Gavage Strategy for Decoction Formula of Traditional Chinese Medicine in Osteosarcoma Model Mice

This study provides a detailed and standardized protocol for about decoction, preparation and the gradual operation that's providing a reference for yin needle. Our research ensures that each animal is dosed equally to produce accurate results. Our study aims to maximize the animal protection and improve the reliability of results from studies about decoction formulations.

To begin, soak 52.5 grams of Hedyotis diffusa and 31.5 grams of the barbed skull cap herb in water for 30 minutes. Heat the herbs to boiling over a strong fire, then heat for 15 to 25 minutes over a mild fire. Next, use rubber bands to fix three layers of gauze over the mouth of a measuring cup.

Filter the decoction through the gauze. Pour the filtered decoction into a container. Similarly, make a second decoction by boiling the herbs and drinking water just enough to submerge the herbs.

After boiling, pour the second decoction into the same container as the first one. Now, soak 42 grams of ginseng in drinking water about three centimeters over the herb's surface. Boil the soaked ginseng over a strong fire for 60 minutes.

Then pour this first decoction into the container with the other decoctions. Make a second ginseng decoction in drinking water just enough to submerge the herb. Heat this decoction over a mild fire for 40 minutes and filter it into the decoction container.

Boil the filtered decoction on a strong fire in a pot. Then lower the flame to a mild fire. Keep boiling the decoction until the desired concentration is obtained.

Pipette 21 milliliters of the concentrated decoction into tubes as per the required dosage for one week. Store the aliquots in a refrigerator at minus 20 degrees Celsius. Place a mouse on the surface of a mouse cage.

Grab its tail with the thumb and index finger of the left hand. Then, hold the neck of the mouse with the thumb, index, and middle fingers of the right hand. Hold the mouse to align the head, neck, and body in a straight line.

Insert the loaded gavage needle into the mouth of the mouse from the corner, and push the tongue against the upper jaw. Keep the mouse body in the same alignment and gently push the gavage needle toward the stomach along the esophagus. Inject the decoction formula into the stomach.

Then remove the needle and release the mouse back into the cage. Administration of the medicinal decoction reduced the osteosarcoma tumor volume in mice.