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Objective:To investigate the molecular mechanism of Qiyu Sanlong prescription （QYSL） in inhibiting the "addiction" of lung cancer A549 cells to miRNA21. Method:The human lung cancer A549 cells were routinely passaged and divided into the blank group， blank serum group， QYSL-containing serum group， and siRNA group. The prepared QYSL-containing serum was used for intervention， with the optimal concentration and action time determined in previous studies. The protein and mRNA expression levels of miRNA21 and related molecules in its target phosphatase and tensin homolog deleted on chromosome 10 （PTEN）/phosphatidylinositol 3-kinase （PI3K） signaling pathway were detected by real-time polymerase chain reaction （Real-time PCR） and Western blot assay. Result:The comparison with the blank serum group revealed that the mRNA expression levels of miRNA21 in the QYSL-containing serum group and the siRNA group were decreased， while the PTEN mRNA expression in the QYSL-containing serum group was increased， showing significant differences （<italic>P</italic><0.01）. Compared with the blank serum， the QYSL-containing serum and siRNA significantly down-regulated PI3K and mammalian target of rapamycin （mTOR） mRNA expression （<italic>P</italic><0.01）， whereas the QYSL-containing serum did not change the mRNA expression of protein kinase B （Akt）. The protein expression levels of PTEN in the QYSL-containing serum group and the siRNA group were obviously elevated in contrast to that in the blank serum group （<italic>P</italic><0.05）. Meanwhile， the protein expression levels of phosphorylated Akt （p-Akt） and phosphorylated mTOR （p-mTOR） evidently declined in the QYSL-containing serum group （<italic>P</italic><0.05）， but there was no significant reduction in total Akt and mTOR protein expression. The PI3K protein expression was slightly down-regulated， with no statistical significance. Conclusion:QYSL inhibits the transcription of miRNA21， increases the expression of PTEN， and reduces the expression of key molecules in PI3K/Akt/mTOR signaling pathway， thus mildly inhibiting the "addiction" of lung cancer cells to oncogenes and blocking their proliferation.
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Objective:To evaluate the effect of Mashao Pingchuan decoction on traditional Chinese medicine <bold>（</bold>TCM<bold>）</bold> symptoms， quality of life， peripheral blood eosinophils （Eos） and serum inflammatory factors interleukin-6 （IL-6） and interleukin-8 （IL-8） in patients with cold asthma syndrome of bronchial asthma. Method:A total of 67 patients with cold asthma who attended the Respiratory Clinic of Anhui Provincial Hospital of TCM from January 2018 to December 2019 were selected and randomly divided into a control group and an observation group. The control group was given basic treatments such as budesonide formoterol powder inhalation， and the observation group was given Mashao Pingchuan decoction on the basis of the control group. Both groups intervened for 7 consecutive days. Observe and record the general condition， TCM symptom score， asthma control questionnaire （ACQ）-7 score， Marks-asthma quality of life questionnaire （Marks-AQLQ） score of the two groups of asthma patients， the peripheral blood Eos count was measured by hematology analyzer， and the serum IL-6 and IL-8 levels of the subjects were measured by enzyme-linked immunosorbent assay （ELISA）. Result:The total effective rate of TCM symptoms in the two groups was 100%（30/30）， and the effect in the observation group was more obvious （<italic>Z</italic>=-2.169，<italic>P</italic><0.05<italic>）</italic>. After treatment， the scores of TCM symptoms in the two groups were significantly lower than those before treatment （<italic>P</italic><0.01）. The scores of phlegm in the throat， expectoration， cough， and chills in the observation group were lower than those in the control group （<italic>P</italic><0.05）. After treatment， the ACQ-7 scores and Marks-AQLQ scores of the two groups were significantly lower than before treatment （<italic>P</italic><0.01）. There was no statistically significant difference between the observation group and the ACQ-7 scores and Marks-AQLQ scores. After treatment， the peripheral blood Eos counts and serum IL-6 and IL-8 levels in the two groups were significantly lower than before treatment （<italic>P</italic><0.01）. Peripheral blood Eos count and serum IL-6 and IL-8 contents in observation group were lower than those in control group （<italic>P</italic><0.05）. Conclusion:Mashao Pingchuan decoction combined with budesonide formoterol powder inhalation can effectively improve the clinical effectiveness of asthma （cold asthma）， improve the symptoms of asthma in TCM， ACQ-7 score， Marks-AQLQ score， peripheral blood Eos count， serum inflammatory factor content.
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Objective:To observe the effect of Qiyu Sanlong decoction （QYSL） on the expressions of key molecules in signal axis of mammalian rapamycin target protein （mTOR）/yeast Atg6 homologous （Beclin1）/ microtubule-associated protein1 light chain3 （LC3） in A549 cells. Method:With A549 cells as the research object， the effect of QYSL medicated serum on cell viability of A549 cells were detected by cell counting kit-8 （CCK-8） method. The effect of QYSL decoction on A549 cell apoptosis， autophagosome formation and the expression of autophagy markers were detected by Terminal-deoxynucleoitidyl transferase mediated nick end labeling （TUNEL） method， transmission electron microscope （TEM）， Real-time polymerase chain reaction （Real-time PCR） and Western blot. Result:QYSL medicated serum could inhibit the viability of A549 cells in a concentration-dependent manner. Compared with the blank serum group， the number of apoptotic A549 cells in the QYSL medicated serum group was significantly increased （P<0.01）， and the formation of autophagosome was significantly increased. Compared with the blank serum group， the mRNA and protein expressions of mTOR in A549 cells in the QYSL serum group were significantly decreased （P<0.01）， while mRNA and protein expressions of Beclin-1， autophagy related genes 5 （ATG5）， autophagy related genes 13 （ATG13） were significantly increased （P<0.01）. Conclusion:QYSL decoction can induce autophagy in A549 cells， and its specific mechanism may be related to the down-regulation of mTOR expression， the up-regulation of Beclin1， ATG5， ATG13 and LC3 expression， and the promotion of LC3Ⅰ conversion to LC3Ⅱ.
