最新刊期
卷 26 ， 期 8 ， 2020
- 摘要：Diabetes mellitus (DM) is a chronic metabolic disease characterized by hyperglycemia. Its main complications of diabetes, such as diabetic angiopathy, have seriously affected the quality of life for patients, and have become an important cause of death and disability. The underlying pathological changes include macrovascular lesions and microvascular lesions. Diabetic macrovascular lesions mainly involve thoracic aorta, coronary artery, carotid artery, cerebral artery and peripheral blood vessels, etc., and the common clinical diseases include coronary heart disease, stroke, peripheral neuropathy, lower extremity arteriosclerosis, etc. Diabetic microvascular lesions mainly involve the heart, brain, kidney and other microvessels. Nowadays, various new oral hypoglycemic agents and insulin have emerged in the society and are widely used in clinical practice. However, traditional Chinese medicines(TCMs) have stable curative effect, less side effect, and can improve glucose metabolism, lipid metabolism, insulin resistance, oxidative stress, expression of inflammatory cytokines, vascular endothelial injury, microcirculation disorders, balance of fibrinolysis system and blood coagulation system, and improve the syndromes of TCMs, etc. They have been widely recognized and applied in the prevention and treatment of diabetic angiopathy. A profound understanding on the etiology, pathogenesis and treatment of diabetic angiopathy has been formed in Chinese medicine. Therefore, in this paper, we would summarizes the understanding on Chinese medicine for diabetic angiopathy and the mechanism of Yiqi Huoxue prescription in the treatment of diabetic angiopathy in the past three years.关键词：Yiqi Huoxue prescription;diabetic angiopathy;Danggui Buxue decoction;Danqi Jiangtang capsule;Yiqi Huoxue Tongluo decoction;Qi deficiency and blood stasis
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发布时间：2021-02-09 - 摘要：Objective:To investigate the protective effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts on vascular calcification induced by high glucose in mice by observing the expression of osteopontin (OPN) and smooth muscle 22α (SM22α) as well as vascular calcium deposition in the common carotid artery and thoracic aorta of mice.Method:Totally 130 male C57BL/6 mice were randomly divided into normal control group and high glucose group. The mice in high glucose group were intraperitoneally injected with streptozotocin(STZ), and fed on a high-fat diet for 7 months. Then, the mice were randomly divided into model group, low-dose and high-dose Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts groups (0.819, 1.638 g·kg-1), and metformin group (150 mg·kg-1). Each group was intragastrically administered once a day for 9 weeks. The changes in blood glucose were measured. Seven days before the end of the administration, a group of 4-week old male C57BL/6 mice were purchased and fed normally for one week as a youth group. At the end of the administration, the common carotid artery and thoracic aorta tissues of the mice were collected. Von Kossa staining was used to determine the degree of calcium deposition in the common carotid artery and thoracic aorta. The expression levels of OPN and SM22α protein in the common carotid artery and thoracic aorta were detected by immunohistochemistry. The expression of OPN and SM22α protein in the common carotid artery of mice was determined by Western blot.Result:As compared with the young group, the blood glucose of the normal control group was slightly increased without statistical difference, the common carotid artery and thoracic aorta were uniformly stained, and no black granular precipitate was observed. As compared with the normal control group, the blood glucose of the model group was increased (P<0.01), with a large amount of brown-black particles deposited in the intimal elastic fibers, showing obvious calcium salt deposition. As compared with the model group, blood glucose was significantly decreased in each administration group (P<0.05, P<0.01), and the degree of vascular calcium salt deposition was significantly reduced. There were no significant changes in expression levels of OPN protein and SM22α protein in the common carotid artery and thoracic aorta between the youth group and normal control group. As compared with the normal control group, the expression of intimal OPN protein in the common carotid artery and thoracic aorta of the model group was positive, SM22α protein expression was weakly positive, and the gray value of OPN protein expression in the common carotid artery was significantly increased (P<0.01), while the gray value of SM22α protein was decreased significantly (P<0.01). As compared with the model group, the expression levels of intimal OPN protein and SM22α protein in the common carotid artery and thoracic aorta of each administration group were significantly improved, and the gray value of OPN protein expression in the common carotid artery was reduced (P<0.05, P<0.01), while SM22α protein expression was significantly increased (P<0.01).Conclusion:High glucose can induce calcification of common carotid artery and thoracic aorta in mice and accelerate vascular aging. This formation process may be related to the expression of OPN and SM22α. Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts can reduce vascular calcification and delay vascular aging by regulating the expression of OPN and SM22α.关键词：vascular calcification;osteopontin (OPN);smooth muscle 22α (SM22α);high glucose;Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts;vascular aging
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发布时间：2021-02-09 - 摘要：Objective:To investigate the effects of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts (GNC) on the protein expression of α-smooth muscle actin (α-SMA) and runt-related transcription factor2(Runx2) after high glucose-induced vascular aging in mice, and elucidate the protective mechanism of GNC in delaying vascular aging.Method:Totally 130 male C57BL/6 mice were randomly divided into normal control group and high glucose group. The mice in high glucose group were intraperitoneally injected with streptozotocin (STZ). After successful modeling, the mice received high-fat diet for 7 months, and then they were randomly divided into model group, GNC low-dose and high-dose groups (0.819, 1.638 g·kg-1), and metformin group (150 mg·kg-1). The drug was given by intragastric administration once a day for 9 weeks. Seven days before tissues collection, a new batch of 4-week-old male C57BL/6 mice were purchased and fed normally for 1 week as a youth group. The general condition of the mice was observed. Morphological changes of the common carotid artery in mice were determined by hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM). Masson trichromatic staining was used to observe the fibrosis of common carotid artery in mice. The expression levels of matrix metalloproteinases-2 (MMP-2), cyclin-dependent kinase inhibitor 2A (p16), cyclic-dependent kinase inhibitor 1A (p21), α-SMA and Runx2 in the common carotid arteries of mice were detected by immunohistochemistry.Result:The results of HE, TEM and Masson showed that there was almost no change in the inimal and adventitial thickness, ultrastructure and relative contents of collagen and elastic fibers in the common carotid arteries of mice between the youth group and normal control group. As compared with the normal control group, the intima of the common carotid artery in the model group was not smooth, the endothelial cells were almost completely detached, the cytoplasm was lysed, the inner elastic membrane became thinner, fractured, or even detached, and the proliferating collagen fibers sneaked into the tunica media. The hyperplasia of tunica media and tunica adventitia was obvious and disordered (P<0.01). The vascular smooth muscle cells showed deformations, protuberances, bifurcations, and even fragmentation, and focal necrosis was observed. There were significantly more vacuoles, lysosomes, and obvious autophagy vesicles. The relative content of collagen and elastic fibers in vascular walls increased significantly (P<0.01). Compared with the model group, the above situation was relieved in each administration group (P<0.01). The results of immunohistochemistry showed that high glucose induced high expression of MMP-2, p16, p21 and Runx2 in the common carotid arteries(P<0.01), low expression of α-SMA(P<0.01), and the protein expression tended to be normal after drug intervention(P<0.05, P<0.01).Conclusion:High glucose can induce the aging of common carotid artery in mice and change the expression of α-SMA and Runx2 proteins. The Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts can delay vascular aging by regulating the protein expression of α-SMA and Runx2.关键词：vascular aging;α-smooth muscle actin (α-SMA);runt-related transcription factor-2(Runx2);high glucose;Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts;common carotid artery
