Construction of Lactobacillus curvularius ATCC55221 and application thereof in treatment of parturient nipple rhagadia
A technology of ATCC55221, Lactobacillus crispatus, applied in the field of biopharmaceuticals, can solve problems such as the inability to meet tissue repair
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Embodiment 1
[0061] 1. Establishment of mouse nipple chapped model
[0062] (1) The experimental groups were divided into negative control group (group C), model group (group M), low-dose treatment group (LT group) of engineering strains of Lactobacillus crispatus, and high-dose treatment group (group HT) of engineering strains of Lactobacillus crispatus. Group of 8;
[0063] (2) Take 32 Balb / c mice, numbered 1-32 at random. No food or water was allowed 12 hours before the operation. The mice were fixed with a fixed frame, and the hair on both sides of the back of the experimental site was roughly cut off with an automatic shaving machine close to the skin, and then the fine hair was removed with a depilatory cream. Wash thoroughly with warm water immediately after epilation, leaving skin bare for later use. On the day of operation, they were anesthetized by intraperitoneal injection of 3.6% chloral hydrate 10mL / kg. Immerse the metal disc in liquid nitrogen at -196°C, and immediately s...
Embodiment 2
[0068] 1. Appearance evaluation of chapped nipple model in animal experiments after treatment
[0069] 1. Appearance chapped degree score
[0070] The scoring criteria were evaluated based on the depth of the wound and the degree of tissue damage, and the scores were all evaluated by an experimenter who was blind to the experiment. The higher the score, the more severe the degree of chapping; 0-1 points represent normal or healed. See the table below for the scoring criteria of the degree of chapping.
[0071] Rating criteria for the degree of chapping
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[0073]
Embodiment 3
[0075] 1. Histopathological detection of chapped nipple model in animal experiments after treatment
[0076] The evaluation was carried out from three aspects: regeneration of epidermis and dermis, thickness of granulation tissue, and angiogenesis. HE staining was used to observe the pathological changes of the wound. Masson staining was used to observe the repair of skin scar after injury.
[0077] 1. Histological score - HE staining of wound tissue
[0078] (1) Dehydration. Take out the wound tissue that has been fixed, rinse it well with PBS, and then dehydrate with ethanol, respectively with 70% ethanol, 80% ethanol, 90% ethanol, 95% ethanol, and 95% ethanol for 150 minutes; then dehydrate with absolute ethanol twice , respectively 60min;
[0079] (2) Embedding. The dehydrated tissues were treated with xylene for 30 minutes, and embedded in paraffin;
[0080] (3) Slicing and sealing. Cut out a flat plane with a rotary machine, set the thickness of the section to 55 ...
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