Abstract：Objective:The effect of Liuwei Dihuangwan on blood metabolism during growth and development of rats was investigated by taking the overall metabolic profile and biomarkers of metabolomics as indexes.Method:Ultra performance liquid chromatography-high definition mass spectrometry(UPLC-HDMS) was employed to establish a blood metabolomics study method for characterizing the blood metabolic profile of rats at different time before and after administration. The pattern recognition method was used to integrate and analyze the metabolic profiles, and the differential metabolic markers related to drug action were searched. Based on metabolomics pathway analysis(MetPA) and Kyoto encyclopedia of genes and genomes(KEGG) and other databases, the metabolic pathways related to differential markers were analyzed to study the effect of Liuwei Dihuangwan on blood metabolism during growth and development of rats.Result:Liuwei Dihuangwan significantly affect the blood metabolism profiles during growth and development of rats by regulating 30 blood metabolic markers and 12 related target metabolic pathways, and 8 key metabolic markers were delineated.Conclusion:Liuwei Dihuangwan can significantly regulate the blood metabolic network during the growth and development of rats, thereby affecting the growth and development of rats.
Keywords：Liuwei Dihuangwan;growth and development;blood metabolism;metabolic markers;amino acid metabolism;3-methoxy-4-(sulfooxy)-benzeneacetic acid;glycocholic acid
Abstract：Objective:Through metabonomics research methods, the effect of Siwutang on metabolites and metabolic pathways in natural aging mice were observed.The related targets and mechanism of Siwutang intervention in natural aging mice were analyzed.Method:Taking 20-month-old natural aging model mice(equivalent to 60-65 years old of human beings) as the experimental subjects, at the same time, mice aged 3 months were established as the youth group.UPLC-Q-TOF-MS technique was employed to analyze the mouse plasma with mobile phase of acetonitrile(containing 0.1%formic acid)-0.1%formic acid solution for gradient elution and positive ion mode of electrospray ionization, and the metabolic markers were analyzed by principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA), and their metabolic pathways were summarized.Result:Siwutang had obvious reversal effect on the expression levels of 16 aging-related metabolites, among which 9 metabolic markers were statistically significant(P<0.01), mainly affecting glutathione metabolism, pyrimidine metabolism, arginine and proline metabolism, selenium amino acid metabolism and other metabolic pathways.Conclusion:Siwutang can affect the metabolites in the plasma of 20-month-old natural aging mice, and the metabolic disorder during the aging process of mice can be improved by glutathione metabolism, pyrimidine metabolism, selenium amino acid metabolism and other pathways, and this paper can provide biological information for the study of material basis of this compound for aging.
Abstract：Objective:To investigate the therapeutic effect and immune mechanism of Baitouweng Tang on ulcerative colitis(UC) rats.Method:The mode of UC rats was made of 2, 4-dinitrochlorobenzene (DNCB)/ethanol enema. Rats were randomly divided into control group, model group, mesalazine group, and high-dose, middle-dose and low-dose Baitouweng Tang groups. The mesalazine group were administered with mesalazine (0.5 g·kg-1). Baitouweng Tang groups were given Baitouweng Tang (10, 5, 2.5 g·kg-1), while the other groups were given double steaming water. After 7 days of continuous administration, the general condition and disease activity index of rats in each group were observed. After anesthesia in rats, blood was taken from the abdominal aorta. Then the rats were put to death, and the length and morphological observation of the colon were measured. Ultraviolet spectrophotometry detection was used to detect the activities of myeloperoxidase (MPO) in blood and colon tissue. The levels of P-selectin, macrophage migration inhibitory factor (MIF) and thromboxane B2 (TXB2) in blood and colon tissues were detected by enzyme-linked immunosorbent assay(ELISA). Immunohistochemistry and Western blot methods were undertaken to determine the expressions of Toll-like receptor 4 (TLR4) and nuclear transcription factor-κB (NF-κB) proteins in colon tissue.Result:Compared with the model group, the rats in model group showed severe symptoms, such as loose stools, diarrhea and bloody stools, while Baitouweng Tang obviously ameliorated them. Moreover, Baitouweng Tang significantly reduced DAI, colon general and pathological scores, which were high in model group(P<0.01). The levels of MPO, P-selectin, MIF and TXB2 in the serum and colon tissues of the model group were obviously increased(P<0.01), and Baitouweng Tang could reverse them. Similarly, the expressions of TLR4 and NF-κB in colons of model group were markedly higher than those in control group(P<0.01). However, Baitouweng Tang group showed lower expressions than the model group (P<0.01).Conclusion:Baitouweng Tang could inhibit TLR4/NF-κB signaling pathway in treatment of ulcerative colitis, and reduce the expressions of P-selectin, MPO, MIF and TXB2, and thus promoting intestinal mucosal repair and improving intestinal function.
Abstract：Objective:To investigate the neuroprotective effect of modified Yuejuwan and Ganmai Dazao Tang (YJGZ) on glutamate-induced cell injury of mouse hippocampal neuron cell line (HT22).Method:The HT22 cell was cultured aseptically, and the cell injury model was established with high concentration of glutamate acid, YJGZ aqueous extract and drug-containing serum were prepared. Then experiment was divided into normal group, model group, YJGZ drug-containing serum groups (1%, 5%, 10%), YJGZ aqueous extract group (166 mg·L-1), and nimodipine group (100 μmol·L-1). Methye thiazolye telrazlium (MTT) was used to detect the cell survival rate of each group, the level of lactate dehydrogenase (LDH) release and intracellular reactive oxygen species (ROS) were detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the protein expression of hippocampal N-methyl-D-aspartate receptor subunit 2B protein (NR2B), cyclic adenosine response element binding protein (CREB), phosphorylated cyclic adenosine response element binding protein (p-CREB), extracellular regulated protein kinase (ERK), and phosphorylated extracellular regulated protein kinase (p-ERK).Result:As compared with normal group, the cell survival rate was significantly decreased in model group (P<0.01), LDH release rate was significantly increased (P<0.05), intracellular reactive oxygen species (ROS) level was increased (P<0.05), expression of NR2B protein was increased (P<0.05), and the protein expression levels of CREB, p-CREB, ERK and p-ERK were decreased (P<0.05). As compared with model group, YJGZ aqueous extract group and nimodipine group can significantly improve the survival rate of HT22 cells under glutamate model conditions (P<0.01), reduce cell damage, reduce LDH release rate and intracellular ROS levels (P<0.05, P<0.01), decrease the expression level of NR2B protein (P<0.05), and increase the protein expression levels of CREB, p-CREB, ERK and p-ERK (P<0.05). However, the survival rate of HT22 cells was not increased in the YJGZ-containing serum group as compared with model group.Conclusion:YJGZ aqueous extract has a significant protective effect on glutamate-induced HT22 cell injury.
Keywords：Yuejuwan;Ganmai Dazao Tang;hippocampal neuron cell line (HT22);glutamate;neuroprotection
Abstract：Objective:To investigate the protective mechanism of Liuwei Dihuangwan on diabetes mellitus with liver injury in terms of inflammation.Method:The 18 db/db mice were selected from animal model of spontaneous type 2 diabetes, mice were randomly divided into model group, Liuwei Dihuangwan group(9.75 g·kg-1·d-1, once a day), resveratrol group(42.6 mg·kg-1·d-1, once a day)based on fasting blood-glucose (FBG). The 12 litter wild db/m mice were randomly divided into normal group, Liuwei Dihuangwan group(9.75 g·kg-1·d-1, once a day). The normal group and the model group were given the same amount of distilled water by gavage. The mice in each group were treated for 16 weeks. FBG, triglyceride (TG) and alanine aminotransferase (ALT) were examined. Histopathological changes were observed by liver biopsy hematoxylin-eosin (HE) staining. Silent information regulator 6 (SIRT6), nuclear factor-kappaB p65 (NF-κB p65), monocyte chemoattractant protein-1 (MCP-1), vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) in the liver tissue were detected by Western blot.Result:Compared with normal group, FBG, TG and ALT in model group increased significantly(P<0.01), protein expression levels of NF-κB p65, MCP-1, VCAM-1 and ICAM-1 in model group were significantly up-regulated(P<0.01), the protein expression level of SIRT6 in model group was significantly down-regulated(P<0.01). Compared with model group, Liuwei Dihuangwan and resveratrol could significantly reduce the levels of serum FBG and TG(P<0.01), the protein expression of SIRT6 was significantly increased(P<0.01), NF-κB p65, MCP-1 and VCAM-1 were significantly decreased (P<0.05, P<0.01), and liver steatosis and inflammatory cells infiltration were alleviated.Conclusion:Liuwei Dihuangwan can protect the diabetes mice from liver injury, and its mechanism is related to the improvement of lipid metabolism and the inhibition of inflammatory response.
