; observe the morphology of intrahepatic and extrahepatic bile ducts in mice by pathological s...

Embodiment 3

[0090] Example 3 In vitro experiments to explore the impact of CD177+ cells on biliary epithelial cells (BEC)

[0091] 1. CD177+ cells were co-cultured with normal fetal BECs in vitro:

[0092] (1) CD177+ cell sorting in children with BA: collect peripheral blood samples from children with BA, infantile hepatitis syndrome, and hemangioma at the same age, and sort out CD177+ cells in peripheral blood by flow cytometry (see the implementation for detailed steps) example 1).

[0093] (2) Co-cultivation of sorted cells and normal fetal BECs in vitro: Place human embryonic biliary epithelial cells (BECs) in Poly-L-Lysine (PLL)-coated T75 culture flasks at 37°C, 5% CO 2 They were cultured and expanded in an incubator, and the biliary epithelial cells were purchased from ScienCell. Add the CD177+ cells from the liver or blood samples of the BA children and the disease control group extracted in the above steps to BEC medium for resuspension until the number of cells is 1×10 5 cell...

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Abstract

The invention provides application of CD177 in preparation of a product for diagnosing biliary atresia, and relates to the technical field of molecular diagnosis. According to the invention, by detecting a large number of biliary atresia child patient samples and combining animal experiments, it is proved that the number of CD177+ neutrophile granulocytes in biliary atresia peripheral blood and liver tissue can serve as a powerful index for evaluating the liver injury degree at first; and therefore, detection of CD177 genes, CD177 protein or the CD177+ neutrophile granulocytes can serve as a more convenient, direct and accurate method for clinically evaluating the illness state of biliary atresia child patients. The invention also proves that CD177 has a pro-inflammatory effect in biliaryatresia; and the inhibitor of CD177 can be used for research and development of drugs for treating biliary atresia.

Description

technical field [0001] The invention relates to the technical field of molecular diagnosis, in particular to an application of CD177 in preparing a product for diagnosing biliary atresia. Background technique [0002] Biliary atresia (BA) is a bile duct obstructive disease mainly manifested by neonatal jaundice, with poor prognosis and high mortality. The etiology and pathogenesis are still unclear. The basic pathological features of BA are progressive inflammation of intrahepatic and extrahepatic bile ducts, biliary atresia, and liver fibrosis. The progression of hepatic fibrosis is faster and more aggressive than other adult diseases. Even though Kasai surgery can alleviate the symptoms of cholestasis in some children with BA, most of them still suffer from the subsequent progressive destruction of intrahepatic bile ducts, and even liver failure. , becoming a life-threatening serious disease in children. my country is an area with a high incidence of BA, with an incidenc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/68C12Q1/02A61K39/395A61K45/00A61P1/16
CPCC12Q1/6883G01N33/5091G01N33/6893C07K16/2896A61K45/00A61P1/16C12Q2600/118C12Q2600/158G01N2333/70596G01N2800/52G01N2800/08G01N2800/38A61K2039/505
Inventor 张锐忠苏亮付铭谭乐东童燕陆陈严夏慧敏
Owner GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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