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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 investigate the effect of Jianpi Bufei prescription （JPBFP） on airway inflammation， airway hyperresponsiveness （AHR）， and cyclic adenosine monophosphate （cAMP） signaling pathway activity in ovalbumin （OVA）-sensitized and challenged juvenile asthma rats. Method:Seventy-five male SD rats were randomly divided into a blank group （<italic>n</italic>=15） and an experimental group （<italic>n</italic>=60）. The rats in the experimental group were sensitized by aluminum hydroxide gel containing 0.2% OVA and stimulated by aerosol inhalation of normal saline containing 1% OVA to induce an asthma model， followed by assignment into the following groups： a model group （<italic>n</italic>=15）， a JPBFP group （<italic>n</italic>=15， 8.37 g·kg<sup>-1</sup>·d<sup>-1</sup>）， an aminophylline group （<italic>n</italic>=15， 40 mg·kg<sup>-1</sup>·d<sup>-1</sup>）， and a dexamethasone group （<italic>n</italic>=15， 0.1 mg·kg<sup>-1</sup>·d<sup>-1</sup>）. AHR was detected by the pulmonary function analyzer， changes in inflammatory cells by white blood cell （WBC） count and differential blood count in bronchoalveolar lavage fluid （BALF）， and pathological changes of lung tissues by hematoxylin-eosin （HE）， Masson， and periodic acid-schiff （PAS） staining. The interleukin （IL）-4， IL-5， IL-13， interferon （IFN）-<italic>γ</italic>， and tumor necrosis factor （TNF）-<italic>α</italic> levels in serum and the cAMP level in plasma were tested by the enzyme-linked immunosorbent assay （ELISA）. Protein kinase A （PKA） expression in lung tissues was detected by immunohistochemistry. The cAMP-response element-binding protein （CREB） mRNA and protein expression in lung tissues was detected by the real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. Result:Compared with the blank group， the model group showed increased lung resistance， decreased pulmonary compliance （<italic>P</italic><0.05）， elevated WBC count and proportion of eosinophils in BALF （<italic>P</italic><0.05）， up-regulated levels of IL-4， IL-5， IL-13， and TNF-<italic>α</italic> in peripheral blood， declining IFN-<italic>γ</italic> level （<italic>P</italic><0.01）， severe pathological changes of lung tissues， dwindled cAMP， and down-regulated PKA and CREB expression （<italic>P</italic><0.01）. Compared with the model group， JPBFP inhibited AHR， reduced WBC count and proportion of eosinophils in BALF and lung resistance （<italic>P</italic><0.05）， improved pathological changes of lung tissues， increased pulmonary compliance， and up-regulated cAMP in serum and PKA and CREB expression in lung tissues （<italic>P</italic><0.01）. Conclusion:JPBFP can improve AHR， inhibit airway inflammation， and alleviate lung injury in asthma rats. Its mechanism may be related to the up-regulation of the activity of the cAMP/PKA/CREB signaling pathway.
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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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To investigate the ATP-sensitive potassium channel (KATP channel) protein expressions during different periods under hypoxia condition and explore the effect of Qibai Pingfei capsule medicated serum (hereinafter referred to as QBPF) on the correlation between the protein expressions of KATP channel and nitric oxide in rat pulmonary arterial smooth muscle cells(PASMCs). Qibai Pingfei capsules were given to SD rats via continuous gavage for 10 days to obtain QBPF. Primary rats PASMCs were cultured by the direct adherent culture method. Western blot was applied to detect the protein expression levels of KATP channel (Kir6.1 and SUR2B) in PASMCs. Then the noncompetitive inhibitor of NO synthase--Nω-nitro-L-arginine methyl ester(L-NAME) and KATP channel inhibitor--glyburide(GLYB) were applied respectively to evaluate the effect of QBPF on the protein expressions of KATP channel. The protein expressions of Kir6.1 and SUR2B were increased after 6-hour hypoxia treament, peaked at the 24-hour hypoxia treament, and decreased in both 48-hour and 72-hour hypoxia groups. Especially, QBPF could further up-regulate the Kir6.1 and SUR2B protein expressions under 24-hour hypoxia condition; however, such up-regulation effect could be blocked by KATP channel inhibitor GLYB and NO specific inhibitor L-NAME, indicating that QBPF played the role of opening KATP channel. The regulatory mechanism was probably associated with up-regulating KATP channel protein expression via NO relative pathway, involving pulmonary vasodilation, and thus relieving the occurence and development of COPD.