Abstract：Objective:To study the effects of modified Buzhong Yiqitang on cardiac function and fat acid binding protein3 (FABP3)and peroxisome proliferator-activated receptor gamma(PPARγ) protein of cardiac myocytes in type 2 diabetic mellitus (T2DM) rats, and to explore the mechanism of this prescription to protect T2DM cardiomyopathy.Method:After 6 weeks of high-glucose and high-fat diet, streptozotocin (STZ) was intraperitoneally injected (50 mg·kg-1) to establish a diabetic rat model, and then randomly divided into normal group, model group, modified Buzhong Yiqitang group (21 g·kg-1) and metformin hydrochloride group (2 g·kg-1). After continuous administration for 6 weeks, echocardiography was used to detect the cardiac structure and function of the rats in each group. Combined with the measured blood glucose level, it was judged that the rat model building of diabetic cardiomyopathy was successful.Rat heart tissue was dissected, and rat myocardial morphology was observed by hematoxylin-eosin(HE)staining. Western blot was used to detect the expression levels of FABP3 and PPARγ proteins in rat myocardium.Result:Compared with normal group, levels of blood sugar, total triglyceride(TC), triglyceride(TG), insulin (INS) in model group rats increased significantly (P<0.01), slow heart rate, diastolic and systolic left interior diameter (LVIDd)increases, the reduced systolic left ventricular posterior wall thickness, the ratio of peak A to peak E (E/A) ratio, short axial shortening rate(FS)and ejection fraction (EF)were different degree reduced (P<0.01), FABP3 protein expression were significantly elevated, PPARγ protein expression levels were significantly lower (P<0.01). Compared with model group, the situation of blood glucose, TC, TG, INS in modified Buzhong Yiqitang group was significantly improved (P<0.01), cardiomyopathy was significantly improved (P<0.01), FABP3 protein expression was significantly decreased, and PPARγ protein expression was significantly increased (P<0.01).Conclusion:Modified Buzhong Yiqitang can improve glucose and lipid metabolism disorder in diabetic cardiomyopathy rats, improve cardiac function in diabetic cardiomyopathy rats, increase the expression of PPARγ in cardiomyocytes, and reduce the expression of FABP3 protein.
Keywords：Buzhong Yiqitang;diabetic cardiomyopathy;traditional Chinese medicine treatment;type 2 diabetes mellitus;fat acid binding protein-3;peroxisome proliferator-activated receptor gamma
Abstract：Objective:To evaluate the effect of Chuankezhi injection on mouse model of pneumonia induced by influenza A (H1N1) FM1 strain.Method:ICR mice were randomly divided into normal group, model group, tamiflu control group (27.5 mg·kg-1·d-1) and Chuankezhi injection group (1.5 mL·kg-1·d-1). In the death protection experiment, mice were infected with 2×half lethal dose (LD50) of influenza virus FM1.The Chuankezhi injection was given once a day for 4 days. The number of death animal within 14 days was counted. The mortality and the death protection rate were calculated. In the treatment experiment, mice were infected with 0.8×LD50 of influenza virus, and the Chuankezhi injection was given once a day for 4 days. On the 5th day after the infection, the levels of interleukin-8 (IL-8) in lung, prostaglandin E2 (PGE2) and vasopressin (AVP) in brain were tested by enzyme-linked immunosorbent assay (ELISA). The viral load of influenza virus in lung was tested by Real-time PCR. In the pre-treatment experiment, mice were given Chuankezhi injection once a day for 5 days. 1 hour after the last treatment, mice were infected with 0.8×LD50 influenza virus. 4 days after the infection, the lung index, spleen index, thymus index, and viral load in lung tissue were calculated.Result:Compared with normal group, the IL-8, PGE2 content, lung index and viral load in the lung tissue of model group were significantly increased (P<0.01). Compared with model group, Treatment of Chuankezhi injection could reduce the rate of deaths. Significantly inhibit the level of IL-8, PGE2, and the viral load of influenza(P<0.05, P<0.01). Pre-treatment of Chuankezhi injection could significantly increase the thymus index of infected mice(P<0.01), reduce lung index and the viral load of influenza(P<0.05). Both administration methods can significantly reduce the infiltration of inflammatory cells in the bronchial, perivascular and alveolar interstitium and reduce the exudation of red blood cells in the lumen.Conclusion:Chuankezhi injection could effectively prevent the mouse model of pneumonia induced by influenza A (H1N1) virus. The mechanism might be related to the reduction of inflammation and inhibiting viral replication.
Keywords：Chuankezhi injection;influenza A H1N1 virus;viral pneumonia;pharmacodynamic effect
Abstract：Objective:To investigate the protective effect of Perillae Folium with aqueous extract (PFAE) on some key factors of Adriamycin (ADR)-induced oxidative injury in human renal tubular epithelial cells(HK-2), including the survival rate, oxidative injury indexes and cell apoptosis, in order to define the underlying mechanism.Method:A model of ADR-induced HK-2 cells oxidative injury was established in vitro, then cell viability was detected by cell counting kit-8 (CCK-8) after intervention with positive reference N-acetylcysteine (NAC) or PFAE (5, 15, 45 g·L-1) at different concentrations. According to the morphological changes under microscopy, the optimum concentration of PFAE was screened out for the follow-up experiments. Then, the experiments were divided into six groups: blank group, ADR (0.05 g·L-1) group, PFAE (15 g·L-1) group, ADR+ PFAE (0.05+ 15) g·L-1 group, NAC (0.81 g·L-1) group, and ADR+ NAC (0.05+ 81) g·L-1 group. After that, malondialdehyde (MDA), superoxide dismutase (SOD), total antioxidant capacity(TAC) were measured in the cell homogenate after 24 h administration. The level of reactive oxygen species (ROS) was detected by 2′, 7′-dichloroflurescin diacetate (DCFH-DA) fluorescence probe. Flow cytometry and TdT-mediated dUTP Nick-End Labeling (TUNEL) were used to monitor the cell apoptosis. Western blot was used to observed the expressions of mitochondrial apoptosis-associated proteins, like B lymphocyte tumor-2 gene (Bcl-2), Bcl-2 related X protein (Bax), cysteine aspartate protease-9 (Caspase-9), cysteine aspartate protease-3 (Caspase-3) and poly ADP-ribose polymerase (PARP), as well as their shear bodies. In addition, the phosphorylation protein expressions of p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK), c-Jun amino-terminal kinase (JNK) in mitogen-activated protein kinase (MAPK) signaling transduction pathway were detected by Western blot.Result:Compared with blank group, ADR group showed a decreased cell viability (P<0.01), and lower SOD level (P<0.01), but higher expressions of MDA and ROS (P<0.01), and an increased apoptotic rate (P<0.01). The ADR group also increased in rate of Bax/Bcl-2, cleaved Caspase-9/Caspase-9, cleaved Caspase-3/Caspase-3, and cleaved PARP/PARP (P<0.01), as well as the phosphorylation protein expressions of p38 MAPK, ERK and JNK (P<0.05, P<0.01). Compared with the ADR group, both ADR+ PFAE groups and ADR+ NAC group had higher cell proliferation rates (P<0.01). In addition, the protective effect of PFAE on cells was the most obvious at the concentration of 15 g·L-1. The ATC and SOD levels were increased in ADR+ PFAE group and ADR+ NAC group (P<0.01), while their content of MDA and ROS, cell apoptosis, relative ratio of apoptotic protein expression, and phosphorylation protein expressions of p38 MAPK and ERK were all decreased (P<0.01). However, there was no effect on the expression of phosphorylated JNK protein.Conclusion:PFAE could alleviate the oxidative injury of HK-2 cells induced by ADR, and have an antioxidant effect, which inhibited cell apoptosis through mitochondrial apoptotic pathway and ERK/p38 MAPK signaling pathway.
