Application of domesticated strain of Clostridium gordii in preparation of radiotherapy sensitizer
A technology of Clostridium gordii and sensitizer, which is applied in the field of preparation of radiotherapy sensitizers by domesticated strains of Clostridium gordii, can solve the problem of no use of bacterial radiotherapy sensitizers, and achieve a significant sensitization effect
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Embodiment 1
[0032] The preparation method of the spore freeze-dried powder of Gordia domesticated strain is as follows:
[0033] Add 50g / L aseptic mannitol cryoprotectant to the obtained clostridium gordii domesticated strain spore liquid, detect the total viable spore amount (5 × 10 7 cfu). The spore liquid was placed in -20°C, -40°C, and -80°C refrigerators for pre-freezing, respectively, for 0.5h, 1.0h, 2.0h, and 4.0h. Under the conditions of cold trap temperature -45°C and vacuum degree of 20 Pa, freeze-dry for 8 hours to obtain lyophilized spore powder of the domesticated strain of Clostridium gordii, and count the number of spores.
[0034] result
[0035] It can be seen from Table 1 that the number of spores pre-frozen at -20°C, -40°C and -80°C for 0.5 h decreased less than that before pre-freezing, but the effect of pre-freezing was lower than that before pre-freezing. Poor, in the form of mucus, which is not conducive to subsequent vacuum freeze-drying, and the effect of pre-f...
Embodiment 2
[0040] Research methods:
[0041] (1) Effect on CEN-2Z cell proliferation
[0042] The lyophilized powder (0mg / ml, 3mg / ml, 5mg / ml, 10mg / ml, 13mg / ml, 17mg / ml and 20mg / ml) of the metabolites of the acclimatized strain of Gordia with different final concentrations were mixed with CEN-2Z cells After co-cultivation for 24h, the MTT assay was carried out to detect the dose-dependent relationship of cell proliferation. For each concentration, 6 replicate wells and zero-adjustment wells were set.
[0043] The lyophilized powder of the metabolites of the domesticated strain of Gordia with a final concentration of 10mg / ml was co-cultured with CEN-2Z cells for 0h, 6h, 12h, 18h, 24h, and 48h, respectively, and the time-dependent relationship on cell proliferation was detected by MTT assay. Each group has 6 replicate holes, and zero-adjustment holes are set at each time.
[0044] result
[0045] The results of the study showed that with the increase of the action time and concentration...
Embodiment 3
[0058] Research methods:
[0059] ①Establishment of tumor-bearing mouse model
[0060] Slowly inject nasopharyngeal carcinoma CEN-2Z cells into female Balb / c-nu / nu nude mice (5 weeks old, 20g) subcutaneously in the lateral femur of the right back by subcutaneous injection, until the tumor volume grows to 300mm 3 , an in vivo test was performed.
[0061] ② Animal grouping and handling
[0062] Control group: n=10 (n represents the number of animals in each group), tail vein injection PBS damping fluid 0.2ml;
[0063] Clostridium gordii domesticated strain spore group: n=10, the 1st day, the 5th day, the 8th day and the 11th day tail vein injected the clostridium gordii domesticated strain spore, the dose was 3.0×10 5 cfu / (kg·body weight).
[0064] X-ray group alone: n=10 rats, received radiotherapy on the 4th day, 7th day, 10th day, and 13th day, once / day, with a dose of 400cGy / time.
[0065] Clostridium gordii domesticated strain spores combined with X-ray group: n=10 r...
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