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发布时间：2021-02-09 - 摘要：Objective:To investigate the protective effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma (GNC) extracts on myocardial fibrosis in diabetic mice by observing the degree of myocardial fibrosis and collagen types I (Collagen Ⅰ), collagen types Ⅲ (Collagen Ⅲ) and transforming growth factor-β1 (TGF-β1) protein expression in myocardial tissues.Method:A diabetic mice model was induced by streptozotocin (STZ) and high-fat diet. A normal control group was established. According to random number table method, diabetic mice were divided into model group, GNC low-dose and high-dose groups (0.819, 1.638 g·kg-1), and metformin group (150 mg·kg-1). Intragastrical administration was given in all groups, and the mice in normal control group received an equal dose of deionized water once a day for 9 weeks. The myocardial interstitial fibrosis in mice was observed by Masson trichromatic staining. Image-pro plus 6.0 analysis software was used to calculate the ratio of collagen area to total area. Immunohistochemistry was used to detect Collagen I, Collagen Ⅲ and TGF-β1 protein expression in myocardial tissues. The protein expression electrophoresis and gray value levels of Collagen I, Collagen Ⅲ and TGF-β1 in the myocardial tissues were detected by Western blot.Result:The results of Masson staining showed that as compared with the normal control group, the myocardial cells of diabetic mice were hypertrophic and disordered, and the myocardial stroma, especially the blue-stained collagenous fibers around the blood vessels, were heavily deposited and connected to each other in a network (P<0.01). As compared with the model group, the arrangement of myocardial cells was significantly improved in GNC low-dose and high-dose groups and metformin group, and the collagenous fibers in the myocardial stroma were significantly decreased (P<0.05). Immunohistochemistry and Western blot results showed positive expression of Collagen Ⅰ, Collagen Ⅲ and TGF-β1 in myocardial tissues, with significantly increased content of protein expression in diabetic mice (P<0.05, P<0.01). As compared with the model group, the positive protein expression decreased and the protein content tended to be normal in each administration group (P<0.05, P<0.01).Conclusion:High-fat diet combined with STZ can induce myocardial fibrosis in diabetic mice, and increase Collagen I, Collagen Ⅲ and TGF-β1 protein expression. Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts can improve myocardial fibrosis in diabetic mice by regulating the expression of Collagen I, Collagen Ⅲ and TGF-β1 protein.关键词：diabetes;myocardial fibrosis;streptozotocin(STZ);Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma (GNC) extracts;collagen types I (Collagen Ⅰ);collagen types Ⅲ (Collagen Ⅲ);transforming growth factor-β1 (TGF-β1)
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发布时间：2021-02-09 - 摘要：Objective:To explore the protective mechanism of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma (GNC) extracts on cardiac aging in diabetic mice by observing the activation of AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway, changes of cardiac pathomorphological and related senescent proteins.Method:C57BL/6 male mice, SPF level, were randomly divided into normal control group and high-glucose group. The mice in high-glucose group were intraperitoneally injected with streptozotocin (STZ) and fed with high-fat diet. After successful modeling, they were randomly divided into model group, low-dose GNC group (0.819 g·kg-1), high-dose GNC group (1.638 g·kg-1) and metformin group (150 mg·kg-1). The drug was administered by gavage once a day for a continuous period of 9 weeks. 4-week-old male C57BL/6 mice were normally fed for 1 week as a youth group. General conditions of mice were observed. Hematoxylin-eosin (HE) staining combined with transmission electron microscope (TEM) was used to observe the cardiac pathomorphology in mice. Von Kossa staining was used to determine the degree of calcium salt deposition in cardiac micro vessels. Western blot was used to detect the activation of signaling pathways in myocardial tissue of mice, as well as the expression levels of matrix metalloproteinases-2 (MMP-2), tumor suppressor p53 (p53), and phospho-tumor suppressor p53 (p-p53).Result:As compared with the normal group, the blood glucose in the model group increased (P<0.01), as compared with the model group, the blood glucose in each administration group decreased significantly (P<0.05, P<0.01). The results of three pathological morphology experiments (HE, TEM, and Von Kossa) showed that as compared with the normal control group, the mice in model group showed cardiomyocytes hypertrophy, disordered arrangement of myocardial fibers, focal dissolving and necrosis, mitochondria swelling, degeneration, crest fracture, vacuolar alteration, disordered microvascular structure of the heart, uneven staining, and a large amount of calcium deposition in tunica media and intima. As compared with the model group, the pathomorphological changes of mice in each administration group were improved in varying degrees. Compared with the normal group, the expression levels of MMP-2, p53 and p-p53 protein in the model group were significantly increased (P<0.05, P<0.01), the protein ratios of p-liver kinase B2(LKB1)/LKB1, p-AMPK/AMPK were significantly decreased (P<0.05, P<0.01), and the average gray level of p-mTOR/mTOR and p-p70S6 kinase(p70S6k)/p70S6k protein was significantly increased (P<0.05, P<0.01), while the protein ratios of p-mTOR/mTOR, p-p70S6k/p70S6k were increased (P<0.01). As compared with the model group, the expression levels of MMP-2, p53 and p-p53 protein in each administration group were significantly decreased (P<0.05, P<0.01), the protein ratios of p-LKB1/ LKB1, p-AMPK/AMPK were significantly increased (P<0.05, P<0.01), while the protein ratios of p-mTOR/mTOR and p-p70S6k/p70S6k were decreased (P<0.05, P<0.01).Conclusion:STZ combined with high-fat diet can induce cardiac aging in mice, and GNC can improve cardiac aging in diabetic mice, which may be related to the inhibition of AMPK/mTOR pathway related protein expression.关键词：AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway;diabetes;Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma (GNC) extract;high glucose;heart aging;p53;senescence
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发布时间：2021-02-09
Topic on Intervention of TCMs with Replenishing Qi and Activating Blood Circulation in Vascular Senescence Induced by Hyperglycemia
- 摘要：Pinelliae Rhizoma is a toxic traditional Chinese medicine, and its dose differs greatly between at ancient and modern times, so that it is difficult for researchers to determine its dosage during research and development of famous classical formulas. According to the literature research, it is found that the dose of Pinelliae Rhizoma in famous classical formulas has undergone a process from large to small. Based on the principle of " following the classic and conforming to pharmacopoeia" , and in combination with the modern actual situation, the reasonable dose of Pinelliae Rhizoma in the 17 formulas from the Catalogue of Ancient Famous Classical Formulas (The First Batch) is determined. According to the records of the original book, the evolution of past dynasties and the consideration of the current clinical application, dosage regimen and administration methods of the 17 famous classical formulas and so on, the authors suggested that the dosages of Pinelliae Rhizoma in Banxia Houpotang and Maimendong Tang are 15.0 g, its dosages in Wendantang and Sangbaipi Tang are 6.0 g, dosages of Pinelliae Rhizoma in Xuanfu Daizhe Tang, Zhuye Shigaotang, Banxia Xiexintang, Gancao Xiexintang, Huangliantang, Gualou Xiebai Banxiatang, Houpo Mahuangtang and Jinshui Liujunjian are 7.5 g, the dosages of Pinelliae Rhizoma in Zhurutang, Shengyang Yiweitang and Yangweitang are 6.7, 4.5, 5.0 g, respectively, dosages of Pinelliae Rhizoma in Banxia Baizhu Tianma Tang and Huopo Xialing Tang are 5.6 g.关键词：Pinelliae Rhizoma;dosage;historical evolution;famous classical formulas;dosage regimen;processing of traditional Chinese medicine;taking methods
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发布时间：2021-02-09 - 摘要：Objective:To observe the effect of modified Dihuang Yinzi on the learning and memory ability and on the neurons in CA1 area of hippocampus of rats suffering from vascular dementia.Method:The 84 male SD rats were randomly selected to form the sham operation group of 12 rats, and the other 72 rats were chosen for the vascular dementia model by means of Bivascular occlusion, and among which 60 were chosen randomly into 6 groups of 12 rats, namely, the model group, the nimodipine group (0.011 g·kg-1), and the high, medium and low dose modified Dihuang Yinzi (4.54, 2.27, 1.14 g·kg-1) respectively. After 30 days of continuous gavage, Morris water maze was used to detect the learning and memory ability of rats, hematoxylin eosin (HE) was used to observe the morphological changes of hippocampal CA1 neurons, transmission electron microscopy was used to observe the ultrastructural changes of hippocampal CA1 neurons, TUNEL was used to detect the apoptosis level of hippocampal CA1 neurons, immunohistochemical(IHC) was used to detect the expression level of phosphatidylinositol-3(PI3K), protein kinase B (Akt), Caspase-3 in hippocampal CA1 tissues.Result:Compared with sham operation group, the escape latency of the model group was significantly prolonged, the number of times of crossing the original platform was significantly reduced (P<0.01), the neuronal morphology of hippocampal CA1 area was damaged to varying degrees, the apoptosis rate was significantly increased (P<0.01), the integral optical density and average optical density of PI3K, Akt were significantly reduced (P<0.01), and the integral optical density and average optical density of Caspase-3 were significantly reduced (P<0.01). Compared with model group, the escape latency was shortened (P<0.05, P<0.01), the number of crossing the original platform was increased (P<0.05, P<0.01), the integrated optical density and average optical density of PI3K and Akt were significantly increased (P<0.01), and the integrated optical density and average optical density of Caspase-3 were decreased in different degrees (P<0.05, P<0.01).Conclusion:Modified Dihuang Yinzi can improve the learning and memory ability of vascular dementia model rats and the damaged neurons in the hippocampal CA1 area. The potential mechanism may be related to the activation of PI3K/Akt signal transduction pathway and the inhibition of apoptosis of neurons in hippocampal CA1 area of rats.关键词：modified Dihuang Yinzi;vascular dementia;neurons;apoptosis;phosphatidylinositol-3 kinase/protein kinase B