Keywords：aqueous extract of Perillae Folium;renal tubular epithelial cells;adriamycin;oxidative injury;mitochondrial apoptosis;mitogen-activated protein kinase (MAPK) signaling pathway
Abstract：Objective:To explore the protective effect of formula of Gougancai decoction (FGD) on acute liver injury induced by carbon tetrachloride (CCl4) in rats, in order to provide basis for the development of pharmaceutical preparations or healthcare products.Method:Sixty rats were randomly divided into normal group, Silymarin group (120 mg·kg-1) and FGD groups (475, 950, 1 900 mg·kg-1). The normal group and the model group were given equal volume of saline by gavage, while the other groups were administered with the corresponding dose of drugs according to the body weight. After 10 days, the acute liver injury model was established with 12%carbon tetrachloride peanut oil solution (5 mL·kg-1), except the normal group. All of the rats were put to death to collect serum and liver tissues. The contents of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total bilirubin (TBIL), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected by biochemical methods, the levels of tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) in liver tissues were determined by enzyme-linked immunosorbnent assay(ELISA). Nuclear factor-κB (NF-κB) and peroxisome proliferator-activated receptor-γ (PPAR-γ) protein expression in liver tissues were detected by Western blot, and htoxylin eosin (HE) staining was used to observe the variation of liver histopathological.Result:Compared with the normal group, the serum activities of AST, ALT, ALP and the content of TBIL, MDA in the model group were significantly increased (P<0.01), the levels of TNF-α, IL-1β, IL-6 in liver tissue were remarkably increased (P<0.01), but the serum activities of SOD, GSH-Px were significantly decreased (P<0.01), the expression of NF-κB was enhanced in liver tissue (P<0.01), and PPAR-γ was down-regulated (P<0.01), indicating the successful modeling of acute liver injury. Compared with the model group, FGD could reduce the activities of AST, ALT, ALP and the contents of TBIL, MDA (P<0.05, P<0.01), decease the level of TNF-α, IL-1β, IL-6 (P<0.05, P<0.01), and down-regulate the expression of NF-κB (P<0.05, P<0.01), but up-regulate the activities of SOD, GSH-Px and the expression of PPAR-γ (P<0.05, P<0.01). The liver tissue lesions were alleviated to varying degrees.Conclusion:FGD has a protective effect on CCl4-induced acute liver injury in rats, and its mechanism may be related to the activation of PPAR-γ and the inhibition of NF-κB signaling pathway, with anti-inflammatory and anti-oxidative effects.
Keywords：formula of Gougancai decoction;acute liver injury;nuclear factor-κB (NF-κB);peroxisome proliferator-activated receptor-γ(PPAR-γ);anti-inflammation;anti-oxidation
Abstract：Objective:To study the effect of Rosae Chinensis Flos total flavones(RCTF) on the focal cerebral ischemia-reperfusion model in rats, in order to preliminarily explore the mechanism of action.Method:Rats were randomly divided into sham-operated group, model group, large, medium, and low-dose RCTF group(200, 100, 50 mg·kg-1) and positive group [Nimodipine group(20 mg·kg-1) and Naoluotong group (500 mg·kg-1)] . After 7 days of continuous administration, 1 hour later after the last administration, the middle cerebral artery middle cerebral artery occlusion (MCAO) model was duplicated. After 2 hours of modeling, perfusion was performed for 22 hours. Mortality and neurological deficits were scored. Serum S-100β was detected; brain tissue malondialdehyde(MDA), superoxide dismutase (SOD), nitric oxide (NO), nitric oxide synthase (NOS), tumour necrosis factor-α(TNF-α), interleukin-1β(IL-1β), intercellular adhesion molecule-1(ICAM-1), adenosine triphosphate (ATP)ase were measured. The brain tissue morphological changes were observed.Result:The rat model of focal cerebral ischemia and reperfusion was successfully replicated. Compared with the model group, RCTF in large, medium, and low-dose RCTF group significantly decreased the score of neurological deficit in rats (P<0.01), significantly decreased the content of S-100β in serum (P<0.01), significantly reduce the levels of MDA, NO and NOS in brain tissue (P<0.01), significantly increase the level of SOD in brain tissue (P<0.01), and significantly increase Na+ K+ -ATPase, Mg2+ -ATPase, and Ca2+ in brain tissue (P<0.01), significantly reduced TNF-α content, IL-1β, ICAM-1 content in brain tissue (P<0.01), and significantly improved brain tissue damage (P<0.01).Conclusion:RCTF have a protective effect on cerebral ischemia-reperfusion injury in rats. The mechanism may be related to the resistance of anti-free radicals, the reduction of inflammation in brain tissue and the improvement of brain energy metabolism after cerebral ischemia reperfusion injury.
Abstract：Objective:To clarify the antitussive, expectorant, antipyretic and anti-inflammatory effects of Tanreqing inhalation solution, and provide basis and data support for further research and development of this preparation.Method:The methods of cough induced by ammonia and tracheal phenol red excretion were used to observe the antitussive and expectorant effects of Tanreqing inhalation solution in mice. The fever model of rats was established by intraperitoneal injection of bacterial lipopolysaccharide(LPS) to observe the antipyretic effect of the Tanreqing inhalation solution, the acute pneumonia model of rats was established by atomizing LPS inhalation, and the anti-inflammatory effect of Tanreqing inhalation solution was observed.Result:Tanreqing inhalation solution could reduce the number of coughs in mice induced by ammonia water, increase the amount of phenol red excretion in mouse trachea, decrease the levels of body temperature and its related regulatory factors of prostaglandin E2(PGE2) and cyclic adenosine monophosphate(cAMP) of rats induced by LPS, decrease the white blood cell(WBC) count and the neutrophil ratio(NEUT) in bronchoalveolar lavage fluid(BALF) of rats with LPS-induced acute pneumonia, and reduce the levels of nuclear transcription factor-κB(NF-κB) and interleukin-1β(IL-1β) in lung tissue.Conclusion:Tanreqing inhalation solution has obvious antitussive, expectorant, antipyretic and anti-inflammatory effects, which is worthy of further development and promotion.
Abstract：Objective:To observe the effect of puerarin on phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and glycogen synthase kinase-3β (GSK-3β) in insulin resistant HepG2 cells.Method:HepG2 cells were treated with palmitic acid 0.5 mmol·L-1 and insulin 9×10-4 U·L-1 to induce insulin resistant condition for 24 h. Cell viability was detected by methyl thiazolyl tetrazolium (MTT) assay to determine the concentration of puerarin. This experiment included normal control group, model control group and puerarin groups of different doses (40, 80, 160, 320 μg·L-1). Glucose detection kit was used to detect the content of glucose in cell culture supernatant. Tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) levels in supernatant of cell culture medium were detected by enzyme-linked immunosorbent assay (ELISA). Hepatic glycogen assay kit was used for detecting the hepatic glycogen content in HepG2 cells. Western blot was applied to detect protein expression levels of PI3K, Akt, p-Akt, GSK-3β and p-GSK-3β.Result:Compared with those in the normal control group, the glucose consumption rate was significantly down-regulated in HepG2 cells in the model control group (P<0.05), the intracellular glycogen content was decreased (P<0.05), the content of TNF-α and IL-6 were increased in supernatant of cell culture medium (P<0.05), PI3K protein level was decreased, Akt phosphorylation level was down-regulated (P<0.05), and GSK-3β protein expression was up-regulated (P<0.05). Compared with those in the model control group, in the puerarin groups, the glucose consumption rate was increased in a dose-dependent manner (P<0.05), the intracellular glycogen content was increased, the contents of TNF-α and IL-6 were reduced in supernatant of cell culture medium (P<0.05), PI3K protein expression was enhanced, Akt phosphorylation level was up-regulated (P<0.05), GSK-3β protein expression was down-regulated, but its phosphorylation inactivation was increased (P<0.05).Conclusion:Puerarin alleviates the insulin resistance of HepG2 cells by strengthening the PI3K/Akt/GSK-3β signal transduction process and increasing the glycogen content in hepatocytes.
