Experimental Procedure 1. Separation and preparation of single cell suspension:
1) Cell line cultured in vitro: digested with trypsin and blown into a single cell suspension;
2) In vivo organ cells: The animals were sacrificed, the organs were removed, and a single cell suspension was prepared in Hanks' solution.
2. Rubber sheet preparation:
1) Take 20~50μl of 0.5% common melting point agarose kept in a 56°C water bath and spread on a frosted glass slide to form a primer.
2) Take 100~150μl of 0.5% common melting point agarose and add it to the primer. Then add a cover slip to it and condense at 4 °C for 10 minutes.
3) Remove the cover slip and mix 50-100 μl of 1.0% low-melting agarose in a 37 ° C water bath with 50-100 μl cell suspension (105 cells/ml), immediately spread, and cover glass. The sheet was condensed at 4 ° C for 10 minutes.
4) Remove the coverslip and take 70~100μl of 0.5% low-melting agarose plate in a 37°C water bath, cover the slide and condense at 4 °C.
3. Cell lysis and electrophoresis:
1) After removing the coverslip, the prepared rubber plate was immersed in a cell lysate precooled at 4 ° C, and lysed at 4 ° C for 1 hour.
2) Remove the rubber plate, place it in an electrophoresis tank, and immerse it in the electrophoresis solution for 20 minutes.
3) Electrophoresis at 4 ° C for 20 minutes (25 V, 300 mA).
4. Neutralization and dyeing:
1) After the electrophoresis is finished, the rubber plate is immersed in the neutralizing solution for 15 minutes each time, and the mixture is neutralized twice, taking care to replace the neutralizing solution.
2) Remove the rubber plate, place it on the staining rack, add 5 μg/ml of PI, and stain for 20 minutes in the dark.
3) Decolorize the distilled water for 15 minutes.
5. Microscopic examination and analysis:
1) Observation under a fluorescence microscope, green light excitation absorbing filter 590 nm. Take a photo record if necessary.
2) Count the observed cells, record the frequency of occurrence of comet cells, measure the length and length of the micrometer probe with an eyepiece, and calculate the migration distance of nuclear DNA.
A wet agarose gel sheet was obtained by a two-layer gel method by lysis, DNA unwinding, electrophoresis, and neutralization. The wet agarose gel pieces were dehydrated in ice-cold absolute ethanol for 10 minutes and then spontaneously dried in the air. All operations were completed within 8 hours after blood collection, and the operation was protected from light. The cells were stained and photographed using 50 μl of 30 μM ethidium bromide solution. All photographs were analyzed using single-cell gel electrophoresis software, and the tail length and the olive tail moment of more than 100 cells were randomly measured, and the arithmetic mean of the tail length and the olive tail moment represented the individual DNA damage.
Medical Cold Patch
Patch for diarrhea
[Name] Medical Cold Patch
[Package Dimension] 5cm 4pieces/box
The pain relief patch is composed of three layers, namely, backing lining, middle gel and protective film. It is free from pharmacological, immunological or metabolic ingredients.
[Scope of Application] For cold physiotherapy, closed soft tissue only.
[Indications]
The patches give a fast relief for diarrhea.
[How To Use a Patch]
Please follow the Schematic Diagram. One piece, one time.
The curing effect of each piece can last for 6-8 hours.
[Attention]
Do not apply the patch on the problematic skin, such as wounds, eczema, dermatitis,or in the eyes. People allergic to herbs and the pregnant are advised not to use the medication. If swelling or irritation occurs, please stop using and if any of these effects persist or worsen.notify your doctor or pharmacist promptly. Children using the patch must be supervised by adults.
[Storage Conditions]
Store below 30c in a dry place away from heat and direct sunlight.
Patch For Diarrhea,Medicated Patches For Arthiritis,Plaster For Diarrhea,Pad For Diarrhea
Shandong XiJieYiTong International Trade Co.,Ltd. , https://www.xjplaster.com