Method of detecting influence on human oral cell [gamma] H2AX level due to tobacco chewing gum

Method of detecting influence on human oral cell [gamma] H2AX level due to tobacco chewing gum

  • CN 106,367,468 A
  • Filed: 08/26/2016
  • Published: 02/01/2017
  • Est. Priority Date: 08/26/2016
  • Status: Active Application
First Claim
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1. a kind of method that detection gum base type tobacco product affects on human mouth cell γ

  • h2ax level is it is characterised in that includeFollowing steps;

    (1) described gum base type tobacco product to be measured is placed in freezing 20-40min in liquid nitrogen;

    (2) the described gum base type tobacco product freezing step (1), is pulverized with pulverizer immediately, collects broken after pulverizingLast gum base type tobacco product;

    (3) again described unqualified gum base type tobacco product is placed in freezing in liquid nitrogen;

    (4) 20-40min is soaked in the described unqualified gum base type tobacco product artificial saliva that step (3) obtains, every 2-5minAgitation is once;

    The temperature of described artificial saliva is 36-38 DEG C;

    (5) the extracting solution centrifugation 5-10min that step (4) is obtained;

    (6) supernatant being obtained with osmotic tester determination step (5), it is consistent with cell culture medium to adjust its osmotic pressure,Obtain gum base type tobacco product extracting solution;

    (7) measure the nicotine content in extracting solution;

    (8) culture human mouth mucosal epithelial cells are as detection object;

    (9) the gum base type tobacco product extracting solution obtaining in step (6) is added to cultured human mouth mucosa in step (8)In epithelial cell;

    (10) after the cultivation of 4 hours, detect cell γ

    h2ax situation of change with high intension imager.

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