Abstract：Objective:To investigate the anticancer effect of isoliquiritigenin (ISL) on human clear cell renal cell carcinoma 786-O cells, and explore its possible molecular mechanism.Method:Thiazolyl blue tetrazolium bromide (MTT) assay was used to detect effect of ISL (0, 10, 25, 50, 75, 100 μmol·L-1) on proliferation of 786-O cells. The effect of ISL on migration and invasion of 786-O cells was detected by cell scratch test and Transwell assay. The autophagy was observed under the fluorescence microscope through acridine orange staining and Ad-GFP-LC3 transfection experiment. Western blot was used to detect the expression of autophagy related protein and analyze the changes of phosphatidylinositol-3-kinase (PI3K)/protein kinase B(Akt)/mammalian target of rapamycin (mTOR) signaling pathway to explore the possible mechanism.Result:MTT results showed that ISL could significantly inhibit the proliferation of 786-O cells in a time-dose dependent manner (P<0.05). The results of scratch and Transwell experiments indicated that ISL could inhibit the migration and invasion of 786-O cells (P<0.05). Acridine orange staining and ad-GFP-LC3 transfection showed that ISL can induce autophagy in 786-O cells. Besides, ISL can induce LC3-Ⅱ, Beclin1, Atg5 protein expressions (P<0.05), and significantly reduce p62 protein expression (P<0.05); autophagy inhibitor 3-methyl adenine (3-MA) can be added to reverse the above phenomena (P<0.05). Meanwhile, ISL can inhibit phosphorylation of Akt, mTOR levels (P<0.05), inhibitor LY294002 and rapamycin (Rap) can be added to significantly re-regulate LC3-Ⅱ, Beclin1, Atg5 protein expressions (P<0.05) and significantly down-regulate p62 protein expression (P<0.05).Conclusion:ISL can inhibit the proliferation, migration and invasion of clear cell renal carcinoma 786-O cells, and induce autophagy by inhibiting the PI3K/Akt/mTOR signaling pathway.
Keywords：isoliquiritigenin (ISL);clear cell renal cell carcinoma;786-O;autophagy;phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway
Abstract：Objective:To study inhibitory effect of total flavonoids from Ampelopsis grossedentata (TF) on transplanted tumors of human hepatocellular carcinoma in nude mice, and predict that its mechanism may be related to relevant factors regulating phosphatidylinositol 3 kinase (PI3K)/protein kinases B(Akt)/p53 pathway in apoptosis.Method:The nude mice transplanted BEL-7404 hepatoma model was established and divided into model group, 5-fluorouracil (5-FU) group (1.0 g·L-1) and TF (30, 15, 7.5 g·L-1) groups. Nude mice were put to death after two weeks of administration. The tumor tissues were excised, and tumor inhibition rate (IR) and relative tumor proliferation rate (T/C) were calculated. Reverse transcription PCR(RT-PCR) was used to detect PI3K, Akt1, p53 gene(p53), Caspase-3, B cell lymphoma/lewkmia-2 (Bcl-2), Bcl-2 associated X protein (Bax) mRNA expressions, immunohistochemical method was used to detect expressions of relevant proteins PI3K, Akt1, p53, Caspase-3, Bcl-2, Bax.Result:The establishment of xenograft tumor in mice showed that TF was administered orally once per day for two consecutive weeks. IRs were 53.26%, 35.94%, and 26.74%, respectively. T/Cs were 59.74%, 69.66%, and 84.82%, respectively. RT-PCR experiments showed that compared with model group, when TF concentration was 30 g·L-1, mRNA expressions of PI3K, Akt1, and Bcl-2 were significantly down-regulated, and mRNA expressions of tumor suppressor genes p53, Capsase-3, and Bax were significantly up-regulated. Immunohistochemical method results showed that compared with model group, at TF concentrations of 30, 15 g·L-1, all PI3K, Akt1, Bcl-2 protein expressions were significantly down-regulated, while p53, Capsase-3, Bax protein expressions were significantly increased.Conclusion:TF has an obvious anti-liver cancer activity in vivo. Its mechanism may be correlated with up-regulation of expressions of p53, Caspase-3, and activation of apoptosis PI3K/Akt/p53 pathway, thereby inhibiting Bcl-2, increasing expression of Bax, and promoting hepatocellular apoptosis.
Keywords：total flavonoids from ampelopsis grossedentata;anti-liver cancer;phosphatidylinositol 3 kinase (PI3K)/protein kinases B(Akt)/p53 signaling pathway;mechanism of action
Abstract：Objective:To detect the effect of modified Shengjiangsan on the expression of reactive oxygen species(ROS) in mitochondria of renal foot cells of rats, in order to study the mechanism of modified Shengjiangsan.Method:After be fed for 7 days, the 60 SD male rats were randomly divided into four groups: blank control group, model group, positive medicine group and traditional Chinese medicine(TCM) treatment group. After establishment of the rat model of membranous nephropathy, model group, positive medicine group and TCM treatment group were treated differently. After 4 weeks, all of the rats were put to death, and the expressions of ROS, 24-hour urinary protein quantity, total cholesterol, triglyceride, total protein, albumin, urea nitrogen, creatinine were detected by Real-time fluorescence quantitative polymerase chain reaction Real-time PCR.Result:The expression of 24-hour urinary protein quantity, total cholesterol, triglyceride in positive medicine group and TCM treatment group were reduced, and the expressions of total protein, albumin in positive medicine group and TCM treatment group were reduced compared with those of model group (P<0.05). The expressions of 24-hour urinary protein quantity, total cholesterol, triglyceride, total protein, albumin in TCM treatment group have no difference with those of positive medicine group. After treatment, the expression of ROS mRVA in mitochondria of renal foot cells of rats of positive medicine group and TCM treatment group were increased compared with that of model group (P<0.05), but with no difference in the ROS mRNA expression between positive medicine group and TCM treatment group.Conclusion:Modified Shengjiangsan can effectively control the development of ROS in mitochondria of renal foot cells of rats, and repair the renal function of membranous nephropathy rats by recovering foot cells.
Abstract：Objective:To observe the effect of acupoint application therapy of modified Wuzhuyu Binglang Tang on chronic non-atrophic gastritis (CNAG) and the mechanism of action.Method:Two hundred and seventeen patients were randomly divided into control group (107 cases) and observation group (110 cases) by random number table. Both groups’ patients got Omeprazole enteric-coated tablets half an hour before meals, 20 mg/time, 2 times/day, bismuth potassium citrate tablets half an hour before meals, 2 tablets/time, 2 times/day, amoxicillin capsules after meals, 1.0 g/time, 2 times/day, two dimensional furazolidone tablets after meals, 1 tablet/time, 2 times/day, mosapride citrate tablets, 5 mg/time, 3 times/day. The treatment lasted for 2 weeks. Patients in observation group got acupoint application of modified Wuzhuyu Binglang Tang at Zhongwan, Weiyu, Geyu, Shenyu and Piyu for 6 h/day, 1 time/day. And the patients in control group got model drug at the same acupoints. Before and after treatment, epigastric fullness and epigastric pain were scored. Levels of motilin, gastrin, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and prostaglandin E2 (PGE2) were detected. In 1-month follow-up visit, Hp and adverse reaction were detected under gastroscope.Result:The clinical effect in observation group under gastroscope was better than that in control group (Z=2.015, P<0.05), and the scores of traditional Chinese medicine symptoms in observation group was better than those in control group (Z=2.663, P<0.01). And scores of epigastric fullness, epigastric pain, post-food abdominal distention, early satiety, nausea, hiccup, pantothenic acid, heating, nausea and vomiting were lower than those in control group (P<0.01). The negative conversion rate of Hp in observation group was 93.64%, which was higher than 84.11%in control group (χ2=5.002, P<0.05). And levels of TNF-α and IL-6 were lower than those in control group, PGE2 was higher than that in control group (P<0.01). And mild erythema and itching disappeared after symptomatic treatment.Conclusion:The acupoint application therapy of modified Wuzhuyu Binglang Tang can relieve the clinical symptoms, promote the negative conversion rate of Hp, improve the clinical effect, regulate gastrointestinal hormones, inhibiting release of inflammatory factors, protect gastric mucosa, and improve gastrointestinal function, with safety in clinic use.
