Medicinal preparation for preventing from pyloric helicobacterium inflammation

A technology of Helicobacter pylori and pharmaceutical preparations, which is applied in the field of immunity, can solve problems such as toxicity restrictions, and achieve the effect of convenient operation, simple process, and strong immune stimulation

Inactive Publication Date: 2004-12-08
RENJI HOSPITAL ATTACHED TO SHANGHAI NO 2 MEDICALUNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A large number of effective and non-toxic antigens can be obtained through genetic engineering technology, and only cholera toxin (CT) and E. further applied to the human body

Method used

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  • Medicinal preparation for preventing from pyloric helicobacterium inflammation
  • Medicinal preparation for preventing from pyloric helicobacterium inflammation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Extraction of H.pylori genomic DNA:

[0012] H. pylori strain NCTC11637 (gifted by Professor Andren Lee, School of Microbiology and Immunology, University of New South Wales, Sydney, Australia) was grown on agar plates, cultured under microaerophilic conditions at 37°C for 48 hours, and the bacteria were collected in TE buffer at pH 8.0 In the method, after being treated with lysozyme, 10% SDS and 20mg / ml proteinase K, the whole genome DNA of Hp was extracted by phenol: chloroform extraction method, and kept at -20°C for later use.

Embodiment 2

[0014] Preparation of urease B subunit (UreaB) gene

[0015] Urease B subunit (UreaB) gene: The complete sequence of this gene was obtained from EMBL sequence No. M60398. Use the GCG gene analysis software system to analyze the restriction site, reading frame, and mRNA secondary structure of the target gene. The primers are designed as follows:

[0016] sense: 5'-AGGAAGATGCACGAAGACTGGGGCACC-3';

[0017] Antisense: 5'-GAAGATCTGGACGAAAATGCTAAATAGTTG-3'. PCR reaction conditions: heat denaturation at 94°C for 4min, 30 cycles of 30sec at 94°C, 30sec at 45°C and 1.5min at 72°C, and extension at 72°C for 10min. After the PCR products were identified by 1% Agarose electrophoresis, they were recovered by tapping the gel.

Embodiment 3

[0019] Preparation of outer membrane protein (OMP27) gene

[0020] The isolated and purified H. pylori genomic DNA was used as a template. Nucleic acid sequence design primers are as follows:

[0021] sense 5'-AAAAAAAGGTACCGAAGAGAGCGCGGCTTTTGTGGGA-3';

[0022] Antisense: 5'-AAAAAAAAGCTTTCATTAAAAATCCCTCAAGTAACTGAT-3'. PCR reaction conditions: heat denaturation at 94°C for 3 min, 30 cycles at 94°C for 30 sec, 50°C for 30 sec, and 72°C for 1 min, and extension at 72°C for 5 min. After the PCR products were identified by 1% Agarose electrophoresis, they were recovered by tapping the gel.

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Abstract

A medicine for preventing the infection of pylorospirobacillus is prepared from 3 kinds of antigen protein and CpG-ODN through dissolving them in aseptic distilled water at 4 deg.C and mixing. It is applied by dropping it in nasal cavity.

Description

Technical field: [0001] The invention relates to the field of immunization, in particular to a medicine for preventing and treating Helicobacter pylori infection, in particular to a medicine preparation for preventing Helicobacter pylori (Hp) infection. Background technique: [0002] Helicobacter pylori is a Gram-negative helical bacterium that settles in the human stomach. Since Warren and Marshall discovered Hp in 1982, a large number of studies have shown that Hp infection is the main cause of chronic active gastritis and peptic ulcer. It is closely related to the occurrence of gastric malignant tumors, and was recognized as a primary carcinogen by the World Health Organization in 1996. The eradication of Hp is very important to the prevention and treatment of the above diseases. The current main method is drug eradication (antibiotics plus proton pump inhibitors or bismuth). Affected, in addition, due to the high natural infection rate of Hp in the population, reaching ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/72A61K38/48A61K39/39A61P1/04A61P31/02A61P37/02
Inventor 萧树东史彤郑青庞智朱红音刘文忠楼屹
Owner RENJI HOSPITAL ATTACHED TO SHANGHAI NO 2 MEDICALUNIV
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