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发布时间：2021-02-09
Classic Prescriptions
- 摘要：Objective:To investigate the effect of ancient recipe Yanghetang and its drug-contained serum on apoptosis of triple negative breast cancer cell line MDA-MB-231 based on the signal pathway of early growth response gene 1 (Egr1)/cyclin dependent inhibitor (p21).Method:The drug-containing serum of Yanghetang was prepared from rats, and MDA-MB-231 cells were cultured in vitro. The blank control group was set, and MDA-MB-231 cells in Yanghetang high, middle, and low dose groups (23.4, 11.7, 5.85 g·kg-1) were intervened for 24, 48, 72 h, respectively. After that, the cell counting kit-8(CCK-8) method was used to detect the cell proliferation of each group. The blank control group was set, while MDA-MB-231 cells in Yanghetang high, middle, and low dose groups (23.4, 11.7, 5.85 g·kg-1) were treated for 48 h, and then flow cytometry was used to detect the apoptosis of each group and the distribution of cell cycle. The expression of Egr1 and p21 mRNA in each group was detected by quantificational Real-time polymerase chain reaction (Real-time PCR), while the expression of Egr1 and p21 protein in each group was detected by Western blot.Result:After MDA-MB-231 cells were intervened by Yanghetang for 24, 48, 72 h, MDA-MB-231 cell proliferation was significantly inhibited in Yanghetang high and middle dose groups as compared with the blank control group (P<0.01). After MDA-MB-231 cells were intervened by Yanghetang for 48 h, the apoptosis was significantly increased in Yanghetang high and middle dose groups as compared with the blank control group (P<0.05, P<0.01). In the Yanghetang high, middle dose groups, the proportion of cell cycle G0/G1 phase decreased, and the proportion of G2/M phase increased (P<0.05, P<0.01). The mRNA and protein expressions of Egr1, p21 were increased in Yanghetang high and middle groups (P<0.05, P<0.01).Conclusion:Yanghetang can activate Egr1/p21 signaling pathway in MDA-MB-231 cells, increase the expression of Egr1 and p21, and cause G2/M cell cycle arrest, thereby inducing apoptosis of MDA-MB-231 cells.关键词：triple negative breast cancer;MDA-MB-231 cells;Yanghetang;early growth response gene 1 (Egr1);cyclin dependent inhibitor (p21)
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发布时间：2021-02-09 - 摘要：Objective:To investigate the protective effect of salvianolic acid B on HepaRG hepatocyte injury induced by arsenic trioxide (As2O3 ) and its mechanism.Method:HepaRG cells were incubated with 5μmol·L-1 As2O3 for 24 h to induce hepatocyte injury. The cells were divided into control group, model group, salvianolic acid B 10 μmol·L-1 group, salvianolic acid B 10 μmol·L-1+ As2O3 group, salvianolic acid B 5 μmol·L-1+ As2O3 group, and salvianolic acid B 2.5 μmol·L-1+ As2O3 group. HepaRG cells were preincubated with salvianolic acid B for 2 h and then incubated with As2O3 for 24 h. At the end of the incubation, cell viability was detected by thiazolyl blue tetrazolium bromide assay, apoptosis was observed by Hoechst33342 fluorescence staining, apoptosis rate was detected by annexin V-FITC/propidium iodide double staining flow cytometry, and mitochondrial membrane was observed by JC-1 fluorescence staining. Western blot was used to detect the protective effect of expressions of relevant proteins Bcl-2, Bax, Akt, p-Akt on salvianolic acid B in the liver.Result:As2O3 concentration-dependently reduced the survival rate of HepaRG cells(P<0.01), salvianolic acid B had no effect on normal cell viability for 2 h, pre-incubation with salvianolic acid B(5, 10 μmol·L-1) for 2 h significantly increased the decreased cell survival rate caused by As2O3 (P<0.01). As2O3 significantly increased hepatocytes apoptosis rate(P<0.01), while pre-incubation with salvianolic acid B(10 μmol·L-1) deceased apoptosis rate(P<0.01). Incubation with As2O3 for 24 h caused decrease of mitochondrial membrane potential, pre-incubation with salvianolic acid B maintained mitochondrial membrane potential, indicating that the anti-apoptotic effect of salvianolic acid B were related to the mitochondrial pathway modulation. Western blot analysis showed that salvianolic acid B promoted the ratio of Bcl-2/Bax and promoted p-Akt/Akt compared with As2O3 group(P<0.01).Conclusion:Salvianolic acid B has a protective effect on hepatocyte injury induced by As2O3, and its mechanism is related to maintenance of mitochondrial function and inhibition of hepatocyte apoptosis.关键词：salvianolic acid B;arsenic trioxide;liver cell injury;liver protection
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发布时间：2021-02-09 - 摘要：Objective:To observe the effect of realgar nanoparticles (a representative drug in toxin eliminating therapeutics) targeting hypoxia-inducible factors (HIF), which act as effector molecules on metabolic reprogramming of lung cancer stem cells, and to explore the effect mechanism of lung cancer stem cells and metabolic reprogramming in the process of lung cancer metastasis, so as to verify the effectiveness of toxin eliminating therapeutics in the prevention and treatment of lung cancer metastasis.Method:Lung cancer A549 cells were cultured in vitro, and lung cancer stem cells were then identified and selected. The stem cells were divided into blank control group, cisplatin group (5 mg·L-1), realgar nanoparticles low, medium and high dose groups (100, 200, 400 mg·L-1). After intervention, glucose oxidase method was used to detect the effect of realgar nanoparticles on the glucose metabolism of lung cancer stem cells, real-time polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of hypoxia-inducible factors-1α (HIF-1α), C-myc and p53, while Western blot was used to detect the expression of related proteins HIF-1α, phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and mammalian target of rapamycin (mTOR), and enzyme linked immunosorbent assay (ELISA) was used to detect the glucose transporter 1 (GLUT1), pyruvate dehydrogenase kinase 1 (PDK1), pyruvate kinase M (PKM), phosphofructokinase(PFK), pyruvate dehydrogenase (PDH) and lactic dehydrogenase (LDH) expression.Result:As compared with the blank control group, realgar nanoparticles can reduce the glucose consumption of lung cancer stem cells, and the glucose consumption was reduced with the increase of dose in a time-and dose-dependent manner (P<0.01). Realgar nanoparticles can inhibit the mRNA expression of HIF-1α, a key factor in metabolic reprogramming of lung cancer stem cells (P<0.05, P<0.01), down-regulated C-myc mRNA and up-regulated the p53 mRNA expression (P<0.05, P<0.01), down-regulated protein expressions of PI3K, Akt, mTOR(P<0.05, P<0.01), and inhibited the expression of related enzymes GLUT1, PDK1, PFK, PKM, PDH, and LDH levels (P<0.05, P<0.01). With the increase of dose, the regulation and control ability of realgar nanoparticles gradually increased.Conclusion:Toxin eliminating therapeutics can drive the metabolic reprogramming of lung cancer stem cells by targeting HIF effector molecule, and then inhibit the invasion and metastasis of lung cancer.关键词：lung cancer;stem cells;metabolic reprogramming;toxin eliminating therapeutics;realgar nanoparticles;hypoxia-inducible factors
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发布时间：2021-02-09 - 摘要：Objective:To observe the effect of Bimin decoction(BMD) on nuclear factor-kappa B(NF-κB) signaling pathway and aquaporin 5(AQP5) expression in allergic rhinitis (AR) rats with lung and spleen Qi deficiency syndrome(LSQDS), in order to study the mechanism in treating AR.Method:Fifty-six SD rats were randomly divided into seven groups: control group, AR group, LSQDS AR group, BMD low dose two weeks group and four weeks group, BMD high dose two weeks group and four weeks group. The control group did not intervened, the AR group established the AR disease model with ovalbumin (OVA) as the allergen, the other five groups established the LSDQS model with smoke and senna gavage, and also established the AR disease model with OVA sensitization at the same time as the AR group. After the model was established successfully, four BMD intervention groups were separately given low dose BMD (11.3 g·kg-1) for 2 weeks and 4 weeks, and high dose BMD (22.6 g·kg-1) for 2 weeks and 4 weeks. To observe the general situation of the rats, hematoxylin eosin (HE) staining was used to observe the pathological changes of the nasal mucosa, immunohistochemistry and Western blot were used to detect the expression levels of NF-κB and AQP5, real-time fluorescent quantitative polymerase chain reaction technique (Real-time PCR)was used to detect mRNA levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6).Result:The typical AR symptoms were found in AR rats, the AR symptoms and lung and spleen Qi deficiency symptoms were found in AR rats with LSQDS at the same time, and the AR symptoms and lung and spleen Qi deficiency symptoms were significantly improved after the intervention of BMD. Compared with the control group, the typical histopathological changes of nasal mucosa were found in AR group and LSQDS AR group, with a higher behavioral score (P<0.05), and the expression of NF-κB and AQP5 protein increased (P<0.05), the expression of IL-1β, TNF-α, IL-6 and AQP5 mRNA increased (P<0.05). Compared with AR group, the pathological changes of nasal mucosa in LSQDS AR group were more serious, and the expression of NF-κB protein in nucleus increased (P<0.05), the expression of TNF-α and AQP5 mRNA increased (P<0.05). Compared with LSQDS AR group, the pathological changes of nasal mucosa in the groups which interfered by BMD were improved, and the expression of NF-κB and AQP5 protein decreased (P<0.05), the expression of IL-1β, TNF-α, IL-6 and AQP5 mRNA decreased (P<0.05). Compared with BMD low-dose two-week group, the expression of NF-κB protein in nucleus decreased (P<0.05) in BMD high dose four week group.Conclusion:Compared with AR group, the AR condition of the rats with LSQDS is more serious under the same allergen stimulation, BMD can treat AR and reduce the over secretion of glands, which may be related to inhibit the expression of AQP5 by inhibiting the NF-κB signaling pathway.关键词：lung and spleen Qi deficiency syndrome;allergic rhinitis;Bimin decoction;nuclear factor-kappa B;aquaporin 5