Abstract：Objective:To observe the clinical efficacy of Hegan granule in the treatment of non-erosive reflux disease with liver stagnation and spleen deficiency syndrome and its effect on quality of life, 5-hydroxytryptamine (5-HT) and vasoactive intestinal peptide (VIP) level.Method:Patients with non-erosive reflux disease of liver stagnation and spleen deficiency were randomly divided into treatment group (32 cases) and control group (33 cases). The treatment group was orally given Hegan granules after meal, 10 g/bag, 1 bag/time, 2 times/day. The control group was orally given with omeprazole enteric-coated tablets twice daily, 20 mg/grain. Both groups were treated for 8 weeks. The total scores of traditional Chinese medicine (TCM) symptoms, SF-36 health scale scores, 5-HT and VIP levels were observed before and after treatment.Result:The medical efficacy of the two groups was 87.5%and 81.8%in the treatment group and the control group, respectively. The treatment group was superior to control group (P<0.05). The treatment group had special symptoms and secondary symptoms after treatment, with better improvements than control group (P<0.05). The efficacy of the two groups was comparable in the main symptoms, after the treatment, the SF-36 health scale of treatment group in physiological function, overall health, vitality, social function, emotional function and mental health were higher than those in control group (P<0.05). The levels of 5-HT and VIP in two groups were lower after treatment, and treatment group was significantly lower than control group (P<0.05).Conclusion:Hegan granule has a good clinical efficacy in treating patients with non-erosive reflux disease and liver stagnation and spleen deficiency. It can significantly alleviate the symptoms of TCM, improve the quality of life of patients, and reduce the sensitivity of esophageal viscera.
Abstract：Objective:To investigate the clinical efficacy of Xiaoer anorexia granule combined with Saccharomyces brassicus in the treatment of infantile anorexia and its effect on serum cytokines.Method:Totally 240 cases of infantile anorexia treated in Jinhua People's Hospital from October 2015 to October 2017 were randomly divided into three groups. Group A (combination group) orally took Xiaoer anorexia granule, 1 pack/time for 1-3 years old, 2 packs/time for 4-6 years old, 3 times a day, and combined with saccharomyces brasiliensis, 0.5 g/time, 2 times a day, 15-30 min before meal. Group B took saccharomyces brasiliensis by the same method as the combination group. Group C took Xiaoer anorexia granules by the same method as above, with 80 cases in each group. 8 weeks is a course of treatment. Serum inflammatory cytokines leptin, SP and Ghrelin were also detected. The clinical efficacy, body weight, height improvement and serum cytokine levels were observed in 3 groups.Result:The total improvement rate of group A was 91.25%, which was significantly higher than 71.25%of group B and 73.75%of group C, with statistically significant differences (P<0.01). The total effective rate of group A was 95.00%, which was significantly higher than 71.25%of group B and 70.005%of group C, with statistically significant differences (P<0.01). After 8 weeks of treatment, the serum leptin level in group A was significantly lower than that in group B and group C (P<0.01). The levels of serum SP and Ghrelin in group A were higher than those in group B and C (P<0.01). However, there was no significant difference in cytokine between group B and group C. The 3 groups of patients who received effective treatment were followed up for 6 months (excluding missing cases). There was no significant difference in body weight and height among three groups before treatment. After treatment, the body weight and height of three groups were higher than those before treatment (P<0.05, P<0.01). However, group A was superior to group B and group C (P<0.01). There was no significant difference between group B and group C.Conclusion:Treatment of infantile anorexia with Xiaoer anorexia granules combined with Saccharomyces cerevisiae can improve the total clinical effective rate, adjust serum cytokine levels and increase height and body mass.
Keywords：child with anorexia;Saccharomyces boulardii;Xiaoer anorexia granules;cytokines
Abstract：Objective:To discuss the clinical effect of Dahuang Zhechong Wan on pelvic pain caused by endometriosis with Qi stagnation and blood stasis syndrome and study the mechanism of action.Method:One hundred and twenty-six patients were randomly divided into control group (64 cases) and observation group (62 cases) by random number table. Both groups’ patients got Duphaston from the 5th to 25th days of menstrual cycle, 1 tablet/day, 2 times/days. Patients in control group got Sanjie Zhentong Jiaonang at the first day of menstruation, 4 grains/time, 3 times/days. Patients in observation group got Dahuang Zhechong Wan, 3 g/time, 2 times/days. The treatment in two groups continued for 3 menstrual cycles. Before treatment and at the first, second and third menstrual cycles after the treatment, visual analogue score (VAS) of pain was used for dysmenorrheal. Before AND after the treatment, scores of symptoms, signs, Qi stagnation and blood stasis syndrome and endometriosis were scored by endometriosis health profile-5 (EHP-5). And hemorheology was detected, and levels of matrix metalloproteinases-2 (MMP-2), MMP-9, tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), prostaglandin E2 (PGE2), prostaglandin F2α (PGF2α) and substance P (SP) were detected.Result:By rank sum test, the clinical effect of disease in observation group was better than that in control group (Z=2.198, P<0.05). At the first, second and third menstrual cycles, score of VAS was lower than that in control group (P<0.01). After treatment, scores of symptoms, signs, Qi stagnation and blood stasis syndrome and EHP-5 were lower than those in control group (P<0.01). And the ameliorate of hemorheological indices, such as the whole blood viscosity (high shear and low shear), plasma viscosity, erythrocyte sedimentation rate and platelet aggregation rate, were all better than those in control group (P<0.01). And levels of MMP-2, MMP-9, TNF-α, IL-1, PGF2α and SP were lower than those in control group (P<0.01), whereas level of PGE2 was higher than that in control group (P<0.01).Conclusion:In addition to treatment of progestogen, Dahuang Zhechong Wan can relieve pelvic pain, improve quality of life and clinical effect, and regulate levels of prostaglandins, matrix metalloproteinases and proinflammatory factors.
Abstract：Objective:To observe the effect of modified Yinchen Wuling San in treating liver function and blood lipid of non-alcoholic fatty liver disease (NALFD) with moisture and heat implication and the mechanism on intestinal microflora.Method:One hundred and thirteen patients were randomly divided into control group (57 cases) and observation group (56 cases) by random number table according to SAS. Both groups′patients got non drug intervention. Patients in control group got polyene phosphatidylcholine capsules, 2 grains/time, 3 times/days, and live combined Bacillus subtilis and Enterococcus faecium Enteric-coated capsules, 2 grains/time, 3 times/days. In addition to therapy of control group, patients in observation was also given modified Yinchen Wuling San, 1 dose/day. The treatment in two groups continued for 12 weeks. Before and after treatment, levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and intestinal microflora were detected. And CT and B-mode ultrasonography of abdomen were detected. And traditional Chinese medicine (TCM) syndromes were scored.Result:After treatment, levels of ALT and AST in observation group were lower than those in control group (Z=2.002, P<0.05), and score of TCM symptoms were lower than those in control group, whereas level of CT liver/spleen was higher than that in control group (P<0.01). And effect of laboratory indicators was better than that in control group (Z=1.955, P<0.05), CT was better than that in control group (Z=2.002, P<0.05), and B-mode ultrasonography of abdomen was better than that in control group (Z=1.961, P<0.05). Compared with healthy control group, enterobacteriaceae, staphylococcus of patients in two groups increased, while bacteroid, bacillus bifidus and bacillus lactis decreased (P<0.01). After treatment, enterobacteriaceae, staphylococcus in observation group were lower than those in control group, whereas bacteroid, bacillus bifidus and bacillus lactis were higher than those in control group (P<0.01).Conclusion:Modified Yinchen Wuling San can protect liver function of NAFLD, regulate lipid metabolism, ameliorate intestinal microflora, and recover the ecological equilibrium of intestine, with a good clinical effect.
