Application of bacteroides fragilis in preparing composition for preventing and treating colon cancer
A technology of Bacteroides fragilis and a composition is applied in the application field of Bacteroides fragilis in the preparation of compositions for preventing and treating colon cancer, and achieves the effects of promoting apoptosis, inhibiting tumor growth and having strong pharmacological effects.
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[0017] Example 1 Preparation of Bacteroides fragilis bacterial liquid
[0018] (1) Isolate Bacteroides fragilis from baby feces
[0019] According to the "normal flora inspection method", weigh 0.5 g of stool and place it in a test tube containing 4.5 ml of normal saline, and dilute it to 10 -1 ~10 -6 , Drop it on the Bd medium plate from high dilution to low dilution, place it in an anaerobic gas production box and add an anaerobic gas production bag for anaerobic culture. Put it at 37°C, open the can for inspection after 2~7 days. Select a single colony for pure culture, staining microscopy, culture characteristics, physiological and biochemical tests.
[0020] (2) Enrichment culture of Bacteroides fragilis and preparation of bacterial liquid
[0021] A single purified colony was selected and inoculated in modified tryptone broth for enrichment culture. The resulting bacterial solution was centrifuged at 3000r / min for 15min, and the supernatant was removed. After washing the precip
Example Embodiment
[0022] Example 2 Bacteroides fragilis virulence test
[0023] The following experiments were carried out using the Bacteroides fragilis strain prepared in Example 1:
[0024] (1) Acute toxicity test in mice
[0025] A total of 60 C57BR / C mice weighing 22±2 grams (male and female) were divided into high-dose group, low-dose group and control group, with 20 mice in each group. Each mouse in the high-dose group was given 10 per day 12 0.5ml of cfu / ml bacterial liquid, the low-dose group is given 10 per day 9 0.5ml of cfu / ml bacterial solution, and 0.5ml of normal saline per day in the control group. Observe for 14 consecutive days.
[0026] Experimental results: The mice grew normally without death, and the LD50 was undetectable.
[0027] (2) Rat toxicity test
[0028] Thirty SD rats of 150±20 grams (half male and female) were divided into 3 groups: high and low dose group and control group. Each rat in the high-dose group was given 10 per day 12 cfu / ml bacterial liquid 2ml, low-
Example Embodiment
[0033] Example 3 The preventive effect of Bacteroides fragilis on colon cancer
[0034] The following experiments were carried out using the Bacteroides fragilis strain prepared in Example 1:
[0035] Animal model establishment: 60 ICR mice were randomly divided into 2 groups, namely: saline group (10) and model group (50). 1,2-Dimethylhydrazine (DMH) was dissolved in physiological saline to a final concentration of 20mg / kg, the pH was adjusted to 6.5 with 0.25mol / L NaOH, and the mice were injected intraperitoneally. One week later, dextran sodium (DSS) was added to the drinking water of the model group at a concentration of 20 g / L, and the mice were given continuous drinking for 7 days. When the mice had the second bloody stool, and at the 9th week after the administration of DMH, the model mice were randomly divided into 4 groups according to their body mass, namely: model group (20) and Bacteroides fragilis group: high-dose group ( 10 9 cfu / ml), middle dose group (10 8 c
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