Application of probiotics in preparation of medicine for treating progeny injury caused by pregnancy stress
A technology of probiotics and stress, applied in the field of biomedicine, can solve the problems of susceptibility to anxiety, depression, obsessive-compulsive disorder, developmental delay affecting fetuses and newborns, premature infants and low body weight, etc., to improve damage and aging death, reduce activation, and relieve inflammation
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Embodiment 1
[0028] Nissl cell count test
[0029] Perform Nissl staining on the hippocampus of the adult 60-day-old offspring: take frozen sections of the hippocampus of the offspring rats of each age group, three in each group, stick the frozen sections on a slide, put the slide in an ultra-clean bench to dry for one day, and Submerged in ethanol (100%, 95%, 75%) in sequence. After the surface was dry, put it into Nissl staining solution for 5-20min, observe the color depth of the tissue at any time under the microscope, gently wash the slide with double distilled water, put it in Anhydrous ethanol is used for color separation in a few seconds, and the color separation can be observed in real time. Gently rinse the slides with tap water, degrease them with xylene Ⅰ for 3 minutes, then change to xylene Ⅱ for 3 minutes, seal the stained brain tissue slices with resin, drop an appropriate amount of neutral resin onto the slides, and remove the coverslip from the Put it from bottom to top, ...
Embodiment 2
[0032] Detection of Microglial Activation in the DG Region of the Hippocampus of Progeny Rats
[0033] The activation of microglial cells in the DG area of the hippocampus was detected by IBA immunohistochemistry, 3 mice in each group, and the specific steps were as follows: using a coronal section with a thickness of 20ul, serially sectioned with a frozen microtome, and transferred to a 24-well plate after the tissue was cut Inside. Using immunohistochemical method, tissue slides were washed three times in 0.01M PBS solution, each time for 10 min. The hippocampal slide samples were then placed in 3% H 2 o 2 Inside, fully remove the enzyme in the tissue, put the hippocampal slide in PBS and wash three times, 10min each time, completely remove the residual H 2 o 2 . Blocking: Place hippocampal slide samples in blocking solution. IBA-1 overnight at room temperature. The hippocampal slide samples were taken out, and the brain hippocampal slides were washed with PBS three...
Embodiment 3
[0036] Detection of the expression levels of inflammatory factors IL-1β and TNF-α in peripheral blood plasma of offspring
[0037] The specific detection method is: (1) Dilution of standard substance. (2) Pour 40ul of sample buffer into the well in sequence, then add 10ul of sample (sample diluted 5x to 10x), drop the sample into the bottom of the well, gently shake the 96-well plate to mix well. (3) After sealing the 96-well plate with a parafilm, place it in a temperature box, and the incubation program is 30min at 37°C. (4) The washing solution needs to be used after reducing its concentration by 30 times, and dilute it with double distilled water. (5) After incubation for 30 minutes, tear off the sealing film and pour out the solution in the 96-well plate, dry it, then add washing solution to each channel for rinsing, stay at room temperature for 30 seconds, pour off the washing solution, and repeat this step for 5 rounds , pat dry on filter paper. (6) Fill each sample ...
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