Keywords：non-alcoholic fatty liver;moisture and heat implication;Yinchen Wuling San;intestinal microflora
Abstract：Objective:Taking single Liuwei Dihuangwan(LDW) as the research object, the moisture content change and volumetric shrinkage characteristics in its drying process were investigated, which provided the theoretical basis for improving the drying efficiency of the pills and reducing the quality problems of the pattern pills, crusts and crack pills.Method:The drying characteristics of LDW at drying temperature of 50, 75, 100, 125 ℃ were studied by constant temperature hot air drying and vacuum drying.Based on sphere model of Fick's second law and Arrhenius equation, the effective diffusion coefficient and activation energy of the drying process were obtained.The volumetric shrinkage characteristics of the pills during the drying process were studied by the projected area method.The Weibull function was used to fit the drying dynamics curve of single LDW.Result:Hot air drying and vacuum drying of LDW both belonged to the decreasing drying processes, and the time required to achieve the same moisture content in vacuum drying was shorter than that in hot air drying.The moisture ratio in the drying process of single LDW obeyed the Weibull function distribution(R2=0.994 5-0.999 7), the scale parameter(α) decreased with the increase of temperature, and drying temperature had significant influence on the shape parameter (β). The effective diffusion coefficients of hot air drying and vacuum drying were 2.626×10-3-7.823×10-2, 3.782×10-3-9.042×10-2 m2·s-1, their activation energy were 47.18, 42.69 kJ·mol-1, respectively.The volume ratio of hot air drying and vacuum drying of LDW ranged from 0.638 to 0.741 and 0.607 to 0.689, respectively.Conclusion:Weibull function can be adopted to predict the drying and dehydration law of LDW.Under the condition of low temperature drying, slow-down of shrinkage rate of the pills is helpful to prevent the formation of splitting pills, this study provides theoretical and technical basis for dying of LDW.
Abstract：Objective:To investigate the effect of Rhei Radix et Rhizoma on gastrointestinal function of normal rats and explore its therapeutic mechanism, and compare the difference of this herb before and after simmering.Method:Healthy SD rats were randomly divided into 7 groups, including the blank group, the high, medium and low dose groups of raw Rhei Radix et Rhizoma(1.5, 3.0, 6.0 g·kg-1·d-1) and the high, medium and low dose groups of simmered Rhei Radix et Rhizoma(1.5, 3.0, 6.0 g·kg-1·d-1), eight rats in each group were continuous intragastric administration for 10 days, the effect of Rhei Radix et Rhizoma on gastrointestinal function of rats were evaluated by body weight, food intake, gastric remnant rate, small intestine propelling rate, stomach weight coefficient and gastrointestinal tissue morphology; enzyme-linked immunosorbent assay(ELISA) was used to detect contents of motilin(MTL) in rat plasma and gastrin(GAS), tumor necrosis factor-α(TNF-α), interleukin-10(IL-10) in rat serum, the mechanism of effect of Rhei Radix et Rhizoma on gastrointestinal function was explored and the effect of Rhei Radix et Rhizoma on gastrointestinal function of normal rats before and after simmering was compared.Result:In each administration group, weight growth of rats was slowed down, gastric emptying and intestinal propulsion were inhibited, inflammatory reaction was triggered, and gastric mucosal injury was increased.Compared with the blank group, except for the low-dose and medium-dose groups of simmered Rhei Radix et Rhizoma, stomach weight coefficient, gastric remnant rate and content of TNF-α of rats were significantly increased(P<0.05, P<0.01), and the content of IL-10 was significantly reduced, in addition to the low-dose group of simmered Rhei Radix et Rhizoma, the weight, small intestine propelling rate, contents of GAS and MTL of rats were significantly decreased(P<0.05, P<0.01). Compared with the isodose raw Rhei Radix et Rhizoma group, the weight, small intestine propelling rate, contents of GAS, IL-10 and MTL were all increased with different degrees, the gastric weight coefficient, gastric remnant rate and content of TNF-α of rats were all decreased to different degrees.Conclusion:Gastrointestinal dysfunction caused by Rhei Radix et Rhizoma is related to inhibiting the expression of gastrointestinal hormones and promoting the expression of inflammatory cytokines, in addition, the effect of " stomach injured by bitterness and cold property" of Rhei Radix et Rhizoma is weakened after simmering, which provides scientific basis for clinical rational use of Rhei Radix et Rhizoma.
Keywords：Rhei Radix et Rhizoma;simmering;gastrointestinal motility;gastrointestinal hormones;inflammatory factors;enzyme-linked immunosorbent assay
Abstract：Objective:Based on the color space technology of CIE-LAB, the color of vinegar-processed Corydalis Rhizoma decoction pieces was digitized, combining with the contents of 10 major alkaloids in the decoction pieces, to discuss the correlation between the color and contents of main ingredients of vinegar-processed Corydalis Rhizoma decoction pieces, and investigate the intrinsic quality difference in the decoction pieces with different color.Method:The precision colorimeter was used to determine the color parameters of vinegar-processed Corydalis Rhizoma decoction pieces; HPLC was employed to determine contents of main chemical components in the decoction pieces, which was performed on Agilent ZORBAX SB-C18 column(4.6 mm×250 mm, 5 μm) with mobile phase of acetonitrile(A)-0.1% potassium dihydrogen phosphate aqueous solution(B) for gradient elution(0-10 min, 5%-22%A; 10-30 min, 22%-25%A; 30-50 min, 25%-60%A; 50-70 min, 60%-95%A), detection wavelength of 280 nm, column temperature at 30 ℃ and flow rate of 1.0 mL·min-1.Result:The quality of vinegar-processed Corydalis Rhizoma decoction pieces with different color was in line with the requirement of the 2015 edition of Chinese Pharmacopoeia, but there were differences in the intrinsic quality between the decoction pieces.The total content of chemical components in the samples showed a positive correlation with the a*(green-red axis) and total chromatic aberration value(ΔE) in the CIE-LAB color space, and it was significantly negative correlated with L*(lightness) and b*(blue-yellow axis). In the 10 tested components, except for D-tetrahydrojatrorrhizine and tetrahydrocoptisine, contents of protopine and other 6 components were positively correlated with color, and only the content of corydaline was negatively correlated with color.Conclusion:Color analysis technology can objectively quantify the color of the decoction pieces, and can achieve a quick evaluation of quality of the decoction pieces by analyzing correlation between the color and the contents of main active ingredients.
Abstract：Objective:Ganoderma sinense is one of the most famous medicinal fungi in China. Because it is a model medicinal fungus of basidiomycete, the functional identification of its sesquiterpene synthase is of great significance for the biosynthesis and regulation studies of fungal sesquiterpene.Method:A sesquiterpene synthase gene was discovered by mining the genome data of G. sinense. The sesquiterpene's conservative motifs was analyzed through multiple sequence alignment with two identified sesquiterpene synthases of G. sinense and three terpenoid synthases in the Nr database, which have the highest similarity to it. Subsequently, heterologous expression was observed in Escherichia coli, and protein expression was detected by SDS-PAGE. Volatile compounds were collected by solid phage microextraction (SPME) and detected by GC-MS.Result:The enzyme containing sesquiterpene conserved motifs DDXXDE and NSE/DTE were efficiently expressed in E. coli, and 11 sesquiterpenes were synthesized with endogenous FPP as the substrate. The product α-cadinol at 18.6 min was considered to be the main product, and previous studies showed a significant anticancer activity. According to the comparison with chemical standards, three products were identified as γ-muurlene, α-muurolene and δ-cadinene, respectively.Conclusion:The functional identification of multi-product sesquiterpene synthase from G. sinense can promote the study on the mechanism underlying its product diversity, and lay a foundation for the production of rare or novel sesquiterpenes by improving the catalytic activity of enzymes with enzyme engineering technology.
