The Study On Professor Zhou Zhongying’s Clinical Experience In The Treatment Of Primary Lung Cancer And The Experimental Study On Antitumor Effect Of YiFeiJieDu Decoction

Research objective:Based on the analysis of multi angle and multi link data mining and knowledge discovery,the clinical experience of Professor Zhou Zhongying and the dialectical thinking on the treatment of primary lung cancer were sumed up.The formation of the clinical process,and the refinement of his academic ideas Which can give an effective guidance for the application of traditional Chinese medicine in the treatment of lung cancer.Further study on anti-tumor of YiFeiJieDu Decoction in vitro.Research methods:Nearly 10 years professor Zhou Zhongying treatment of lung cancermedical records were collected and arranged.In strict accordance with the inclusion criteria,exclusion criteria selection,We conform to 646 cases(2451 cases)of medical record information which meet the standard to establish the medical record database.Operating association rules,frequency method,clustering analysis,multi-level mining analysis of syndrome,pathogenesis,and prescription,combined with medical interview,treatise learning,in-depth typical case studies,summarizes Professor Zhou’s academic thoughts.The experimental part:Human lung adenocarcinoma A549 cell was used as the research object.The A549 cells were cultured and treated with basic culture,low middle and high concentrations of Lung cancer basic prescription(YiFeiJieDu Decoction).Using MTT method to investigate the inhibitory effect on A549 cell proliferation,Using flow cytometry to investigate apoptosis and cell cycle.Western blot method was used to observe the expression of proteins related with apoptosis;The A549 cells were cultured and treated with 150μmol·L-1 CoCl2,150μmol·L-1 CoC12 +15mg·mL-1 Yifei Jiedu Decoction,15mg·mL-1 YiFeiJieDu Decoction,respectively.Untreated cells were used for the normal control.Morphological changes were observed under inverted microscope.Wound scratch assay were performed to further identify if YiFeiJieDu Decoction could reduce the migration ability of A549 cells.Transwell was used to explore the interven-tion effect of YiFeiJieDu Decoction on the invasion and movement of A549 cells.The expressions of E-cadherin,Vimentin,Snail,β-catenin,P-gsk-3β,P-smad2/3 were examined by Western blot.Results:(1)General information:There were 646 cases of primary lung cancer,which 332 are male and 314 are female.The average age is(61.75±0.05)years old.(2)Syndrome frequency distribution:324 kinds of symptoms were included.Cough,little phlegm,dry mouth,chest tightness,chest pain were common.61 kinds of tongue and pulse were involved.Dark purple tongue,dark red tongue,fissured tongue,yellow thin greasy coating,thin yellow greasy coating,rolling,thready,taut pulse were common.(3)Pathogenesis frequency distribution:133 kinds of Pathogenesis were found.The main Pathogenesis in the order were:Qi and Yin deficiency,the the phlegm blood stasis qi stagnation and toxin resistance,the heat toxin phlegm and blood stasis resistance,cancerous toxin transfer.Through the analysis of the pathogenesis of syndrome factors,the main disease factors followed were phlegm,blood stasis,cancerous toxin,heat,qi depression.deficiency of Yin,deficiency of Qi.The main disease location factors were lung,stomach,spleen,kidney,liver.(4)The distribution of commonly used herbs:Herb use frequency statistics:there are 345 herbs in the medical record in all,the main herbs followed by:Coasial Glehnia Root,Adenophora tetraphylla,Ophiopogon japonicus,Mountain arrowhead,Wartwort,Agrimonia pilosa Ledeb,Pseudostellaria heterophylla,Ranunculus ternatus,Hedyotis diffusa,Batryticated silkworm,Glabrous Sarcandra Herb,CoixSeed,Arisaema heterophyllum,Trionyx sinensis Wiegmann,Scutellaria barbata,Pinellia ternate.Herb efficacy analysis:Commonly used herbs(refers to the frequency of more than 400 times)involving 46 kinds of herbs,analysis by classification of herb efficacy,the attack evil herb to clearing away heat and toxin transforming phlegm and dissipating masses,promoting blood circulation to remove blood,dispel the wind,detoxicating and activating the collaterals and attacking toxin,dissipating phlegm and dissipating phlegm,blood stasis,clearing damp and promoting diuresis,respectively accounting for 10.93%,12.74%,9.45%,7.86%,5.09%.The centralizer herbs mainly in tonifying qi and nourishing yin and tonifying qi and blood,tonifying the liver and kidney,respectively accounted for 16.48%,6.54%,4.80%.The coordinating herbs mainly in invigorating and activating spleen.