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发布时间：2021-02-09 - 摘要：Objective:To investigate the role and mechanism of Testudinis Carapax et Plastrum aqueous extract in promoting osteogenic differentiation of mouse preosteoblast cell line(MC3T3-E1) by regulating nuclear transcription factor-κB(NF-κB) inflammation microenvironment.Method:MC3T3-E1 cells were cultured in vitro, and osteogenic induction (OI) was performed. Testudinis Carapax et Plastrum was prepared and treated the cells. Cells were devided into control group, osteogenic induction group and Testudinis Carapax et Plastrum (20 mg·L-1)with osteogenic induction group. The proliferation of MC3T3-E1 was detected by cell counting kit-8 (CCK-8), and the optimum concentration of intervention was determined. MC3T3-E1 differentiation and osteogenic mineralization were assayed using alkaline phosphatase (ALP) and Alizarin red staining (ARS), respectively. The expressions of NF-κB p65, NF-κB p105, interleukin-6(IL-6), ALP and Collagen-Ⅰ(COL-Ⅰ) mRNA were detected by Real-time PCR.Result:The results of CCK-8 showed that the proliferation of MC3T3-E1 did not change statistically with time, but it showed an upward trend, while the proliferation at 20 mg·L-1 was more obvious than other groups. The ALP and ARS showed that the positive staining rate of osteogenic induction group and Testudinis Carapax et Plastrum with osteogenic induction group were higher than control group.Real-time PCR results showed that on the 7th day in culture, the expression of NF-κB p105 and IL-6 mRNA in Testudinis Carapax et Plastrum with osteogenic induction group was significantly lower than that in control group (P<0.01), and the expression of ALP and COL-Ⅰ mRNA was significantly upregulated(P<0.05), on the 14th day, the expression of NF-κB p65, NF-κB p105 and IL-6 mRNA in Testudinis Carapax et Plastrum with osteogenic induction group was significantly lower than that in control group (P<0.01). The expression of ALP and COL-Ⅰ mRNA was significantly increased (P<0.05, P<0.01).Conclusion:Testudinis Carapax et Plastrum aqueous extract can promote osteogenic differentiation of MC3T3-E1 via a mechanism associated with the regulation of inhibition of NF-κB inflammatory microenvironment.关键词：Testudinis Carapax et Plastrum aqueous extract;mouse preosteoblast cell line(MC3T3-E1);osteogenic induction;nuclear transcription factor-κB;inflammatory microenvironment
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发布时间：2021-02-09
Pharmacology
- 摘要：Objective:To explore the effect of modified Qingyitang combined with continuous blood purification in the adjuvant treatment of severe acute pancreatitis (SAP) complicated with multiple organ dysfunction (MODS) caused by heat accumulation of viscera.Method:Totally 100 cases of patients of SAP complicated with MODS, who were diagnosed as heat accumulation of viscera by traditional Chinese medicine(TCM) and treated in ICU of the First Affiliated Hospital of Hunan University of Chinese Medicine during May 2015 and May 2019, were randomly divided into two groups, namely control group and observation group, with 50 cases in each group. The patients in control group were treated with fasting and abstinence, gastrointestinal decompression, inhibition of trypsin secretion, gastric mucosal protection, early jejunal nutrition, reduction of inflammatory reaction, continuous blood purification (CBP), mechanical ventilation and circulatory support. The patients in observation group were treated by nasojejunal tube according to syndrome differentiation in addition to routine comprehensive therapy. Modified Qingyitang was injected for 7 days. The remission time of abdominal pain and distention, the time of first exhaust and defecation, the time of ICU residence, the number of samples falling off, the cause of death and the number of cases were recorded. Relevant indexes were measured before treatment, on the 3rd and 7th day of treatment, including the evaluation indexes of pancreatitis: blood amylase (AMS), blood lipase (LPS), and modified computed tomography severity index (MCTSI), inflammatory response indexes were interleukin-6 (IL-6) and hypersensitive C-reactive protein (hs-CRP). Organ function indexes included APACHE-Ⅱ, arterial partial pressure of oxygen (PaO2), oxygenation index (PaO2/FiO2), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), glutamyltransferase (γ-GGT), urine volume, creatinine (CREA), urea nitrogen (UREA), glomerular filtration rate (GFR), creatine kinase (CK), creatine kinase isoenzymes (CKMB), lactate dehydrogenase (LDH), myoglobin (Mb). Tissue perfusion evaluation indexes included acute physiology and chronic health score, serum lactic acid (Lac) and central venous pressure (CVP). TCM treatment score was based on the syndrome score of acute pancreatitis with heat accumulation of viscera syndrome.Result:The total effective rate of TCM syndromes was 86.67%(39/45) in observation group and 73.91%(34/46) in control group (χ2 =13.524, P<0.01). On the 7th day of treatment, the symptoms and indicators of the two groups were improved. Compared with before treatment, AMS, LPS, IL-6, hs-CRP, MCTSI, APACHE-Ⅱ, Lac, CVP, PaO2, PaO2/FiO2, ALT and AST were improved on the 3rd and 7th day after treatment in observation group and control group. The levels of AMS, LPS, IL-6, hs-CRP, MCTSI, APACHE-Ⅱ, Lac, CVP, PaO2, PaO2/FiO2, ALT, AST, ALP, γ-GGT, urine volume were significantly improved (P<0.05). Compared with control group on the 3rd and 7th day, the levels of AMS, LPS, IL-6, hs-CRP, MCTSI, APACHE-Ⅱ, Lac, CVP, PaO2, PaO2/FiO2, ALT, AST, ALP, γ-GGT, urine volume were significantly improved (P<0.05). CREA, UREA, GFR, CK, CKMB, LDH and Mb were significantly improved (P<0.05). Compared with control group, the abdominal pain, abdominal distension relief time, first exhaust/defecation time, ICU stay time in observation group were significantly shortened (P<0.05), and the mortality rate in observation group was significantly reduced (P<0.05).Conclusion:Patients of SAP accompanied with MODS can be treated with blood purification combined with modified Qingyitang by promoting pancreas repair, inhibiting inflammation and improving organ function. It plays an important role in improving symptoms, alleviating TCM syndromes, delaying progression of disease, reducing hospital stay and reducing mortality.关键词：severe acute pancreatitis;multiple organ dysfunction (MODS);blood purification;heat accumulation of viscera;modified Qingyitang
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发布时间：2021-02-09 - 摘要：Objective:To observe the clinical efficacy of Gandouling decoction on hepatic function of patients with phlegm and blood stasis type Wilson's disease.Method:From January 2015 to December 2017, totally 72 cases of phlegm and blood stasis type Wilson's disease admitted to Encephalopathy Center in the First Affiliated Hospital of Anhui University of Chinese Medicine were randomly divided into treatment group and control group, with 36 cases in each group. Patients in both groups were injected with sodium dimercaptopropane sulfonate for routine treatment. At the same time, patients in control group received Hugan Tablets, and patients in treatment group received Gandouling decoction for a total of 6 treatment courses. Before and after treatment, traditional Chinese medicine(TCM)syndrome effective rate, serum enzyme index [alanine aminotransferase(ALT), aspartate aminotransferase (AST), alkaline phosphatase(AKP)], bilirubin metabolism index [total bilirubin(TBIL)], liver fibrosis index [laminin(LN), hyaluronic acid(HA), collagen type IV(CⅣ), procollagen type Ⅲ peptide(PⅢP)]and blood coagulation index [fibrinogen (FBG), prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) ]were observed.Result:TCM syndrome effective rates of treatment group and control group were 86.11%(31/36) and 63.98%(23/36) respectively, with a significant difference between two groups (P<0.05). ALT, AST, AKP, TBIL decreased in two groups after treatment (P<0.01), and the effects of ALT, AST, AKP, TBIL in treatment group were significantly better than those in control group (P<0.01), liver fibrosis index decreased in both groups after treatment (P<0.01), and the effect in treatment group was significantly better than that in control group (P<0.01). Blood coagulation indexes were improved to different degrees in both groups (P<0.05, P<0.01), and there were significant differences between treatment group and control group in decreasing PT, APTT, TT levels (P<0.05, P<0.01) and increasing FBG level (P<0.01).Conclusion:Gandouling decoction can significantly improve hepatic function of patients with phlegm and blood stasis type Wilson's disease, which is manifested in improving serum enzymes and bilirubin indexes, reversing liver fibrosis, promoting clotting factors and reducing bleeding tendency, in order to delay the progress of the disease and improve the life quality of patients.关键词：Gandouling decoction;Wilson's disease;phlegm and blood stasis type;sodium dimercaptopropane sulfonate;liver function
