| 1 Purpose1.1 Clinical study: Based on the data mining platform of R language,we summarize the academic experience and drug use characteristics of Professor Nianzhi Zhang in the treatment of non-small cell lung cancer and analyze the rules of compounding and principles of formulas to provide reference for clinical treatment.1.2 Experimental study: To observe the effect of the empirical formula "Fei-ji-fang" on the malignant progression of A549 lung cancer in mice,and to explore the possible mechanism of action of Fei-ji-fang in non-small cell lung cancer.2 Methods2.1 Clinical study: Relevant case data were retrospectively collected,and with the help of R language software,the common herbs,drug pairs and core ideas of Professor Nianzhi Zhang for the treatment of non-small cell lung cancer were summarized.2.2 Experimental study: Using human A549 non-small cell lung cancer cells,30 BALB/cnude nude mice were constructed as a holotype.After successful membrane construction,they were randomly divided into model group,Fei-ji-fang low-dose group,Fei-ji-fang medium-dose group,Fei-ji-fang high-dose group,cisplatin group,and Fei-ji-fang highdose combined with cisplatin group,and were given corresponding interventions,and tumor length diameter,short diameter and mouse weight were recorded every 3 days during the intervention period,and relevant specimens were kept on the 15 th day.The tumors were weighed after stripping and the curves of weight and tumor volume changes in mice were drawn;the pathological changes of tumor tissues,lung,liver and kidney were observed by HE staining;simultaneous detection of IFN-γ and IL-6 levels of expression in mouse serum by ELISA;using immunohistochemistry to detect the expression of E-cadherin,Vimentin,β-catenin,c-Myc,Ki-67 and P53 proteins in tumor tissues;WB to detect the expression of E-cadherin,Vimentin,β-catenin and c-Myc proteins.3 Results3.1 Clinical study3.1.1 Patient baseline data: 82 patients with non-small cell lung cancer were included,of whom male:female = 1.48:1,mean age(60.74 ±13.76)years,history of smoking was62.20%,and history of secondhand smoke or environmental fume inhalation was 29.27%.3.1.2 Data mining results: All patients had a total of 328 prescriptions,involving 160 herbal medicines,with a cumulative frequency of 4884 times.25 herbal medicines were used more frequently than 1% of the cumulative frequency and were set as high-frequency herbal medicines.The four qi frequencies of HF herbal medicines were more often flat,cold,warm,slightly cold,and cool;the five tastes were more often sweet,bitter,light,pungent,and slightly bitter;and the frequencies of meridians were more often lung meridians,spleen meridians,heart meridians,stomach meridians,and kidney meridians,respectively.The subcategories of the efficacy of high-frequency herbal medicines were counted,and the most frequent were qi tonics,heat-clearing and detoxifying medicines,yin tonics,water-relieving and swelling-reducing medicines,and heart-nourishing and tranquilizing medicines.69 correlation rules were mined for 160 herbal medicines,including one first-order,21 second-order,35 third-order,and 12 fourth-order,involving11 kinds of drugs.The correlation analysis of the high-frequency drugs showed that:Chaoguya-Chaomaiya,Banzhilian-Baihuasheshecao,Zhebeimu-Kuxingren,DangshenBaizhu,Suanzaoren-Baiziren were the top five with larger correlation coefficients,and the differences were all statistically significant.High-frequency drugs were classified into clusters based on K-means algorithm and PAM algorithm,which can be divided into 4and 9 classes,respectively,while the hierarchical clustering is based on the common nodes in the genealogical chart and can be broadly divided into 4 categories based on the clinical experience of Prof.Nianzhi Zhang.3.2 Animal experiments3.2.1 Mice weight,tumor volume and weight: With the extension of the intervention time,the body mass of mice in the cisplatin group and high-dose combined cisplatin group showed a trend of first increasing and then decreasing,while the body weight of the remaining groups gradually increased;at the time of specimen retrieval on day 15,the tumor volume and weight of mice in all groups were decreased compared with the model group,and the difference between the high-dose and cisplatin groups was not statistically significant,but all were higher than the combined group(P < 0.05).3.2.2 HE staining results: All groups showed a large number of tumor cells with bigger nuclei and nuclear schizophrenia on microscopic examination of tumor tissues;the lung tissues of the model group,low-dose group and middle-dose group were incomplete in structure and surrounded by heterogeneous proliferating cells and inflammatory cells;the liver and kidney tissues of each group did not show any obvious abnormalities.3.2.3 ELISA results: As compared with the model group,the lower expression of serum IL-6 and higher expression of IFN-γ were observed in all groups,and the decrease or increase in both groups was most obvious in the combined group,and both were superior to the high-dose group and the cisplatin group(P < 0.05).3.2.4 IHC results: Expression of E-cadherin protein was increased in all groups compared with the model group,and was higher in the combination group than in the cisplatin and high-dose groups(P < 0.05);expression of Vimentin and P53 protein was declined in all groups in comparison with the model group,and was significantly different in the cisplatin,high-dose and combination groups(P < 0.05);as compared with the model group,expression of β-catenin,c Myc and Ki-67 protein were reduced in all groups compared with the model group(P < 0.05),and the expression of β-catenin in the combination group was lower than that in the cisplatin group(P < 0.05).3.2.5 WB results: E-cadherin protein expression was increased(P < 0.05)and Vimentin,c-Myc,and β-catenin protein expression was decreased(P < 0.05)in all groups compared with the model group,and the differences in expression of the above proteins were higher in the combination group than in the cisplatin and high-dose groups(P < 0.05).4 Conclusion4.1 The data mining study found that Professor Zhang Nianzhi used mainly qi tonics,heat-clearing and detoxifying drugs,and yin tonics in the clinical treatment of non-small cell lung cancer patients and focused on both attacking and tonifying because the clinical evidence was often mixed with deficiency and reality.The medicine used is mostly calm,good at using cold and warm,and the treatment focuses on the lung and takes into account the whole body.The treatment is always based on benefiting the qi and nourishing the yin,detoxifying and dispersing the nodules,and the clinical evidence is combined with moistening the lung to stop coughing,nourishing the heart and calming the mind,and strengthening the spleen and stomach.4.2 Animal experiments showed that the low,medium and high dose groups of Fei-ji-fang could inhibit the tumor growth and metastasis of A549 lung cancer mice to some extent,and the anti-tumor effect of the high dose combined with cisplatin was generally better than that of cisplatin alone,and the intrinsic mechanism of action might be related to the regulation of body immunity and inhibition of epithelial mesenchymal transformation. |