Abstract：Objective:To explore the receiuer operating characteristic(ROC) curve mathematical statistics method to establish the content limit of isoflavones glucoside contents in Astragali Radix, in order to distinguish the difference in isoflavones glucoside content between Hengshan wild-simulated Astragali Radix in Shanxi and cultivated Astragali Radix in Gansu.Method:A total of 225 samples of 45 batches of wild-simulated Astragali Radix in Hengshan and 210 samples of 42 batches of cultivated Astragali Radix in Gansu were selected as research objects, and isoflavones glucoside contents of these samples were determined by using the method in the edition 2015 Chinese Pharmacopoeia. Data was statistically analyzed, and the ROC curve was used to determine the critical value of wild-simulated Astragali Radix in Hengshan and transplanted Astragali Radix in Gansu.Result:The average contents of isoflavones glucoside in imitative wild Astragali Radix in Henshan and cultivated Astragali Radix in Gansu were 0.107%and 0.039%respectively, with statistically significant differences(P<0.01). The ROC curve had a critical value of 0.069%, and its sensitivity and specificity was 100%and 93.33%, and its model diagnostic accuracy was 0.978, which indicated accurate prediction results.Conclusion:ROC curve can be used to distinguish the critical value of the wild-simulated Astragali Radix in Hengshan and cultivated Astragali Radix in Gansu. The results reflected the regularity of " high quality and good price" in the market. The research provided a theoretical basis for the establishment of standard of high-quality Chinese herbal medicines.
Abstract：Objective:To discuss the phenotypic character and the HPLC fingerprints of radial striations from different germplasms Rehmanniae Radix.Method:The changes in the shape and column diameter of the radial striations of Rehmanniae Radix were observed and measured in the whole growth period. Besides, the HPLC fingerprints of the root, radial and un-radial striations were established to sign the chemical quality and analyzed by principal component analysis(PCA)and systematic cluster analysis.Result:There were significantly differences and regularities in the shape and proportion of the radial striations of different germplasms Rehmanniae Radix. The fingerprints showed the consistency between different types of chemical ingredients, and the differences in chemical quality characteristics mainly lay in the content of chemical compositions and theirs relative ratio. The results of PCA indicated that active ingredients, such as acteoside, catalpol, rehmaionoside D, rehmaionoside A and leonuride, were involved in the quality expression of different parts from various germplasms of Rehmanniae Radix, but each ingredient had a distinctive contribution rate to the differential quality expression between different parts from various germplasms of Rehmanniae Radix. However, the other components involved in the differential quality expression had different contribution rates in different germplasms.The systematic cluster analysis indicated that great differences in the chemical quality between the radial striations and un-radial striations of Beijing-1, Qinhuai, Qinhuai Zheng and 1706 germplasms, but with small differences in 85-5 and Baixuan germplasms.Conclusion:There are differences in phenotypic character of the radial striations and HPLC fingerprints between different germplasms Rehmanniae Radix.
Abstract：Objective:The aim of this study was to investigate high performance liquid chromatography (HPLC) fingerprint, anti-inflammatory activity as well as the correlation between them in Speranskiae Tuberculatae Herba.Method:Fingerprint by HPLC was established on Speranskiae Tuberculatae Herba from different sources. The common peaks were evaluated on the basis of similarity evaluation together with hierarchical cluster analysis (HCA) and principal component analysis (PCA). Ear swelling induced by xylene in mice model was used to study the anti-inflammatory activity. Grey relational analysis (GRA) and partial least square regression analysis (PLSR) were used to study the relationship between the HPLC fingerprint and the anti-inflammatory activity.Result:The HPLC fingerprint of Speranskiae Tuberculatae Herba was established and 24 common peaks were determined, with the similarity above 0.907 (except for S2 and S5). Four peaks were identified by comparing to reference substances. The result of HCA showed that Speranskiae Tuberculatae Herba from different sources were clustered into four categories, in consistent with the result of PCA. Combined with discriminant analysis of partial least squares discrimination analysis (PLS-DA), three signature compounds represented by 5, 6, 7 peaks were responsible for the differences between groups. Different sources of Speranskiae Tuberculatae Herba differed in anti-inflammatory activity. The sectrum-activity relationship showed that the peaks 1, 4, 5, 6, and 10 were positively correlated with the anti-inflammatory activity.Conclusion:The HPLC fingerprint of Speranskiae Tuberculatae Herba was established, and 5 components closely related to the anti-inflammatory activity were determined. The present study provide more comprehensive reference for quality control of the Speranskiae Tuberculatae Herba.
Abstract：Objective:To explore the change rules of active ingredients in Phyllanthi Fructus of different storage years, in order to provide theory basis for storage.Method:Seven Phyllanthi Fruatus samples of different storage years were collected. HPLC-UV detection method was established to determine the contents of gallic acid, corilagin, chebulagic acid, ellagic acid and quercetin. Samples were fingerprinted by FT-NIR and identified by PLS-DA model.Result:Gallic acid, which was the bioactive marker in Chinese Pharmacopoeia, had the highest content. It was followed by ellagic acid and chebulagic acid, and corilagin and quercetin had the least content. The components had significant differences between samples of different storage years (P<0.05). Gallic acid and quercetin reached the maximum value at 6 years of storage, which were 79.36 and 1.68 mg·g-1 respectively. The contents of chebulagic acid, corilagin and ellagic acid reached a maximum at 4 years of storage, which were 18.85, 7.97, 21.46 mg·g-1, respectively. FT-NIR data was optimized by MSC+ SG (second derivative, the window parameter as 11, and the polynomial order as 3). The classification accuracy was 84.5%. Spectral data reduced to several important potential variables, and was fused with 5 active components based on minimum cross-validation root mean square error, and the classification accuracy increased to 98.8%.Conclusion:The analysis of PLS-DA by HPLC-UV and FT-NIR could effectively explain the accumulation characteristics of active components in Phyllanthi Fruatus. According to the data fusion strategy, PLS-DA model could distinguish samples of different qualities. The results provide a scientific basis for the quality evaluation and identification of Phyllanthi Fruatus.
Abstract：Objective:To study the origin of Nanyang Chrysanthemi Flos materia medica and its historical origin, in order to evaluate the quality of Nanyang Chrysanthemi Flos by HPLC method, and define the advantages of Nanyang Chrysanthemi Flos with the origin of Nanyang.Method:Records of Nanyang Chrysanthemi Flos in the " Chinese Medical Code" and related ancient documents were studied to explain the origin and application. The genetic relationship between Nanyang Chrysanthemi Flos and other pieces was revealed in ancient literatures. Then 8 chemical constituents in 14 batches of Chrysanthemi Flos were determined by HPLC multi-components quantitation, and the comprehensive weighted score analysis was performed based on the results. The HPLC fingerprints were established, and the similarity analysis and clustering analysis were made to comprehensive evaluation the quality of Nanyang Chrysanthemi Flos and define the genetic relationship between Nanyang and other pieces at the chemical composition level.Result:The results of the herbal textual research show that Nanyang Chrysanthemi Flos spread from place to place since the Han dynasty, and impact many medicinal chrysanthemums of later generations. HPLC fingerprints similarity and cluster analysis also indicated the genetic relationship between Nanyang Chrysanthemi Flos and other species at the chemical level. The comprehensive score analysis results show that Nanyang Chrysanthemi Flos is of good quality and very suitable for medicinal purposes.Conclusion:Nanyang has been a quality production area of Chrysanthemi Flos since ancient times to present. Nanyang Chrysanthemi Flos is very suitable for medicinal purposes and worthy of promotion.
Abstract：Objective:To screen out active ingredients, and predict the anti-cancer targets of Ligustri Lucidi Fructus-Astragali Radix based on the " herbs-active ingredient-action targets" network.Method:The traditional Chinese medicine (TCM) system pharmacology platform (TCMSP) was employed to screen out the active ingredients and putative targets of anti-cancer of glossy privet fruit and astragalus. DisGeNET database and Online Mendelian Inheritance in Man (OMIM database) were employed to predict the targets for treating cancer, and then " herbs-active ingredients-key targets" network was constructed by using Cytoscape software. The omicshare platform was employed to match the putative targets of ingredients and the targets for treating cancer. Finally, the protein interaction network of key targets was constructed by using String database, and the analysis of biological functions and pathways of them was carried out by using DAVID database.Result:Totally 33 drug active ingredients were screened out, involving a total of 203 targets, and 14 of them were related to cancer. These 14 key targets played an therapeutic role in treating cancer by regulating target proteins, such as ERBB2, AR, SRC, EGFR, ESR1, as well as proteoglycans in cancer, cancer pathways, microRNAs in cancer and other pathways.Conclusion:The therapeutic mechanism of Ligustri Lucidi Fructus-Astragali Radix reflects the multi-component, multi-target, and multi-pathway characteristics of TCMs, providing the scientific basis for further study and the material basis of Ligustri Lucidi Fructus-Astragali Radix against cancer.