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<p><b>OBJECTIVE</b>To evaluate the efficacy of Bufei Yishen Granule BFYSG) combined with Shufei Tie acupoint sticking therapy on quality of life of patients with stable chronic obstructive pulmonary disease (COPD).</p><p><b>METHODS</b>A multi-center, double-blinded, double-dummy and randomized controlled method was adopted in this trial. A total of 244 patients were randomly assigned to a trial group and a control group according to the random number, each with 122 patients; treatment allocation occurred when the participants met the inclusion criteria and signed the informed consent form. In the trial group, patients were treated with BFYSG combined with "Shufei Tie" acupoint sticking therapy and sustained-release theophylline dummy, and in the control group patients were treated with oral sustained-release theophylline and BFYSG dummy combined with "Shufei Tie" acupoint sticking therapy dummy. The therapeutic course for two groups was 4 months and the follow-up was 6 months. The frequency and duration of acute exacerbation calculated by adding up each frequency and duration of acute exacerbation in treatment and follow-up time respectively, the quality of life measured by the World Health Organization Quality of Life (WHOQOL)-BREF scale and adult COPD quality of life (COPD-QOL) scale were observed.</p><p><b>RESULTS</b>Among the 244 enrolled patients, 234 were screened for full analysis set (FAS); 221 were screened for per-protocol analysis set (PPS). After 4-month treatment and 6-month follow-up there were differences between the trial group and the control group in frequency of acute exacerbation (FAS: P=0.013; PPS: P=0.046); duration of acute exacerbation (FAS: P=0.005; PPS: P=0.006); scores of physiological, psychological and environment aspects of the WHOQOL-BREF questionnaire (FAS: P=0.002, P=0.006, P=0.000; PPS: P=0.00, P=0.001, P=0.000); scores of daily living ability, social activity, depression symptoms aspects of the COPD-QOL questionnaire (FAS: P=0.000, P=0.000, P=0.006; PPS: P=0.002, P=0.001, P=0.001).</p><p><b>CONCLUSION</b>BFYSG combined with acupoint sticking therapy could improve the quality of life of patients with stable COPD.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Acupuncture Points , Anxiety , Psychology , Combined Modality Therapy , Depression , Psychology , Disease Progression , Drugs, Chinese Herbal , Therapeutic Uses , Pulmonary Disease, Chronic Obstructive , Drug Therapy , Psychology , Quality of Life , Surveys and Questionnaires , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>By using the metabonomics method, to study the plasma metabonomics of the stable phase chronic obstructive pulmonary disease (COPD) patients of Fei-qi deficiency syndrome (FQDS), and of Chinese materia medica (CMM) intervention, thus exploring possibly existent biomarkers.</p><p><b>METHODS</b>Forty stable phase COPD patients of FQDS were recruited as Group A. Liuwei Buqi Capsule (LWBQC) was given to them as intervention. A healthy control group (Group B, 37 cases) was set up. The pulmonary function test was performed on patients in Group B and Group A before and after intervention. The plasma metabolites were detected using high performance liquid chromatography-mass spectrometry (HPLC-MS/MS). Statistical data were analyzed using principal component analysis (PCA) and partial least squares (PLS). The original spectrum and data of plasma metabonomics were compared between the two groups. The whole spectrum amino acid metabonomics tests were performed in the two groups.</p><p><b>RESULTS</b>Compared with Group B, the pulmonary function significantly decreased before intervention in Group A (P < 0.05). The pulmonary function was more mildly improved after 30 days' treatment than before treatment, but still lower than it in Group B (P < 0.05). The metabolic spectrum before treatment in Group A was significantly different from Group B, but showing regressive trend to Group B after treatment. Fifteen possible disease markers were found in COPD patients of FQDS. Results of the whole spectrum of amino acid metabolomics showed different features.</p><p><b>CONCLUSIONS</b>The changes of metabolic spectrum and amino acids could be found in the stable phase COPD patients of FQDS using plasma metabonomics, and potential markers could be detected. The intervention of the stable phase COPD patients of FQDS by Chinese medicine could brought positive changes in the metabolic profiling and amino acid metabolism.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Amino Acids , Metabolism , Case-Control Studies , Drugs, Chinese Herbal , Therapeutic Uses , Medicine, Chinese Traditional , Metabolomics , Phytotherapy , Plasma , Metabolism , Pulmonary Disease, Chronic Obstructive , Diagnosis , Drug Therapy , MetabolismABSTRACT
<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>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Diagnosis, Differential , Gene Expression Profiling , Medicine, Chinese Traditional , Oligonucleotide Array Sequence Analysis , Methods , Pulmonary Disease, Chronic Obstructive , Genetics , Allergy and Immunology , Syndrome , T-Lymphocytes , Metabolism , Yang Deficiency , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To establish an experimental animal model of Alzheimer's disease (AD) with clinical and pathological specificity conformed to AD in human, and to observe the effect of Zhinao Capsule (ZNC) on learning, memory and patho-morphological parameters in the model.</p><p><b>METHODS</b>The experimental AD model of rats was improved and established by combined injection of beta-amyloid protein (beta-AP) to lateral ventricle and transforming growth factor beta 1 (TGF beta 1) to brain. The learning and memory function of the model rats were tested by step-through test and water-maze test, and the number and cross area of beta-AP deposited macula in cerebral cortex and CA1 region of hippocampus were estimated quantitatively using immunohistochemical method and image pattern analysis.</p><p><b>RESULTS</b>The improved AD animal model showed both the specificity of behavior (learning and memory impairments) and the typical pathological specificity (beta-AP deposited macula). ZNC could effectively improve the impairment of learning and memory and reduce the number and cross area of beta-AP deposited macula in cerebral cortex and hippocampus CA1 region in the AD model rats.</p><p><b>CONCLUSION</b>The AD rat model induced by the combined injection of beta-AP and TGF beta 1 is a good animal model simulated to the clinical reality, which could be used to screen and evaluate the anti-dementia agents. ZNC could display anti-dementia effect of the AD model rats.</p>