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发布时间：2021-02-09 - 摘要：Objective:To explore the clinical efficacy of Tongqiao Huoxuetang plus acupuncture on post-stroke cognitive impairment (PSCI) and its effect on neurotransmitters and neurological function of patients.Method:Totally 122 PSCI patients admitted in our hospital from January 2017 to March 2019 were randomly divided into observation group and control group by simple random number table method, with 61 patients in each group. Both groups were given oral donepezil hydrochloride tablets. The control group was given acupuncture treatment, while the observation group was given Tongqiao Huoxuetang plus acupuncture in addition to therapy of control group. All patients were treated continuously for 4 weeks. The clinical efficacy was compared between two groups. Before and after treatment, the general cognitive function of two groups was tested by mini-mental state examination (MMSE), changes of their neurologic impairment and daily living ability were evaluated by National institutes of health stroke scale (NIHSS) and Barthel index (BI) respectively, and euzymelinked immunosorbent assay was adopted to test the levels of neurotransmitters [acetyl choline (ACh), dopamine (DA), noradrenaline (NE), 5-hydroxytryptamine (5-HT)] and neurologic impairment markers [neuronspecific enolase (NSE), visinin-like protein-1 (VILIP-1), myelin basic protein (MBP)]. And the occurrence of adverse reactions was recorded.Result:The overall effective rate of the observation group was 91.8% (56/61), which was much higher than 78.7% (48/61) of control group (P<0.05). After treatment, both groups saw much higher MMSE and BI scores but lower NIHHS scores than before treatment (P<0.05), and above indicators in observation group were improved more significantly than those in control group over the same period after treatment (P<0.05). The levels of serum ACh, DA, NE and 5-HT after treatment were obviously higher than those before treatment (P<0.05), and observation group had significantly higher ACh, DA, NE and 5-HT levels than control group over the same period (P<0.05). After treatment, both groups witnessed a great decrease in NSE, VILIP-1, MBP (P<0.05), while the observation group saw dramatically lower NSE, VILIP-1, MBP levels than control group over the same period after treatment (P<0.05). Neither of two groups saw serious adverse reactions.Conclusion:In treating PSCI, Tongqiao Huoxuetang plus acupuncture can significantly improve neurological function of the patients, which may be due to increase of neurotransmitters. Besides, Tongqiao Huoxuetang plus acupuncture is proved to be generally effective in promoting the recovery of patients' neurological function and enhancing their abilities in daily life, with a good tolerance among patients.关键词：post-stroke cognitive impairment;Tongqiao Huoxuetang;acupuncture;neurotransmitter;neurological function
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发布时间：2021-02-09 - 摘要：Objective:To discuss the effect of Feiyiliu mixture as an adjuvant method for chemotherapy and immune support on progression-free survival (PFS) and survival rate of patients with Ⅲb and Ⅳ stage squamous cell carcinoma of lung (SQCC), and to investigate its intervention effect on tumor markers and cytokines in peripheral blood.Method:One hundred and thirty-two patients were randomly divided into control group (66 cases) and observation group (66 cases) by random number table. Patients in control group received programme of gemcitabine hydrochloride combined with cisplatin (alternative paclitaxel combined with cisplatin) and they also got thymopolypeptides for injection, while the patients in observation group received Feiyiliu mixture, with a treatment course of 4 cycles in both groups. PFS and 12 months' survival rate were recorded during follow-up. Scores of European quality of life collaboration cancer core scale (EORTC QLQ-C30) were graded before and after treatment. Levels of CD3+, CD4+, CD8+, CD4+ /CD8+, cytokeratin 19 fragment 21-1 (CYFRA21-1), squamous cell carcinoma antigen (SCC), carbohydrate antigen 125 (CA125), vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and MMP-9 were detected, and size of solid tumor was evaluated before and after treatment.Result:Clinical remission rate of solid tumor was (35/66)53.03% in observation group, higher than(23/66) 34.85% in control group (χ2=4.286, P<0.05). PFS in observation group was 9.8 months (95% CI, 4.58-22.37), more than 5.7 months in control group (95% CI, 4.69-20.41) (P<0.05). 12 months’ survival rate was 45.45% (30/36) in observation group, more than 27.27% (18/48) in control group (χ2=4.714, P<0.05). Scores of physical function, social function and overall quality of life in observation group were higher than those in control group (P<0.01), and scores of fatigue, nausea and vomiting, shortness of breath, loss of appetite, constipation and diarrhea were lower than those in control group (P<0.01). Levels of CD3+, CD4+ and CD4+ /CD8+ in observation group were higher than those in control group (P<0.05), while levels of CD8+, CYFRA21-1, SCC, CA125, VEGF, MMP-2 and MMP-9 were lower than those in control group (P<0.01).Conclusion:Adjuvant Feiyiliu mixture combined with chemotherapy and immune support can prolong PFS, improve survival rate, stabilize immune function, improve quality of life, inhibit the expression of cytokines such as CYFRA21-1 and vascular endothelial growth factor, and improve prognosis for patients with Ⅲb and Ⅳ stage SQCC. Therefore, it is worth further study and use关键词：non-small cell lung cancer;squamous cell carcinoma;Feiyiliu mixture;progressive survival;survival rate;quality of life;tumor markers;cytokines
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发布时间：2021-02-09 - 摘要：Objective:To observe the clinical efficacy of modified Buyang Huanwu Tang on patients with lumbar disc herniation (LDH) after percutaneous foramen endoscopy by the collateral disease theory, and its effect on inflammatory factors and pain-inducing mediators.Method:One hundred and forty-four patients were randomly divided into control group (72 cases) and observation group (72 cases) by random number table. Both groups’ patients were treated with percutaneous endoscopic lumbar discectomy(PELD), and symptomatic treatment dehydration and anti-inflammation were also given to those patients after the operation. Patients in observation group got modified Buyang Huanwu Tang, 1 dose/day. The course of treatment was 4 weeks, and a 12-week fellow-up was recorded. Before the operation and at the 3rd day, the 4th and 16th week after the operation, scores of visual analogue score of pain degree (VAS) were recorded. And before the operation and at the 1st, 4th and 16th week after the operation, scores of dysfunction index (ODI) of Oswestry were recorded. Failed back surgery syndrome of LDH was recorded during 16 weeks after the operation. And Japanese orthopaedic association (JOA) and Qi deficiency and blood stasis syndrome were scored. And the levels of prostaglandin E2 (PGE2), thromboxane B2 (TXB2), interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and serotonin (5-HT) were all detected, and the effect was assessed by improved Macnab.Result:At the 1st day and the 1st, 4th and 16th week after treatment, scores of VAS were all lower than those in control group (P<0.01). And at the 1st, 4th and 16th week after treatment, scores of ODI were lower than those in control group (P<0.01). The rate of incidence was 18.06%(13/72), which was lower than 37.5%(27/72) in control group (χ2=6.784, P<0.01). Scores of the total JOA and subjective symptoms, objective signs and daily activities were all higher than those in control group (P<0.01). And scores of symptom scores and total scores of deficiency and blood stasis syndrome were all lower than those in control group (P<0.01). After treatment, levels of TNF-α, IL-1β, PGE2, TXB2 and 5-HT were lower than those in control group (P<0.01). According to the rank sum test, the effect of modified Macnab was better than that in control group (Z=2.151, P<0.05).Conclusion:Based on Luobing theory, modified Buyang Huanwu Tang can alleviate pain and other symptoms, promote the recovery of lumbar vertebral function, inhibit the expressions of inflammatory factors and pain-causing mediators, alleviate the residual symptoms after recent operation, reduce the incidence of FBSS, promote the rehabilitation of patients after operation, and improve the efficacy.关键词：lumbar disc herniation;endoscopic discectomy;collateral pathology theory;Buyang Huanwu Tang;inflammatory factors;painful mediators
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发布时间：2021-02-09
Clinic
