Use of fam134b in the preparation of medicines for treating sepsis

A technology of drugs and reagents, applied in the fields of molecular biology and biomedicine, which can solve the problems of persistent infection, superinfection, immune dysfunction, etc.

Active Publication Date: 2019-12-03
FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Immune dysfunction in the state of sepsis is mainly manifested as the response disorder of the innate immune system and the adaptive immune system, and then cannot effectively limit and kill pathogenic bacteria, resulting in the persistence of infection and even the occurrence of superinfection

Method used

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  • Use of fam134b in the preparation of medicines for treating sepsis
  • Use of fam134b in the preparation of medicines for treating sepsis
  • Use of fam134b in the preparation of medicines for treating sepsis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1 Correlation between FAM134B and sepsis

[0071] 1. In vivo experiment

[0072] Take 6-8 weeks old wild-type male C57BL / 6 mice (body weight: 20-22g), carry out numbering, weigh, and divide into three groups randomly (random number method): normal group, sham injury group and sepsis group, The mice in the sepsis group were subjected to CLP to construct a sepsis model, and the mice in the sham injury group were opened to expose the cecum, and then the abdomen was closed after feeding. The animals were sacrificed at 6h, 24h, and 72h after the operation, and the spleens of the mice were removed. DCs were isolated and purified by immunomagnetic beads method (CD11c Microbeads), DC function was evaluated, and the expression of FAM134B in each group was detected and compared at different time points.

[0073] (1) Establishment of sepsis animal model

[0074] Male C57BL / 6 mice (20-22 g) aged 6-8 weeks were fasted for 12 hours before the operation and allowed to drink ...

Embodiment 2

[0093] Example 2 Effect of Interfering with FAM134B Gene Expression on DC Functional Activation

[0094] 1. In vivo experiment

[0095] 1.1 Obtaining of FAM134B knockout mice (FAM134B- / -)

[0096] Nanjing University-Nanjing Institute of Biomedicine is the earliest research institution in China that has passed AAALAC certification and has maintained the certification for the longest time. It is the only national genetic engineering mouse seed center certified by the Ministry of Science and Technology. The important model animals required for this application (FAM134B knockout mice, FAM134B- / -) have been produced and provided by Nanjing University-Nanjing Institute of Biomedicine:

[0097] a. Design and construction of sgRNA: design two sgRNAs between intron 3-4 and 3'-UTR position of Fam134b gene, and synthesize and construct them in vitro;

[0098]b. Microinjection: Inject the constructed sgRNA and Cas9 mRNA together into 0.5-day fertilized eggs, and transplant 20-25 injecte...

Embodiment 3

[0141] Example 3 Effect of Overexpression of FAM134B on DC Functional Activation

[0142] 1. Steps

[0143]The upstream and downstream sequences of the FAM134B gene were queried from Genbank, and specific amplification primers were designed using VectorNTI software. Introducing a restriction site, digesting the expression vector with a restriction endonuclease, obtaining the target gene, and preparing a linearized expression vector. The gene of interest was constructed into a linearized expression vector using T4 DNA ligase. Correct clones were verified by sequencing, and high-purity plasmid extraction was carried out. After the DC2.4 cells were transfected with the lentiviral vector containing overexpressed FAM134B for 9 hours, the medium of the cells was changed, and the DC2.4 cell line with stable genetic overexpression of FAM134B was obtained after continuing to culture for 72 hours. According to 2×10 6 DC2.4 cells were inoculated at the ratio of cells / ml and cultured ...

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Abstract

The invention discloses a novel use of FAM134B. As proved by experiments in vivo and in vitro, promotion of FAM134B expression can promote the maturation of DC or increase DC activity, thereby realizing the purpose of treating sepsis. In addition, according to the research results of the present invention, those skilled in the art will know that the expression level of FAM134B is correlated with sepsis, and therefore FAM134B can be used as a molecular marker for diagnosis of sepsis. According to the correlation between FAM 134B and DC cells, the invention also provides a method for preparing highly active DC cells in vitro, thereby realizing the industrial production of DC cells.

Description

technical field [0001] The invention belongs to the field of molecular biology and biomedicine, and relates to the application of FAM134B in the treatment of sepsis. Background technique [0002] Sepsis is a fatal organ dysfunction caused by the body's uncontrolled response to infection, and it is the leading cause of death for patients in the intensive care unit (ICU). According to the latest epidemiological data, there are about 31.5 million sepsis patients in countries around the world every year, and the death toll is as high as 5.3 million. Due to the high cost of treatment and a large amount of medical consumption, it has caused a huge economic burden. Taking the United States as an example, there are more than 750,000 sepsis patients in the United States every year, with a mortality rate of 28.6%, and the total annual cost of sepsis treatment is 16.7 billion US dollars. The incidence and mortality of severe sepsis in my country are also at a relatively high level. Ac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K48/00A61P31/04A61P37/02A61K49/00C12Q1/6883A61D1/00
CPCA61D1/00A61K45/00A61K48/0008A61K48/005A61K49/0008A61P31/04A61P37/02C12Q1/6883C12Q2600/158
Inventor 姚咏明任超姚人骐董宁
Owner FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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