Research Of The Clinical Experiences Of Professor Li Jiageng In Syndrome Differentiation And Treatment For Pulmonary Diseases Based On Data Mining And Verification

Objective:This study screened Professor Li Jiageng for differential treatment of lung diseases(lung cancer,chronic bronchitis,allergic rhinitis),explored the compatibility of drug pairs by data mining,and refined Professor Li Jiageng's prescription characteristics and academic thoughts.Through the verification of data mining lung cancer drug pairs through network pharmacology and experimental research,we can more accurately apply the clinical experience gained by data mining to clinical practice in the future,and open ideas for follow-up research.Method1.We select the outpatient medical records of Professor Li Jiageng in the Phoenix Clinic of Hubei Hospital of traditional Chinese Medicine and the National Medical School of Hubei University of traditional Chinese Medicine from 2016 to 2019.Through the medical record entry system developed by Access2010 database,we input the screened medical records and clean the input data,import the cleaned data into Weka3.6 data mining software,and use the methods of frequency analysis,association rules and cluster analysis to carry out data mining.Next,we use Pajek2.0complex network software to distinguish frequency by node size,drug efficacy by node color,calculate node connection parameters based on the results of drug pair analysis of association rules,and use the results of cluster analysis as core circle drugs to draw complex network diagram.2.We selected Coix seed-Weilingxian medicine pair as the research object,and searched the effective components and their targets in TCMSP database with "Coix seed" and "Weilingxian" as keywords,and finally obtained the genes of effective component targets.We use "lung cancer" as the keyword to search the lung cancer related genes and targets in Uin Prot database,Gene Cards database,Drug Bank database and TTD database,eliminate the duplicate genes and targets,and finally obtain the genes of the disease target.The effective component targets and disease targets were mapped by R software,and the possible targets of Coix seed and Weilingxian in interfering with lung cancer were obtained.The active component targets and lung cancer related targets were introduced into the component-target network construction tool Cytoscape 3.7.2 to construct the network diagram of Coix seed-Weilingxian and targets,and the active components and disease targets were represented by node(node)with different shapes and colors.The effective components of the target of Coix seed and Weilingxian in interfering with lung cancer were imported into STRING database to obtain PPI network map,the "Multiple proteins" function in STRING database was selected,and the species was set to "homo sapiens".We export the PPI diagram and the "string_interactions" file by checking the "hide disconnected nodes in the network" option in the PPI network diagram and setting the value of "minimum required interaction score" to 0.4.The core proteins enriched in the top 30 in the "string_interactions" file were imported into the DAVID database,and the species was set to "homo sapiens" for GO enrichment and KEGG enrichment analysis.3.We selected 100 mice and inoculated Lewis tumor cells under their right axilla,0.2 m L/.The size of subaxillary tumor in mice was observed 4-5 days after transplantation of tumor cells.The mice which grew well and the tumor volume under the right axilla was about100 mm were divided into 5 groups: model group(Model,MOD),Coix seed-Weilingxian low dose(Yiyiren-Weilingxian,YW-L)group,Coix seed-Weilingxian middle dose(Yiyiren-Weilingxian,YW-M)group,Coix seed-Weilingxian high dose(Yiyiren-Weilingxian,YW-M)group and cyclophosphamide(Cyclophosphamide,CTX)group.There are 15 rats in each group.Coix seed 20 g and Weilingxian 15 g were made into water decoction,which was concentrated into a concentrated solution containing crude drug 0.45 g