- 摘要：Objective:To explore the effect of Dioscoreae Nipponicae Rhizoma extract (DNRe) on rats with acute gouty arthritis (AGA) based on urine metabolomics and to search for the related potential biomarkers and metabolic pathways.Method:Rat model of AGA induced by monosodium urate (MSU) was selected, 40 rats were randomly divided into the blank group (k), the DNRe group (g), the model group (m), and the DNRe treatment group (gm), with 10 rats in each group. The drug-administered group was administered with DNRe at a dose of 0.48 g·kg-1 once a day for 5 days. The urine was gathered after the last administration, and analyzed with UPLC-Q-TOF/MS coupled with pattern recognition techniques, electrospray ionization (ESI) under positive and negative ion scanning mode was adopted, data collection range was m/z 100-1 500 with full scanning mode.Result:A total of 12 common potential biomarkers were identified as sarcosine, dimethylglycine, deoxycytidine, uric acid, 5-hydroxytryptamine (5-HT), L-cystathionine, 4-pyridoxic acid, deoxyuridine, melatonin, 5-methoxytryptamine, fumaric acid and cytidine. Compared with the blank group, the 12 potential biomarkers in the DNRe group were significantly down-regulated. Compare with the model group, 10 metabolites were up-regulated and 2 metabolites were down-regulated in the 12 potential biomarkers of the DNRe treatment group, the abnormal expression of 10 markers including sarcosine, uric acid, L-cystathionine, 4-pyridoxic acid, deoxyuridine, 5-methoxytryptamine, cytidine, dimethylglycine, melatonin, fumaric acid could be modulated by DNRe. The strongest metabolic pathways associated with AGA were cysteine and methionine metabolism, and tryptophan metabolism.Conclusion:The effect of DNRe on AGA may be related to the promotion of conversion level from cystathionine to cysteine in the cysteine and methionine metabolism, and the up-regulating melatonin level in tryptophan metabolism.关键词：Dioscoreae Nipponicae Rhizoma extract (DNRe);monosodium urate;acute gouty arthritis;urine metabolomics;cysteine and methionine metabolism;tryptophan metabolism;L-cystathionine
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发布时间：2021-02-09 - 摘要：Objective:To establish differential metabolites between different varieties of Angelica sinensis, and provide reference for breeding, introduction, regional cultivation and ecological cultivation of new varieties of A. sinensis.Method:Comprehensive non-target metabonomics analysis was conducted for five new varieties of A. sinensis collected at the same time from the same origin: Mingui No. 1 (MG1), Mingui No. 2 (MG2), Mingui No. 4 (MG4), Mingui No. 5 (MG5), and Mingui No. 6 (MG6). The 50% methanol extract of each variety was taken, and then the differential metabolites among varieties were found by using ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS), software Progenesis QI, principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and non-targeted metabonomics analysis. Differential metabolites were identified based on precise molecular weight, secondary fragments, KEGG database, HMDE database and related literature information.Result:The results of UPLC-Q-TOF-MS and multivariate statistical analysis showed that there were significant differences in the metabolites of five Angelica varieties. As compared with MG1, the contents of chlorogenic acid, ferulic acid, tryptophan and ferulic aldehyde were significantly lower in MG2, MG4, MG5 and MG6, while the contents of ligustilide, coumarin, bovine keratin, palmitin, protocatechualdehyde and linolenic acid were significantly higher (P<0.05). The results of multivariate statistical analysis showed that the metabolites of MG2 and MG5 were similar with those of MG6, but were significantly different from those of MG4.In addition, 38 distinct metabolites were identified, involving 7 potential targeted metabolic pathways. Different varieties of A. sinensis could regulate the synthesis of their metabolites through phenylpropanoid biosynthesis and sesquiterpene-like compounds metabolism, lipid metabolism, amino acid metabolism, carbohydrate metabolism, nucleotide metabolism, carotenoids, linolenic acid, linoleic acid and some other metabolic pathways.Conclusion:UPLC-Q-TOF-MS and Progenesis QI metabonomics techniques were used to compare the chemical constituents of different varieties of A. sinensis from the overall level. The differences and their regularities were found, which could provide reference for quality control, variety sorting, identification, breeding and ecological planting of A. sinensis.关键词：Angelica sinensis;variety;UPLC-Q-TOF-MS;metabolites;chlorogenic acid;ferulic acid;tryptophan
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发布时间：2021-02-09 - 摘要：Objective:To investigate the processing purpose of Morindae Officinalis Radix (MO), Euodiae Fructus (EF) and Polygalae Radix (PR) processed by Glycyrrhizae Radix et Rhizoma (Gly).Method:The content of dapsone in rat plasma was determined by high performance liquid chromatography (HPLC), the mobile phase was acetonitrile (A)-water (B) for gradient elution (0-5 min, 10%-25%A; 5-20 min, 25%A) and detection wavelength was set at 292 nm. PK Solution 2.0 software was used to simulate pharmacokinetic parameters.Result:Within 300 min after dapsone was administrated, compared with the control (CTL) group, the elimination of dapsone was slowed down and its plasma concentration was increased in the unprocessed product of MO (UMO) group. The elimination of dapsone was accelerated and its peak concentration (Cmax) was decreased in the processed products of MO with Gly (GMO) groups, and they had positive correlation with proportion of Gly in GMO. Compared with the CTL group, the elimination of dapsone was slowed down, and its plasma concentration was increased and its peak time (Tmax) was postponed in the unprocessed product of EF (UEF) group, while their Cmax and Tmax were changed in the processed products of EF with Gly (GEF) groups. Compared with the CTL group, the elimination of dapsone was slowed down and its plasma concentration was increased in the unprocessed product of PR (UPR) group, while the elimination was accelerated and its plasma concentration was decreased in the processed products of PR with Gly (GPR) groups.Conclusion:The elimination of dapsone is slowed down in rats administered with UMO, UEF and UPR, while its elimination is accelerated in rats administered with the processed products of these three herbs with different proportions of Gly. Among the proportions, effect of processed products of these three herbs with 100∶6 (ratio of unprocessed product-Gly) on pharmacokinetics of dapsone is not significant.关键词：Morindae Officinalis Radix;Euodiae Fructus;Polygalae Radix;Glycyrrhizae Radix et Rhizoma;processing;cytochrome P450 enzyme;dapsone
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发布时间：2021-02-09
Drug Metabolism
- 摘要：Objective:Because traditional methods are difficult to identify the fermentation mycelium, DNA barcoding technology was used to quickly identify the raw material strain Paecilomyces hepiali of Jinshuibao capsules and related products.Method:A total of 168 samples of 8 species of P. hepiali and its confusable species were identified by internal transcribed spacer (ITS) sequences, and based on the ITS sequences, P. hepiali specific primers were designed to quickly identify the related products.Result:The length of ITS sequences in 44 P. hepiali samples from different sources was 499 bp and there was no mutation site. It was shown that P. hepiali could be distinguished from 7 confusable species based on ITS sequences. The specific primer (ITS-BF/ITS-BR) of P. hepiali designed by ITS sequences could be amplified to obtain a short fragment of 102 bp in length, which could be used to rapidly identify P. hepiali from other confusable species, and to distinguish relevant products in the market.Conclusion:The rapid identification of P. hepiali and its related products can be achieved through the ITS sequences and specific primers, which provides a reference for the production and quality control of Jinshuibao capsules.关键词：Paecilomyces hepiali;DNA barcoding;internal transcribed spacer (ITS);specific primers;Cordyceps sinensis;Jinshuibao capsules;fermentation
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发布时间：2021-02-09 - 摘要：Objective:To investigate the effects of black bean juice with different stewing times on the appearance character and the content changes of effective components of Polygoni Multiflori Radix Praeparata.Method:HPLC was employed with Agilent ZORBAX Extend-C18 column (4.6 mm×250 mm, 5 μm), a gradient mobile phase of methanol (A)-water (B) was eluted (0-30 min, 5%-100%A; 30-40 min, 100%A), the flow rate was 1.0 mL·min-1, the injection volume was 10 μL and the column temperature was 35 ℃, detection wavelength was set at 280 nm. The contents of stilbene glycoside, emodin, emodin methyl ether, emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside in samples prepared at different processing times were simultaneously determined by HPLC.Result:The content of stilbene glycoside decreased gradually with the increase of stewing time, compared with 8 h, its content decreased by 76% at 64 h. The contents of emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside increased first, and then decreased, reaching the highest value at 24 h, and then decreased to the level similar to the content of 8 h after 40 h, and then fluctuated slightly. The contents of emodin and emodin methyl ether increased first, and then decreased, reached the maximum when stewed for 32 h, then decreased slowly and tended to be stable.Conclusion:The stewing time has significant influence on the content of various components in Polygoni Multiflori Radix Praeparata, and the changing trend is different, the processing time of Polygoni Multiflori Radix Praeparata shall be standardized. At the same time, it is not sufficient to take stilbene glycoside and anthraquinones as the indicator ingredients for this decoction pieces, the quality control indicators such as polysaccharides shall be considered.关键词：Polygoni Multiflori Radix;processing method;black bean juice;stew;stilbene glycoside;free anthraquinones;combined anthraquinones
