Method for making animal model simulated human cornea fungal infection

A fungal infection and animal model technology, applied in the field of medical models, can solve the problems of difficult to grasp the depth of scratches, corneal perforation, unsuitable infection, etc., and achieve the effect of economical and simple operation method, easy material and high success rate

Inactive Publication Date: 2008-04-30
王丽娅
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the experiment, the amount of fungus in the cornea decreased rapidly after inoculation in mice without immunosuppressive agents
In 2004, they used the same method to inoculate Fusarium solani and found that in the pathological sections, BALB / C mice without immunosuppressants could not be detected or occasionally had a small amount of fungal hyphae in the corneal stroma on the fourth day after inoculation. , so this method is based on making animals in a state of immunocompromise in advance, but most patients with fungal keratitis are not in an immunosuppressed state when they develop clinically. Easy to cause corneal perforation, not suitable for research on the occurrence, development and mechanism of infection
These two methods have a high success rate of infection, and both avoid the influence of hormones on the experimental results, but they do not conform to the clinical course of natural infection. The operation process of the method is relatively complicated, and there are certain requirements for the experimenter's surgical skills.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Preparation of fungal strains: select the required fungal strains and inoculate them on potato dextrose medium or Sandcastle medium, and cultivate them at 15-35°C for 3-5 days. The fungi are all filamentous fungi such as Fusarium, Aspergillus, Curvularia and Penicillium.

[0024] Inoculum preparation: Because fungal keratitis is mostly caused by plant trauma, and fungal keratitis is often caused by plant epiphytes, various plant supports that fungi are prone to epiphyte, such as corn , sorghum, cotton, leguminous, wheat, rice and other crops and woody plants such as poplar, phoenix, elm, locust tree, etc. Trim the roots, stems, leaves or fruit shells to appropriate sizes, and put them in after autoclaving Put it into a fungal culture tube, let the fungus grow on the surface, and cultivate it at 15-35°C for 3-6 days, so that more than half of the surface is covered with fungi.

[0025] Animal preparation: choose murine animals without eye diseases, such as various outbr...

Embodiment 2

[0034] In the post-inoculation treatment step, in addition to applying antibiotic eye drops, immunosuppressant eye drops are added every day after inoculation, 2-5 times / day, and the others are the same as in Example 1. Immunosuppressants can be adrenocortical hormones such as prednisolone, prednisone, dexamethasone, etc., or cyclosporine.

[0035] It has been observed that the success rate of model making is close to 100%, and typical fungal keratitis can be observed on the third day after inoculation.

[0036] Local drip application of immunosuppressants avoids systemic use, does not change the overall immune status of animals, and is close to the clinical natural disease process. The penetrability of various fungi varies, and 30% of the animals may have corneal perforation on the 3rd to 6th day after inoculation. The pathological section shows that the cornea becomes thinner, the whole layer of the cornea is covered with fungal hyphae, and the hyphae can be seen penetrating...

Embodiment 3

[0037] Example 3: The strain of Fusarium solani was inoculated in potato dextrose medium and cultured at 28° C. for 3 days.

[0038]Trim the corn stalk to a suitable size, put it into a fungal culture tube after high-pressure sterilization, make the fungus grow on the surface, and cultivate it at room temperature for 3 days, so that more than half of the surface is covered with fungus.

[0039] 60 outbred Kunming mice without eye diseases were selected, and antibacterial drugs were instilled in the eyes three days before inoculation, three times a day.

[0040] After the mice were anesthetized, under strict aseptic conditions, a fungus-bearing corn stalk was used to scratch the corneal surface of the animal to the depth of the superficial stromal layer. After the scratch, palpebral fissure was performed.

[0041] After 24 hours, open the eyelids and use antibacterial eye drops every day to prevent bacterial infection. Observe the condition changes under the slit lamp microsco...

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PUM

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Abstract

The invention discloses a making method of an animal model for imitating fungal infection of porcine cornea, composed of steps of preparing fungi strains, preparing inoculum, preparing animals, inoculating animals and processing after the inoculating. The invention has the advantages that model-making success rate is high, generally the success rate of outbred animals reaches over 85% and the success rate of inbred animals reaches over 90%, immunosuppressant is not necessary, fungal hyphaes are observed in pathological section of cornea among the second day and thirteenth day after inoculating, operational method is economical, simple and easy to apply, materials are easy to get, inoculating method is easy to master, researchers need no special technology, the invention is suitable for the process of clinical natural infection of fungal keratitis, and is adaptable for the research of the generation, development and the mechanism of infection.

Description

technical field [0001] The invention belongs to medical models, in particular to a method for making an animal model simulating human corneal fungal infection. Background technique [0002] Fungal keratitis is an infectious corneal ulcer with a high rate of blindness. With the wide application of antibiotics and glucocorticoids and the improvement of the understanding and diagnosis of this disease, its incidence rate has been increasing. Common pathogenic fungi include Fusarium, Aspergillus, Penicillium, and Candida. Most of the patients in our country are farmers, and they often occur after vegetative corneal trauma. At present, there are very few basic studies on corneal fungal infection, and its pathogenesis is unclear. The lack of animal models simulating human corneal fungal infection has seriously hindered the basic research of fungal keratitis. The following methods for making animal models of fungal keratitis are currently disclosed: [0003] 1. Corneal stromal inj...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K38/13A61K31/573A61P31/10
Inventor 王丽娅王璐璐吴细丕孙声桃张俊杰魏秋彩
Owner 王丽娅
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