Abstract：With the implementation of the national " The Belt and Road" strategy, the international exchange of Chinese medicine is increasing day by day, and foreign drug resources provide a vast space for the development of traditional Chinese medicine(TCM). The introduction of foreign high-quality resources to promote the development of TCM has become the mission of the times to TCM practitioners.In this paper, the successful cases of herbalization of foreign drugs were analyzed.From the three aspects of the image thinking, the original application, the clinical and experimental validation, TCM was recognized and the preliminary research method of herbalization of the foreign medicines was put forward.Through the repeated exploration mode of " theoretical discussion(literature research+ field investigation)→experimental verification→clinical practice" , the performance of foreign drugs was investigated, providing reference for the research method of herbalization of foreign drugs.
Keywords：" The Belt and Road ";exotic drugs;herbalization;theory of traditional Chinese medicine;clinical practice;pharmacological experiments;literature research
Abstract：Objective:To study the potential anti-inflammatory active ingredient of Chinese herbal Polygoni Cuspidati Rhizoma et Radix.Method:The inflammatory model of peritoneal macrophages (RAW264.7 cells) induced by lipopolysaccharide (LPS) in mice was used to screen out the anti-inflammatory activity of 95% ethanol extract of Polygoni Cuspidati Rhizoma et Radix and its macroporous resin elution site (30%, 60%, 95% ethanol eluting site). The characteristic fingerprints were established by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) technology, and then partial least squares method (partial least squares, PLS) was used to study the spectrum-effect relationship between the peak area of the characteristic components and the inhibition rate of nitric oxide (NO), and potential anti-inflammatory active ingredients were identified according to variable important in projection (VIP).Result:The ethanol extract macroporous resin 60% ethanol elution site of Polygoni Cuspidati Rhizoma et Radix had the strongest inhibition ability of nitric oxide formation, with a certain dose-dependent relationship. The study of spectrum-effect relationship showed that 3 components had potential anti-inflammatory activity, namely Emodin-8-O-β-D-glucoside (E-8-G), Emodin-1-O-β-D-giucoside, and Emodin-8-O-(6′-O-malonyl)-glucoside. In addition, the anti-inflammatory activity of E-8-G was further validated at the cell level through molecular docking analysis.Conclusion:Three potential anti-inflammatory active ingredients were found base on the spectrum-effect relationship. This study strategy is helpful to find the active ingredients (group) of traditional Chinese medicine, and provides new research ideas and methods for studying the material basis of Chinese herbal medicine.
Keywords：Polygoni Cuspidati Rhizoma et Radix;anti-inflammation;spectrum-effect relationship;UPLC-Q-TOF/MS;emodin -8-O-β-D-glucoside(E-8-G)
Abstract：Hypertensive renal damage is one of the most serious complications of hypertension, and it is also the main cause of end-stage renal disease. Renal damage can further promote the rise of blood pressure and difficult to control, forming a vicious circle. Traditional Chinese medicine(TCM) considers that deficiency of the original and excess of the standard is its basic pathogenesis, and insufficient kidney-Qi and blood stasis are one of the most common syndromes of hypertensive kidney damage. Astragali Radix membranaceus is praised as the most important medicine for invigorating Qi, and Salviae Miltiorrhizae Radix is likened to " four things with the same function" . Based on the theory of invigorating Qi and activating blood circulation, the effective ingredients of Astragali Radix membranaceus-Salviae Miltiorrhizae Radix are mainly astragaloside, Astragali Radix polysaccharide, mulberry isoflavone, salvianolic acid and tanshinone. Many studies have shown that in the process of hypertensive kidney damage, Astragali Radix membranaceu Its active ingredients, whether effective monomers, monomer compatibility or direct compatibility of drug pairs, can regulate blood pressure, reduce urinary protein, protect renal tubules, protect glomerular filtration barrier, improve renal hemodynamics and protect renal function by regulating multiple signal transduction pathways related to hypertensive renal damage. Lowering blood pressure and protecting renal function are two pronged functions. Based on the theory of Invigorating Qi and activating blood circulation, this paper reviews the research progress of Astragali Radix-Salviae Miltiorrhizae Radix in the treatment of hypertensive renal damage, with a view to providing scientific basis for the further study and clinical application of Radix astragali-Salviae Miltiorrhizae Radix in hypertensive renal damage.
Abstract：Acute pancreatitis (AP) is a common clinical acute abdominal disease, which can be divided into three types: mild, moderate and severe. Severe acute pancreatitis still has a high mortality rate, but there is no specific drug for this disease, and non-operative symptomatic supportive treatment is the main therapy in the early stage of the disease. In recent years, a large number of clinical studies have shown that traditional Chinese medicine has a unique advantage in prevention and treatment of AP. According to the characteristics of the disease, such as abdominal pain, no stool, treatment based on syndrome differentiation can alleviate a series of symptoms, especially the formulations of " removing stasis by purgation" represented by Da Chengqitang, including the formula evolved based on Da Chengqitang. Specifically, Chaishao Chengqitang and Chaiqin Chengqitang can improve intestinal function, alleviate gastrointestinal symptoms, such as intestinal paralysis combined with AP patients, prevent intestinal flora ectopia, protect intestinal mucosal barrier function, and help activate enteral nutrition in early stage, regulate immune inflammatory response, so as to avoid systemic inflammatory response syndrome. In addition, categorized formulas Da Chengqitang can also promote pancreatic microcirculation, promote the recovery of damaged parts of pancreas during AP pathogenesis, reduce blood amylase, and improve systemic coagulation function and treatment effect. In China, integrated traditional Chinese and Western medicine treatment has become an important part of the treatment of AP at present. It can prevent AP from aggravating, reduce complications and shorten hospitalization time. To summarize the therapeutic effect of Da Chengqitang and its categorized formulas on AP in clinic, the authors review clinical studies of categorized formulas Da Chengqitang on AP in recent five years based on " intestinal function, intestinal nutrition, inflammatory mediators and pancreatic microcirculation" .
Keywords：Da Chengqitang;categorized formulas;acute pancreatitis;clinical study
Abstract：Cognitive dysfunction of diabetes mellitus is one of the complications of diabetes mellitus. Its clinical manifestations include decreased memory ability, learning ability, language expression and comprehension ability, and inattention. The pathogenesis is complex, which is related to various factors such as abnormal glucose metabolism, abnormal lipid metabolism, insulin resistance, abnormal phosphorylation of Tau protein, inflammatory reaction, oxidative stress, cholinergic system injury, neuronal cell apoptosis, et al. Although traditional hypoglycemic drugs such as metformin and glibenclamide have a little effect in controlling diabetic cognitive impairment, but there has no effective drugs for this disease which causes a heavy burden on families and society. The concepts of holism and preventing disease of traditional Chinese medicine(TCM)have obvious advantages in delaying the occurrence and development the complications.TCM believes that the basic pathogenesis of diabetic cognitive dysfunction is based on deficiency complicated with excess. The therapeutic principles includes tonifying the kidney essence, reinforcing Qi and nourishing Yin, dissipating phlegm for resuscitation, and promoting blood circulation to remove blood stasis.In recent years, there have been a lot of research achievements on the prevention and treatment of cognitive dysfunction in diabetes mellitus with TCM. Researchers of TCM do abundant researches, and find that TCM can improve cognitive dysfunction by lowering blood glucose, promoting intracellular cholesterol excretion, improving insulin resistance, lowering p-Tau, anti-oxidative stress, inhibiting inflammatory reactions, improving the function of cholinergic nerve, inhibiting neuronal apoptosis. The article reviews the experimental research on the prevention and treatment of cognitive dysfunction of diabetes mellitus by TCM and its extracts and reveals the target and mechanism, in order to provide sufficient scientific basis for TCM to prevent cognitive dysfunction of diabetes mellitus.
Keywords：traditional Chinese medicine;cognitive dysfunction in diabetes mellitus;pathogenesis;advance of studies