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Objective::To study the effect of Qiyu Sanlong decoction on the growth of subcutaneous tumor in lung cancer mice and the expressions of key autophagy molecule, yeast Atg6 homologous (Beclin1), autophagy related genes5 (Atg5), and microtubule-associated protein1 light chain3 (LC3B). Method::Lewis lung carcinoma cells (LLC) were used to reproduce the lung cancer mice transplanted model. After the modeling, the mice were randomly divided into model group, Qiyu Sanlong decoction group, chemotherapy group and combination group, with 18 transplanted mice in each group. In model group, mice were fed with 0.9% saline 20 mL·kg-1 daily. In Qiyu Sanlong decoction group, mice were fed with Qiyu Sanlong decoction 80.48 g·kg-1 daily. The chemotherapy group was intraperitoneally injected with 0.4 mL cisplatin solution (DDP) at the 1st, 3rd and 5th day. The combination group was orally given the drugs at the concentration of 80.48 g·kg-1, and 0.4 mL DDP solution was intraperitoneally injected at the 1st, 3rd and 5th day. After 21 days of continuous treatment, tumor tissue was exfoliated and weighed, and the tumor inhibition rate was calculated. Hematoxylin-eosin (HE) staining was used to observe the histological changes of tumor. The expressions and localizations of Beclin1 and LC3B in tumor tissues were detected by immunohistochemical staining. Protein expressions of Beclin1, Atg5, LC3B-Ⅰand LC3B-Ⅱ were determined by Western blot, and the ratio of LC3B-Ⅱ/LC3B-Ⅰ was calculated. The transcription levels of Beclin1, Atg5 mRNA in tumor tissues were detected by Real-time PCR. Result::Qiyu Sanlong decoction had a mild inhibitory effect on transplanted tumor, with an inhibitory rate of 31.2%. Under microscope, patchy necrotic tumor cells were observed in the tumor tissues of Qiyu Sanlong decoction group. Immunohistochemical staining and Western blot analysis showed that Qiyu Sanlong decoction could up-regulate the expressions of Beclin1, Atg5 and LC3B protein (P<0.01), and promote the conversion from LC3B-Ⅰ into LC3-Ⅱ compared with the model group. Real-time PCR results showed that Qiyu Sanlong decoction could promote the transcription of Beclin1 mRNA and Atg5 mRNA compared with the model group (P<0.01). Conclusion::Qiyu Sanlong decoction has a mild inhibitory effect on lung tumors, and its mechanism may be related to up-regulating the expressions of autophagy key proteins Beclin1, Atg5 and LC3B, and promoting the conversion from LC3B-Ⅰ to LC3B-Ⅱ.
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Objective:To summarize medication law of prescription and clinical experience on the treatment of chronic cough through data mining technology. Method:The formula information of 650 cases of chronic cough cases diagnosed and treated by professor HAN Ming-xiang was processed with data mining technology, and system cluster analysis and factor analysis were conducted by SPSS Statistics 22.0. Result:High-frequency medicines of professor HAN Ming-xiang for treating chronic cough were Stemonae Radix, Citri Reticulatae Pericarpium, Asteris Radix et Rhizoma and other 37 herbs, their frequency of occurrence accounted for 80.52%. According to the efficacy, the medicines could be classified into 10 categories, such as expectorant cough antiasthmatic drugs, tonifying deficiency drugs, and diaphoretic drugs. Cluster analysis showed 5 combinations of drugs. Factor analysis yielded 13 common factors, such as Stemonae Radix-Asteris Radix et Rhizoma-Farfarae Flos-Cynanchi Stauntonii Rhizoma et Radix, Asari Radix et Rhizoma-Schisandrae Chinensis Fructus-Zingiberis Rhizoma-Fritillariae Thunbergii Bulbus. Conclusion:Combined with clinical experience, it can be concluded that clinical treatment of professor HAN Ming-xiang for chronic cough attaches great importance to the pathogenesis of wind-evil, Yang deficiency and cold phlegm underlying lung, phlegm and blood stasis and Qi stagnation. The treatment mainly starts from the lung and spleen, with the method of warm and moist, or with some traditional Chinese medicines with the function of removing phlegm by warming the lung, dispersing cold by thinning the surface, strengthening the spleen and resolving phlegm, strengthening the lung and fixing the surface, and relieving spasmolysis by wind.
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<p><b>OBJECTIVE</b>To study T lymphocyte related genes with differential expression in patients with chronic obstructive pulmonary disease (COPD) of Fei-qi deficiency (FQD) syndrome type by gene chips.</p><p><b>METHODS</b>Lymphocytes in peripheral blood were isolated by Ficoll technique from blood samples collected from COPD patients of FQD syndrome type, Fei-yin deficiency (FYD) syndrome type, and also from healthy subjects for control. They were sorted and purified by flow cytometry, and the different expressed genes were screened from them by gene chip technique.</p><p><b>RESULTS</b>There were 15 genes with high differential expression between patients of FQD type and those of FYD syndrome type, and between patients of FQD type and healthy subjects.</p><p><b>CONCLUSION</b>Gene chip technique could be used for studying the gene expression profiles of TCM syndrome, and the T-lymphocyte related genes with differential expression in COPD patients with FQD were screened preliminarily.</p>