per milliliter.MOD group was given 20 m L/kg/d saline,CTX group was given cyclophosphamide 30mg/kg/d,YW-L group,YW-M group and YW-H group were given 4.55g/kg/d,9.1 g/kg/d,18.2 g/kg/d Coix seed and Weilingxian concentrated water solution respectively,with a total volume of20 m L/kg.The mice in each group were given continuous intragastric administration for 30 days,once a day.After intragastric administration,the samples were taken,the chest cavity of mice was carefully opened,the lung tumor was removed and weighed,and part of the tumor tissue was cut and fixed in 4% paraformaldehyde solution for HE staining,Tunel staining and immunohistochemical detection.The rest of the tumor tissue was preserved in liquid nitrogen for Real-time PCR and Western Blot detection.Result1.Clinical regularity of Professor Li Jiageng in the treatment of Pulmonary Diseases This study selected 653 cases of lung diseases diagnosed and treated by Professor Li Jiageng from 2016 to 2019(207cases of lung cancer,244 cases of chronic bronchitis,202 cases of allergic rhinitis).There were 314 males,accounting for 48.1% of the total,and 339 females,accounting for 51.9% of the total.Professor Li believes that the common syndromes of lung cancer are yin deficiency and toxin heat syndrome,phlegm turbidity obstructing the lung syndrome,lung depression and phlegm stasis syndrome,qi stagnation and blood stasis syndrome,qi and yin deficiency syndrome.The core prescriptions of its treatment are:Sangxing decoction for the treatment of yin deficiency toxin-heat syndrome;Zhisou San combined with Sanao decoction for the treatment of phlegm stagnation and lung stasis syndrome;Chaihu Shugan Power for the treatment of lung stagnation and phlegm stasis syndrome;Siwu decoction for the treatment of qi stagnation and blood stasis syndrome;Shengmai Powder combined with Zhisou Powder for the treatment of Qi and Yin deficiency syndrome.The commonly used core drugs are: Hedyotis diffusa with Chong Lou has the effect of heat-clearing and detoxification,which is often used in anti-tumor therapy;Diffusa alone with steamed Baibu,grilled ephedra with almonds and perilla leaves with almonds has the effect of relieving cough and asthma;Schizonepeta tenuifolia with Fangfeng and Huoxiang has the effect of dispelling wind and relieving surface;Coix seed with Weilingxian has the effect of reducing swelling and relieving dyspnea.Radix Salviae Miltiorrhizae with Radix Paeoniae Miltiorrhizae,Radix Salviae Miltiorrhizae with Radix Paeoniae Miltiorrhizae and fried Hawthorn with fried Hawthorn have the effect of nourishing blood and promoting blood circulation.Professor Li believes that the common syndrome types of chronic bronchitis are wind-cold attacking the lung,wind-heat invading the lung,phlegm-heat obstructing the lung,liver-fire invading the lung,deficiency of lung and spleen qi,kidney-yang deficiency.Among them,the core prescriptions for treatment are: Sanao decoction for the treatment of wind-cold attacking the lung syndrome;Yinqiao Powder for the treatment of wind-heat invading the lung syndrome;Huangqin decoction combined with Zhisou Power for the treatment of phlegm-heat obstruction of the lung syndrome;Daige Powder for the treatment of liver-fire invading the lung syndrome;Sijunzi decoction for the treatment of lung and spleen qi deficiency syndrome;Zhenwu decoction for the treatment of kidney-yang deficiency syndrome.Among them,the core drugs used for treatment are: winter flower with steamed hundred parts,fried ephedra with almonds for relieving cough and resolving phlegm,lung and asthma;Schizonepeta tenuifolia with Fangfeng,patchouli with Fangfeng,Huoxiang with Schizonepeta for the treatment of dampness and relieving cold;Honeysuckle with forsythia and dandelion with forsythia for heat-clearing and table-clearing treatment;Salvia miltiorrhiza with red peony and salvia miltiorrhiza with Chuanxiong for blood-nourishing and blood-activating therapy.Pueraria lobata with Uncaria for calming the liver and