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发布时间：2021-02-09 - 摘要：Objective:To study the chemical structure and morphological features of a polysaccharide BHP-1 isolated and purified from the bulbs of Lilium davidii var. unicolor.Method:Thermogravimetry (TG) and scanning electron microscopy (SEC) was used to determine the thermal characteristics and the surface morphology changes of the polysaccharide BHP-1, respectively. The chemical structure of BHP-1 was analyzed by gas chromatography-mass spectrometer (GC-MS), partial acid hydrolysis, sodium periodate oxidation-Smith degradation and nuclear magnetic resonance (NMR).Result:The results showed that the backbone of the polysaccharide BHP-1 mainly contained α-(1→4)-linked D-glucopyranosyl and β-(1→4)-linked D-mannopyanosyl, and the branches were probably linked at the O-2 and/or O-3 of the mannosyl and glucosyl residues, with T-α-D-glucopyranosyl as a terminal structure. It was a natural mannoglucose containing a small amount of O-acetyl group. BHP-1 began to degrade at 220 °C, and the degradation ended at 520 ℃, indicating a good thermal stability, smooth surface and a large number of depressions. The depressions were formed by closely interlaced and sunken sheet layers with irregular holes.Conclusion:BHP-1 was a natural mannoglucose with a good thermal stability, smooth surface, a large number of depressions and irregular holes, and contained a small amount of O-acetyl group. Its chemical structure was reported for the first time.关键词：Lilium davidii var. unicolor;polysaccharide;structural characterization;thermogravimetric and topographic analysis;mannoglucose
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发布时间：2021-02-09 - 摘要：Objective:To investigate the differences and correlations of active ingredients and principal ingredients of steroid saponins in rhizomes of 9 kinds of Paris.Method:The differences and correlations of active ingredients and principal ingredients of steroid saponins in rhizomes of 9 kinds of Paris. were qualitatively analyzed by liquid chromatography-mass spectrometry (LC-MS).Result:PolyphyllinⅠ, polyphyllin Ⅱ, polyphyllin Ⅵ, and polyphyllin Ⅶ, the quality control indexes of medicinal materials in Chinese Pharmacopoeia (2015 edition) were detected in rhizomes of 8 species, namely P. polyphylla var. yunnanensis, P. delavayi, P. mairei, P. polyphylla var. yunnanensis(kuoban), P. veitnamensis, P. axialis, P. thibetica, and P. polyphylla var. polyphylla, polyphyllin Ⅴ was also detected in the study. There were significant differences in active ingredients of polyphyllin Ⅲ (Dioscin), polyphyllin H and gracillin. Meanwhile, P. polyphylla var. yunnanensis (S1), P. delavayi (S2), P. mairei (S3), P. polyphylla var. yunnanensis (kuoban) (S4), P. veitnamensis (S5), P. axialis (S6) and P. polyphylla var. polyphylla (S9) could be classified into one category according to the principal component analysis (PCA) model of the LC-MS data under the positive ion mode of Paris. P. thibetica (S7) and P. forrestii (S8) were clustered as two separate categories, with a longest range from P. polyphylla var. yunnanensi.Conclusion:There are little differences in active ingredients and the principal ingredients of steroid saponins in rhizome of P. polyphylla var. yunnanensis, P. delavayi, P. mairei, P. polyphylla var. yunnanensis, P. veitnamensis, P. axialis and P. polyphylla var. polyphylla. However, further research is required to determine whether these could substitute P. polyphylla var. yunnanensis. as an alternative medicine.关键词：Paris;rhizome;liquid chromatography-mass spectrometry;steroid saponins
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发布时间：2021-02-09 - 摘要：Objective:To determine the chloroplast genomes of Liriope spicata var. prolifera, Ophiopogon japonicus in Sichuan and Zhejiang, analyze their sequence characteristics and complete the screening of specific DNA barcodes.Method:The chloroplast genomes of L. spicata var. prolifera, O. japonicus in Sichuan and Zhejiang were sequenced, spliced and annotated through high-throughtput sequencing technology, and the structural characteristics and phylogenetic relationships of chloroplast genomes were analyzed by bioinformatics.Result:The total length of chloroplast genome of L. spicata var. prolifera was 155 998 bp, the total content of guanine and cytosine (GC) was 37.7%, and 85 protein-coding genes, 37 transfer RNA (tRNA) genes and 8 ribosomal RNA (rRNA) genes were successfully annotated, a total of 274 simple sequence repeats (SSRs) were detected, the number of codons encoding leucine was the most, while the number of codons encoding tryptophan was the least. The total length of chloroplast genome of O. japonicus in Sichuan province was 156 078 bp, the total content of GC was 37.8%, and 85 protein-coding genes, 37 tRNA genes and 8 rRNA genes were successfully annotated, a total of 265 SSRs were detected, the number of codons encoding leucine was the most, while the number of codons encoding tryptophan was the least. The total length of chloroplast genome of O. japonicus in Zhejiang province was 156 207 bp, the total content of GC was 37.7%, and 85 protein-coding genes, 37 tRNA genes and 8 rRNA genes were successfully annotated, a total of 274 SSRs were detected with the highest number of codons encoding leucine and the lowest number of codons encoding tryptophan.Conclusion:The phylogenetic trees show that compared with O. japonicus in Sichuan province, L. spicata var. prolifera is more closely related to O. japonicus in Zhejiang province. The variation of non-coding regions of L. spicata var. prolifera, O. japonicus in Zhejiang and Sichuan is greater than that in the coding region. The entire chloroplast genome can be used as a super barcode for identifying species of Ophiopogon and Liriope.关键词：chloroplast;Liriope spicata var. prolifera;Ophiopogon japonicus in Zhejiang province;O. japonicus in Sichuan province;chloroplast genome;molecular identification;chloroplast transformation
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发布时间：2021-02-09 - 摘要：Objective:To study the spectrum-effect relationship between HPLC fingerprints and anti-hepatoma activity of Paris polyphylla var. yunnanensis, P. forrestii and P. vietnamensis, and to elucidate its effective substance.Method:HPLC was used to establish the fingerprint of three extracts from the plant. The mobile phase was consisted of acetonitrile (A)-water (B) for gradient elution (0-10 min, 20%A; 10-20 min, 20%-25%A; 20-30 min, 25%-30%A; 30-40 min, 30%-35%A; 40-50 min, 35%-40%A; 50-60 min, 40%A; 60-75 min, 40%-45%A; 75-80 min, 45%-60%A), and the flow rate was 0.9 mL·min-1. The UV detection wavelength was 203 nm. Thiazolyl blue tetrazolium bromide (MTT) array was used to detect the inhibitory effects of three extracts on the proliferation of HepG2 cells, and the half inhibitory concentration (IC50) was calculated. Cluster analysis and grey relational analysis were used to analyze the data of spectrum and efficacy, and to find out the components that contributed a lot to the anti-liver cancer effect.Result:A total of 11 common peaks were identified as common peaks among HPLC fingerprints of three kinds of Paris. After treated 72 h, P. forrestii has the highest inhibitory effect on the HepG2 cells, the IC50 of P. forrestii, P. polyphylla var. yunnanensis and P. vietnamensis were 148.33, 178.87, 208.09 mg·L-1, respectively. According to the grey relational analysis, the common peaks 1-10 from P. polyphylla var. yunnanensis had great correlation to anti-tumor effect, and the common peaks 1-7 for P. forrestii, the common peaks 1-4, 6-10, N1 for P. vietnamensis, all the correlation degrees with IC50 were >0.7.Cluster analysis of variables in each Paris showed that peaks with correlation degree >0.7 could cluster with IC50.Conclusion:The established HPLC fingerprint method is reliable with good reproducibility. The peaks 1-4, 6 and 7 from three kinds of Paris have the greatest contribution to the anti-hepatoma effect.关键词：Paridis Rhizoma;fingerprint;anti-hepatoma effect;cluster analysis;grey correlation degree;spectrum-effect relationship;HepG2 cells
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发布时间：2021-02-09 - 摘要：Objective:The processing method of red ginseng was determined by comparing the effects of different steaming time and pressure on the total content of six ginsenosides.Method:The contents of ginsenoside Rg1, Re, Rf, Rb1, Rc and Rb2 were determined by ultra high performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QqQ-MS/MS). The Waters ACQUITY UPLC BEH C8 column (2.1 mm×100 mm, 1.7 μm) was used. The mobile phase was 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile solution (B) for gradient elution (0-4 min, 81%-79%A; 4-6.3 min, 79%-75%A; 6.3-6.5 min, 75%-71%A; 6.5-9.5 min, 71%A; 9.5-16.5 min, 71%-68.5%A; 16.5-16.6 min, 68.5%-60%A; 16.6-19 min, 60%-100%A). The flow rate was set at 0.4 mL·min-1 and the column temperature was set at 35 ℃. The mass spectrographic analysis employed electrospray ionization (ESI) and negative ion collection mode with capillary ionization voltage of 2.5 kV, desolvation temperature of 350 ℃, desolvation gas flow of 700 L·h-1 and cone gas flow of 50 L·h-1. Multiple reaction monitoring (MRM) mode was used to collect information, the collection range was m/z 100-1 500, detection was performed by MRM mode at m/z 799.59-637.49 for ginsenoside Rg1, m/z 945.54-475.79 for ginsenoside Re, m/z 799.59-475.49 for ginsenoside Rf, m/z 1 107.59-783.97 for ginsenoside Rb1, m/z 1 077.58-783.96 for ginsenoside Rc, m/z 1 077.75-191.19 for ginsenoside Rb2.Result:When the steaming time was 3 hours, the total mass fraction of six ginsenosides in each sample group was 7.099 8-16.768 5 mg·g-1, and the total amount of the six ginsenosides in atmospheric steaming was 2.5-12.6 times of that in pressurized steaming, which was obviously better than that in pressurized steaming.Conclusion:Under the conditions of this experiment, the best processing method of red ginseng is atmospheric steaming for 3 hours with fresh ginseng.关键词：fresh ginseng;sun-dried ginseng;steaming;ginsenosides;pressure;time;ultra high performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QqQ-MS/MS)
