By vantal Ten days after observing the estrous cycle of mouse fixator, the vaginal exfoliated cells of mouse fixator were smeared continuously for two sexual cycles (5 days each cycle) to observe the changes of estrous cycle. The characteristics of the pre-estrous smear were: small, round and nucleated epithelial cells (squamous epithelial cells) existed; Estrus: characterized by irregular (keratinized) squamous epithelial cells; Late estrus: characterized by white blood cells and keratinocytes; Estrous period: the appearance of white blood cells and round epithelial cells is characterized. According to the literature, if the mouse fixator loses its regular estrous cycle or remains in estrous period, it indicates anovulation.In view of the actual needs of society, suture kit We need to change some original problems to better serve the society and benefit people. https://pinnaclemedics.com/
2.4 On the 20th day of specimen collection and modeling, fasting began at 8 pm. The next day, blood was collected from the tail vein, and the serum was separated for T, FSH and LH detection. Then, two anesthetics (10% chloral hydrate, 3 ml/kg) were taken from the normal group and the model group, and the ovaries were injected intraperitoneally for observation. At 8 o’clock on the evening of the 48th day, people stopped fasting. The next day, they were weighed and anesthetized, and 4 ml of blood was taken from abdominal aorta, and the serum was separated for sex hormone detection. Both ovaries were taken out completely, and the adipose tissue around the ovaries was cut off, weighed and observed. The bilateral ovaries of mouse fixator were washed with PBS and placed in EP tube, and 4% paraformaldehyde was added to soak them. After pathological section, the ovaries were used for morphological observation.
Mouse fixator
2.5 Observation indicators and methods
2.5.1 After the last administration, the ovarian index began to fast at 8 pm and was weighed the next morning. Both ovaries were taken out completely, weighed and the ovarian index was calculated.
Ovarian index = ovarian weight (mg)/body mass (g)〜100%
2.5.2 Observation of ovarian tissue morphology Observe the appearance of ovarian tissue under pose microscope; The largest plane of the ovary was embedded in paraffin, and the wax blocks were sliced continuously with a thickness of 4 μ m, and stained with hematoxylin-eosin (HE) and observed.
2.5.3 Detection of serum sex hormones The contents of T, FSH and LH in serum were detected by enzyme-linked immunosorbent assay (ELISA), and the specific operation was carried out in strict accordance with the instructions of the kit. Detect the antibody or antigen in the sample by enzyme-linked immunosorbent assay (ELISA), and judge the concentration of antibody or antigen in the sample according to the color depth after enzyme-linked immunosorbent assay (ELISA).
2.6 Statistical methods were analyzed by SPSS21.0 statistical software. If the measurement data conforms to the normal distribution, the data is expressed as (x s), the two groups are compared by independent sample T test, and the multi-groups are compared by one-way analysis of variance. When the variance homogeneity test is met, the LSD method is used to analyze and compare the differences between the groups. When the variance is uneven, Dunnett TT 3 method is selected for pairwise comparison. If it does not conform to the normal distribution, two samples or more groups of rank sum tests are carried out, and the data are described by median and quartile.
3 results
3.2 The estrous cycle changes of the normal group and the model mice fixator. The estrous cycle changes in the normal group according to the rules of estrous interval, estrous prophase, estrous period and estrous anaphase. The estrous cycle of modeling group loses its regularity.
3.3 Comparison of body mass and serum sex hormone levels.
Mouse mouth gag
3.4 On the 21st day of HE staining in ovarian tissue, follicles at all levels can be seen under the microscope in the normal group. The mature follicles have complete structures (indicated by arrows), with cumulus, egg cells and radial crowns, and the ovarian granular cell layers are mostly 8-9 layers. It results in mature follicles (arrow), no oocytes and cumulus, and the granular cell layer of ovary is reduced and sparsely arranged. On the 49th day, in the normal group, follicles at all levels can be seen under microscope, and the mature follicles have complete structure (indicated by arrows), cumulus, egg cells and radiation crowns, and the ovarian granular cell layers are mostly 8-9 layers. Microscopically, in model group ◆ and model group □, the ovary showed polycystic change, no mature follicles (indicated by arrows), no oocytes and cumulus, and most of the ovarian granular cells were 1-3 layers, which were sparsely arranged and rarely corpus luteum. Compared with model group I, model group II has less corpus luteum and deformed follicles.
3.5 Observation on the Histomorphology of Ovary on the 49th day; The ovary of the fixator in the normal group was normal in shape and ruddy in color; In model group ◆, the surface of ovary was pale, and single or more cystic dilated follicles could be seen on the surface of ovary. Model group □ showed milky white color change.
3.6 On the 49th day, the ovarian index and corpus luteum number of three groups of mice were compared.