The scope of this study was the therapeutic effect of the Chinese medicine xiaoyao pills on mice with postmenopausal osteoporosis. In this study, we wanted to confirm whether the xiaoyao pills could exert a therapeutic effect and the specific mechanism. In this study, we combine the traditional Chinese medicine with modern pharmacology and the normal genus and authorized by informations to search and medically analyze the protein dose event will show your cure.
Also process compared with single pharmacology our genome just analyze our scheme is more comprehensive and may provide new idea for the treatments of osteoporosis. This idea will promote the development of the field of traditional Chinese medicine for osteoporosis, promote and advance understanding of the mechanism and revolution of traditional Chinese medicine, its primary research idea, and the direction of the provider practical guidance and efficacious evaluation visit of our clinical application. To begin disease associated gene merging, place the previously saved text files from five databases containing the herbal name, ingredient ID, gene ID, and the script disease dot R in the same folder.
Open the R code. Copy and paste the path where the text file is stored into the line where the set WD is located in the R code. Open the R software.
Run the modified R code and save. Set the gene and name the file disease dot text. Open the Venn diagram website.
In the input section, copy and paste the already-saved text content into the list. Name the content with the respective database and click submit. Click save images PNG below the image and save the text result in the same folder.
Place the all targets dot symbol dot text file and the disease dot text file in the same folder. Open the R code. Copy and paste the path where the text file is stored into the line where the set WD is located in the R code.
Open the R software, run the modified R code and save. Set the gene and name the file drug disease dot text. Open the Venn diagram website.
In the input section, copy and paste all targets dot symbol dot text and disease dot text into the list and name the files. Click save image as PNG below the image and save the text results in the same folder. Open the String website and click multiple protein.
Click browse and upload the drug disease dot text file. Then select Homo sapiens in organisms. Click search, followed by continue.
Now click settings and set the minimum required interaction score to the highest confidence. From the network display option, check hide disconnected nodes in the network. Then click update.
Adjust the nodes in the network diagram so that there is no overlap or occlusion. Click exports, followed by download as a high resolution bitmap to get the protein inter-working network map in PNG format. Download the TSV file as a short tabular text output.
Save PNG and TSV files in a unified folder. To begin prepare wax blocks from euthanized mice injected with xiaoyao pills. Cut the wax blocks into six micrometer thick continuous sections and bake the slices at 70 to 72 degrees Celsius for 30 to 60 minutes.
Transfer the slices to the dewaxing solution for eight minutes, then dehydrate twice in 75%anhydrous ethanol for five minutes each before rinsing in PBS for five minutes. Now perform EDTA high pressure repair for 15 minutes. After rinsing with PBS, drop-wise, add 3%hydrogen peroxide to the tissue and incubate for 10 minutes.
Wash the tissue three times with PBS for five minutes each. Using an immunohistochemistry pen, draw a circle around the tissue. After washing the tissue with PBS, stain overnight in primary antibody solution.
The following day, rinse the tissue three times for five minutes each in PBS. Drop-wise, add the secondary antibody and incubate for 60 minutes. After rinsing with PBS, incubate the tissue in DAB ready to use solution for three to five minutes.
Now stain the tissue in hematoxylin for three minutes. Again, rinse the tissue with PBS before placing it in differentiation solution for 10 seconds. After PBS rinse, add the return solution for 10 seconds.
Finally, perform 75%anhydrous ethanol treatment for five minutes and seal the slide with neutral resin. Hematoxylin eosin staining showed tightly arranged trabeculae in sham mice, while OVX mice displayed typical osteoporotic changes, with sparse trabeculae and enlarged marrow cavities. Micro-CT comparison showed incomplete trabecular meshwork and thinning of the bone cortex in OVX mice compared to sham mice.
After drug intervention, hematoxylin eosin staining showed restored trabeculae structure and reduced marrow cavity compared to the OVX group. Imaging indices showed fewer fractured trabeculae and restored reticulation in the high dose xiaoyao pills group compared to the OVX group. Immunohistochemistry results showed increased alkaline phosphatase and collagen type one and decreased interleukin-17, ACT one and interleukin-6 in the drug group.