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发布时间：2021-02-09 - 摘要：Objective:To study the components of Ginseng Radix et Rhizoma of different origins and growth years.Method:Rapid liquid chromatography-time of flight mass spectrometry (RRLC-Q-TOF-MS) was applied to detect the raw data of Ginseng Radix et Rhizoma.After peak extraction, alignment, and normalization, the multivariate statistical analysis was made for the resulted dataset to find out the different compounds.The compounds were identified by using accurate molecular weight and tandem mass spectra, and the standard references were used to further confirm the identification.The changing trends of these components in different ginseng samples were observed.Result:The ginseng samples of different growth years and different origins were divided into different groups in the score plot of PLS-DA, and the variables with the variable importance in projection (VIP) value of more than 1 were considered to contribute more to the separations, then the t-test was applied to determine whether potential biomarkers were statistically significant (P<0.05) between the two groups.The contents of eleven compounds, including ginsenoside Rb1, Rh4, and Rk2, were significantly different between ginseng samples aged 3 and 5 years, and the contents of these compounds increased as the rise of the ginseng growth years.Ginsenosides Rg1, Rf, Rh1, Rb1, and other six compounds were significantly different in ginseng samples from Jilin and Heilongjiang province.Conclusion:LC-MS is a rapid and accurate method for the analysis of ginseng samples, and could help to find out the different components among samples.关键词：Ginseng Radix et Rhizoma;different origins;different growth years;3 years;5 years
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发布时间：2021-02-09 - 摘要：Objective:To establish an LC-MS/MS method for the determination of alcohol-soluble components and water-soluble components in Arnebiae Radix(L-shikonin, β, β′-dimethylacrylshikonin, β-acetoxy-isovalerylshikonin, lithospermic acid, caffeic acid, rosmarinic acid), for the purpose of quality control of the herb.Method:Chromatographic separation was carried out at 35 ℃ on Agilent ZORBAX SB C18 (4.6 mm×50 mm, 1.8 μm), with 0.1% formic acid acetonitrile (A)-0.1% formic acid solution (B) as the mobile phase for gradient elution (0-3 min, 5%-95%A; 3-7 min, 95%A; 7-7.01 min, 95%-5%A; 7.01-8.5 min, 5%A). The flow rate was 0.2 mL·min-1, and the injection volume was 5 μL. In a negative ionization mode, MRM scanning mode was adopted for quantification.Result:The calibration curves of L-shikonin, β, β′-dimethylacrylshikonin, β-acetoxy-isovalerylshikonin, lithospermic acid, caffeic acid, rosmarinic acid showed a good linearity, and the linear ranges of the above six compounds were 10.12-1 012 μg·L-1(r=0.998 1), 10.88-1 088 μg·L-1(r=0.991 2), 10.08-806.4 μg·L-1(r=0.997 6), 20.32-1 016 μg·L-1(r=0.996 6), 10.37-1 037 μg·L-1(r=0.999 6), 10.26-1 026 μg·L-1(r=0.997 8), respectively. The average recoveries of the analysts were 95.8%(RSD 3.2%), 103.5%(RSD 2.3%), 105.3%(RSD 2.1%), 96.1%(RSD 3.3%), 98.9%(RSD 2.7%), 100.8%(RSD 3.4%). The contents of six components in 13 batches of samples showed significant differences.Conclusion:The established method is feasible and simple, and provides a basis for comprehensive quality evaluation of Arnebiae Radix.关键词：Arnebiae Radix;alcohol-soluble component;water-soluble component;L-shikonin;β, β′-dimethylacrylshikonin;β-acetoxy-isovalerylshikonin;lithospermic acid
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发布时间：2021-02-09
Pharmacy Fundamentals
- 摘要：Objective:To explore the potential active ingredients and possible anti-breast cancer mechanism of Glycyrrhizae Radix et Rhizome and Aurantii Fructus based on the method of network pharmacology.Method:The main potential targets of Glycyrrhizae Radix et Rhizome and Aurantii Fructus on breast cancer were summarized by comparing the Glycyrrhizae Radix et Rhizome-Aurantii Fructus active ingredients screened from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and breast cancer targets searched in Therapeutic Target Database (TTD). Cytoscape 3.7.1 software was used to establish a Glycyrrhizae Radix et Rhizome-Aurantii Fructus active ingredients-target-disease network and perform topology analysis based on the network.Result:According to related conditions of drug-like (DL) and oral bioavailability (OB), the network of Glycyrrhizae Radix et Rhizome-Aurantii Fructus active ingredients-breast cancer target was obtained, covering a total of 133 nodes, 116 chemical components and 17 breast cancer drug targets, 109 active components of Glycyrrhizae Radix et Rhizome interacting with breast cancer drug target, 6 active ingredients of Aurantii Fructus interacting with breast cancer drug targets, and 1 common active ingredient of Aurantii Fructus and Glycyrrhizae Radix et Rhizome interacting with breast cancer targets. There were 400 breast cancer target-interaction target pairs in the network diagram.Conclusion:The anti-breast cancer effect of Glycyrrhizae Radix et Rhizome and Aurantii Fructus is based on the overall pharmacodynamic effect of multi-component, multi-pathway and multi-target, the investigation of its potential anti-breast cancer mechanism provides theoretical basis for further experimental research.关键词：network pharmacology;Glycyrrhizae Radix et Rhizome;Aurantii Fructus;breast cancer;mechanism
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发布时间：2021-02-09
Data Mining
- 摘要：Non-psychoactive medicinal cannabis is the cannabis that contain less than 0.3% of tetrahydrocannabinol (THC) and high content of cannabidiol (CBD) (more than 2.0% is reasonable), and mainly used for medicinal purpose. Cannabis have a long medical history, in this review, we retrospected the medicinal history of cannabis in China and the world. We summarized the cannabinoid especially CBD application of medicine in epilepsy, mental disease, tumor, analgesic and inflammatory, also the widely application in food, healthy products, cosmetics and other fields. Based on the problems like shortage of high CBD cultivars, weakness of fundamental research existed in non-psychoactive medicinal cannabis industry. We should fully utilize the genetic information of medicinal cannabis, establish the comprehensive identification system of non-psychoactive medicinal cannabis varieties, accelerate the breeding process of non-psychoactive medicinal cannabis cultivars, to ensure the safety and effectiveness of non-psychoactive medicinal cannabis. Strengthen the basic research on CBD and other cannabinoids to promote the application of non-psychoactive medicinal cannabis products and sustainable development of medical cannabis industry.关键词：non-psychoactive medicinal cannabis;herbal textual research;medical history;cannabidiol (CBD);tetrahydrocannabinol;medicine application;CBD oil
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发布时间：2021-02-09 - 摘要：The chemical components (groups) contained in traditional Chinese medicine(TCM) and its compound preparations are the material basis for its curative effect, because of the integrity of the action of TCM and the complexity of its compositions and mechanism. The separation and analysis of chemical constituents in TCM and its compound prescriptions by various methods is always a key problem to be solved in the research of disease prevention and treatment of TCM. The binding of drug molecules to receptors at the cellular level was explored to provide a new idea for the screening of active components of TCM or its compound preparations. Traditional methods have some drawbacks, such as cumbersome operation, time-consuming, waste of solvents and irreversible adsorption of samples. The basic information of pharmacological parameters cannot be given in the separation process, and the active ingredients cannot be efficiently and accurately located. At present, cell membrane chromatography (CMC), one of the methods, is used to study the interaction between drug molecules and receptors, and can combine the existing chromatography and mass spectrometry technology, cell biology and receptor pharmacology, and correctly reflect the interaction between active parts, active components and cell membrane and membrane receptors, so it has unique advantages in screening effective parts, separation of active components and high-throughput screening from complex TCM system. The principles and characteristics of CMC, the cell membrane model in the field of active ingredient selection of TCM and its research status in combination with gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) were reviewed, and its development prospects and future research methods were discussed, which provides theoretical basis and practical guidance for the research and utilization of CMC in the field of TCM.关键词：cell membrane chromatography;mass spectrometry;combination;traditional Chinese medicines;active compounds;screening
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发布时间：2021-02-09
Academic Discussion
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