(5)The results of cluster analysis:according to cluster analysis of the 345 kinds of herbs,the herbs of lung cancer basic prescription are Coasial Glehnia Root,Adenophora tetraphylla,Ophiopogon j aponicus,Mountain arrowhead,Wartwort,Agrimonia pilosa Ledeb,Hedyotis diffusa,Batryticated silkworm,Glabrous Sarcandra Herb,Hedyotis diffusa.(6)The results of association analysis;analysis of the common pathological factors or the merge pathogenesis’s relationship with the herbs through the association rules.Such as herbs for associated with the heat toxin phlegm and blood stasis resistance:Coasial Glehnia Root,Adenophora tetraphyll a,Ophi opogon j aponi cus,Mountain arrowhead,Wartwort,Agrimonia pilosa Ledeb,Pseudostellaria heterophylla,Ranuncu-lus ternatus,Hedyotis diffusa,CoixSeed,Polistes mandarinus Saussure;herbs for associated with wind.Rhizoma typhonii,Centipede,Scorpion,Batryticated silkworm,Arisaema heterophyllum,Coasial Glehnia Root,Adenophora tetraphylla,Ophi opogon j aponi cus,Mountain arrowhead,Wartwort,Agrimonia pilosa Ledeb.(7)YiFeiJieDu Decoction can inhibit the proliferation of lung cancer cells with a dose and time dependent manner,can reduce the synthesis of DNA in A549 cells,arrest the cell in G1 phase and showed a dose dependent,induce the apoptosis,apoptosis rate was(11.6±3.07%,22.70±2.33%,26.67±3.69%vs 8.07±2.62%),the expression of cleaved-caspase 3,cleaved-caspase9 showed a concentration dependent increase,significant difference between the high dose group(P<0.05),the expression of p-JNK,p-p38MARK,p-ERK protein is a concentration dependent increase,significant difference between the high dose group(P<0.05).CoCl2 can makeA549cells show typical EMT morphological changes,which were accompanied by up-regulation of E-cadherin expression and down-regulation of Vimentin.Wound scratch assay indicated that YiFeiJieDu Decoction could reduce the migration ability ofA549 cells.Transwell assay showed that YiFeiJieDu Decoction could inhibit invasion of A549 cells.Westem Blot showed that YiFeiJieDu Decoction could up-regulate the expression of E-cadherin,P-gsk-3β protein(P<0.05)and down-regulate the expressions of Vimentin,Snai 1,β-catenin,P-Smad2/3 protein(P<0.05)respectively.Research Conclusions:(1)The basic pathogenesis of lung cancer is "The the phlegm blood stasis and heat,Qi and Yin deficiency".The main pathological factors of primary lung cancer are "phlegm,blood stasis,heat and cancer toxin".This disease is closely related to the lung,involving the spleen,stomach,liver,kidney.(2)Professor Zhou treating lung cancer common treatment is to clearing away heat and toxin,transforming phlegm and dissipating masses,promoting blood circulation and dissipating blood stasis,dispel the wind and activating the collaterals,tonifying qi and nourishing yin,tonifying qi and blood,tonifying the liver and kidney.The herbs of lung cancer basic prescription are Coasial Glehnia Root,Adenophora tetraphylla,Ophiopogon japonicus,Mountain arrowhead,Wartwort,Agrimonia pilosa Ledeb,Hedyotis diffusa,Batryticated silkworm,Glabrous Sarcandra Herb,Hedyotis diffusa.Treatment is mainly based on the basic prescription,primary and secondary pathogenesis,or main symptoms.(3)YiFeiJieDu Decoction probably inhibited the proliferation and promote apoptosis of A549 cells.YiFeiJieDu Decoction probably inhibited the migration,invasion,and EMT of human adenocarinoma A549 cells.The mechanism may be related to the inhibition of Wnt/beta-catenin pathway.