relieving wind;stir-fried malt with fried grain sprout for digestion and stomach treatment.Professor Li believes that the use of drugs for pulmonary diseases can be divided into the following categories.The commonly used antipyretic drugs include Radix Paeoniae Rubra,Radix scrophulariae,Forsythia suspensa,Hedyotis diffusa,Radix Scutellariae,Radix S and so on.The commonly used drugs for tonifying deficiency include Radix Astragali,Radix Paeoniae Alba,fried Atractylodes macrocephala,Radix Ophiopogonis,Radix Pseudostellari and so on.The commonly used drugs for resolving phlegm,relieving cough and relieving asthma include Pinellia ternata,almond,roasted ephedra,Fructus Rehmanniae,steamed Baibu,Platycodon grandiflorum and so on.The commonly used drugs of anti-food drugs include fried Hawthorn,fried malt,fried grain malt,fried chicken inner gold,fried divine koji.The commonly used drugs for promoting blood circulation and removing blood stasis include Salvia miltiorrhiza,Ligusticum chuanxiong,Rhizoma Corydalis,fried zedoary,chicken blood.The commonly used drugs for regulating qi include tangerine peel,Rhizoma Cyperi,fried Fructus Aurantii,Allium macrostemon,bergamot and so on.The commonly used drugs for dehumidifying include patchouli,Peran,Amomum villosum,Magnolia officinalis and Atractylodes.The commonly used drugs for diuresis and dampness include Poria cocos,Coix seed,alisma alisma,plantain,money herb and so on.The commonly used drugs for calming the liver and relieving wind include fried earth dragon,scorpion,Uncaria,calcined oyster,stiff silkworm and so on.The commonly used drugs for astringent medicine include Schisandra chinensis,Wumei,floating wheat,cuttlebone and Cornus officinalis.The commonly used drugs for calming nerves include Poria,Radix Polygalae,calcined keel,albizzia bark,and wild jujube kernel.The commonly used drugs for dispelling wind and dampness include Weilingxian,mulberry twig,mulberry parasite,solitary living and so on.The commonly used hemostatic drugs include white thatch root,fairy crane grass,Baiji,Platycladus orientalis charcoal,alizarin and so on.In addition,the commonly used drugs for resuscitation include Acorus tatarinowii,the commonly used drugs for purgative include hemp kernel and Yu Li Ren,and the commonly used drugs for attacking poison,killing insects and relieving itching are Fructus Rehmanniae.2.Study on the Therapeutic Mechanism of Coix seed-Weilingxian against Lung Cancer based on Network Pharmacology TCMSP database shows that there are 9 and 11 active ingredients in Coix seed and Weilingxian,6 in Coix seed in intervention of lung cancer,and 3 in Weilingxian.Coix seed plays a leading role in the intervention of Coix seed-Weilingxian drug on lung cancer.The "component-target" network diagram shows that the effective components of Coix seed-Weilingxian medicine on lung cancer intervention are Stigmasterol(stigmasterol),beta-sitosterol(?-sitosterol)and Sitosterol alpha1(sitosterol ? 1).PPI network map showed that ADRA2 A,CSAP3,JUN,MAOA,ADRA1 B,SLC6A4,HSP90AA1,MAOB,PTSG2 and ADRA1 A were the core target genes of Coix seed-Weilingxian intervention on lung cancer,and KEGG enrichment showed that Coix seed-Weilingxian interfered lung cancer mainly by regulating PI3K-AKT signal pathway,calcium(calcium)signal pathway,estrogen(Estrogen)signal pathway,p53 signal pathway, IL-17 signal pathway and AGE-RAGE signal pathway.3.Study on the Anticancer effect and Mechanism of Coix seed-Weilingxian on Lewis Lung Cancer mice The results of tumor weight and tumor inhibition rate in mice showed that compared with MOD group,the tumor weight in lung tissue of YW-L group,YW-M group,YW-H group and CTX group decreased(P<0.01,P< 0.05),and the tumor inhibition rate increased.Compared with YW-L group,the tumor weight in lung tissue of CTX group decreased,while the tumor inhibition rate increased.The results of HE staining showed that the cells in the MOD group were disordered and tight,with deformed and necrotic cells and obvious nuclear