Tetrastigma　hemsleyanum　Diels　et　Gilg　(THDG)　is　used　as　a　Chinese　traditional　anti-inflammatory　medicine　for　about　thousands　of　years.　In　this　work,　Tetrastigma　hemsleyanum　Diels　et　Gilg＇s　polysaccharide　(TP)　can　inhibit　E.　coli＇s　growth　in　initial　dosing　period.　Compared　with　the　antibacterial　effect　of　Achyranthe＇s　polysaccharide　(AP)　from　their　metabolic　profile,　it＇s　obviously　that　their　metabolic　sites　for　E.　coli　were　inconsistent.　Moreover,　TP　could　not　only　increase　the　level　of　fructose-6-phosphate　(F6P),　decrease　the　level　of　fructose-1,6-diphosphate　(FBP),　but　also　charge　the　amount　of　the　two　differential　metabolic　with　the　change　of　the　concentration　and　the　dosing　time.　Actually,　F6P　could　transform　into　FBP　by　catalyze　of　6-phosphofructokinase-1(6-PFK-1),　which　is　an　important　process　in　glycolysis.　Furthermore,　FBP　was　considered　have　positively　correlated　with　E.　coli＇s　growth　rate.　Therefore,　TP　can　inhibit　the　E.　coli＇s　proliferation　by　interfering　with　the　process　for　glycolysis　and　gluconeogenesis.　Based　on　the　experimental　result,　we　proposed　a　new　mouthwash　method　to　evaluate　the　anti-bacterial　activity.　Compared　with　AP,　TP　can　inhibit　the　E.　coli＇s　growth　within　2　h　with　a　low　concentration　(0.5%)　and　a　short　dosing　time　(5　min).　This　study　extends　the　applications　of　THDG　and　establishes　a　new　assessment　method　for　the　pharmacology　activity　of　Chinese　herbal　medicine.　©　2019