pyknosis in some areas.In YW-L group,YW-M group,YW-H group and CTX group,the arrangement of cells was loose,the intercellular space was enlarged,there were a large number of deformed and necrotic cells,nuclear pyknosis was obvious,and there were lymphocytes infiltration on the edge of the tumor,among which the cell deformation and necrosis in CTX group was the most obvious.The results of Tunel staining showed that the number and density of green fluorescence in lung tissues of YW-L group,YW-M group,YW-H group and CTX group were higher than those in MOD group.Compared with YW-L group,the number and density of green fluorescence in lung tissue of CTX group increased significantly.Compared with MOD group,the apoptosis index of lung tissue in YW-L group,YW-M group,YW-H group and CTX group increased(P< 0.01).Compared with YW-L group,the apoptosis index of lung tissue in CTX group increased(P< 0.05).The results of Bcl-2,Bax and Caspase-3 immunohistochemistry showed that compared with MOD group,the number of Bax and Caspase-3positive cells in lung tissue of mice in YW-L group,YW-M group,YW-H group and CTX group increased(P< 0.05,P< 0.01),while the number of Bcl-2 positive cells decreased(P< 0.05,P < 0.01).Compared with YW-L group,the number of Bax and Caspase-3 positive cells in lung tissue of CTX group increased,while the number of Bcl-2 positive cells decreased(P< 0.01).The results of Real-time PCR showed that compared with MOD group,the m RNA expression levels of Bax and Caspase-3 in YW-L group,YW-M group,YW-H group and CTX group increased,while the m RNA expression level of Bcl-2 decreased(P< 0.05,P < 0.01).Compared with YW-L group,the m RNA expression level of Bax and Caspase-3 in lung tissue of CTX group increased,while the m RNA expression level of Bcl-2decreased(P< 0.01).The results of Western Blot assay showed that compared with MOD group,the protein expression levels of Bax and Caspase-3 in lung tissue of YW-L group,YW-M group,YW-H group and CTX group increased(P < 0.05,P< 0.01),while the protein expression level of Bcl-2decreased(P < 0.05,P< 0.01).Compared with YW-L group,the protein expression level of Bax and Caspase-3 in lung tissue of CTX group increased(P< 0.05,P < 0.01),while the protein expression level of Bcl-2 decreased(P< 0.05,P< 0.01).Compared with MOD group,the protein expression levels of p-PI3K-Tyr607,p-AKT-Thr308 and p-m TOR-Ser2448 in lung tissue of YW-L group,YW-M group,YW-H group and CTX group decreased in different degrees(P< 0.05,P< 0.01).Compared with YW-L group,the expression levels of p-PI3K-Tyr607,p-AKT-Thr308 and p-m TOR-Ser2448 in lung tissue of mice in CTX group were decreased in different degrees(P< 0.05,P < 0.01).Conclusion1.Professor Li Jiageng's clinical experience in differential diagnosis and treatment of lung diseases includes:(1)attaching importance to protecting Wei qi;(2)attaching importance to regulate qi smoothly and treating lung from the liver;(3)attaching importance to the treatment of lung disease from phlegm and blood stasis;(4)attaching importance to protect spleen and stomach;(5)flexible application,light and stable.Professor Li Jiageng's academic thoughts on differential diagnosis and treatment of pulmonary diseases include:(1)the combination of syndrome differentiation and disease differentiation,which complement each other;(2)the classics are suitable for use and complement each other.2.Coix seed-Weilingxian medicine mainly interferes with lung cancer by regulating inflammation,glucose metabolism and estrogen.Coix seed-Weilingxian medicine has multi-channel and multi-level characteristics in the intervention of lung cancer.On the one hand,it can directly regulate the cell cycle,on the other hand,it can also interfere with the occurrence and development of tumor by regulating inflammation and estrogen levels.3.Coix seed-Weilingxian can inhibit the proliferation and promote the apoptosis of Lewis lung cancer cells,and its mechanism may be related to its inhibition of PI3K/AKT/m TOR signal pathway.